RU2806709C2 - Lactase enzymes with improved properties - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: peptide exhibiting the enzymatic activity of beta-galactosidase. The invention relates to the use of a peptide exhibiting beta-galactosidase enzymatic activity for the production of a lactose-reduced dairy product.

EFFECT: using the enzyme in a wide temperature and pH range for effective lactose reduction.

14 cl, 16 dwg, 2 tbl, 21 ex

Description

FIELD OF THE INVENTION

The present invention relates to a new and improved peptide or dimeric peptides exhibiting beta-galactosidase enzymatic activity, as well as improved methods for reducing lactose in compositions such as dairy products.

BACKGROUND ART

For the growth of lactic acid bacteria in milk, the optimal way for them to obtain glucose and galactose as a carbon source is the hydrolysis of lactose. Lactase (beta-galactosidase; EC 3.2.1.23) is an enzyme that carries out the step of hydrolysis of the milk sugar lactose to monosaccharides. Commercially, lactase is used to break down lactose in dairy products. Individuals with lactose intolerance have difficulty digesting dairy products that are high in lactose. It is estimated that approximately 70% of the world's population has a limited ability to digest lactose. Accordingly, there is a growing demand for dairy foods that do not contain or contain only low concentrations of lactose.

Lactases are isolated from a wide range of organisms, including microorganisms such as Kluyveromyces and Bacillus. Common sources of fungal lactases are the yeasts Kluyveromyces, particularly K. fragilis and K. lactis, and other fungi, such as the genera Candida, Torula and Torulopsis, with B. coagulans and B. circulans being well-known sources of bacterial lactases. Several commercial preparations derived from these organisms are commercially available, such as Lactozym® (from Novozymes, Denmark), HA-Lactase (from Chr. Hansen, Denmark) and Maxilact® (from DSM, the Netherlands), all from K. lactis. All of these lactases are so-called neutral lactases, having optimal pH values from pH 6 to pH 8, as well as an optimal temperature value of approximately 37°C. When using such lactases in the production of, for example, low lactose yoghurt, the enzyme treatment either needs to be carried out in a separate step prior to fermentation, or, most often, high doses of enzymes must be used since their activity will decrease as the pH decreases during the fermentation process. In addition, these lactases are also not suitable for the hydrolysis of lactose in milk carried out at high temperatures, which in some cases would be useful to maintain microorganisms in low numbers and therefore guarantee high quality milk. In addition, known lactases are not suitable for use if an ultra-pasteurized milk production process (UHT) in which enzymes are added prior to UHT processing is desired.

WO2010092057 and WO0104276 refer to beta-galactosidases active at low temperatures. WO07110619 refers to a beta-galactosidase with high trans-galactosylation activity, where WO2009071539 refers to a beta-galactosidase with reduced trans-galactosylation activity.

PURPOSE OF THE INVENTION

The purpose of embodiments of the present invention is to develop beta-galactosidases with properties that provide improved lactose-free or lactose-low products.

It is a further object of embodiments of the invention to provide beta-galactosidases with properties that provide improved, for example, simpler, faster, more reliable or less expensive production methods for reducing the lactose content of a product, such as lactose-free or low-lactose products.

SUMMARY OF THE INVENTION

The present inventors have identified beta-galactosidases having previously undescribed properties that provide improved lactose-free or low-lactose products, as well as improved methods for producing such lactose-free or low-lactose products. In particular, these beta-galactosidases have been shown to be highly stable with relatively high activity over very wide ranges of temperature as well as pH. They can also be used at certain temperatures, such as high temperatures, and pH values not typically considered for these enzymes. First of all, this makes it possible to use beta-galactosidases at certain pH and temperature values that were not previously known to be possible. This also allows the same specific enzyme to be used in several different applications with high industrial demand.

Thus, in a first aspect, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity, which has the amino acid sequence represented by SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, or their amino acid sequence, having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions.

In a second aspect, the present invention relates to a dimeric peptide exhibiting beta-galactosidase enzymatic activity, which consists of two peptides having the amino acid sequence represented by SEQ ID NO: 2 and 3; 5 and 6; 20 and 21; 23 and 24; 26 and 27; or 28 and 29, or enzymatically active fragments thereof, or the amino acid sequence of any of them having not more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions.

In a third aspect, the present invention provides a nucleotide sequence that encodes a peptide or dimeric peptide exhibiting beta-galactosidase enzymatic activity in accordance with the invention.

In a further aspect, the present invention relates to a host cell containing a nucleotide sequence that encodes a peptide or dimeric peptide in accordance with the invention exhibiting beta-galactosidase enzymatic activity.

In a further aspect, the present invention relates to a method for producing a peptide or dimeric peptide in accordance with the invention exhibiting beta-galactosidase enzymatic activity, wherein the method comprises expressing a vector containing a nucleotide sequence in accordance with the invention in a suitable host cell; and purifying said peptide or dimeric peptide from expression products of said host cell.

In a further aspect, the present invention relates to a method of reducing the lactose content of a composition containing lactose, for example in dairy products, comprising the step of contacting said composition with a peptide or dimeric peptide exhibiting beta-galactosidase enzymatic activity, which has the amino acid sequence represented by SEQ ID NO: 1-33; or where the dimeric peptide consists of two peptides having the amino acid sequence represented by SEQ ID NO: 2 and 3, 5 and 6, 20 and 21, 23 and 24, 26 and 27, or 28 and 29; or a sequence with at least 80% sequence identity to any one of the specified sequences; or with a host cell expressing any one of these peptides at a pH in the range of 3-10 and at a temperature in the range of 0°C-140°C.

In a further aspect, the present invention relates to the use of a peptide or dimeric peptide exhibiting beta-galactosidase enzymatic activity, which has the amino acid sequence represented by SEQ ID NO: 1-33, or wherein the dimeric peptide consists of two peptides having the amino acid sequence represented by SEQ ID NO: 2 and 3, 5 and 6, 20 and 21, 23 and 24, 26 and 27, or 28 and 29; or a sequence with at least 80% sequence identity to any one of the specified sequences; or a host cell expressing any one of these peptides to produce a lactose-reduced dairy product.

In some embodiments of the invention, the lactose-containing composition or the dairy product is selected from the group consisting of lactose-free milk, low-lactose milk, yogurt, including unpasteurized as well as pre- and post-pasteurized yogurt, cheese, fermented milk products, dietary supplements, and probiotic dietary products. . In some other embodiments of the invention, the host cell is any cell selected from bacteria of the genus Bifidobacterium, such as Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium longum, or from the genus Lactobacillus, such as L. sakei, L. amylovorus, L. delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, L. delbrueckii subsp. Indicus, L. crispatus, L. reuteri, L. helveticus, or from Streptococcus thermophilus. In some other embodiments of the invention, the lactose concentration is reduced to less than about 1%, such as less than about 0.1% or less, such as less than about 0.01%.

In a further aspect, the present invention relates to a method for producing a dairy product, comprising the steps of: a) providing a lactose-containing milk-based substrate; b) adding to said lactose-containing milk-based substrate a peptide or dimeric peptide exhibiting beta-galactosidase activity having the amino acid sequence represented by SEQ ID NO: 1-33; or where the dimeric peptide consists of two peptides having the amino acid sequence represented by SEQ ID NO: 2 and 3, 5 and 6, 20 and 21, 23 and 24, 26 and 27, or 28 and 29; or a sequence with at least 80% sequence identity to any one of the specified sequences; and c) treating said milk-based substrate with said peptide or dimeric peptide exhibiting beta-galactosidase activity.

In a further aspect, the present invention relates to a dairy product obtained by the method in accordance with the invention.

In a further aspect, the present invention provides a food product, such as a dairy product, containing a peptide or dimeric peptide exhibiting beta-galactosidase enzymatic activity, which has the amino acid sequence represented by SEQ ID NO: 1-33, or wherein the dimeric peptide consists of two peptides having the amino acid sequence represented by SEQ ID NO: 2 and 3, 5 and 6, 20 and 21, 23 and 24, 26 and 27, or 28 and 29; or a sequence with at least 80% sequence identity to any one of the specified sequences.

In a further aspect, the present invention provides a food product, such as a dairy product, comprising a host cell expressing a peptide or dimeric peptide exhibiting beta-galactosidase enzymatic activity, which has the amino acid sequence represented by SEQ ID NO: 1-33, or wherein dimeric the peptide consists of two peptides having the amino acid sequence represented by SEQ ID NO: 2 and 3, 5 and 6, 20 and 21, 23 and 24, 26 and 27, or 28 and 29; or a sequence with at least 80% sequence identity to any one of the specified sequences. In some specific embodiments of the invention, the food product is selected from beverages, baby foods, cereals, breads, cookies, confectionery, cakes, nutritional supplements, dietary supplements, probiotic foods, prebiotic foods, animal feed, feed and poultry medicinal products or a dairy product selected from the group consisting of lactose-free milk, low-lactose milk, powdered milk, baby milk, yogurt, ice cream, cheese, fermented milk products, dietary supplement and probiotic dietary food products.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

Fig. 1. Specific activity of purified enzymes determined at pH 6.7 at 37° C. with lactose as substrate, described as SUAL-1, discussed in Example 6. Under these conditions, the measured standard deviation was less than 6%.

Fig. 2. Specific activity of purified enzymes determined at pH 6.7 at 37° C. in the presence of galactose, described as SUAG, discussed in Example 7. Under these conditions, the measured standard deviation was less than 15%.

Fig. 3. Specific activity of purified enzymes determined at pH 6.7 at 4°C with lactose as substrate, described as SUAL-2, discussed in example 8. Under these conditions, the measured standard deviation was less than 5%.

Fig. 4. Specific activity of purified enzymes determined at pH 6.7 at 43°C with lactose as substrate, described as SUAL-3, discussed in example 9. Under these conditions, the measured standard deviation was less than 5%.

Fig. 5. Specific activity of purified enzymes determined at pH 5.5 at 4°C with lactose as substrate, described as SUAL-4, discussed in example 10. Under these conditions, the measured standard deviation was less than 5%.

Fig. 6. Specific activity of purified enzymes determined at pH 5.5 at 37°C with lactose as substrate, described as SUAL-5, discussed in example 11. Under these conditions, the measured standard deviation was less than 5%.

Fig. 7. Specific activity of purified enzymes determined at pH 5.5 at 43°C with lactose as substrate, described as SUAL-6, discussed in example 12. Under these conditions, the measured standard deviation was less than 5%.

Fig. 8. Specific activity of purified enzymes determined at pH 4.5 at 4°C with lactose as substrate, described as SUAL-7, discussed in example 13. Under these conditions, the measured standard deviation was less than 5%.

Fig. 9. Specific activity of purified enzymes determined at pH 4.5 at 37°C with lactose as substrate, described as SUAL-8, discussed in example 14. Under these conditions, the measured standard deviation was less than 5%.

Fig. 10. Specific activity of purified enzymes determined at pH 4.5 at 43° C. with lactose as substrate, described as SUAL-9, discussed in example 15. Under these conditions, the measured standard deviation was less than 5%.

Fig. 11. Percentage of residual lactose in pasteurized milk after treatment with a fixed amount of enzyme after 24 hours at 5°C, determined by high-performance liquid chromatography (HPLC).

Fig. 12. Percentage of residual lactose in UHT milk after treatment with a fixed amount of enzyme after 24 hours at 25°C, determined by HPLC.

Fig. 13. Percentage of residual activity of purified enzymes at elevated temperatures, determined using lactose as substrate. Activity at pH 6.7 at 37°C was considered 100%.

Fig. 14. Specific activity of purified enzymes determined at pH 6.7 at 4°C, 37°C and 43°C. The measured specific activity is described in µmol of glucose formed per minute per mg of enzyme. The method for determining inhibition by galactose is described in example 7, and the calculation is performed in µmol of glucose formed per minute per mg of enzyme.

Fig. 15. Specific activity of purified enzymes, determined at pH 5.5 at 4°C, 37°C and 43°C. The measured specific activity is described in µmol of glucose formed per minute per mg of enzyme.

Fig. 16. Specific activity of purified enzymes determined at pH 4.5 at 4°C, 37°C and 43°C. The measured specific activity is described in µmol of glucose formed per minute per mg of enzyme.

DETAILED DESCRIPTION OF THE INVENTION

The present inventors have surprisingly discovered that certain peptides and dimeric peptides exhibiting beta-galactosidase enzymatic activity are stable under a variety of physical conditions and have relatively high activity outside the ranges generally considered optimal for this class of enzymes.

Accordingly, the present inventors have identified enzymes that have relatively high activity at a temperature of about 4°C or 5°C and can therefore be used for the hydrolysis of lactose in the production of, for example, fresh milk. In addition, these enzymes also have relatively high activity in the range of 10°C-25°C, and therefore exactly the same enzymes can be used to hydrolyze lactose in UHT milk. The applicability of these enzymes even over wider temperature ranges is highly relevant since milk can be stored at room/ambient temperatures, which may vary in different areas of the globe and also depending on the time of year. For UHT processing, the temperature is typically about 135°C or about 140°C. It is highly desirable that enzymes can be active in a temperature range up to 140° C. so that the enzyme can be added to raw milk prior to the UHT stage. In current practice, the enzyme is added after the UHT step due to a significant reduction in the functional activity of enzymes known in the art after the high temperature step, for example to a value below the measured activity. Milk is also stored at room temperature, which can vary significantly in different areas of the world.

New and improved peptides exhibiting beta-galactosidase enzymatic activity have also been found to be active in the temperature range typically used for pasteurization. Accordingly, these enzymes can be added to raw milk prior to pasteurization. It should be clear that the functional activity of enzymes known in the art after the pasteurization step is significantly reduced, for example to a value below the measured activity.

An additional advantage of these new and improved peptides exhibiting beta-galactosidase enzymatic activity is that they have a relatively low degree of inhibition by galactose. The reduced galactose inhibition of these new enzymes is highly relevant for applications where low lactose concentrations are desired.

With regard to applicability to fermented milk products, a significant advantage is that the enzymes described herein have high beta-galactosidase enzymatic activity over a relatively wide temperature range from 4°C to 43°C, for example about 37°C, where fermentation is normal should be optimal, but this beta-galactosidase enzymatic activity is also present at low pH, for example, lowered to 4.5, or lowered to 4.0, or lowered to 3.5, or even lowered to pH 3.

In summary, the present inventors have discovered that certain peptides exhibiting beta-galactosidase enzymatic activity are active over a wide temperature range, active over a wide pH range, have overall high hydrolytic activity without side activities, and that there is no or little inhibition of these peptides by galactose, for example, less than 60%, and that they are stable over long periods of storage.

Beta-galactosidase activity can be determined by measuring the amount of glucose released after incubation with lactose under specified conditions. The released glucose can be detected using a color reaction.

Definitions

As used herein and in the context of the present invention, the term "milk" is to be understood as the secreted milk resulting from the milking of any mammal such as a cow, sheep, goat, buffalo or camel.

As used herein, the term “lactose-containing composition” refers to any composition, such as any liquid, containing lactose to a sufficient extent to be measured, for example, having a lactose content greater than 0.002% (0.002 g/100 ml). This term covers milk and milk-based substrates.

In the context of the present invention, the term "milk-based substrate" can mean any raw and/or processed milk material. Useful milk-based substrates include, but are not limited to, solutions/suspensions of any milk or milk-like products containing lactose, such as whole or low-fat milk, skim milk, buttermilk, low-lactose milk, reconstituted milk powder, evaporated milk, solutions milk powder, UHT milk, whey, whey permeate, acid whey, cream, fermented milk products such as yogurt, cheese, dietary supplement and probiotic dietary products. Generally, the term "milk-based substrate" refers to raw and/or processed milk material that is further processed to produce a dairy product.

As used herein, the term “pasteurization” refers to the process of reducing or eliminating the presence of living organisms, such as microorganisms, in a milk-based substrate. Preferably, pasteurization is achieved by maintaining a certain temperature for a certain period of time. A certain temperature is usually achieved by heating. The temperature and length of time can be chosen to kill or inactivate certain bacteria, such as harmful bacteria, and/or inactivate enzymes in the milk. This may be followed by a rapid cooling step.

As used herein, the term “dairy product” can mean any food product that has a milk base as one of its main components. Typically, the main component is a milk base, and in some embodiments of the invention the main component is a milk-based substrate treated in accordance with the method of the present invention with an enzyme having beta-galactosidase activity.

The dairy product according to the invention may be, for example, skim milk, low fat milk, whole milk, cream, UHT milk, extended shelf life milk, fermented milk product, cheese, yogurt, butter, milk spread, buttermilk, fermented milk drink , sour cream, whey drink, ice cream, condensed milk, boiled condensed milk or flavored milk drink.

The dairy product may further contain components other than milk, for example plant components such as, for example, vegetable oil, vegetable protein and/or vegetable carbohydrates. Dairy products may also contain additional additives such as, for example, enzymes, flavorings, microbial cultures such as probiotic cultures, salts, sweeteners, sugars, acids, fruits, cooked fruits, fruit juices or any other component known in the art as a component or additive in a dairy product.

As used herein, the terms “fermented milk product” or “fermented milk product” should be understood to mean any dairy product whose production process includes any type of fermentation. Examples of fermented milk products include foods such as yogurt, buttermilk, crème fraiche, soft cottage cheese (quark), and fresh cheese. The fermented milk product can be produced by any method or method including stages known in the art.

As used herein, the term "fermentation" refers to the conversion of carbohydrates into alcohols or acids by microorganisms. In some embodiments of the invention, fermentation in accordance with the present invention involves converting lactose to lactic acid. In the context of the present invention, "microorganism" may include any bacterium or fungus capable of fermenting a milk substrate.

As used herein, the term “increased beta-galactosidase enzyme activity” refers to relatively increased activity of the beta-galactosidase enzyme compared to the reference sequence.

As used herein, the term “peptide exhibiting enhanced beta-galactosidase enzymatic activity” refers to any peptide that has the enzymatic activity of catalyzing the hydrolysis of the disaccharide lactose to its monosaccharide components glucose and galactose. This peptide may also be called lactase or simply beta-galactosidase (EC: 3.2.1.23).

As used herein, the terms “peptide” and “oligopeptide” are generally considered synonymous, and each term can be used interchangeably as the context requires as indicating a chain of at least two amino acids linked by peptidyl bonds. The word "polypeptide" is used herein for chains containing more than ten amino acid residues. All peptide and polypeptide formulas or sequences given herein are written from left to right and in the direction from amino terminus to carboxy terminus. As used herein, the term “proteins” refers to peptide sequences produced by a host organism and may include post-translational modifications such as attached glycans.

As used herein, the terms “amino acid” or “amino acid sequence” refer to an oligopeptide, peptide, polypeptide or protein sequence or any fragment thereof and natural or synthetic molecules. As used herein, “fragment” refers to peptide fragments exhibiting beta-galactosidase enzymatic activity that retain some enzymatic activity. When an “amino acid sequence” referred to herein refers to the amino acid sequence of a naturally occurring protein molecule, “amino acid sequence” and other similar terms are not intended to limit the amino acid sequence to the full-length native amino acid sequence associated with the peptide molecule.

Examples of peptides of the invention also include fragments of at least about 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800 or more residues in length, or greater than the full length enzyme. Accordingly, a “peptide fragment” or an “enzymatically active fragment” of the invention are fragments that retain at least some functional enzymatic activity. Typically, the peptide fragment of the invention will still contain a functional catalytic domain or other essential active sites of the peptide exhibiting beta-galactosidase enzymatic activity. Other domains may be deleted.

Typically, the specific enzyme activity of beta-galactosidase will be measured and reported in µmol glucose produced/min/mg enzyme used. However, this specific value will vary depending on the conditions applied, such as temperature and pH. Accordingly, beta-galactosidase enzyme activity values may also be reported as relative to a known reference enzyme, such as the beta-galactosidase enzyme defined by SEQ ID NO: 34 or SEQ ID NO: 35.

Alternatively, the specific enzyme activity of beta-galactosidase can be measured and reported in micromoles (µM) of glucose produced per second per µM of enzyme used. However, this specific value will vary depending on the conditions applied, such as temperature and pH.

Unless otherwise noted, as used herein, the term “sequence identity” for amino acids refers to sequence identity calculated as (n _ref -n _dif )⋅100/n _ref , where n _dif is the total number of non-identical residues in the two sequences in the alignment , and where n _ref represents the number of residues in one of the sequences.

In some embodiments of the invention, sequence identity is determined by traditional methods, for example, Smith and Waterman, 1981, Adv. Appl. Math. 2: 482, by searching to apply the similarity method Pearson & Lipman, 1988, Proc. Natl. Acad. Sci. USA 85:2444, applications of the CLUSTAL algorithm W Thompson et al., 1994, Nucleic Acids Res 22:467380, using computer applications of these algorithms (GAP, BESTFIT, FASTA and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group). The BLAST algorithm (Altschul et al., 1990, Mol. Biol. 215: 403-10) can also be used, for which software is available through the National Center for Biotechnology Information, www.ncbi.nlm.nih. gov/). When using any of the algorithms mentioned above for length window, gap penalty, etc. use default settings.

A peptide with a specific amino acid sequence, as described herein, may differ from the sequence of the reference peptide by substitution, addition/insertion or deletion of any of the amino acids.

Some embodiments in accordance with the present invention relate to the use of a peptide with the amino acid sequence represented by SEQ ID NO: 1-33, or a sequence with at least 80% identity to any one of these sequences. In some embodiments of the invention, this sequence identity may be at least about 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93 %, 94%, 95%, 96%, 97%), 98%), or 99%), for example, a peptide having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 substitutions, additions or deletions, compared to any one reference amino acid sequence represented by SEQ ID NO: 1-33. Elements of the invention are also biologically active fragments of the peptides in accordance with the invention. Biologically active fragments of a peptide of the invention include peptides containing amino acid sequences substantially identical to or derived from the amino acid sequence of a peptide of the invention that comprise fewer amino acids than a full-length protein but exhibit a substantial portion of the biological activity of the corresponding full-length peptide. Typically, biologically active fragments contain a domain or motif with at least one activity of a protein variant of the invention. The biologically active fragment of the peptide of the invention may be a peptide whose length is, for example, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000 or more amino acids.

As used herein, the term “host cell” includes any type of cell subject to transformation, transfection, transduction, or the like. a nucleic acid construct or expression vector that contains a polynucleotide encoding the peptides of the present invention. The host cell may be any type of cell from which a particular enzyme is obtained, or alternatively a cell type that is prone to produce a particular enzyme. This term includes both wild-type and attenuated strains.

A suitable host cell may be any bacterium including lactic acid bacteria within the order Lactobacillales, which includes Lactococcus spp., Streptococcus spp., Lactobacillus spp., Leuconostoc spp., Pseudoleuconostoc spp., Pediococcus spp., Brevibacterium spp., Enterococcus spp. and Propionibacterium spp. It also includes lactic acid producing bacteria belonging to the group of anaerobic bacteria, bifidobacteria, i.e. Bifidobacterium spp., often used as food crops alone or in combination with lactic acid bacteria. This definition also includes Lactococcus lactis, Lactococcus lactis subsp.cremoris, Leuconostoc mesenteroides subsp.cremoris, Pseudoleuconostoc mesenteroides subsp. cremoris, Pediococcus pentosaceus, Lactococcus lactis subsp. lactis biovar. diacetylactis, Lactobacillus casei subsp.casei and Lactobacillus paracasei subsp. Paracasei and species of thermophilic lactic acid bacteria, including as examples Streptococcus thermophilus, Enterococcus faecium, Lactobacillus delbrueckii subsp. lactis, Lactobacillus helveticus, Lactobacillus delbrueckii subsp.bulgaricus and Lactobacillus acidophilus. Other specific bacteria within this definition include bacteria of the family Bifidobacteriaceae, for example from the genus Bifidobacterium, for example strains of bifidobacterium animalis or bifidobacterium longum, bifidobacterium adolescentis, bifidobacterium bifodum, bifidobacterium breve, bifidobacterium catenulatum, bifidobacterium infantus, or from the genus Lactobacillus, for example L. sakei, L. amylovorus, L. delbrueckii subsp. Lactis and L. helveticus.

This definition of host cell also includes the Agaricus strain, e.g.

A. bisporus; Ascovaginospora; Aspergillus, for example A. niger, A. awamori, A. foetidus, A. japonicus, A. oryzae; Candida; Chaetomium; Chaetomastia; Dictyostelium, for example D. discoideum; Kluveromyces, for example K. fragilis, K. lactis; Mucor, eg M. javanicus, M. mucedo, M. subtilissimus; Neurospora, eg N. crassa; Rhizomucor, for example R. pusillus; Rhizopus, for example R. arrhizus, R. japonicus, R. stolonifer; Sclerotinia, for example S. libertiana; Torula; Torulopsis; Trichophyton, for example T. rubrum; Whetzelinia, for example W. sclerotiorum; Bacillus, for example B. coagulans, B. circulans, B. megaterium,

B. novalis, B. subtilis, B. pumilus, B. stearothermophilus, B. thuringiensis; Bifidobacterium, for example B. Iongum, B. bifidum, B. animalis; Chryseobacterium; Citrobacter, for example C. jreundii; Clostridium, for example C. perfringens; Diplodia, for example D. gossypina; Enterobacter, for example E. aerogenes, E. cloacae Edwardsiella, E. tarda; Erwinia, for example E. herbicola; Escherichia, for example E. coli; Klebsiella, for example K. pneumoniae; Miriococcum; Myrothesium; Mucor; Neurospora, eg N. crassa; Proteus, for example P. vulgaris; Providencia, eg P. stuartii; Pycnoporus, for example Pycnoporus cinnabarinus, Pycnoporus sanguineus; Ruminococcus, for example R. torques; Salmonella, for example S. typhimurium; Serratia, for example S. liquefasciens, S. marcescens; Shigella, for example S. flexneri; Streptomyces, for example S. antibioticus, S. castaneoglobisporus, S. violeceoruber; Trametes; Trichoderma, for example T. reesei, T. viride; Yersinia, for example Y. enterocolitica.

Specific embodiments of the invention

As described above, the present invention relates to a peptide exhibiting beta-galactosidase enzymatic activity, which has the amino acid sequence represented by SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 , 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, or their amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions .

Accordingly, in one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 1, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7 , 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta enzymatic activity -galactosidase, which has the amino acid sequence represented by SEQ ID NO: 2, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 3, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 4, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity which has the amino acid sequence represented by SEQ ID NO: 5, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 6, or the amino acid a sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 substitutions, additions or deletions of amino acids. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 7, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 8, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 9, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity which has the amino acid sequence represented by SEQ ID NO: 10, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 11, or the amino acid a sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 substitutions, additions or deletions of amino acids. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 12, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity which has the amino acid sequence represented by SEQ ID NO: 13, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 14, or the amino acid a sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 substitutions, additions or deletions of amino acids. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 15, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity which has the amino acid sequence represented by SEQ ID NO: 16, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 17, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 18, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 19, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 20, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity which has the amino acid sequence represented by SEQ ID NO: 21, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 22, or the amino acid a sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 substitutions, additions or deletions of amino acids. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 23, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 24, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 25, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 26, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 27, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 28, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 29, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 30, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity which has the amino acid sequence represented by SEQ ID NO: 31, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity that has the amino acid sequence represented by SEQ ID NO: 32, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions. In one embodiment, the present invention provides a peptide exhibiting beta-galactosidase enzymatic activity which has the amino acid sequence represented by SEQ ID NO: 33, or an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 8 described herein, at a temperature of about 4°C and a pH of 6.7, wherein the activity exceeds the activity of the specified beta-galactosidase enzyme SEQ ID NO: 34 or SEQ ID NO: 35 by at least about 20%, for example, at least about 30%, for example, at least about 40%, for example , at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example , at least about 100%, for example at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of about 4°C and a pH of 6.7, and the activity exceeds the activity defined by SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase enzyme by at least about 20%, for example at least about 30%, for example at least about 40%, for example at least about 50%, for example at least by at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example, at least by at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%. In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of about 4°C and a pH of 6.7, where this activity is higher than about 2, for example, higher than about 4, e.g., higher than about 6, e.g., higher than about 8, e.g., higher than about 10, e.g., higher than about 12, e.g., higher than about 14, e.g., higher than about 16 µmol of glucose formed per second per µmol of enzyme.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 10 described herein, at a temperature of about 4°C and a pH of 5.5, at this activity exceeds the activity of the specified beta-galactosidase enzyme SEQ ID NO: 34 or SEQ ID NO: 35 by at least about 20%, for example, at least about 30%, for example, at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of about 4°C and a pH of 5.5, and the activity exceeds the activity defined by SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase enzyme by at least about 20%, for example at least about 30%, for example at least about 40%, for example at least about 50%, for example at least by at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example, at least by at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%. In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in μmol of glucose produced per second per μmol of enzyme at a temperature of about 4°C and a pH of 5.5, where this activity is greater than about 1, e.g. than about 2, for example, higher than about 3, for example, higher than about 4, for example, higher than about 5, for example, higher than about 6, for example, higher than about 7, for example, higher than about 8 µmol of glucose produced per second per µmol of enzyme.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 13 described herein, at a temperature of about 4°C and a pH of 4.5, at this activity exceeds the activity of the specified beta-galactosidase enzyme SEQ ID NO: 34 or SEQ ID NO: 35 by at least about 20%, for example, at least about 30%, for example, at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of about 4°C and a pH of 4.5, and the activity exceeds the activity defined by SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase enzyme by at least about 20%, for example at least about 30%, for example at least about 40%, for example at least about 50%, for example at least by at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example, at least by at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%. In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of about 4°C and a pH of 4.5, where this activity is greater than about 0.5, for example, higher than about 1.0, e.g., higher than about 1.5, e.g., higher than about 2.0, e.g., higher than about 2.5, e.g., higher than about 3.0, e.g. higher than about 3.5, for example, higher than about 4.0 µmol of glucose produced per second per µmol of enzyme.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 9 described herein, at a temperature of approximately 43°C and a pH of 6.7, at this activity exceeds the activity of the specified beta-galactosidase enzyme SEQ ID NO: 34 or SEQ ID NO: 35 by at least about 20%, for example, at least about 30%, for example, at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of approximately 43°C and a pH of 6.7, and the activity exceeds the activity defined by SEQ ID NO:34 or SEQ ID NO:35 beta-galactosidase enzyme by at least about 20%, for example at least about 30%, for example at least about 40%, for example at least about 50%, for example at least by at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example, at least by at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%. In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of about 43°C and a pH of 6.7, where this activity is higher than about 10, for example, higher than about 20, e.g., higher than about 40, e.g., higher than about 60, e.g., higher than about 80, e.g., higher than about 100, e.g., higher than about 120, e.g., higher than about 140, for example, is higher than about 160 µmol of glucose produced per second per µmol of enzyme.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in μmol of glucose produced per second per μmol of enzyme under the conditions described in Example 12 described herein, at a temperature of about 43°C and a pH of 5.5, at this activity exceeds the activity of the specified beta-galactosidase enzyme SEQ ID NO: 34 or SEQ ID NO: 35 by at least about 20%, for example, at least about 30%, for example, at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of about 43°C and a pH of 5.5, and the activity exceeds the activity defined by SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase enzyme by at least about 20%, for example at least about 30%, for example at least about 40%, for example at least about 50%, for example at least by at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example, at least by at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%. In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of about 43°C and a pH of 5.5, where this activity is higher than about 10, for example, higher than about 20, e.g., higher than about 25, e.g., higher than about 35, e.g., higher than about 40, e.g., higher than about 45, e.g., higher than about 50, e.g., higher than about 55, for example, is higher than about 60 µmol of glucose produced per second per µmol of enzyme.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 15 described herein, at a temperature of approximately 43°C and a pH of 4.5, at this activity exceeds the activity of the specified beta-galactosidase enzyme SEQ ID NO: 34 or SEQ ID NO: 35 by at least about 20%, for example, at least about 30%, for example, at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of approximately 43°C and pH 4.5, and the activity exceeds the activity defined by SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase enzyme by at least about 20%, for example at least about 30%, for example at least about 40%, for example at least about 50%, for example at least by at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example, at least by at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%. In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of about 43°C and a pH of 4.5, where this activity is greater than about 1, e.g. than about 2, for example, higher than about 3, for example, higher than about 4, for example, higher than about 5, for example, higher than about 6, for example, higher than about 7, for example, higher than about 8, for example, higher than about 9, for example, higher than about 10, for example, higher than about 11, for example, higher than about 12 µmol of glucose produced per second per µmol of enzyme.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 6 described herein, at a temperature of approximately 37°C and a pH of 6.7, at this activity exceeds the activity of the specified beta-galactosidase enzyme SEQ ID NO: 34 or SEQ ID NO: 35 by at least about 20%, for example, at least about 30%, for example, at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of approximately 37°C and a pH of 6.7, and the activity exceeds the activity defined by SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase enzyme by at least about 20%, for example at least about 30%, for example at least about 40%, for example at least about 50%, for example at least by at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example, at least by at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%. In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of about 37°C and a pH of 6.7, where this activity is higher than about 10, for example, higher than about 20, e.g., higher than about 30, e.g., higher than about 40, e.g., higher than about 50, e.g., higher than about 60, e.g., higher than about 70, e.g., higher than about 80, for example, higher than about 90, for example, higher than about 100, for example, higher than about 110, for example, higher than about 120, for example, higher than about 130 µmol of glucose produced per second per µmol of enzyme.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 11 described herein, at a temperature of approximately 37°C and a pH of 5.5, at this activity exceeds the activity of the specified beta-galactosidase enzyme SEQ ID NO: 34 or SEQ ID NO: 35 by at least about 20%, for example, at least about 30%, for example, at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in μmol of glucose produced per second per μmol of enzyme at a temperature of approximately 37°C and pH 5.5, and the activity exceeds the activity defined by SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase enzyme by at least about 20%, for example at least about 30%, for example at least about 40%, for example at least about 50%, for example at least by at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example, at least by at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%. In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of about 37°C and a pH of 5.5, where this activity is higher than about 5, for example, higher than about 10, for example, higher than about 15, for example, higher than about 20, for example, higher than about 25, for example, higher than about 30, for example, higher than about 35, for example, higher than about 40, for example, higher than about 45, for example, higher than about 50, for example, higher than about 55, for example, higher than about 60 µmol of glucose produced per second per µmol of enzyme.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 14 described herein, at a temperature of about 37°C and a pH of 4.5, at this activity exceeds the activity of the specified beta-galactosidase enzyme SEQ ID NO: 34 or SEQ ID NO: 35 by at least about 20%, for example, at least about 30%, for example, at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%.

In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in μmol of glucose produced per second per μmol of enzyme at a temperature of approximately 37°C and pH 4.5, and the activity exceeds the activity defined by SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase enzyme by at least about 20%, for example at least about 30%, for example at least about 40%, for example at least about 50%, for example at least by at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example, at least by at least about 200%, for example at least about 300%, for example at least about 400%, for example at least about 500%. In some embodiments of the invention, the peptide in accordance with the invention has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme at a temperature of about 37°C and a pH of 4.5, where this activity is higher than about 3, for example, higher than about 6, e.g., higher than about 7, e.g., higher than about 8, e.g., higher than about 9, e.g., higher than about 10, e.g., higher than about 11, e.g., higher than about 12, for example, higher than about 13, for example, higher than about 14, for example, higher than about 15, for example, higher than about 16, for example, higher than about 18 µmol of glucose produced per second per µmol of enzyme.

In some embodiments of the invention, the peptide in accordance with the invention is derived from bacteria of the genus Bifidobacterium, such as Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium longum, or from the genus Lactobacillus, such as L. sakei, L. amylovorus, L. delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, L. delbrueckii subsp. Indicus, L. crispatus, L. reuteri, L. helveticus, or from Streptococcus thermophilus.

In some embodiments of the invention, the peptide in accordance with the invention exhibits galactose inhibition of less than 60%, for example, less than 55%, for example, less than 50%, for example, less than about 45%, for example, less than about 40%.

As described above, in part, the present invention relates to a method for producing a dairy product, comprising the steps of a) producing a lactose-containing milk-based substrate; b) adding to said lactose-containing milk-based substrate a peptide exhibiting beta-galactosidase activity and having the amino acid sequence represented by SEQ ID NO: 1-33, or a sequence with at least 80% identity to any one of these sequences; and c) treating said milk-based substrate with said peptide exhibiting beta-galactosidase activity.

In some embodiments of the present invention, the peptide is derived from any one of the genus Bifidobacterium, such as Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium longum, or from the genus Lactobacillus, such as L. sakei, L. amylovorus , L. delbrueckii subsp. bulgaricus, L. delbrueckii subsp.lactis, L. delbrueckii subsp. Indicus, L. crispatus, L. reuteri, L. helveticus, or from Streptococcus thermophilus.

In some embodiments according to the present invention, step c) occurs at a pH in the range of 3-10, for example in the range of 3-9, for example in the range of 3-8, for example in the range of 3-7, for example in the range of 3- 6, for example, in the range 3-5, for example, in the range 3-4, for example, in the range 4-10, for example, in the range 4-9, for example, in the range 4-8, for example, in the range 4-7, for example, in the range 4-6, for example, in the range 4-5, for example, in the range 5-10, for example, in the range 5-9, for example, in the range 5-8, for example, in the range 5-7, for example, in the range of 5-6, for example in the range of 6-10, for example in the range of 6-9, for example in the range of 6-8, for example in the range of 6-7.

In some embodiments in accordance with the present invention, step c) or part of step c) occurs at a temperature of no more than about 25°C, for example, no more than about 20°C, for example, no more than about 18°C, for example, not more than about 16°C, for example, no more than about 14°C, for example, no more than about 12°C, for example, no more than about 10°C, for example, no more than about 8°C, for example, not more than about 7°C, for example, not more than about 6°C, for example, not more than about 5°C, for example, not more than about 4°C, for example, not more than about 3°C, for example, not more than about 2°C.

In some embodiments in accordance with the present invention, step c) or part of step c) occurs at a temperature of at least about 25°C, for example, at least about 30°C, for example, at least about 35°C, for example, at least about 40°C, for example at least about 45°C, for example at least about 50°C, for example at least about 55°C, for example at least about 60°C, for example at least about 65°C, for example at least about 70°C, for example at least about 75°C, for example at least about 80°C, for example at least about 85°C, for example at least about 90°C, for example at least about 95°C, for example at least about 100°C, for example at least about 110°C, for example at least about 120°C, for example at least about 130°C, for example at least about 120°C, for example at least about 130°C, for example at least about 135°C, for example at least about 140°C.

In some embodiments in accordance with the present invention, the dairy product is selected from the group consisting of lactose-free milk, low-lactose milk, yogurt, cheese, fermented milk products, dietary supplement and probiotic dietary products.

In some embodiments of the present invention, the milk-based substrate is selected from fresh milk or raw milk obtained directly from a pasteurization step, milk obtained directly from an ultra-pasteurization (UHT) step, or milk obtained directly from a fermentation step.

In some embodiments of the present invention, the galactose inhibition of the peptide used is less than 60%, such as less than 55%, such as less than 50%, such as less than about 45%, such as less than about 40%.

In some embodiments in accordance with the present invention, the dairy product is a fermented milk product, and said step b) is performed during or before fermentation.

In some embodiments of the present invention, the method does not require the addition of additional enzyme after fermentation.

In some embodiments according to the present invention, the dairy product is a fermented milk product, and said step b) is performed immediately after fermentation.

In some embodiments according to the present invention, the dairy product is fresh milk, and said step b) is performed before, in combination with, or immediately after the fermentation step.

In some embodiments in accordance with the present invention, the dairy product is ultra-pasteurized (UHT) milk, and said step b) is performed before, in combination with, or immediately after the UHT step.

In some embodiments according to the present invention, step c) is started at a temperature of from 40°C to 100°C, for example, at a temperature from 50°C to 100°C, for example, at a temperature from 60°C to 100°C, for example , at a temperature from 70°C to 100°C, for example, at a temperature from 80°C to 100°C, for example, at a temperature from 40°C to 90°C, for example, at a temperature from 40°C to 80°C , for example, at a temperature from 40°C to 70°C, for example, at a temperature from 40°C to 60°C, for example, at a temperature from 40°C to 50°C.

In some embodiments of the present invention, the peptide, when hydrolyzing lactose in a milk-based substrate, has a ratio of lactase activity to transgalactosylase activity greater than 1:1.

In some embodiments of the present invention, upon completion of step c), less than 80% of the lactose is hydrolyzed, and more than 90% of the lactose is hydrolyzed after a week.

Numbered embodiments of the invention:

1. A peptide exhibiting beta-galactosidase enzymatic activity, which has the amino acid sequence represented by SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 , 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, or enzymatically active fragments thereof, or the amino acid sequence of any of them , having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions.

2. A dimeric peptide exhibiting beta-galactosidase enzymatic activity, which consists of two peptides having the amino acid sequence represented by SEQ ID NO: 2 and 3; 5 and 6; 20 and 21; 23 and 24; 26 and 27; or 28 and 29, or enzymatically active fragments thereof, or the amino acid sequence of any of them having not more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acid substitutions, additions or deletions.

3. A peptide or dimeric peptide according to embodiments 1 or 2, which has beta-galactosidase activity measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 8 described herein, at a temperature of about 4°C and a pH of 6.7, wherein the activity exceeds the activity of the specified beta-galactosidase enzyme by at least about 20%, e.g., at least about 30%, e.g., at least about 30%, e.g. by about 40%, for example, by at least about 50%, for example, by at least about 60%, for example, by at least about 70%, for example, by at least about 80%, for example, by at least by about 90%, for example, at least about 100%, for example, at least about 200%, for example, at least about 300%, for example, at least about 400%, for example, at least by about 500%.

4. A peptide or dimeric peptide according to any of embodiments 1-3, which has beta-galactosidase activity measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 10 described herein, at a temperature of about 4 °C and a pH of 5.5, wherein the activity exceeds the activity of the specified enzyme SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase by at least about 20%, for example, at least about 30%, for example, according to by at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, by at least about 90%, for example, at least about 100%, for example, at least about 200%, for example, at least about 300%, for example, at least about 400%, for example, by at least by about 500%.

5. A peptide or dimeric peptide according to any of embodiments 1-

4, which has beta-galactosidase activity, measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 13 described herein, at a temperature of approximately 4° C. and a pH of 4.5, and the activity exceeds the activity of a certain SEQ ID NO: 34 or SEQ ID NO: 35 of beta-galactosidase enzyme by at least about 20%, for example at least about 30%, for example at least about 40%, for example at least about 40% 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, at least about 90%, for example, at least about 100%, for example, at least about 200%, for example, at least about 300%, for example, at least about 400%, for example, at least about 500%.

6. A peptide or dimeric peptide according to any of embodiments 1-5, which has beta-galactosidase activity measured in µmol of glucose produced per second per µmol of enzyme under the conditions set forth in Example 9 herein at a temperature of approximately 43 °C and a pH of 6.7, wherein the activity exceeds the activity of the specified enzyme SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase by at least about 20%, for example, at least about 30%, for example, according to by at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, by by at least about 90%, for example, at least about 100%, for example, at least about 200%, for example, at least about 300%, for example, at least about 400%, for example, at least by about 500%.

7. A peptide or dimeric peptide according to any of embodiments 1-6, which has beta-galactosidase activity measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 12 described herein, at a temperature of approximately 43 °C and a pH of 5.5, wherein the activity exceeds the activity of the specified enzyme SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase by at least about 20%, for example, at least about 30%, for example, according to by at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, by at least about 90%, for example, at least about 100%, for example, at least about 200%, for example, at least about 300%, for example, at least about 400%, for example, by at least by about 500%.

8. A peptide or dimeric peptide according to any of embodiments 1-7, which has beta-galactosidase activity measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 15 described herein, at a temperature of approximately 43 °C and a pH of 4.5, wherein the activity exceeds the activity of the specified enzyme SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase by at least about 20%, for example, at least about 30%, for example, according to by at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, by at least about 90%, for example, at least about 100%, for example, at least about 200%, for example, at least about 300%, for example, at least about 400%, for example, by at least by about 500%.

9. A peptide or dimeric peptide according to any of embodiments 1-8, which has beta-galactosidase activity measured in µmol of glucose produced per second per µmol of enzyme under the conditions described in Example 6 described herein, at a temperature of approximately 37 °C and a pH of 6.7, wherein the activity exceeds the activity of the specified enzyme SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase by at least about 20%, for example, at least about 30%, for example, according to by at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, by at least about 90%, for example, at least about 100%, for example, at least about 200%, for example, at least about 300%, for example, at least about 400%, for example, by at least by about 500%.

10. A peptide or dimeric peptide according to any of embodiments 1-9, which has beta-galactosidase activity measured in µmol of glucose produced per second per µmol of enzyme under the conditions set forth in Example 11 described herein, at a temperature of approximately 37 °C and a pH of 5.5, wherein the activity exceeds the activity of the specified enzyme SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase by at least about 20%, for example, at least about 30%, for example, according to by at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, by at least about 90%, for example, at least about 100%, for example, at least about 200%, for example, at least about 300%, for example, at least about 400%, for example, by at least by about 500%.

11. A peptide or dimeric peptide according to any of embodiments 1-10, which has beta-galactosidase activity measured in μmol of glucose produced per second per μmol of enzyme under the conditions set forth in Example 14 described herein, at a temperature of approximately 37 °C and a pH of 4.5, wherein the activity exceeds the activity of the specified enzyme SEQ ID NO: 34 or SEQ ID NO: 35 beta-galactosidase by at least about 20%, for example, at least about 30%, for example, according to by at least about 40%, for example, at least about 50%, for example, at least about 60%, for example, at least about 70%, for example, at least about 80%, for example, by by at least about 90%, for example, at least about 100%, for example, at least about 200%, for example, at least about 300%, for example, at least about 400%, for example, at least by about 500%.

12. The peptide or dimeric peptide according to any of embodiments 1-11, obtained from bacteria of the genus Bifidobacterium, such as Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium longum, or from the genus Lactobacillus, such as L. sakei, L. amylovorus, L. delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, L. delbrueckii subsp. Indicus, L. crispatus, L. reuteri, L. helveticus, or from Streptococcus thermophilus.

13. Peptide or dimeric peptide according to any of embodiments 1-

12, wherein said peptide or dimeric peptide exhibits galactose inhibition of less than 60%, such as less than 55%, such as less than 50%, such as less than about 45%, such as less than about 40%.

14. A nucleotide sequence encoding a peptide or dimeric peptide as defined in any of embodiments 1-13.

15. A host cell containing the nucleotide sequence as defined in embodiment 14.

16. A method of producing a peptide or dimeric peptide, as defined in any of embodiments 1-13, exhibiting beta-galactosidase enzymatic activity, where the method comprises expressing in a suitable host cell a vector containing a nucleotide sequence in accordance with the invention; and purifying said peptide or dimeric peptide from expression products of said host cell.

17. A method of reducing the lactose content in a composition containing lactose, such as dairy products, comprising the step of contacting said composition with a peptide or dimeric peptide exhibiting beta-galactosidase enzymatic activity, wherein the peptide has the amino acid sequence represented by SEQ ID NO: 1- 33; or where the dimeric peptide consists of two peptides having the amino acid sequence represented by SEQ ID NO: 2 and 3, 5 and 6, 20 and 21, 23 and 24, 26 and 27, or 28 and 29; or enzymatically active fragments thereof, or any sequence with at least 80% sequence identity to any of the above sequences; or with a host cell expressing any of these peptides at a pH in the range of 3-10 and at a temperature in the range of 0°C-140°C.

18. The method according to embodiment 17, wherein said composition is a dairy product selected from the group consisting of lactose-free milk, low-lactose milk, yogurt, cheese, fermented milk products, dietary supplement and probiotic dietary products.

19. The method according to any one of embodiments 17-18, wherein the host cell is any cell selected from bacteria of the genus Bifidobacterium, such as Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium longum, or from the genus Lactobacillus, such as L. sakei, L. amylovorus, L. delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, L. delbrueckii subsp. Indicus, L. crispatus, L. reuteri, L. helveticus, or from Streptococcus thermophilus.

20. The method according to any of embodiments 17-19, wherein the lactose concentration is reduced to less than about 1%, such as less than about 0.1% or less, such as less than about 0.01%.

21. Use of a peptide or dimeric peptide exhibiting beta-galactosidase enzymatic activity, which has the amino acid sequence represented by SEQ ID NO: 1-33, or where the dimeric peptide consists of two peptides having the amino acid sequence represented by SEQ ID NO: 2 and 3 , 5 and 6, 20 and 21, 23 and 24, 26 and 27, or 28 and 29; or a sequence with at least 80% sequence identity to any of the above sequences; or a host cell expressing any of the above peptide or dimeric peptide to produce a lactose-reduced dairy product.

22. Use according to embodiment 21, wherein said dairy product is selected from the group consisting of lactose-free milk, low-lactose milk, yogurt, cheese, fermented milk products, dietary supplement and probiotic dietary products.

23. Use according to any of embodiments 21-22, wherein said host cell is any cell selected from bacteria of the genus Bifidobacterium, such as Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium longum, or from the genus Lactobacillus , such as L. sakei, L. amylovorus, L. delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, L. delbrueckii subsp. Indicus, L. crispatus, L. reuteri, L. helveticus, or from Streptococcus thermophilus.

24. A method for producing a dairy product, including the stages:

a) providing a lactose-containing milk-based substrate;

b) adding to said lactose-containing milk-based substrate a peptide or dimeric peptide exhibiting beta-galactosidase activity, wherein the peptide has the amino acid sequence represented by SEQ ID NO: 1-33; or where the dimeric peptide consists of two peptides having the amino acid sequence represented by SEQ ID NO: 2 and 3, 5 and 6, 20 and 21, 23 and 24, 26 and 27, or 28 and 29; or a sequence with at least 80% sequence identity to any of the above sequences; And

c) treating said milk-based substrate with said peptide or dimeric peptide exhibiting beta-galactosidase activity.

25. The method according to embodiment 24, wherein said peptide or dimeric peptide is derived from any one bacterium of the genus Bifidobacterium, such as Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium longum, or from the genus Lactobacillus, such as L. sakei , L. amylovorus, L. delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, L. delbrueckii subsp. Indicus, L. crispatus, L. reuteri, L. helveticus, or from Streptococcus thermophilus.

26. The method according to any of embodiments 24-25, wherein step (c) is carried out at a pH in the range of 3-10, for example in the range of 3-9, for example in the range of 3-8, for example in the range of 3-7 , for example, in the range 3-6, for example, in the range 3-5, for example, in the range 3-4, for example, in the range 4-10, for example, in the range 4-9, for example, in the range 4-8, for example , in the range 4-7, for example, in the range 4-6, for example, in the range 4-5, for example, in the range 5-10, for example, in the range 5-9, for example, in the range 5-8, for example, in range 5-7, for example, in the range 5-6, for example, in the range 6-10, for example, in the range 6-9, for example, in the range 6-8, for example, in the range 6-7.

27. The method according to any of embodiments 24-26, wherein step c) or part of step (c) is carried out at a temperature of no more than about 25°C, for example, no more than about 20°C, for example, no more than about 18°C, for example, not more than about 16°C, for example, not more than about 14°C, for example, not more than about 12°C, for example, not more than about 10°C, for example, not more than about 8°C, for example, not more than about 7°C, for example, not more than about 6°C, for example, not more than about 5°C, for example, not more than about 4°C, for example, not more than about 3°C, for example, no more than about 2°C.

28. The method according to any of embodiments 24-27, wherein step (c) or part of step (c) is carried out at a temperature of at least about 25°C, for example at least about 30°C, for example at least about 35°C, for example at least about 40°C, for example at least about 45°C, for example at least about 50°C, for example at least about 55°C, for example at least about 60°C, for example at least about 65°C, for example at least about 70°C, for example at least about 75°C, for example at least about 80°C, for example at least about 85°C, for example, at least about 90°C, for example, at least about 95°C, for example, at least about 100°C, for example, at least about 110°C, for example, at least about 120°C, for example, at least about 130°C, for example, at least about 120°C, for example, at least about 130°C, for example, at least about 135°C, for example, at least approximately 140°C.

29. Use in accordance with any of embodiments 24-28, wherein said dairy product is selected from the group consisting of lactose-free milk, low-lactose milk, yogurt, cheese, fermented milk products, dietary supplement and probiotic dietary products.

30. The method according to any one of embodiments 24-29, wherein said milk-based substrate is selected from fresh milk or raw milk obtained directly from a pasteurization step, milk obtained directly from an ultra-pasteurization (UHT) step, or milk obtained directly from fermentation stages.

31. The method according to any of embodiments 24-30, wherein the galactose inhibition of said peptide or dimeric peptide is less than 60%, for example less than 55%, for example less than 50%, for example less than about 45%, for example less than approximately 40%.

32. The method according to any of embodiments 24-31, wherein said dairy product is a fermented milk product and said step b) is performed during or before fermentation.

33. The method according to embodiment 32, which does not require the addition of additional enzyme after fermentation.

34. The method according to any one of embodiments 24-31, wherein said dairy product is a fermented milk product and said step (b) is carried out immediately after fermentation.

35. The method according to any one of embodiments 24-31, wherein said dairy product is fresh milk and said step (b) is carried out before, in combination with or immediately after the pasteurization step.

36. The method according to any one of embodiments 24-31, wherein said dairy product is ultra-pasteurized (UHT) milk, and said step (b) is carried out before, in combination with, or immediately after the UHT step.

37. The method in accordance with any of embodiments 24-36, in which step c) is started at a temperature of from 40°C to 100°C, for example at a temperature from 50°C to 100°C, for example at a temperature from 60°C C to 100°C, for example, at a temperature from 70°C to 100°C, for example, at a temperature from 80°C to 100°C, for example, at a temperature from 40°C to 90°C, for example, at a temperature from 40°C to 80°C, for example, at a temperature from 40°C to 70°C, for example, at a temperature from 40°C to 60°C, for example, at a temperature from 40°C to 50°C.

38. The method according to any one of embodiments 24-37, wherein the peptide or dimeric peptide, when hydrolyzing lactose in a milk-based substrate, has a ratio of lactase activity to transgalactosylase activity of greater than 1:1.

39. The method according to any one of embodiments 24-38, wherein at the end of step (c) less than 80% of the lactose is hydrolyzed, and more than 90% of the lactose is hydrolyzed after a week.

40. A dairy product produced by the process as defined in any one of embodiments 24-39.

41. A food product, such as a dairy product, containing a peptide or dimeric peptide exhibiting beta-galactosidase enzymatic activity, where the peptide has the amino acid sequence represented by SEQ ID NO: 1-33, or where the dimeric peptide consists of two peptides having the amino acid sequence represented by SEQ ID NO: 2 and 3, 5 and 6, 20 and 21, 23 and 24, 26 and 27, or 28 and 29; or a sequence with at least 80% sequence identity to any of the specified sequences.

42. A food product, such as a dairy product, containing a host cell expressing a peptide or dimeric peptide exhibiting beta-galactosidase enzymatic activity, where the peptide has the amino acid sequence represented by SEQ ID NO: 1-33, or where the dimeric peptide consists of two peptides having the amino acid sequence represented by SEQ ID NO: 2 and 3, 5 and 6, 20 and 21, 23 and 24, 26 and 27, or 28 and 29; or a sequence with at least 80% sequence identity to any one of the specified sequences.

43. The food product according to embodiment 42 selected from beverages, baby foods, cereals, breads, cookies, confectionery, cakes, food additives, dietary supplements, probiotic foods, prebiotic foods, animal feeds , poultry feed and medicine or a dairy product selected from the group consisting of lactose-free milk, low-lactose milk, powdered milk, baby milk, yogurt, ice cream, cheese, fermented milk products, dietary supplement and probiotic dietary food products.

SEQUENCES

Examples

General materials and methods

Molecular cloning and genetic techniques

Restriction enzyme digestion, ligation, transformation, and other standard molecular biology procedures were based on methods described in the literature (Maniatis et al. "Molecular cloning: a laboratory manual, 2nd edition" Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY , 1989; Sambrook and Russell "Molecular Cloning: A Laboratory Manual, 3rd edition" Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY 2001; Miller "Experiment in molecular genetics" Cold Spring Harbor Laboratory Press, 1972); or those suggested by the manufacturer. PNR was carried out in a DNA thermal cycler manufactured by Bio-Rad, USA. DNA sequencing was performed by LGC, Berlin, Germany. Proteins were analyzed by polyacrylamide gel electrophoresis (PAGE) under denaturing conditions using sodium dodecyl sulfate on gels containing 10% SDS (Mini-PROTEAN® TGX stain-free™ gel, Biorad, USA). Protein concentrations were determined using the bicinchoninic acid (BCA) method following the protocol supplied with the reagent kit.

Bacterial strains, plasmids and growth conditions

Escherichia coli strain TOP10 (Invitrogen) was used for cloning and isolation of plasmids. To obtain the recombinant protein, we used the E. coli BW25113 strain with beta-galactosidase deficiency (Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ-rph-1, Δ(rhaD-rhaB)568, hsdR514) ( Datsenko KA, Wanner BL; 2000, Proc Natl Acad Sci U.S.A. 97: 6640-6645) in combination with the pBAD/His vector obtained from Invitrogen™ Life Technologies Corporation Europe BV.

Growth media for protein expression

For the production of recombinant protein, a 2xPY nutrient medium containing 16 g/l peptone BD BBL™ Phyton TM, 10 g/l yeast extract, 5 g/l NaCl was used. The growth medium contained ampicillin supplement (100 μg/ml) to maintain the plasmid. Protein production was initiated by adding 0.05% arabinose to the culture medium.

Example 1. Construction of an expression vector for the production of lactases

Genomic DNA of lactic acid bacteria or bifidobacteria was isolated using a commercial genomic DNA isolation reagent kit following the supplier's protocol (DNeasy, Qaigen, Germany). The lactase gene was amplified by polymerase chain reaction (PCR) using two synthetic primers, purified genomic DNA as a biomass source, and reagents supplied in the Phusion U Hot start DNA polymerase kit (Thermo Scientific, USA). The lactase gene was cloned into the start codon of the pBAD/His expression vector using the USER cloning method (Nour-Eldin HH, Geu-Flores F, Halkier BA, Plant Secondary Metabolism Engineering, Methods in Molecular Biology, 643; 2010) resulting in an expression construct . When using the USER method, long complementary overhangs are generated in both the PCR product and the delivery vector. These protruding ends can be annealed to each other to form a stable hybridization product that is used to transform E. coli without ligation. To create overhangs in the PCR product, a single deoxyuridine residue was included in the upstream region of each target DNA amplification primer. The lactase gene was amplified using a forward primer and reverse primer containing a uridine at position 9 (shown in bold) followed by a lactase gene sequence. In parallel, the vector DNA was amplified by PCR using a direct pair and reverse primer containing a single deoxyuracil residue at position 9 (in bold) followed by a vector DNA sequence. PCR products were purified using a commercial PCR product purification kit (Qiagen, Denmark). Purified PCR products (lactase gene and vector DNA) were mixed in equimolar amounts and incubated with a commercially available USER enzyme mixture (New England Biolabs, USA) following the supplier's protocol. These enzymes remove the uracil residue as well as the short fragment upstream of the uridine, resulting in complementary overhangs in the PCR products. These complementary overhangs are annealed to each other to result in the lactase expression vector pBAD. Aliquots of this ligated mixture were transformed into chemically obtained competent E. coli TOP 10 cells. Transformants were selected at 37°C on Petri dishes with LB-Amp medium (LB; Luria-Bertani medium, Amp; 100 μg/ml ampicillin). The next day, PCR of individual colonies was carried out using a small biomass of transformants grown overnight, using primers for the vector (primer 1; and primer 2; PCR-positive clones of individual colonies were cultured in 5 ml of LB-Amp medium and plasmid DNA was isolated from the cells. The cloned lactase gene was sequenced to confirm that no additional mutations were introduced during the gene amplification process. The host expression strain BW25113 was transformed with plasmid DNA.

Example 2 Expression of lactases in an E. coli host expression strain The lactase enzyme was produced in E. coli BW25113 using the pBAD expression system. Freshly transformed E. coli BW25113 cells carrying plasmid DNA were collected from the LB-Amp plate using a sterile loop and used to inoculate 5 ml of LB-Amp medium. The overnight culture (200 μl) was used to inoculate 50 ml of 2x PY medium containing 100 μg/ml ampicillin into a 250 ml flask and placed in a shaker (Innova® 42). The culture was grown at 37°C at 220 rpm until an optical density at 600 nm (OD600) of 0.6-0.8 was achieved. Lactase expression was initiated by adding 0.05% arabinose to the culture medium, and the cells were cultured for an additional 16-20 hours at 18°C at 180 rpm. Cells were collected by centrifugation (5000 rpm, 10 min at 4°C) and stored at -20°C until further use.

Example 3 Protein Purification Using Immobilized Metal Affinity Chromatography

Cells from a 50-mL culture were thawed on ice and lysed using 10 mL of BugBuster® lysis buffer mixture (Novagen) containing 2 mg/mL lysozyme (Sigma Aldrich), 1 unit of benzonase (Sigma Aldrich), and 1× complete inhibitor cocktail protease (EDTA-free, Roche) by incubating cells at room temperature for 30 min. After 30 min, cell debris was removed by centrifugation at 16,000 rpm for 20 min at 4°C. The resulting supernatant was filtered through a filter with a pore diameter of 0.45 μm. A Ni-Sepharose gravity flow column (GE Healthcare) was prepared with 1 mL of suspension by washing with ethanol and water. The column was then equilibrated with a wash buffer of 50 mmol/L NaH ₂ PO ₄ , pH 8.0, containing 300 mmol/L NaCl and 20 mmol/L imidazole. The cell-free extract was applied to the column, and unbound proteins were eluted from the column. The column was washed with 20 ml of wash buffer, and the proteins retained on the column were eluted with 3.5 ml of 50 mmol/L NaH ₂ PO ₄ elution buffer, pH 8.0, containing 300 mmol/L NaCl and 250 mmol/L imidazole. The collected fractions were analyzed by SDS-PAGE on gels containing 10% acrylamide, and the fractions containing purified lactase enzymes were pooled together. Buffer was exchanged for storage buffer (50 mmol/L KH ₂ PO ₄ buffer solution, pH 7.0, containing 10 mmol/L NaCl, 1 mmol/L MgCl ₂ ) using a pre-packed PD-10 desalting gel column. filtration G-25 (GE Healthcare). Purified enzymes were stored at 4°C until further use.

Example 4 Protein Purification Using Gel Filtration Chromatography

Cells from a 50-mL culture were thawed on ice and lysed using 10 mL of BugBuster® lysis buffer mixture (Novagen) containing 2 mg/mL lysozyme, 1 unit of benzonase (Sigma Aldrich), and 1× complete protease inhibitor cocktail (no EDTA). , Roche), by incubating cells at room temperature (25°C) for 30 minutes. After 30 min, cell debris was removed by centrifugation at 16,000 rpm for 20 min at 4°C. The resulting supernatant was filtered through a filter with a pore diameter of 0.45 μm. The clarified cell-free extract was concentrated by filtration through a 30,000 Dalton cutoff filter (Vivaspin 20, GE Healthcare) following the supplier's protocol. A Sephadex G50 superfine gravity flow column (Pharmacia Chemicals, Sweden) was prepared with 1 g of column material prepared by boiling in 100 mL water for 1 h, cooled to room temperature. The column was prepared by applying 20 ml of the cooled suspension to a 30 ml filtration column. The column was washed with MilliQ water and equilibrated with wash buffer B (50 mmol/L NaH ₂ PO ₄ buffer, pH 7.0). 500 μl of concentrated supernatant was applied to the column and allowed to enter the stationary phase. The washing buffer (50 mmol/L NaH ₂ PO ₄ buffer solution, pH 7.0) was applied to the top of the column and the eluent fractions were collected separately. The collected fractions were analyzed on an SDS-PAGE gel containing 10% acrylamide. Protein fractions were pooled together and buffer exchanged for storage buffer (50 mmol/L KH ₂ PO ₄ buffer, pH 7.0, containing 10 mmol/L NaCl, 1 mmol/L MgCl ₂ ) using a desalting column as described. in the previous section. Purified enzymes were stored at 4°C until further use.

Example 5: Measuring protein concentration using the bicinchoninic acid (BCA) method

To determine protein concentration, the Pierce™ Bicinchoninic Acid (BCA) Protein Assay Kit (Thermo Fisher Scientific, Germany) was used following the protocol supplied with the kit.

Example 6 Determination of activity using purified enzymes on lactose as a substrate at pH 6.7 and temperature 37°C

To measure beta-galactosidase activity, purified lactases were diluted to 40-fold concentration in buffer A (buffer solution 50 mmol/L NaH ₂ PO ₄ pH 6.7 containing 100 µmol/L MgSO ₄ ). In a separate reaction, the diluted enzyme was incubated with a lactose solution prepared in buffer B (140 mmol/L lactose solution prepared in 100 mM sodium citrate buffer pH 6.7 containing 100 μmol/L MgSO ₄ ). The reaction mixture was prepared by mixing 13 μL of diluted enzyme and 37 μL of lactose solution in a PCR tube. The reaction mixture was incubated in a DNA thermal cycler with the following incubation parameters: reaction time 10 min at 37°C, enzyme inactivation 10 min at 95°C, cooling to 4°C. The reaction mixtures were stored at -20°C until further use. To determine the amount of glucose formed during the reaction, 10 μl of the reaction mixture was transferred into one well of a standard microtiter plate (Thermo Fischer Scientific, Denmark) containing 80 μl of buffer C (buffer solution 100 mmol/l NaH ₂ PO ₄ , pH 7.0 , containing glucose oxidase, 0.6 g/l (Sigma Aldrich); 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid diammonium salt), ABTS, 1.0 g/l (Sigma Aldrich); horseradish peroxidase , 0.02 g/L (Sigma Adrich)) and incubated at 30°C for 40 min. After 40 min, absorbance was determined at 610 nm using a FLUOStar Omega scanning UV plate reader (BMG Labtech, Germany). For calculations, optical density values from 0.1 to 1.5 were used; if the A610 nm value exceeded 1.5, the reaction mixture was diluted to a 10-fold concentration with buffer A. For each purified enzyme, reactions were performed in triplicate and the average of triplicate measurements was used for calculations. For normalization, protein purification performed on E. coli cells transformed with the pBAD/His vector without insert was used. A standard curve was generated using a known glucose concentration (0–2.5 mmol/L) and the slope of the curve was used to calculate the amount of glucose produced during the reaction. The maximum absorbance value for each lactase was used to determine the amount of micromoles (μmol) of glucose produced per second, described as 1 unit of activity with lactose at pH 6.7 at 37°C (Unit of Activity with Lactose; UAL-1). Specific activity at pH 6.7 at 37°C, described as SUAL-1, is defined as the amount of µmol of glucose produced per second per µmol of enzyme (µmol glucose/s/µmol enzyme) and is calculated by dividing UAL-1 by the protein concentration in µmol Under similar conditions, the specific activity of SEQ ID NO: 34 and SEQ ID NO: 35 was determined. High specific activity at pH 6.7 is highly desirable for the stability of the enzyme when used in fresh and fermented milk. Detailed results of determining the specific activity of enzymes at pH 6.7 at 37°C are described in Fig. 1.

Additionally, activity was described as the amount of micromoles of glucose produced per minute per milligram of added enzyme. The results are presented in Fig. 14.

Example 7 Determination of activity using purified enzymes in the presence of galactose at pH 6.7 and 37°C

Purified lactases were diluted to a 40-fold concentration in buffer A (buffer solution 50 mmol/L NaH ₂ PO ₄ pH 6.7, containing 100 µmol/L MgSO ₄ ). In separate reactions, diluted enzymes were incubated with buffer D (a solution of 140 mmol/L lactose and 140 mmol/L galactose prepared in 100 mM sodium citrate buffer pH 6.7 containing 100 μmol/L MgSO ₄ ). The reaction mixture consisted of 13 μl of diluted enzyme and 37 μl of buffer D in a PCR tube. The reaction mixture was incubated in a thermal cycler under the following incubation parameters (reaction time: 10 min at 37°C, enzyme inactivation: 10 min at 95°C, cooling: 4°C). The reaction mixtures were stored at -20°C until further use. To determine the amount of glucose formed during the reaction, 10 μl of the reaction mixture was transferred into one well of a standard microtiter plate (Thermo Fischer Scientific, Denmark) containing 80 μl of buffer C (buffer solution 100 mmol/l NaH ₂ PO ₄ , pH 7.0 , containing glucose oxidase, 0.6 g/l (Sigma Aldrich); 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid diammonium salt), ABTS, 1.0 g/l (Sigma Aldrich); horseradish peroxidase , 0.02 g/L (Sigma Adrich)) and incubated at 30°C for 40 min. After 40 min, absorbance was determined at 610 nm using a FLUOStar Omega UV plate reader (BMG Labtech, Germany). For calculations, optical density values from 0.1 to 1.5 were used; if the A610 nm value exceeded 1.5, the reaction mixture was diluted to a 10-fold concentration with buffer A. For each purified enzyme, reactions were performed in triplicate and the average of triplicate measurements was used for calculations. For normalization, protein purification performed on E. coli cells transformed with the pBAD/His vector without insert was used. A standard curve was generated using a known glucose concentration (0–2.5 mmol/L) and the slope of the curve was used to calculate the absorbance corresponding to 1 μmol of glucose produced during the reaction. To determine the amount of μmol of glucose produced per second, described as 1 unit of activity with galactose at pH 6.7 at 37°C (Unit Activity with Galactose; UAG), the maximum optical density value for each lactase was used. Specific activity at pH 6.7 at 37°C in the presence of galactose, described as SUAG, is defined as the amount of µmol of glucose produced per second per µmol of enzyme (µmol glucose/s/µmol enzyme) and is calculated by dividing UAG by the protein concentration in µmol .

The percentage of enzyme inhibition by galactose is calculated using the formula % inhibition = 100 * (A-B)/A, where A is the specific activity of enzymes with lactose at pH 6.7 at 37°C (SUAL), as described in example 6, and B means the specific enzyme activity in the presence of galactose at pH 6.7 at 37° C. (SUAG) as described in Example 7. Detailed results of % inhibition by galactose are described in FIG. 2 and Fig. 14. Reduced galactose inhibition is highly relevant for applications where low lactose concentrations are desired. Additionally, activity was described as the amount of micromoles of glucose produced per minute per milligram of enzyme added. The results are presented in Fig. 14.

Note: The relatively high standard deviations of galactose inhibition measurements are due to trace amounts of glucose impurities in the acquired galactose.

Example 8 Determination of activity using purified enzymes on lactose as a substrate at pH 6.7 and 4°C

Purified lactases were diluted to a 40-fold concentration in buffer A (buffer solution 50 mmol/L NaH ₂ PO ₄ pH 6.7, containing 100 µmol/L MgSO ₄ ). In a separate reaction, the diluted enzyme was incubated with a lactose solution prepared in buffer B (140 mmol/L lactose solution prepared in 100 mM sodium citrate buffer pH 6.7 containing 100 μmol/L MgSO ₄ ). The reaction mixture was prepared by mixing 13 μL of diluted purified enzyme and 37 μL of lactose solution in a PCR tube. The reaction mixture was incubated in a DNA thermal cycler using the following incubation parameters: reaction time 60 min at 4°C, enzyme inactivation 10 min at 95°C, storage at 4°C. The reaction mixtures were stored in a freezer at -20°C until further use. The amount of glucose produced during the reaction was determined following the protocol described in Example 6. To determine the amount of micromoles (μmol) of glucose produced per second, described as 1 unit of activity with lactose at pH 6.7 at 4°C (UAL-2 ), the maximum optical density value for each lactase was used. Specific activity at pH 6.7 at 4°C, described as SUAL-2, is defined as the amount of µmol of glucose produced per second per µmol of enzyme (µmol glucose/s/µmol enzyme) and is calculated by dividing UAL-2 by the protein concentration in µmol High specific activity at pH 6.7 at 4°C is highly desirable for lactose hydrolysis in fresh/pasteurized milk applications. Detailed results of determining the specific activity of enzymes at pH 6.7 at 4°C are described in Fig. 3. Additionally, activity was described as the amount of micromoles of glucose produced per minute per milligram of enzyme added. The results are presented in Fig. 14.

Example 9. Determination of activity using purified enzymes on lactose as a substrate at pH 6.7 and a temperature of 43°C

Purified lactases were diluted to a 40-fold concentration in buffer A (buffer solution 50 mmol/L NaH ₂ PO ₄ pH 6.7, containing 100 µmol/L MgSO ₄ ). In a separate reaction, the diluted enzyme was incubated with a lactose solution prepared in buffer B (140 mmol/L lactose solution prepared in 100 mM sodium citrate buffer pH 6.7 containing 100 μmol/L MgSO ₄ ). The reaction mixture was prepared by mixing 13 μL of diluted purified enzyme and 37 μL of lactose solution in a PCR tube. The reaction mixture was incubated in a DNA thermal cycler using the following incubation parameters: reaction time 10 min at 43°C, enzyme inactivation 10 min at 95°C, storage at 4°C. The reaction mixtures were stored in a freezer at -20°C until further use. The amount of glucose produced during the reaction was determined following the protocol described in Example 6. To determine the amount of micromoles (μmol) of glucose produced per second, described as 1 unit of activity with lactose at pH 6.7 at 43°C (UAL-3 ), the maximum optical density value for each lactase was used. Specific activity at pH 6.7 at 43°C, described as SUAL-3, is defined as the amount of µmol of glucose produced per second per µmol of enzyme (µmol glucose/s/µmol enzyme) and is calculated by dividing UAL-3 by the protein concentration in µmol High specific activity at pH 6.7 at 43°C is highly desirable for lactose hydrolysis in fermented milk applications. Detailed results of determining the specific activity of enzymes at pH 6.7 at 43°C are described in Fig. 4. Additionally, activity was described as the number of micromoles of glucose produced per minute per milligram of added enzyme. The results are presented in Fig. 14.

Example 10 Determination of activity using purified enzymes on lactose as a substrate at pH 5.5 and 4°C

Purified lactases were diluted to a 40-fold concentration in buffer A (buffer solution 50 mmol/L NaH ₂ PO ₄ pH 6.7, containing 100 µmol/L MgSO ₄ ). In a separate reaction, the diluted enzyme was incubated with a lactose solution prepared in buffer E (140 mmol/L lactose solution prepared in 100 mM sodium citrate buffer pH 5.5 containing 100 µmol/L MgSO ₄ . The reaction mixture was prepared by mixing 13 µl of diluted purified enzyme and 37 μl of lactose solution in a PCR tube. The reaction mixture was incubated in a DNA thermal cycler using the following incubation parameters: reaction time 60 min at 4°C, enzyme inactivation 10 min at 95°C, storage at 4°C. The substrate solution was prepared in the buffer pH 5.5, the enzyme solution had a pH of 6.7. To initiate the reaction, 13 μl of enzyme was added to 37 μl of the substrate solution. Thus, when these two buffers were mixed, the final pH of the reaction increased from 5.5 to 5.7. The reaction mixtures were stored in a freezer at -20°C until further use.To determine the amount of glucose formed during the reaction, 10 μl of the reaction mixture was transferred to one well of a standard microtiter plate containing 80 μl of buffer C and incubated at 30°C in for 40 minutes. After 40 min, absorbance was determined at 610 nm using a FLUOStar Omega scanning UV plate reader (BMG Labtech, Germany). For calculations, optical density values from 0.1 to 1.5 were used; if the A610 nm value exceeded 1.5, the reaction mixture was diluted to a 5-fold concentration with buffer A. For each purified enzyme, reactions were performed in triplicate and the average of triplicate measurements was used for calculations. The maximum absorbance value for each lactase was used to determine the amount of micromoles (μmol) of glucose produced per second, described as 1 unit of activity with lactose at pH 5.5 at 4°C (UAL-4). Specific activity at pH 5.5 at 4°C, described as SUAL-4, is defined as the amount of µmol of glucose produced per second per µmol of enzyme (µmol glucose/s/µmol enzyme) and is calculated by dividing UAL-4 by the protein concentration in µmol High specific activity at pH 5.5 at 4°C is relevant for the hydrolysis of lactose when used for fermented milk. Detailed results of determining the specific activity of enzymes at pH 5.5 at 4°C are described in Fig. 5. Additionally, activity was described as the amount of micromoles of glucose produced per minute per milligram of enzyme added. The results are presented in Fig. 15.

Example 11 Determination of activity using purified enzymes on lactose as substrate at pH 5.5 and temperature 37°C

Purified lactases were diluted to a 40-fold concentration in buffer A (buffer solution 50 mmol/L NaH ₂ PO ₄ pH 6.7, containing 100 µmol/L MgSO ₄ ). In a separate reaction, the diluted enzyme was incubated with a lactose solution prepared in buffer E (140 mmol/L lactose solution prepared in 100 mM sodium citrate buffer pH 5.5 containing 100 µmol/L MgSO ₄ . The reaction mixture was prepared by mixing 13 µl of diluted purified enzyme and 37 µl of lactose solution in a PCR tube. The substrate solution was prepared in a buffer pH 5.5, the enzyme solution had a pH of 6.7. To initiate the reaction, 13 µl of enzyme was added to 37 µl of the substrate solution. Thus, when mixing these two buffers, the final pH of the reaction increased from 5.5 to 5.7. The reaction mixture was incubated in a DNA thermal cycler using the following incubation parameters: reaction time 10 min at 37°C, enzyme inactivation 10 min at 95°C, storage at 4°C. The reaction mixtures were stored at -20° C. until further use.The amount of glucose formed during the reaction was determined following the protocol described in Example 10. To determine the amount of micromoles (μmol) of glucose formed per second, described as 1 unit of activity with lactose at pH 5.5 at 37°C (Unit of Activity with Lactose; UAL-5), the maximum optical density value for each lactase was used. Specific activity at pH 5.5 at 37°C, described as SUAL-5, is defined as the amount of µmol of glucose produced per second per µmol of enzyme (µmol glucose/s/µmol enzyme) and is calculated by dividing UAL-5 by the protein concentration in µmol High specific activity at pH 5.5 at 37°C is relevant for the hydrolysis of lactose when used for fermented milk and the hydrolysis of lactose in sweet whey. Detailed results of determining the specific activity of enzymes at pH 5.5 at 37°C are described in Fig. 6. Additionally, activity was described as the number of micromoles of glucose produced per minute per milligram of enzyme added. The results are presented in Fig. 15.

Example 12 Determination of activity using purified enzymes on lactose as substrate at pH 5.5 and temperature 43°C

Purified lactases were diluted to a 40-fold concentration in buffer A (buffer solution 50 mmol/L NaH ₂ PO ₄ pH 6.7, containing 100 µmol/L MgSO ₄ ). In a separate reaction, the diluted enzyme was incubated with a lactose solution prepared in buffer E (140 mmol/L lactose solution prepared in 100 mM sodium citrate buffer pH 5.5 containing 100 μmol/L MgSO ₄ ). The reaction mixture was prepared by mixing 13 μL of diluted purified enzyme and 37 μL of lactose solution in a PCR tube. The substrate solution was prepared in a buffer pH 5.5; the enzyme solution had a pH 6.7. To initiate the reaction, 13 μl of enzyme was added to 37 μl of substrate solution. Thus, when these two buffers were mixed, the final pH of the reaction increased from 5.5 to 5.7.

The reaction mixture was incubated in a DNA thermal cycler using the following incubation parameters: reaction time 10 min at 43°C, enzyme inactivation 10 min at 95°C, storage at 4°C. The reaction mixtures were stored at -20°C until further use. The amount of glucose produced during the reaction was determined following the protocol described in Example 10. To determine the amount of micromoles (μmol) of glucose produced per second, described as 1 unit of activity with lactose at pH 5.5 at 43°C (UAL-6 ), the maximum optical density value for each lactase was used. Specific activity at pH 5.5 at 43°C, described as SUAL-6, is defined as the amount of µmol of glucose produced per second per µmol of enzyme (µmol glucose/s/µmol enzyme) and is calculated by dividing UAL-6 by the protein concentration in µmol High specific activity at pH 5.5 at 43°C is relevant for the hydrolysis of lactose when used for fermented milk and the hydrolysis of lactose in sweet whey. Detailed results of determining the specific activity of enzymes at pH 5.5 at 43°C are described in Fig. 7. Additionally, activity was described as the number of micromoles of glucose produced per minute per milligram of enzyme added. The results are presented in Fig. 15.

Example 13 Determination of activity using purified enzymes on lactose as a substrate at pH 4.5 and 4°C

Purified lactases were diluted to a 40-fold concentration in buffer A (buffer solution 50 mmol/L NaH ₂ PO ₄ pH 6.7, containing 100 µmol/L MgSO ₄ ). In a separate reaction, the diluted enzyme was incubated with a lactose solution prepared in buffer F (140 mmol/L lactose solution prepared in 100 mM sodium citrate buffer pH 4.5 containing 100 μmol/L MgSO ₄ ). The reaction mixture was prepared by mixing 13 μL of diluted purified enzyme and 37 μL of lactose solution in a PCR tube. The substrate solution was prepared in a buffer pH 4.5; the enzyme solution had a pH 6.7. To initiate the reaction, 13 μl of enzyme was added to 37 μl of substrate solution. Thus, when these two buffers were mixed, the final pH of the reaction increased from 4.5 to 4.7.

The reaction mixture was incubated in a DNA thermal cycler using the following incubation parameters: reaction time 60 min at 4°C, enzyme inactivation 10 min at 95°C, storage at 4°C. To determine the amount of glucose produced during the reaction, 10 μl of the reaction mixture was transferred to one well of a standard microtiter plate containing 80 μl of buffer C (as described in example 6) and incubated at 30°C for 40 minutes. After 40 min, absorbance was determined at 610 nm using a FLUOStar Omega scanning UV plate reader (BMG Labtech, Germany). For calculations, optical density values from 0.1 to 1.5 were used; if the A610 nm value exceeded 1.5, the reaction mixture was diluted to a 5-fold concentration with buffer A. For each purified enzyme, reactions were performed in triplicate and the average of triplicate measurements was used for calculations. The maximum absorbance value for each lactase was used to determine the amount of micromoles (μmol) of glucose produced per second, described as 1 unit of activity with lactose at pH 4.5 at 4°C (UAL-7). Specific activity at pH 4.5 at 4°C, described as SUAL-7, is defined as the amount of µmol of glucose produced per second per µmol of enzyme (µmol glucose/s/µmol enzyme) and is calculated by dividing UAL-7 by the protein concentration in µmol High specific activity at pH 4.5 at 4°C is relevant for the hydrolysis of lactose when used for fermented milk. Detailed results of determining the specific activity of enzymes at pH 4.5 at 4°C are described in Fig. 8. Additionally, activity was described as the number of micromoles of glucose produced per minute per milligram of enzyme added. The results are presented in Fig. 16.

Example 14 Determination of activity using purified enzymes on lactose as a substrate at pH 4.5 and a temperature of 37°C

Purified lactases were diluted to a 40-fold concentration in buffer A (buffer solution 50 mmol/L NaH ₂ PO ₄ pH 6.7, containing 100 µmol/L MgSO ₄ ). In a separate reaction, the diluted enzyme was incubated with a lactose solution prepared in buffer F (140 mmol/L lactose solution prepared in 100 mM sodium citrate buffer pH 4.5 containing 100 μmol/L MgSO ₄ ). The reaction mixture was prepared by mixing 13 μL of diluted purified enzyme and 37 μL of lactose solution in a PCR tube. The substrate solution was prepared in a buffer pH 4.5; the enzyme solution had a pH 6.7. To initiate the reaction, 13 μl of enzyme was added to 37 μl of substrate solution. Thus, when these two buffers were mixed, the final pH of the reaction increased from 4.5 to 4.7.

The reaction mixture was incubated in a DNA thermal cycler using the following incubation parameters: reaction time 10 min at 37°C, enzyme inactivation 10 min at 95°C, storage at 4°C. The reaction mixtures were stored at -20°C until further use. The amount of glucose produced during the reaction was determined following the protocol described in Example 13. To determine the amount of micromoles (µmol) of glucose produced per second, described as 1 unit of activity with lactose at pH 4.5 at 37°C (Unit of Activity with Lactose; UAL-8), the maximum optical density value for each lactase was used. Specific activity at pH 4.5 at 37°C, described as SUAL-8, is defined as the amount of µmol of glucose produced per second per µmol of enzyme (µmol glucose/s/µmol enzyme) and is calculated by dividing UAL-8 by the protein concentration in µmol High specific activity at pH 4.5 at 37°C is relevant for the hydrolysis of lactose when used for fermented milk and the hydrolysis of lactose in acid whey. Detailed results of determining the specific activity of enzymes at pH 4.5 at 37°C are described in Fig. 9. Additionally, activity was described as the number of micromoles of glucose produced per minute per milligram of enzyme added. The results are presented in Fig. 16.

Example 15 Determination of activity using purified enzymes on lactose as a substrate at pH 4.5 and a temperature of 43°C

Purified lactases were diluted to a 40-fold concentration in buffer A (buffer solution 50 mmol/L NaH ₂ PO ₄ pH 6.7, containing 100 µmol/L MgSO ₄ ). In a separate reaction, the diluted enzyme was incubated with a lactose solution prepared in buffer F (140 mmol/L lactose solution prepared in 100 mM sodium citrate buffer pH 4.5 containing 100 μmol/L MgSO ₄ ). The reaction mixture was prepared by mixing 13 μL of diluted purified enzyme and 37 μL of lactose solution in a PCR tube. The substrate solution was prepared in a buffer pH 4.5; the enzyme solution had a pH 6.7. To initiate the reaction, 13 μl of enzyme was added to 37 μl of substrate solution. Thus, when these two buffers were mixed, the final pH of the reaction increased from 4.5 to 4.7. The reaction mixture was incubated in a DNA thermal cycler using the following incubation parameters: reaction time 10 min at 43°C, enzyme inactivation 10 min at 95°C, storage at 4°C. The reaction mixtures were stored at -20°C until further use. The amount of glucose produced during the reaction was determined following the protocol described in Example 13. To determine the amount of micromoles (μmol) of glucose produced per second, described as 1 unit of activity with lactose at pH 4.5 at 43°C (UAL-9 ), the maximum optical density value for each lactase was used. Specific activity at pH 4.5 at 43°C, described as SUAL-9, is defined as the amount of µmol of glucose produced per second per µmol of enzyme (µmol glucose/s/µmol enzyme) and is calculated by dividing UAL-9 by the protein concentration in µmol High specific activity at pH 4.5 at 43°C is relevant for the hydrolysis of lactose when used for fermented milk and the hydrolysis of lactose in acid whey. Detailed results of determining the specific activity of enzymes at pH 4.5 at 43°C are described in Fig. 10. Additionally, activity was described as the number of micromoles of glucose produced per minute per milligram of enzyme added. The results are presented in Fig. 16.

Example 16 Determination of activity in BLU units

Dilutions of the commercially available enzyme NOLA™ Fit (Chr-Hansen, Denmark) were prepared in the concentration range from 0.5 BLU/ml to 2.5 BLU/ml in buffer G (buffer solution 50 mmol/l NaH ₂ PO ₄ , pH 7 ,0, containing 100 µmol/l MgSO ₄ , 0.045% Bridge dye, Sigma Aldrich). In a separate reaction, the diluted enzyme was incubated with a lactose solution prepared in buffer H (105 mmol/L lactose solution prepared in 100 mM sodium citrate buffer pH 6.7 containing 100 μmol/L MgSO ₄ ). The reaction mixture was prepared by mixing 13 μL of diluted purified enzyme and 37 μL of lactose solution in a PCR tube. The reaction mixture was incubated in a DNA thermal cycler using the following incubation parameters: reaction time 10 min at 37°C, enzyme inactivation 10 min at 95°C, storage at 4°C. Glucose conversion was quantified by transferring 10 μl of the reaction mixture into one well of a standard microtiter plate containing 80 μl of buffer C and incubating at 30°C for 40 min. After 40 min, absorbance was determined at 610 nm using a FLUOStar Omega scanning UV plate reader (BMG Labtech, Germany). A standard curve was generated using the measured absorbance values against BLU/mL. The maximum slope of the curve was used to determine the activity of new enzymes in BLU/ml.

Example 17 Determination of the activity of new lactases in BLU/ml using lactose as substrate

Purified lactases were diluted to a 50-fold concentration in buffer A (buffer solution 50 mmol/L NaH ₂ PO ₄ pH 6.7, containing 100 µmol/L MgSO ₄ ). In a separate reaction, the diluted enzyme was incubated with a lactose solution prepared in buffer H (105 mmol/L lactose solution prepared in 100 mM sodium citrate buffer pH 6.7 containing 100 μmol/L MgSO ₄ ). The reaction mixture was prepared by mixing 13 μL of diluted purified enzyme and 37 μL of lactose solution in a PCR tube. The reaction mixture was incubated in a DNA thermal cycler using the following incubation parameters: reaction time 10 min at 37°C, enzyme inactivation 10 min at 95°C, storage at 4°C. After the reaction, 10 μl of the reaction mixture was transferred to one well of a standard microtiter plate containing 80 μl of buffer C (as described in example 6) and incubated at 30°C for 40 minutes. After 40 min, absorbance was determined at 610 nm using a FLUOStar Omega scanning UV plate reader. For calculations, optical density values from 0.1 to 1.5 were used; if the A610 nm value exceeded 1.5, the reaction mixture was diluted to a 5-fold concentration with buffer A.

Maximum absorbance values were used to calculate enzyme activity in BLU/ml using the standard curve described in Example 16.

Example 18: Measurement of the percentage of residual lactose in fresh milk at refrigeration temperature

2 ml of commercially available pasteurized milk (pasteurized milk 1.5% fat, Arla Food) was mixed in a 10 ml glass tube with 10-125 µl of enzyme (equivalent to 10 BLU/ml) as determined in example 17. Samples were incubated under constant conditions for 24 hours at 4°C. After incubation, the reaction was stopped by heat inactivation at 95°C for 7 min, followed by storage at -20°C until further use. The amount of residual lactose in milk was analyzed using high performance liquid chromatography (HPLC). Samples for analysis were treated with 1.8 ml of protein precipitation solution (0.083 mol/L PCA and 2 mmol/L Na-EDTA) and 2 ml of Milli-Q water (MQW) and then centrifuged at 2800 rpm for 30 min. at 4°C. An aliquot of the supernatant was diluted a total of 200 times using a JANUS automatic dilution system (PerkinElmer, Waltham, MA, USA). Diluted samples were analyzed on a Dionex ICS-5000 system (Thermo Fischer Scientific, Waltham, MA, USA) using a 4 × 250 mm CarboPac SA20 analytical column (Thermo Fischer Scientific, Waltham, MA, USA) and a pulsed amperometric detector. The detector was configured to detect a simple three-stage potential waveform for selective detection of carbohydrates. The eluent was set to 1 mmol/L KOH and continuously regenerated by passing through a precolumn (CR-TC, Thermo Fischer Scientific, Waltham, MA, USA). The eluent flow rate was 1.2 ml/min, and the analysis time per sample injection was 10 min. To quantify lactose in each sample, an external calibration curve was used, constructed from three points by adding known amounts of lactose monohydrate (Sigma-Aldrich, St. Louis, MO, USA) to the MQW. Concentrations were calculated based on the height of chromatographic peaks. The measured percentage of residual lactose in fresh milk is presented in FIG. eleven.

Example 19 Determination of activity in UHT milk at room temperature 2 ml of commercially available UHT milk (UHT milk 1.5% fat, Arla Food) was mixed in a 2 ml glass tube with 2-25 µl of enzyme (equivalent to 10 BLU/ ml) as defined in example 17. Samples were incubated under constant conditions for 24 hours at 25°C. After incubation, the reaction was stopped by heat inactivation at 95°C for 7 min, followed by storage at -20°C until further use. The amount of residual lactose in UHT milk was analyzed by HPLC following the protocol described in Example 18. The measured percentage of residual lactose in fresh milk after hydrolysis is presented in FIG. 12.

Example 20 High Temperature Performance of Enzyme in Buffer

The purified enzyme was diluted to 5 BLU/ml in buffer A (a buffer solution of 50 mmol/l NaH ₂ PO ₄ pH 6.7 containing 100 µmol/l MgSO ₄ ). In a separate reaction, 13 μl of diluted enzyme was incubated in a DNA thermal cycler with a lactose solution (105 mmol/L lactose solution prepared in 100 mM sodium citrate buffer pH 6.7 containing 100 μmol/L MgSO ₄ ). The reaction mixture was prepared by mixing 13 μL of enzyme and 37 μL of lactose solution in a PCR tube. The reaction mixture was incubated in a DNA thermal cycler using the following incubation parameters: reaction time 10 min at 37°C, enzyme inactivation 10 min at 95°C, storage at 4°C. After the reaction, 10 μl of the reaction mixture was transferred to one well of a standard microtiter plate containing 80 μl of buffer C (as described in example 6) and incubated at 30°C for 40 minutes. After 40 min, absorbance was determined at 610 nm using a FLUOStar Omega scanning UV plate reader. For calculations, optical density values from 0.1 to 1.5 were used; if the A610 nm value exceeded 1.5, the reaction mixture was diluted to a 5-fold concentration with buffer A. The measured optical density was designated Abs37°C and was considered the reference value for calculations.

To measure the effect of heat treatment on enzyme activity, in a separate reaction, 13 μl of diluted enzyme (5 BLU/ml) was incubated in a DNA thermal cycler using the following incubation parameters: 72°C for 15 s, or 74°C for 15 s , either at 76°C for 6 s, or 78°C for 6 s, or at 80°C for 4 s, or at 85°C for 5 s, or at 90°C for 5 s, or at 95°C for 5 s, followed by storage at 4°C.

The activity of the heat-treated enzyme was determined by incubation with a lactose solution (a solution of 105 mmol/L lactose prepared in 100 mmol/L sodium citrate buffer at pH 6.7 containing 100 μmol/L MgSO ₄ ) as described above. Optical density measured at different temperatures (for example, at 72°C, 74°C, 76°C, 78°C, 80°C, 85°C, 90°C or 95°C) was designated as Abs72°C, Abs74°C, Abs76°C, Abs78°C, Abs80°C, Abs85°C, Abs90°C, Abs95°C.

The percentage of residual activity at high temperature was determined by the formula

% residual activity=(Abs72°C/Abs37°C)*100.

The percentages of residual activity of various enzymes at different temperatures are described in Fig. 13.

Example 21 Percentage of residual lactose after high temperature treatment

The effect of heat treatment on enzyme characteristics in pasteurized milk was determined by incubating a fixed amount of enzyme in the milk followed by heat treatment. In separate reactions, 50 μl of pasteurized milk was mixed with 10 BLU/ml of purified enzyme (as determined in example 17) in a PCR tube. The milk sample was incubated at 72°C for 15 s or 76°C for 10 s, or 85°C for 5 s and 90°C for 5 s, followed by incubation at 5°C for 24 h. After 24 hours at 5°C, the reaction was stopped by heating at 95°C for 7 minutes, followed by storage at -20°C. The amount of residual lactose was measured using a LactoSens® reagent kit (Chr. Hansen, Denmark) following the supplier's protocol. The measured amount of residual lactose determined at different temperatures was expressed in g/L. The detection limit for the LactoSens® reagent kit is 0.2 g/L to 10 g/L. The results are described in Table 2.

Table 2. Percentage of residual lactose in pasteurized milk treated with a fixed amount of purified enzyme followed by incubation at various temperatures (72°C for 15 s, 76°C for 10 s, 85°C for 5 s and 90°C for for 5 s, followed by incubation at 4°C for 24 h), determined using a LactoSens® reagent kit. The detection limits of the LactoSens® reagent kit range from 0.2 g/L to 10 g/L lactose. In this table, NO means “not determined.”

--->

LIST OF SEQUENCES

<110> Chr. Hansen A/S

<120> LACTASE ENZYMES WITH IMPROVED PROPERTIES

<130> 21101EP00

<160> 41

<170> Patent In version 3.5

<210> 1

<211> 1049

<212>PRT

<213> Bifidobacterium adolescentis

<400> 1

Met Ala Asp Thr Ala Glu Leu Ala Ile Val His Ala Thr Thr Ala Ser

1 5 10 15

Ala Ser Trp Leu Thr Asp Pro Thr Val Phe Ala Ala Asn Arg Lys Pro

20 25 30

Ala His Ser Ser His Arg Tyr Val Ile Gly Glu Thr Ser Glu Pro Lys

35 40 45

Gln Ser Leu Asp Gly Glu Trp Lys Val Arg Ile Glu Gln Ala Arg Asn

50 55 60

Val Asp Val Glu Ser Ala Pro Phe Ala Ala Val Asp Phe Glu Asp Gly

65 70 75 80

Asp Phe Gly Ala Ile Glu Val Pro Gly His Leu Gln Met Ala Gly Tyr

85 90 95

Leu Lys Asn Lys Tyr Val Asn Ile Gln Tyr Pro Trp Asp Gly His Glu

100 105 110

Asp Pro Gln Ala Pro Asn Ile Pro Glu Asn Asn His Val Ala Ile Tyr

115 120 125

Arg Arg Arg Phe Ala Leu Asp Ala Gln Leu Ala Arg Thr Leu Glu Asn

130 135 140

Asp Gly Thr Val Ser Leu Thr Phe His Gly Ala Ala Thr Ala Ile Tyr

145 150 155 160

Val Trp Leu Asp Gly Thr Phe Val Gly Tyr Gly Glu Asp Gly Phe Thr

165 170 175

Pro Ser Glu Phe Asp Val Thr Glu Ala Leu Arg Asn Gly Asn Gly Asn

180 185 190

Ala Ala Asp Ser Pro Glu Ala Glu His Thr Leu Thr Val Ala Cys Tyr

195 200 205

Glu Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe Trp Arg Leu

210 215 220

His Gly Leu Phe Arg Thr Val Glu Leu Ala Ala Gln Pro His Thr His

225 230 235 240

Val Glu Thr Val Gln Leu Glu Ala Asp Tyr Thr Ala Ala Asp Thr Ala

245 250 255

Gly Thr Ala Asp Thr Ala Glu Leu Asn Ala Ala Leu Thr Leu Arg Asn

260 265 270

Ser Ala Asp Ala Met Thr Ile Glu Ser Thr Leu Arg Asp Gly Asp Gly

275 280 285

Asn Val Val Trp Glu Ser Thr Gln Ala Cys Asn Gly Glu Ile Ala Leu

290 295 300

Asn Ser Gly Lys Met Thr Asn Ile Ala Pro Trp Ser Ala Glu Ser Pro

305 310 315 320

Thr Leu Tyr Thr Leu Thr Val Arg Val Val Gly His Asp Gly Ala Ile

325 330 335

Ile Glu Thr Val Thr Gln Lys Ile Gly Phe Arg Thr Phe Arg Ile Glu

340 345 350

Asn Gly Ile Met Thr Leu Asn Gly Lys Arg Ile Val Phe Lys Gly Ala

355 360 365

Asp Arg His Glu Phe Asp Ala Lys Arg Gly Arg Ala Ile Thr Arg Glu

370 375 380

Asp Met Leu Ser Asp Val Val Phe Cys Lys Arg His Asn Ile Asn Ala

385 390 395 400

Ile Arg Thr Ser His Tyr Pro Asn Gln Glu Tyr Trp Tyr Asp Leu Cys

405 410 415

Asp Glu Tyr Gly Leu Tyr Leu Ile Asp Glu Thr Asn Met Glu Thr His

420 425 430

Gly Thr Trp Val Ala Asn Asn Val Glu Arg Pro Glu Asp Gly Ile Pro

435 440 445

Gly Ser Arg Pro Glu Trp Glu Gly Ala Cys Val Asp Arg Ile Asn Ser

450 455 460

Met Met Arg Arg Asp Tyr Asn His Pro Ser Val Leu Ile Trp Ser Leu

465 470 475 480

Gly Asn Glu Ser Ser Ala Gly Glu Val Phe Arg Ala Met Tyr Arg His

485 490 495

Ala His Thr Ile Asp Pro Asn Arg Pro Val His Tyr Glu Gly Ser Val

500 505 510

His Met Arg Glu Phe Glu Asp Val Thr Asp Ile Glu Ser Arg Met Tyr

515 520 525

Ala His Ala Asp Glu Ile Glu Arg Tyr Leu Asn Asp Gly Ser Pro Ala

530 535 540

His Thr Asp Gly Pro Lys Lys Pro Tyr Ile Ser Cys Glu Tyr Met His

545 550 555 560

Ala Met Gly Asn Ser Cys Gly Asn Met Asp Glu Tyr Thr Ala Leu Glu

565 570 575

Arg Tyr Pro Met Tyr Gln Gly Gly Phe Ile Trp Asp Phe Ile Asp Gln

580 585 590

Ala Ile Glu Thr Lys Leu Pro Asp Gly Thr Thr Arg Met Cys Tyr Gly

595 600 605

Gly Asp Phe Gly Asp Arg Pro Ser Asp Tyr Glu Phe Ser Gly Asp Gly

610 615 620

Leu Leu Phe Ala Asp Arg Thr Pro Ser Pro Lys Ala Gln Glu Val Lys

625 630 635 640

Gln Leu Tyr Ala Asn Val Lys Ile Val Val Ser Val Asp Glu Ala Arg

645 650 655

Ile Thr Asn Asp Asn Leu Phe Val Ser Thr Gly Asp Tyr Arg Phe Val

660 665 670

Leu Arg Ile Leu Ala Asp Gly Lys Pro Val Trp Ser Thr Thr Arg Arg

675 680 685

Phe Asp Val Ala Ala Gly Glu Ser Ala Ser Phe Glu Val Asp Trp Pro

690 695 700

Val Asp Asp Tyr Arg Ser Asn Ala Glu Glu Leu Val Leu Glu Val Ser

705 710 715 720

Gln Gln Leu Gly Asn Ala Cys Asp Trp Ala Pro Ala Gly Tyr Glu Leu

725 730 735

Ala Phe Gly Gln Cys Val Val Ala Gly Ala Lys Thr Thr Ala Asp Ala

740 745 750

Val Asp Ala Ala Gly Ala Pro Ala Asp Gly Thr Val Thr Leu Gly Arg

755 760 765

Trp Asn Ala Gly Val Arg Gly Gln Gly Arg Glu Ala Leu Phe Ser Arg

770 775 780

Thr Gln Gly Gly Met Val Ser Tyr Thr Phe Gly Glu Arg Glu Phe Val

785 790 795 800

Leu Arg Arg Pro Ser Ile Thr Thr Phe Arg Pro Leu Thr Asp Asn Asp

805 810 815

Arg Gly Ala Gly His Ala Phe Glu Arg Ala Ala Trp Ala Val Ala Gly

820 825 830

Lys Tyr Ala Arg Cys Val Asp Cys Ala Ile Ala Asn Arg Gly Glu Asn

835 840 845

Ala Val Glu Ala Thr Tyr Thr Tyr Glu Leu Ala Ile Pro Gln Arg Thr

850 855 860

Lys Val Thr Val Arg Tyr Val Ala Asp Thr Ala Gly Leu Val Ser Leu

865 870 875 880

Asp Val Glu Tyr Pro Gly Glu Lys Asn Gly Asp Leu Pro Thr Ile Pro

885 890 895

Ala Phe Gly Ile Glu Trp Ala Leu Pro Val Glu Tyr Ala Asn Leu Arg

900 905 910

Phe Tyr Gly Ala Gly Pro Glu Glu Thr Tyr Ala Asp Arg Arg His Ala

915 920 925

Lys Leu Gly Val Trp Ser Thr Thr Ala Gly Asp Asp Cys Ala Pro Tyr

930 935 940

Leu Leu Pro Gln Glu Thr Gly Asn His Glu Asp Val Arg Trp Ala Glu

945 950 955 960

Ile Thr Asp Asp Ser Gly His Gly Val Arg Val Lys Arg Gly Ala Gly

965 970 975

Ala Lys Pro Phe Ala Met Ser Leu Leu Pro Tyr Ser Ser Thr Met Leu

980 985 990

Glu Glu Ala Leu His Gln Asp Glu Leu Pro Lys Pro Arg His Met Phe

995 1000 1005

Leu Arg Leu Leu Ala Ala Gln Met Gly Val Gly Gly Asp Asp Ser

1010 1015 1020

Trp Met Ser Pro Val His Glu Gln Tyr Gln Leu Pro Ala Asp Gln

1025 1030 1035

Pro Leu Ser Leu Asn Val Gln Leu Lys Leu Phe

1040 1045

<210> 2

<211> 625

<212>PRT

<213> Lactobacillus sakei

<400> 2

Met Gln Pro Asn Ile Gln Trp Leu Asp Thr Pro Ala Val Phe Arg Val

1 5 10 15

Gly Gln Leu Pro Ala His Ser Asp His Arg Tyr Tyr Ala Thr Leu Ala

20 25 30

Glu Met Ala Gln Gln Gln Ser Ser Phe Glu Gln Ser Leu Asn Gly Thr

35 40 45

Trp Gln Phe His Tyr Ser Val Asn Ala Ala Ser Arg Pro Lys Ser Phe

50 55 60

Tyr Glu Leu Ala Phe Asp Ala Gln Asp Phe Glu Pro Ile Thr Val Pro

65 70 75 80

Gln His Ile Glu Leu Ala Gly Tyr Glu Gln Leu His Tyr Ile Asn Thr

85 90 95

Met Tyr Pro Trp Glu Gly His Tyr Tyr Arg Arg Pro Ala Phe Ser Thr

100 105 110

Ser Asp Asp Lys Gln His Leu Gly Met Phe Ser Glu Ala Asp Tyr Asn

115 120 125

Pro Val Gly Ser Tyr Leu His His Phe Asp Leu Thr Pro Ala Leu Arg

130 135 140

Asn Gln Arg Val Ile Ile Arg Phe Glu Gly Val Glu Gln Ala Met Tyr

145 150 155 160

Val Trp Leu Asn Gly Gln Phe Ile Gly Tyr Ala Glu Asp Ser Phe Thr

165 170 175

Pro Ser Glu Phe Asp Leu Thr Pro Tyr Leu Lys Glu Thr Asp Asn Cys

180 185 190

Leu Ala Val Glu Val His Lys Arg Ser Ser Ala Ala Phe Ile Glu Asp

195 200 205

Gln Asp Phe Phe Arg Phe Phe Gly Ile Phe Arg Asp Val Lys Leu Leu

210 215 220

Ala Lys Pro Arg Thr His Leu Glu Asp Leu Trp Val Ile Pro Glu Tyr

225 230 235 240

Asp Val Val Gln Gln Thr Gly Gln Val Lys Leu Arg Leu Gln Phe Ser

245 250 255

Gly Asp Glu Asn Arg Val His Leu Arg Ile Arg Asp Gln His Gln Ile

260 265 270

Ile Leu Thr Ala Asp Leu Thr Ser Ala Ala Gln Val Asn Gly Leu Tyr

275 280 285

Lys Met Pro Glu Leu Val Gln Ala Trp Ser Asn Gln Thr Pro Asn Leu

290 295 300

Tyr Thr Leu Glu Leu Glu Val Val Asp Gln Ala Gly Glu Thr Ile Glu

305 310 315 320

Ile Ser Gln Gln Pro Phe Gly Phe Arg Lys Ile Glu Ile Lys Asp Lys

325 330 335

Val Met Leu Leu Asn Gly Lys Arg Leu Val Ile Asn Gly Val Asn Arg

340 345 350

His Glu Trp His Pro Glu Thr Gly Arg Thr Ile Thr Ala Glu Asp Glu

355 360 365

Ala Trp Asp Ile Ala Cys Met Gln Arg Asn His Ile Asn Ala Val Arg

370 375 380

Thr Ser His Tyr Pro Asp Arg Leu Ser Phe Tyr Asn Gly Cys Asp Gln

385 390 395 400

Ala Gly Ile Tyr Met Met Ala Glu Thr Asn Leu Glu Ser His Gly Ser

405 410 415

Trp Gln Lys Met Gly Ala Val Glu Pro Ser Trp Asn Val Pro Gly Ser

420 425 430

Tyr Asp Glu Trp Glu Ala Ala Thr Leu Asp Arg Ala Arg Thr Asn Phe

435 440 445

Glu Thr Phe Lys Asn His Val Ser Ile Leu Phe Trp Ser Leu Gly Asn

450 455 460

Glu Ser Tyr Ala Gly Ser Val Leu Glu Lys Met Asn Ala Tyr Tyr Lys

465 470 475 480

Gln Gln Asp Pro Thr Arg Leu Val His Tyr Glu Gly Val Phe Arg Ala

485 490 495

Pro Glu Tyr Lys Ala Thr Ile Ser Asp Val Glu Ser Arg Met Tyr Ala

500 505 510

Thr Pro Ala Glu Ile Lys Ala Tyr Leu Asp Asn Ala Pro Gln Lys Pro

515 520 525

Phe Ile Leu Cys Glu Tyr Met His Asp Met Gly Asn Ser Leu Gly Gly

530 535 540

Met Gln Ser Tyr Ile Asp Leu Leu Ser Gln Tyr Asp Met Tyr Gln Gly

545 550 555 560

Gly Phe Ile Trp Asp Phe Ile Asp Gln Ala Leu Leu Val Thr Asp Pro

565 570 575

Val Thr Gly Gln Arg Glu Leu Arg Tyr Gly Gly Asp Phe Asp Asp Arg

580 585 590

Pro Ser Asp Tyr Glu Phe Ser Gly Asp Gly Leu Val Phe Ala Thr Arg

595 600 605

Asp Glu Lys Pro Ala Met Gln Glu Val Arg Tyr Tyr Tyr Gly Glu His

610 615 620

Lys

625

<210> 3

<211> 330

<212>PRT

<213> Lactobacillus sakei

<400> 3

Met Lys Asn Gln Gln Cys Arg Arg Leu Asp Thr Ile Met Ala Asn Thr

1 5 10 15

Asn Lys Arg Leu Ala Val Ile Phe Gly Asp Val Thr Leu Gly Leu Lys

20 25 30

Gly Pro Asp Phe His Tyr Leu Phe Ser Tyr Gln Thr Gly Gly Pro Glu

35 40 45

Ser Leu Arg Ile Gln Gly Lys Glu Trp Leu Tyr Arg Ser Pro Lys Pro

50 55 60

Thr Phe Trp Arg Ala Thr Thr Asp Asn Asp Arg Gly Asn Gln Phe Pro

65 70 75 80

Leu Lys Ser Gly Met Trp Leu Ala Ala Asp Gln Phe Ile Ala Cys Gln

85 90 95

Ser Ile Thr Val Ala Ile Asp Gly Gln Thr Ile Pro Leu Pro Ile Ala

100 105 110

Pro Glu Asn Asn Arg Tyr Ser Gly Gln Glu Thr Ala Gln Glu Val Thr

115 120 125

Val Thr Tyr Thr Tyr Gln Thr Ile Thr Thr Pro Gln Thr Thr Val Glu

130 135 140

Val Ser Tyr Thr Ile Gln Ala Ser Gly Lys Ile Arg Val Ala Val Thr

145 150 155 160

Tyr His Gly Gln Ala Gly Leu Pro Ser Leu Pro Val Phe Gly Leu Arg

165 170 175

Phe Val Met Pro Thr Pro Ala Thr Arg Phe Ile Tyr Gln Gly Leu Ser

180 185 190

Gly Glu Thr Tyr Pro Asp Arg Met Ala Gly Gly Met Ala Gly Glu Tyr

195 200 205

Glu Val Thr Gly Leu Pro Val Thr Pro Tyr Leu Val Pro Gln Asp Cys

210 215 220

Gly Val His Met Ala Thr Asp Trp Val Thr Ile Tyr Arg Gln Ala Val

225 230 235 240

Leu Asp Asn Arg Leu Arg Glu Pro Val Glu Thr Gly Leu Lys Phe Lys

245 250 255

Met Val Asp Gln Pro Phe Ala Phe Ser Cys Leu Pro Tyr Thr Ala Glu

260 265 270

Glu Leu Glu Asn Ala Thr His Ser Glu Leu Pro Ala Pro His Arg

275 280 285

Thr Val Leu Ser Leu Leu Gly Ala Val Arg Gly Val Gly Gly Ile Asp

290 295 300

Ser Trp Gly Ser Asp Val Glu Ala Ala Tyr Gln Ile Asp Ala Thr Gln

305 310 315 320

Asp His His Leu Glu Phe Glu Ile Ser Phe

325 330

<210> 4

<211> 1049

<212>PRT

<213> Bifidobacterium adolescentis

<400> 4

Met Ala Asp Thr Ala Glu Leu Ala Ile Val His Ala Thr Thr Ala Ser

1 5 10 15

Ala Ser Trp Leu Thr Asp Pro Thr Val Phe Ala Ala Asn Arg Lys Pro

20 25 30

Ala His Ser Ser His Arg Tyr Val Ile Gly Glu Thr Ser Glu Pro Lys

35 40 45

Gln Ser Leu Asp Gly Glu Trp Lys Val Arg Ile Glu Gln Ala Arg Asn

50 55 60

Val Asp Val Glu Ser Ala Pro Phe Ala Ala Val Asp Phe Glu Asp Gly

65 70 75 80

Asp Phe Gly Ala Ile Glu Val Pro Gly His Leu Gln Met Ala Gly Tyr

85 90 95

Leu Lys Asn Lys Tyr Val Asn Ile Gln Tyr Pro Trp Asp Gly His Glu

100 105 110

Asp Pro Gln Ala Pro Asn Ile Pro Glu Asn Asn His Val Ala Ile Tyr

115 120 125

Arg Arg Arg Phe Ala Leu Asp Ala Gln Leu Ala Arg Thr Leu Glu Asn

130 135 140

Asp Gly Thr Val Ser Leu Thr Phe His Gly Ala Ala Thr Ala Ile Tyr

145 150 155 160

Val Trp Leu Asp Gly Thr Phe Val Gly Tyr Gly Glu Asp Gly Phe Thr

165 170 175

Pro Ser Glu Phe Asp Val Thr Glu Ala Leu Arg Asn Gly Asn Gly Asn

180 185 190

Ala Ala Asp Ser Pro Glu Ala Glu His Thr Leu Thr Val Ala Cys Tyr

195 200 205

Glu Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe Trp Arg Leu

210 215 220

His Gly Leu Phe Arg Thr Val Glu Leu Ala Ala Gln Pro His Thr His

225 230 235 240

Val Glu Thr Val Gln Leu Glu Ala Asp Tyr Thr Ala Ala Asp Thr Ala

245 250 255

Gly Thr Ala Asp Thr Ala Glu Leu Asn Ala Ala Leu Thr Leu Arg Asn

260 265 270

Pro Ala Asp Ala Met Thr Ile Glu Ser Thr Leu Arg Asp Gly Asp Gly

275 280 285

Asn Val Val Trp Glu Ser Thr Gln Ala Cys Asn Gly Glu Ile Ala Leu

290 295 300

Asn Ser Gly Lys Met Thr Asn Ile Ala Pro Trp Ser Ala Glu Ser Pro

305 310 315 320

Thr Leu Tyr Thr Leu Thr Val Arg Val Val Gly His Asp Gly Ala Ile

325 330 335

Ile Glu Thr Val Thr Gln Lys Ile Gly Phe Arg Thr Phe Arg Ile Glu

340 345 350

Asn Gly Ile Met Thr Leu Asn Gly Lys Arg Ile Val Phe Lys Gly Ala

355 360 365

Asp Arg His Glu Phe Asp Ala Lys Arg Gly Arg Ala Ile Thr Arg Glu

370 375 380

Asp Met Leu Ser Asp Val Val Phe Cys Lys Arg His Asn Ile Asn Ala

385 390 395 400

Ile Arg Thr Ser His Tyr Pro Asn Gln Glu Tyr Trp Tyr Asp Leu Cys

405 410 415

Asp Glu Tyr Gly Leu Tyr Leu Ile Asp Glu Thr Asn Met Glu Thr His

420 425 430

Gly Thr Trp Val Ala Asn Asn Val Glu Arg Pro Glu Asp Gly Ile Pro

435 440 445

Gly Ser Arg Pro Glu Trp Glu Gly Ala Cys Val Asp Arg Ile Asn Ser

450 455 460

Met Met Arg Arg Asp Tyr Asn His Pro Ser Val Leu Ile Trp Ser Leu

465 470 475 480

Gly Asn Glu Ser Ser Ala Gly Glu Val Phe Arg Ala Met Tyr Arg His

485 490 495

Ala His Thr Ile Asp Pro Asn Arg Pro Val His Tyr Glu Gly Ser Val

500 505 510

His Met Arg Glu Phe Glu Asp Val Thr Asp Ile Glu Ser Arg Met Tyr

515 520 525

Ala His Ala Asp Glu Ile Glu Arg Tyr Leu Asn Asp Gly Ser Pro Ala

530 535 540

His Thr Asp Gly Pro Lys Lys Pro Tyr Ile Ser Cys Glu Tyr Met His

545 550 555 560

Ala Met Gly Asn Ser Cys Gly Asn Met Asp Glu Tyr Thr Ala Leu Glu

565 570 575

Arg Tyr Pro Met Tyr Gln Gly Gly Phe Ile Trp Asp Phe Ile Asp Gln

580 585 590

Ala Ile Glu Thr Lys Leu Pro Asp Gly Thr Thr Arg Met Cys Tyr Gly

595 600 605

Gly Asp Phe Gly Asp Arg Pro Ser Asp Tyr Glu Phe Ser Gly Asp Gly

610 615 620

Leu Leu Phe Ala Asp Arg Thr Pro Ser Pro Lys Ala Gln Glu Val Lys

625 630 635 640

Gln Leu Tyr Ala Asn Val Lys Ile Ala Val Ser Val Asp Glu Ala Arg

645 650 655

Ile Thr Asn Asp Asn Leu Phe Val Ser Thr Gly Asp Tyr Arg Phe Val

660 665 670

Leu Arg Ile Leu Ala Asp Gly Lys Pro Val Trp Ser Thr Thr Arg Arg

675 680 685

Phe Asp Val Ala Ala Gly Glu Ser Ala Ser Phe Glu Val Asp Trp Pro

690 695 700

Val Asp Asp Tyr Arg Ser Asn Ala Glu Glu Leu Val Leu Glu Val Ser

705 710 715 720

Gln Gln Leu Gly Asn Ala Cys Asp Trp Ala Pro Ala Gly Tyr Glu Leu

725 730 735

Ala Phe Gly Gln Cys Val Val Ala Gly Ala Lys Thr Thr Ala Asp Ala

740 745 750

Val Asp Ala Ala Gly Ala Pro Ala Asp Gly Thr Val Thr Leu Gly Arg

755 760 765

Trp Asn Ala Gly Val Arg Gly Gln Gly Arg Glu Ala Leu Phe Ser Arg

770 775 780

Thr Gln Gly Gly Met Val Ser Tyr Thr Phe Gly Glu Arg Glu Phe Val

785 790 795 800

Leu Arg Arg Pro Ser Ile Thr Thr Phe Arg Pro Leu Thr Asp Asn Asp

805 810 815

Arg Gly Ala Gly His Ala Phe Glu Arg Ala Ala Trp Ala Val Ala Gly

820 825 830

Lys Tyr Ala Arg Cys Val Asp Cys Ala Ile Ala Asn Arg Gly Glu Asn

835 840 845

Ala Val Glu Ala Thr Tyr Thr Tyr Glu Leu Ala Ile Pro Gln Arg Thr

850 855 860

Lys Val Thr Val Arg Tyr Val Ala Asp Thr Ala Gly Leu Val Ser Leu

865 870 875 880

Asp Val Glu Tyr Pro Gly Glu Lys Asn Gly Asp Leu Pro Thr Ile Pro

885 890 895

Ala Phe Gly Ile Glu Trp Ala Leu Pro Val Glu Tyr Ala Asn Leu Arg

900 905 910

Phe Tyr Gly Ala Gly Pro Glu Glu Thr Tyr Ala Asp Arg Arg His Ala

915 920 925

Lys Leu Gly Val Trp Ser Thr Thr Ala Gly Asp Asp Cys Ala Pro Tyr

930 935 940

Leu Leu Pro Gln Glu Thr Gly Asn His Glu Asp Val Arg Trp Ala Glu

945 950 955 960

Ile Thr Asp Asp Ser Gly His Gly Val Arg Val Lys Arg Gly Ala Gly

965 970 975

Ala Lys Pro Phe Ala Met Ser Leu Leu Pro Tyr Ser Ser Thr Met Leu

980 985 990

Glu Glu Ala Leu His Gln Asp Glu Leu Pro Lys Pro Arg His Met Phe

995 1000 1005

Leu Arg Leu Leu Ala Ala Gln Met Gly Val Gly Gly Asp Asp Ser

1010 1015 1020

Trp Met Ser Pro Val His Glu Gln Tyr Gln Leu Pro Ala Asp Gln

1025 1030 1035

Pro Leu Ser Leu Asn Val Gln Leu Lys Leu Phe

1040 1045

<210> 5

<211> 626

<212>PRT

<213> Lactobacillus amylovorus

<400> 5

Met Lys Ala Asn Ile Lys Trp Leu Asp Asp Pro Glu Val Phe Arg Ile

1 5 10 15

Asn Gln Leu Pro Ala His Ser Asp His Pro Phe Tyr Lys Asp Tyr Arg

20 25 30

Glu Trp Gln Asn His Ser Ser Ser Phe Lys Gln Ser Leu Asn Gly Ala

35 40 45

Trp Gln Phe His Phe Ser Lys Asp Pro Gln Ser Arg Pro Ile Asp Phe

50 55 60

Tyr Lys Arg Ser Phe Asp Ser Ser Ser Phe Asp Thr Ile Pro Val Pro

65 70 75 80

Ser Glu Ile Glu Leu Asn Gly Tyr Ala Gln Asn Gln Tyr Thr Asn Ile

85 90 95

Leu Tyr Pro Trp Glu Ser Lys Ile Tyr Arg Lys Pro Ala Tyr Thr Leu

100 105 110

Gly Arg Gly Ile Lys Asp Gly Asp Phe Ser Gln Gly Lys Asp Asn Thr

115 120 125

Val Gly Ser Tyr Leu Lys His Phe Asp Leu Asn Pro Ala Leu Ala Gly

130 135 140

His Asp Ile His Ile Gln Phe Glu Gly Val Glu Arg Ala Met Tyr Val

145 150 155 160

Tyr Leu Asn Gly His Phe Ile Gly Tyr Ala Glu Asp Ser Phe Thr Pro

165 170 175

Ser Glu Phe Asp Leu Thr Pro Tyr Ile Gln Ala Lys Asp Asn Ile Leu

180 185 190

Ala Val Glu Val Phe Lys His Ser Thr Ala Ser Trp Leu Glu Asp Gln

195 200 205

Asp Met Phe Arg Phe Ser Gly Ile Phe Arg Ser Val Glu Leu Leu Ala

210 215 220

Leu Pro Arg Thr His Leu Met Asp Leu Asp Ile Lys Pro Thr Val Val

225 230 235 240

Asn Asp Tyr His Asp Gly Val Phe Asn Ala Lys Leu His Phe Met Gly

245 250 255

Lys Thr Ser Gly Asn Val His Val Leu Ile Glu Asp Ile Asp Gly Lys

260 265 270

Thr Leu Leu Asn Lys Lys Leu Pro Leu Lys Ser Thr Val Glu Ile Glu

275 280 285

Asn Glu Thr Phe Ala Asn Val His Leu Trp Asp Asn His Asp Pro Tyr

290 295 300

Leu Tyr Gln Leu Ile Ile Glu Val His Asp Gln Asp Gly Lys Leu Val

305 310 315 320

Glu Leu Ile Pro Tyr Gln Phe Gly Phe Arg Lys Ile Glu Ile Thr Lys

325 330 335

Asp His Val Val Leu Leu Asn Gly Lys Arg Leu Ile Ile Asn Gly Val

340 345 350

Asn Arg His Glu Trp Asp Ala Lys Arg Gly Arg Ser Ile Thr Leu Ala

355 360 365

Asp Met Lys Gln Asp Ile Ala Thr Phe Lys His Asn Asn Ile Asn Ala

370 375 380

Val Arg Thr Cys His Tyr Pro Asn Gln Ile Pro Trp Tyr Tyr Leu Cys

385 390 395 400

Asp Gln Asn Gly Ile Tyr Met Met Ala Glu Asn Asn Leu Glu Ser His

405 410 415

Gly Thr Trp Gln Lys Leu Gly Gln Val Glu Ala Thr Ser Asn Val Pro

420 425 430

Gly Ser Ile Pro Glu Trp Arg Glu Val Val Val Asp Arg Ala Arg Ser

435 440 445

Asn Tyr Glu Thr Phe Lys Asn His Thr Ala Ile Leu Phe Trp Ser Leu

450 455 460

Gly Asn Glu Ser Tyr Ala Gly Ser Asn Ile Ala Ala Met Asn Lys Leu

465 470 475 480

Tyr Lys Asp His Asp Ser Ser Arg Leu Thr His Tyr Glu Gly Val Phe

485 490 495

His Ala Pro Glu Phe Lys Lys Glu Ile Ser Asp Leu Glu Ser Cys Met

500 505 510

Tyr Leu Pro Pro Lys Glu Ala Glu Glu Tyr Leu Gln Asn Pro Lys Lys

515 520 525

Pro Leu Val Glu Cys Glu Tyr Met His Asp Met Gly Thr Pro Asp Gly

530 535 540

Gly Met Gly Ser Tyr Ile Lys Leu Ile Asp Lys Tyr Pro Gln Tyr Met

545 550 555 560

Gly Gly Phe Ile Trp Asp Phe Ile Asp Gln Ala Leu Leu Val His Asp

565 570 575

Pro Val Ser Gly Gln Asp Val Leu Arg Tyr Gly Gly Asp Phe Asp Asp

580 585 590

Arg His Ser Asp Tyr Glu Phe Ser Gly Asp Gly Leu Met Phe Ala Asp

595 600 605

Arg Thr Pro Lys Pro Ala Met Gln Glu Val Arg Tyr Tyr Tyr Gly Leu

610 615 620

His Lys

625

<210> 6

<211> 316

<212>PRT

<213> Lactobacillus amylovorus

<400> 6

Met Ala Tyr Thr Asn Asn Leu His Val Val Tyr Gly Glu Ala Ser Leu

1 5 10 15

Gly Val Asn Gly Gln Asp Phe Ala Tyr Leu Phe Ser Tyr Glu Arg Gly

20 25 30

Gly Leu Glu Ser Leu Lys Ile Lys Asp Lys Glu Trp Leu Tyr Arg Thr

35 40 45

Pro Thr Pro Thr Phe Trp Arg Ala Thr Thr Asp Asn Asp Arg Gly Ser

50 55 60

Gly Phe Asn Gln Lys Ala Ala Gln Trp Leu Gly Ala Asp Met Phe Thr

65 70 75 80

Lys Cys Val Gly Ile His Val Gln Val Asp Asp His Arg Phe Asp Glu

85 90 95

Leu Pro Val Ala Pro Ile Asn Asn Gln Phe Ser Asn Gln Glu Phe Ala

100 105 110

His Glu Val Lys Val Ala Phe Asp Tyr Glu Thr Leu Thr Thr Pro Ala

115 120 125

Thr Lys Val Lys Ile Ile Tyr Asn Ile Asn Asp Phe Gly His Met Thr

130 135 140

Ile Thr Met His Tyr Phe Gly Lys Lys Gly Leu Pro Pro Leu Pro Val

145 150 155 160

Ile Gly Met Arg Phe Ile Met Pro Thr Lys Ala Lys Ser Phe Asp Tyr

165 170 175

Thr Gly Leu Ser Gly Glu Thr Tyr Pro Asp Arg Met Ala Gly Ala Glu

180 185 190

Arg Gly Thr Phe His Ile Asp Gly Leu Pro Val Thr Lys Tyr Leu Val

195 200 205

Pro Gln Glu Asn Gly Met His Met Gln Thr Asn Glu Leu Val Ile Thr

210 215 220

Arg Asn Ser Thr Gln Asn Asn Ala Asp Lys Asp Gly Asp Phe Ser Leu

225 230 235 240

Lys Ile Thr Gln Thr Lys Gln Pro Phe Asn Phe Ser Leu Leu Pro Tyr

245 250 255

Thr Ala Glu Glu Leu Glu Asn Ala Thr His Ile Glu Glu Leu Pro Leu

260 265 270

Ala Arg Arg Ser Val Leu Val Ile Ala Gly Ala Val Arg Gly Val Gly

275 280 285

Gly Ile Asp Ser Trp Gly Ser Asp Val Glu Glu Gln Tyr His Ile Asp

290 295 300

Pro Glu Gln Asp His Glu Phe Ser Phe Thr Leu Asn

305 310 315

<210> 7

<211> 1052

<212>PRT

<213> Bifidobacterium bifidum

<400> 7

Met Asn Thr Thr Asp Asp Gln Arg Lys Asn Gly Asp Pro Ile Val Ser

1 5 10 15

Pro Ser Ile Pro Thr Thr Ala Trp Leu Ala Asp Pro Arg Val Tyr Ala

20 25 30

Val His Arg Leu Asp Ala His Ser Asp His Ala Cys Trp Ser Arg Ser

35 40 45

Pro Val Asp Gly Glu Ser Thr Asp Leu Arg Gln Ser Leu Asp Gly Glu

50 55 60

Trp Arg Val Arg Val Glu Thr Ala Pro Thr Gly Arg Phe Pro Asp Gly

65 70 75 80

Thr Ser Asp Gly Pro Asp Trp Ile Ser Asp Val Ser Pro Leu Phe Ala

85 90 95

Ala Pro Gly Phe Asp Asp Ser Ser Phe Ser Arg Val Gln Val Pro Ser

100 105 110

His Leu Glu Thr Ala Gly Leu Leu Ala Pro Gln Tyr Val Asn Val Gln

115 120 125

Tyr Pro Trp Asp Gly His Glu Asp Pro Lys Ala Pro Ala Ile Pro Glu

130 135 140

His Gly His Val Ala Val Tyr Arg Arg Glu Phe Asp Ala Asp Gly Glu

145 150 155 160

Val Ala Gln Ala Val Arg Glu Gly Arg Pro Val Thr Leu Thr Phe Gln

165 170 175

Gly Ala Ala Thr Ala Ile Tyr Val Trp Leu Asn Gly Ser Phe Ile Gly

180 185 190

Tyr Ala Glu Asp Ser Phe Thr Pro Ser Glu Phe Asp Val Thr Asp Ala

195 200 205

Ile Lys Val Asp Gly Asn Val Leu Ala Val Ala Cys Tyr Glu Tyr Ser

210 215 220

Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe Trp Arg Leu His Gly Leu

225 230 235 240

Phe Arg Ser Val Glu Leu Asn Ala Arg Pro Ala Ala His Val Ala Asp

245 250 255

Leu His Ala Asp Ala Asp Trp Asp Leu Ala Thr Ser Arg Gly Ser Leu

260 265 270

Ser Leu Asp Val Leu Ile Asp Gly Ala Ala Asn Ala Ala Thr Ala Asp

275 280 285

Phe Ala Leu Arg Asp Lys Asn Gly Thr Ile Val Trp Arg Thr Ala Thr

290 295 300

Lys Ala Asp Gly Thr Leu His Ala Glu Ala Glu Ile Asp Asp Ala Ala

305 310 315 320

Pro Trp Ser Ala Glu Arg Pro Asp Leu Tyr Glu Leu Ser Val Thr Leu

325 330 335

Leu Asp Ala Asp Gly Lys Val Leu Glu Thr Ala Arg Thr Arg Ile Gly

340 345 350

Phe Arg His Val Ala Ile Glu Asp Gly Ile Leu Lys Leu Asn Gly Lys

355 360 365

Arg Leu Val Phe Arg Gly Val Asn Arg His Glu Phe Asp Cys Arg Arg

370 375 380

Gly Arg Ala Ile Thr Glu Glu Asp Met Leu Trp Asp Ile Arg Phe Met

385 390 395 400

Lys Arg His Asn Ile Asn Ala Val Arg Thr Ser His Tyr Pro Asn Gln

405 410 415

Ser Arg Trp Tyr Glu Leu Cys Asp Glu Tyr Gly Ile Tyr Leu Ile Asp

420 425 430

Glu Thr Asn Leu Glu Thr His Gly Ser Trp Asn Ser Pro Gly Asp Ile

435 440 445

Pro Val Gly Thr Ser Val Pro Gly Asp Asp Glu Ala Trp Leu Gly Ala

450 455 460

Cys Ile Asp Arg Leu Asp Ser Met Ile Leu Arg Asp Arg Asn His Pro

465 470 475 480

Ser Val Leu Val Trp Ser Leu Gly Asn Glu Ser Tyr Ala Gly Glu Val

485 490 495

Leu Lys Ala Met Ser Ala His Ala His Gln Leu Asp Pro Gly Arg Pro

500 505 510

Val His Tyr Glu Gly Val Asn Trp Asn His Ala Tyr Asp Gly Ile Ser

515 520 525

Asp Phe Glu Ser Arg Met Tyr Ala Lys Pro Ala Glu Ile Gln Asp Trp

530 535 540

Leu Glu His Gly Asp Glu Arg Gly Glu Ala Ser Lys Pro Phe Val Ser

545 550 555 560

Cys Glu Tyr Met His Ala Met Gly Asn Ser Cys Gly Gly Leu Ser Glu

565 570 575

Phe Ile Asp Leu Glu Arg Tyr Glu Arg Tyr Ser Gly Gly Phe Ile Trp

580 585 590

Asp Tyr Ile Asp Gln Gly Leu Val Gln Arg Leu Pro Asp Gly Ser Glu

595 600 605

Arg Leu Ser Val Gly Gly Glu Trp Gly Asp Arg Pro Thr Asp Tyr Glu

610 615 620

Phe Val Gly Asn Gly Ile Val Phe Ala Asp Arg Thr Pro Ser Pro Lys

625 630 635 640

Ala Gln Glu Val Lys Gln Leu Tyr Ser Pro Val Lys Leu Ala Pro Asp

645 650 655

Gly His Gly Val Thr Ile Glu Asn Arg Asn Leu Phe Ala Gly Thr Asp

660 665 670

Gly Tyr Val Phe Ala Ala Arg Leu Leu Glu Asp Gly His Glu Ile Trp

675 680 685

His Ala Asp Tyr Arg Phe Asp Val Ala Ala Gly Asp Thr Gln His His

690 695 700

Asp Ile Ala Phe Pro Asp Ile Asp Ala Asp Gly Asp Thr Arg Glu Val

705 710 715 720

Thr Tyr Glu Val Asp Leu Leu Leu Ala Glu Ala Thr Ala Trp Ala Pro

725 730 735

Ala Gly Tyr Glu Leu Ala Phe Gly Gln Leu Thr Gly Thr Leu Asn Pro

740 745 750

Glu Gln Asp Ile Thr Glu Thr Ser His Asp Asp Asp Gly Arg Ala Thr

755 760 765

Arg Thr Leu Ser Arg Trp Asn Ala Gly Ile Arg Arg Asp Asp Glu Glu

770 775 780

Ile Leu Leu Ser Arg Thr Gln Gly Gly Ile Val Ser Trp Lys Arg Asp

785 790 795 800

Asp Arg Glu Met Val Ile Arg Arg Pro Glu Leu Val Thr Phe Arg Pro

805 810 815

Leu Thr Asp Asn Asp Arg Gly Asn His Ser Gly Phe Asp Arg Ala Ala

820 825 830

Trp Phe Ala Ala Gly Arg Tyr Ala Ile Val Thr Glu Thr Lys Ile His

835 840 845

Glu Ser Asp Asp Gly Leu Val Ala Glu Tyr Gln Tyr Glu Leu Ala Asp

850 855 860

Pro Asn His Thr Pro Val Ser Val Thr Tyr His Val Asn Ser Asp Met

865 870 875 880

Arg Met Gln Leu Thr Val Glu Tyr Pro Gly Asn Ala Thr Asp Met Ala

885 890 895

Ser Leu Pro Ala Phe Gly Ile Glu Trp Glu Leu Pro Gly Glu Tyr Asp

900 905 910

Arg Leu Arg Tyr Tyr Gly Pro Gly Pro Glu Glu Thr Tyr Arg Asp Arg

915 920 925

Lys Gln Gly Gly Lys Leu Gly Ile Trp Asp Ala Thr Ala Lys Ala Ser

930 935 940

Met Ala Pro Tyr Leu Met Val Gln Glu Thr Gly Ser His Glu Asp Val

945 950 955 960

Arg Trp Leu Glu Ala Thr Asp Ile Gln Gly His Gly Leu Arg Val Thr

965 970 975

Gln Arg Gly Asp Arg His Phe Thr Ala Ser Leu Leu Pro Trp Asn Thr

980 985 990

Tyr Thr Ile Glu Ala Ala Arg Arg His Glu Asp Leu Pro Lys Pro Arg

995 1000 1005

His Asn Tyr Leu Arg Leu Leu Ala Ala Gln Met Gly Val Gly Gly

1010 1015 1020

Asp Asp Ser Trp Gly Ala Pro Val His Thr Ala Tyr Gln Leu Pro

1025 1030 1035

Ala Gly Arg Pro Leu Thr Leu Asp Val Asn Leu Glu Leu Ile

1040 1045 1050

<210> 8

<211> 1052

<212>PRT

<213> Bifidobacterium bifidum

<400> 8

Met Asn Thr Thr Asp Asp Gln Arg Lys Asn Gly Asp Pro Ile Val Ser

1 5 10 15

Pro Ser Ile Pro Thr Thr Ala Trp Leu Ala Asp Pro Arg Val Tyr Ala

20 25 30

Val His Arg Leu Asp Ala His Ser Asp His Ala Cys Trp Ser Arg Ser

35 40 45

Pro Val Asp Gly Glu Ser Thr Asp Leu Arg Gln Ser Leu Asp Gly Glu

50 55 60

Trp Arg Val Arg Val Glu Thr Ala Pro Thr Gly Arg Phe Pro Asp Gly

65 70 75 80

Thr Ser Asp Gly Pro Asp Trp Ile Ser Asp Val Ser Pro Leu Phe Ala

85 90 95

Ala Pro Gly Phe Asp Asp Ser Ser Phe Ser Arg Val Gln Val Pro Ser

100 105 110

His Leu Glu Thr Ala Gly Leu Leu Ala Pro Gln Tyr Val Asn Val Gln

115 120 125

Tyr Pro Trp Asp Gly His Glu Asp Pro Lys Ala Pro Ala Ile Pro Glu

130 135 140

His Gly His Val Ala Val Tyr Arg Arg Glu Phe Asp Ala Asp Gly Glu

145 150 155 160

Val Ala Gln Ala Val Arg Glu Gly Arg Pro Val Thr Leu Thr Phe Gln

165 170 175

Gly Ala Ala Thr Ala Ile Tyr Val Trp Leu Asn Gly Ser Phe Ile Gly

180 185 190

Tyr Ala Glu Asp Ser Phe Thr Pro Ser Glu Phe Asp Val Thr Asp Ala

195 200 205

Ile Lys Val Asp Gly Asn Val Leu Ala Val Ala Cys Tyr Glu Tyr Ser

210 215 220

Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe Trp Arg Leu His Gly Leu

225 230 235 240

Phe Arg Ser Val Glu Leu Asn Ala Arg Pro Ala Ala His Val Ala Asp

245 250 255

Leu His Ala Asp Ala Asp Trp Asp Leu Ala Thr Ser Arg Gly Ser Leu

260 265 270

Ser Leu Asp Val Leu Ile Asp Gly Ala Ala Asn Ala Ala Thr Ala Asp

275 280 285

Phe Ala Leu Trp Asp Lys Asn Gly Thr Ile Val Trp His Ile Val Thr

290 295 300

Lys Ala Asp Gly Thr Leu His Ala Glu Ala Glu Ile Asp Asp Ala Ala

305 310 315 320

Pro Trp Ser Ala Glu Arg Pro Asp Leu Tyr Glu Leu Ser Val Thr Leu

325 330 335

Leu Asp Ala Asp Gly Lys Val Leu Glu Thr Ala Arg Thr Arg Ile Gly

340 345 350

Phe Arg His Val Ala Ile Glu Asp Gly Ile Leu Lys Leu Asn Gly Lys

355 360 365

Arg Leu Val Phe Arg Gly Val Asn Arg His Glu Phe Asp Cys Arg Arg

370 375 380

Gly Arg Ala Ile Thr Glu Glu Asp Met Leu Trp Asp Ile Arg Phe Met

385 390 395 400

Lys Arg His Asn Ile Asn Ala Val Arg Thr Ser His Tyr Pro Asn Gln

405 410 415

Ser Arg Trp Tyr Glu Leu Cys Asp Glu Tyr Gly Ile Tyr Leu Ile Asp

420 425 430

Glu Thr Asn Leu Glu Thr His Gly Ser Trp Asn Ser Pro Gly Asp Ile

435 440 445

Pro Val Gly Thr Ser Val Pro Gly Asp Asp Glu Ala Trp Leu Gly Ala

450 455 460

Cys Ile Asp Arg Leu Asp Ser Met Ile Leu Arg Asp Arg Asn His Pro

465 470 475 480

Ser Val Leu Val Trp Ser Leu Gly Asn Glu Ser Tyr Ala Gly Glu Val

485 490 495

Leu Lys Ala Met Ser Ala His Ala His Arg Leu Asp Pro Gly Arg Pro

500 505 510

Val His Tyr Glu Gly Val Asn Trp Asn His Ala Tyr Asp Gly Ile Ser

515 520 525

Asp Phe Glu Ser Arg Met Tyr Ala Lys Pro Ala Glu Ile Gln Asp Trp

530 535 540

Leu Glu His Gly Asp Glu Arg Gly Glu Ala Ser Lys Pro Phe Val Ser

545 550 555 560

Cys Glu Tyr Met His Ala Met Gly Asn Ser Cys Gly Gly Leu Ser Glu

565 570 575

Phe Ile Asp Leu Glu Arg Tyr Glu Arg Tyr Ser Gly Gly Phe Ile Trp

580 585 590

Asp Tyr Ile Asp Gln Gly Leu Val Gln Arg Leu Pro Asp Gly Ser Glu

595 600 605

Arg Leu Ser Val Gly Gly Glu Trp Gly Asp Arg Pro Thr Asp Tyr Glu

610 615 620

Phe Val Gly Asn Gly Ile Val Phe Ala Asp Arg Thr Pro Ser Pro Lys

625 630 635 640

Ala Gln Glu Val Lys Gln Leu Tyr Ser Pro Val Lys Leu Ala Pro Asp

645 650 655

Gly His Gly Val Thr Ile Glu Asn Arg Asn Leu Phe Ala Gly Thr Asp

660 665 670

Gly Tyr Val Phe Ala Ala Arg Leu Leu Glu Asp Gly His Glu Ile Trp

675 680 685

His Ala Asp Tyr Arg Phe Asp Val Ala Ala Gly Asp Thr Gln His His

690 695 700

Asp Ile Ala Phe Pro Asp Ile Asp Ala Asp Gly Asp Thr Arg Glu Val

705 710 715 720

Thr Tyr Glu Val Asp Leu Leu Leu Ala Glu Ala Thr Ala Trp Ala Pro

725 730 735

Ala Gly Tyr Glu Leu Ala Phe Gly Gln Leu Thr Gly Thr Leu Asn Pro

740 745 750

Glu Gln Asp Ile Thr Glu Thr Ser His Asp Asp Asp Gly Arg Ala Thr

755 760 765

Arg Thr Leu Ser Arg Trp Asn Ala Gly Ile Arg Arg Asp Asp Lys Glu

770 775 780

Ile Leu Leu Ser Arg Thr Gln Gly Gly Ile Val Ser Trp Lys Arg Asp

785 790 795 800

Asp Arg Glu Met Val Ile Arg Arg Pro Glu Leu Val Thr Phe Arg Pro

805 810 815

Leu Thr Asp Asn Asp Arg Gly Asn His Ser Gly Phe Asp Arg Ala Ala

820 825 830

Trp Phe Ala Ala Gly Arg Tyr Ala Ile Val Thr Glu Thr Lys Ile His

835 840 845

Glu Ser Asp Asp Gly Leu Val Ala Glu Tyr Gln Tyr Glu Leu Ala Asp

850 855 860

Pro Asn His Thr Pro Val Ser Val Thr Tyr His Val Asn Ser Asp Met

865 870 875 880

Arg Met Gln Leu Thr Val Glu Tyr Pro Gly Asn Ala Thr Asp Met Ala

885 890 895

Ser Leu Pro Ala Phe Gly Ile Glu Trp Glu Leu Pro Gly Glu Tyr Asp

900 905 910

Arg Leu Arg Tyr Tyr Gly Pro Gly Pro Glu Glu Thr Tyr Arg Asp Arg

915 920 925

Lys Gln Gly Gly Lys Leu Gly Ile Trp Asp Ala Thr Ala Lys Ala Ser

930 935 940

Met Ala Pro Tyr Leu Met Val Gln Glu Thr Gly Ser His Glu Asp Val

945 950 955 960

Arg Trp Leu Glu Ala Thr Asp Ile Gln Gly His Gly Leu Arg Val Thr

965 970 975

Gln Arg Gly Asp Arg His Phe Thr Ala Ser Leu Leu Pro Trp Asn Thr

980 985 990

Tyr Met Ile Glu Ala Ala Arg Arg His Glu Asp Leu Pro Glu Pro Arg

995 1000 1005

His Asn Tyr Leu Arg Leu Leu Ala Ala Gln Met Gly Val Gly Gly

1010 1015 1020

Asp Asp Ser Trp Gly Ala Pro Val His Thr Ala Tyr Gln Leu Pro

1025 1030 1035

Ala Gly Arg Pro Leu Thr Leu Asp Val Asn Leu Glu Leu Ile

1040 1045 1050

<210> 9

<211> 1055

<212>PRT

<213> Bifidobacterium breve

<400> 9

Met Thr Asn Ser Met Gln Gly Lys Ala Lys Thr Ile Met Thr Asn Leu

1 5 10 15

Gln Ser Ala Gln Gln Phe Ser Gln Ala Trp Leu Thr Asp Pro Arg Val

20 25 30

Phe Ala Val Asn Arg Leu Ala Ala His Ser Ser His Lys Phe Tyr Asp

35 40 45

His Ser Pro Gln Cys Gly Glu Ala Met Asp Leu Lys Gln Ser Leu Asp

50 55 60

Gly Gln Trp Arg Val Gln Met Leu Asp Leu Ala Asp Leu Ala Asp Asn

65 70 75 80

Glu Leu Ala Glu Ala Ala Phe Ala Gln Pro Gly Tyr Asp Ala Ala Gly

85 90 95

Phe Ser Pro Ile Glu Val Pro Ser Ala Leu Glu Thr Lys Gly Phe Leu

100 105 110

Asn His Gln Tyr Val Asn Gln Gln Tyr Pro Trp Ser Gly His Glu Ser

115 120 125

Pro Val Ala Pro Asp Val Pro Lys His Asn His Val Ala Leu Tyr Arg

130 135 140

His Glu Phe Ser Leu Glu Pro Lys Ala Ala Ala Val Leu Glu Ala Asn

145 150 155 160

Lys Thr Ala Ala Asp Asp Ala Ala Lys Arg Arg Val Thr Leu Thr Phe

165 170 175

Gln Gly Ala Ala Thr Ala Ile Val Val Trp Leu Asn Gly Ala Phe Ile

180 185 190

Gly Tyr Ala Glu Asp Ser Phe Thr Pro Ser Glu Phe Asp Val Thr Asp

195 200 205

Val Leu Arg Asp Gly Val Asn Thr Leu Ala Val Ala Cys Phe Glu Phe

210 215 220

Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe Trp Arg Leu His Gly

225 230 235 240

Ile Phe Arg Ser Val Glu Leu Glu Ala Gln Pro Leu Val His Val Asn

245 250 255

Asp Leu Arg Val Leu Ala Asp Tyr Asp His Thr Thr Gly Glu Gly Ser

260 265 270

Leu Asp Val Val Ala Leu Leu Arg Asn Ala Gly Thr Ala Ala Ala Val

275 280 285

Ala Ala Thr Val Leu Asp Ala Ala Gly Asn Thr Val Trp His Ser Lys

290 295 300

Leu Thr Ala Gly Ala Asp Ala Glu Thr Leu Thr Val Lys Ala Asn Val

305 310 315 320

Gly Lys Val Asn Pro Trp Ser Ala Glu Glu Pro Thr Leu Tyr Thr Leu

325 330 335

Gln Val Val Ala Thr Asp Ala Ala Gly Gln Val Ile Glu Ala Ala Leu

340 345 350

Gln Arg Ile Gly Phe Arg His Phe Ala Ile Glu Asp Gly Leu Met Lys

355 360 365

Leu Asn Gly Lys Arg Ile Val Phe Lys Gly Val Asp Arg His Glu Phe

370 375 380

Asp Ala Arg Thr Gly Arg Thr Ile Ala Glu Ala Asp Met Ile Glu Asp

385 390 395 400

Ile His Ser Phe Lys Arg Leu Asn Ile Asn Ala Val Arg Thr Ser His

405 410 415

Tyr Pro Asn Glu Thr Arg Trp Tyr Glu Leu Cys Asp Glu Tyr Gly Ile

420 425 430

Tyr Val Leu Asp Glu Thr Asn Leu Glu Thr His Gly Ser Trp Thr Asp

435 440 445

Pro Gly Asp Val Phe Gln Pro Ala Arg Ala Ile Pro Gly Ser Lys Asp

450 455 460

Glu Trp Arg Ala Ala Cys Val Asp Arg Thr Ala Ser Met Val Arg Arg

465 470 475 480

Asp Tyr Asn His Pro Ser Val Val Ile Trp Ser Leu Gly Asn Glu Ala

485 490 495

Phe Gly Gly Asp Val Phe Tyr Ser Met Arg Asp Phe Val His Glu Asn

500 505 510

Asp Pro Phe Arg Pro Val His Tyr Glu Gly Thr Phe Asn Asp Pro Glu

515 520 525

Phe Ser Ala Ala Thr Asp Ile Met Ser Arg Met Tyr Ala Lys Pro Asp

530 535 540

Glu Ile Val Lys Leu Tyr Leu Gly Glu Asp Gly Lys Lys Pro Tyr Ile

545 550 555 560

Ser Cys Glu Tyr Ser His Ser Met Gly Asn Ser Thr Gly Gly Leu His

565 570 575

Leu Tyr Thr Glu Leu Glu Arg Tyr Pro Leu Tyr Gln Gly Gly Phe Ile

580 585 590

Trp Asp Tyr Val Asp Gln Ala Leu Trp Gln Asp Cys Gly Asn Gly Thr

595 600 605

Glu Arg Leu Ala Tyr Gly Gly Asp Phe Glu Asp Arg Pro Asn Asp Tyr

610 615 620

Glu Phe Ser Gly Asp Gly Val Met Phe Ala Asp Arg Thr Pro Ser Pro

625 630 635 640

Lys Ala Gln Glu Val Lys Gln Leu Tyr Ala Asn Val Lys Leu Val Pro

645 650 655

Asp Glu Ser Gly Val Thr Ile Thr Asn Asp Asn Leu Phe Ile Ser Thr

660 665 670

Ala Ser Ser Leu Phe Thr Ala Arg Val Leu Val Asp Gly Ala Glu Arg

675 680 685

Trp His Ala Asn Tyr Arg Phe Asp Val Pro Ala Gly Glu Thr Val Arg

690 695 700

Glu Pro Ile Ala Phe Pro Lys Val Thr Asp Leu Val Ala Leu Ser Gly

705 710 715 720

Ser Ala Glu Val Thr Tyr Glu Val Asp Gln Arg Leu Ala Glu Ala Thr

725 730 735

Asp Trp Ala Pro Ala Gly Tyr Glu Leu Thr Phe Gly Gln Tyr Val Ala

740 745 750

Ala Val Ser Phe Asp Asp Gly Ala Ala Asp Ala Val Val Ala Gly Asp

755 760 765

Ala Glu Val Ala Ala Asp Gly Phe Asn Ala Gly Ile His Thr Asp Phe

770 775 780

Gly Glu Val Leu Leu Ser Lys Thr Gln Gly Gly Met Val Ser Phe Lys

785 790 795 800

Arg Asp Gly Arg Glu Met Val Ile Arg Arg Pro Asn Leu Thr Thr Phe

805 810 815

Arg Ala Leu Thr Asp Asn Asp Arg Gly Asn Gly Ser Gly Phe Glu Arg

820 825 830

Ala Gln Trp Met Ala Ala Gly Arg Tyr Ala Arg Val Thr Gly Thr Ser

835 840 845

Val Glu Glu Thr Ala Asp Gly Lys Gly Leu Lys Ala Thr Tyr Ser Tyr

850 855 860

Glu Leu Ala Asp Ala Lys His Thr Pro Val Thr Val His Tyr Glu Val

865 870 875 880

Asp Ala Ala Leu Arg Val His Leu Thr Val Glu Tyr Pro Gly Glu Ala

885 890 895

Asp Ala Ala Thr Leu Pro Ala Phe Gly Leu Glu Trp Ile Leu Pro Lys

900 905 910

Gln Tyr Asp Arg Leu Arg Phe Tyr Gly Leu Gly Pro Glu Glu Thr Tyr

915 920 925

Ala Asp Arg Leu His Gly Ala Lys Leu Gly Val Phe Ser Arg Thr Ala

930 935 940

Ala Glu Asp Cys Ala Pro Tyr Leu Leu Pro Gln Glu Thr Gly Asn His

945 950 955 960

Glu Gln Val Arg Trp Ala Glu Ile Thr Asp Glu Tyr Gly His Gly Met

965 970 975

Arg Val Thr Ala Ala Gly Gly Thr Arg Phe Ala Thr Ser Leu Leu Pro

980 985 990

Tyr Ser Ser Leu Met Phe Glu Asp Ala Leu His Gln Asn Glu Leu Pro

995 1000 1005

Lys Pro Arg His Thr Phe Leu Arg Leu Leu Ala Ala Gln Met Gly

1010 1015 1020

Val Gly Gly Asp Asp Thr Trp Gly Ala Pro Val His Asp Glu Phe

1025 1030 1035

Gln Val Pro Ala Asp Gln Pro Leu Lys Leu Asp Val Thr Leu Glu

1040 1045 1050

Leu Ile

1055

<210> 10

<211> 689

<212>PRT

<213> Bifidobacterium catenulatum

<400> 10

Met Thr Gln Arg Arg Ser Tyr Arg Trp Pro Gln Pro Leu Ala Gly Gln

1 5 10 15

Gln Ala Arg Ile Trp Tyr Gly Gly Asp Tyr Asn Pro Asp Gln Trp Pro

20 25 30

Glu Glu Val Trp Asp Asp Asp Val Arg Leu Met Lys Lys Ala Gly Val

35 40 45

Asn Leu Val Ser Val Gly Ile Phe Ser Trp Ala Lys Ile Glu Thr Ser

50 55 60

Glu Gly Val Tyr Asp Phe Asp Trp Leu Asp Arg Ile Ile Asp Lys Leu

65 70 75 80

Gly Glu Ala Gly Ile Ala Val Asp Leu Ala Ser Ala Thr Ala Ser Pro

85 90 95

Pro Met Trp Leu Thr Gln Ala His Pro Glu Val Leu Trp Lys Asp Tyr

100 105 110

Arg Gly Asp Val Cys Gln Pro Gly Ala Arg Gln His Trp Arg Pro Thr

115 120 125

Ser Pro Val Phe Arg Glu Tyr Ala Leu Lys Leu Cys Arg Ala Met Ala

130 135 140

Glu His Tyr Lys Gly Asn Pro Tyr Val Val Ala Trp His Val Ser Asn

145 150 155 160

Glu Tyr Gly Cys His Asn Arg Phe Asp Tyr Ser Glu Asp Ala Glu Arg

165 170 175

Ala Phe Arg Lys Trp Cys Glu Glu Arg Tyr Gly Thr Ile Asp Ala Val

180 185 190

Asn Asp Ala Trp Gly Thr Ala Phe Trp Ala Gln Arg Met Asn Asp Phe

195 200 205

Thr Glu Ile Val Pro Pro Arg Phe Ile Gly Asp Gly Asn Phe Met Asn

210 215 220

Pro Gly Lys Leu Leu Asp Phe Lys Arg Phe Ser Ser Asp Ala Leu Lys

225 230 235 240

Ala Phe Tyr Val Ala Glu Arg Asp Ala Leu Ala Glu Ile Thr Pro Asp

245 250 255

Leu Pro Leu Thr Thr Asn Phe Met Val Ser Ala Ala Gly Ser Val Leu

260 265 270

Asp Tyr Asp Asp Trp Gly Arg Glu Val Asp Phe Val Ser Asn Asp His

275 280 285

Tyr Phe Ile Pro Gly Glu Ala His Leu Asp Glu Leu Ala Phe Ser Ala

290 295 300

Ser Leu Val Asp Gly Ile Ala Arg Lys Asp Pro Trp Phe Leu Met Glu

305 310 315 320

His Ser Thr Ser Ala Val Asn Trp Arg Pro Val Asn Tyr Arg Lys Glu

325 330 335

Pro Gly Gln Leu Val Arg Asp Ser Leu Ala His Val Ala Met Gly Ala

340 345 350

Asp Ala Val Cys Tyr Phe Gln Trp Arg Gln Ser Lys Ala Gly Ala Glu

355 360 365

Lys Phe His Ser Ala Met Val Pro His Thr Gly Glu Asp Ser Ala Val

370 375 380

Phe Arg Asp Val Cys Glu Leu Gly Ala Asp Leu Asn Thr Leu Ala Asp

385 390 395 400

Asn Gly Leu Leu Gly Thr Lys Leu Ala Lys Ser Lys Val Ala Val Val

405 410 415

Phe Asp Tyr Glu Ser Glu Trp Ala Thr Glu His Thr Ala Thr Pro Thr

420 425 430

Gln Lys Val His His Val Asp Glu Pro Leu Gln Trp Phe Arg Ala Leu

435 440 445

Ala Asp His Gly Val Thr Ala Asp Val Val Pro Val Ser Ser Asn Trp

450 455 460

Asp Glu Tyr Glu Val Val Val Leu Pro Ser Val Tyr Ile Leu Ser Glu

465 470 475 480

Glu Thr Thr Arg Arg Val Arg Asp Tyr Val Val Asn Gly Gly Arg Leu

485 490 495

Ile Val Thr Tyr Tyr Thr Gly Leu Ser Asp Glu Lys Asp His Val Trp

500 505 510

Leu Gly Gly Tyr Pro Gly Ser Ile Arg Asp Val Val Gly Val Arg Val

515 520 525

Glu Glu Phe Met Pro Met Gly Asp Asp Phe Pro Gly Val Pro Asp Cys

530 535 540

Leu Gly Leu Ser Asn Gly Ala Val Ala His Asp Ile Ala Asp Val Ile

545 550 555 560

Gly Ser Val Asp Gly Thr Ala Thr Val Leu Glu Thr Phe Arg Asp Asp

565 570 575

Pro Trp Thr Gly Met Asp Gly Ala Pro Ala Ile Val Ala Asn Thr Phe

580 585 590

Gly Glu Gly Arg Ser Val Tyr Val Gly Ala Arg Leu Gly Arg Asp Gly

595 600 605

Ile Ala Lys Ser Leu Pro Glu Ile Phe Glu Ser Leu Gly Met Ala Glu

610 615 620

Thr Gly Glu Asn Asp Ser Arg Val Leu Arg Val Glu Arg Glu Gly Ser

625 630 635 640

Asp Gly Ser Arg Phe Val Phe Ser Phe Asn Arg Thr His Glu Ala Val

645 650 655

Gln Ile Pro Phe Glu Gly Lys Ile Val Val Ser Ser Phe Ala Glu Val

660 665 670

Ser Gly Glu Asn Val Ser Ile Lys Pro Asn Gly Val Ile Val Thr Lys

675 680 685

Gln

<210> 11

<211> 1023

<212>PRT

<213> Bifidobacterium catenulatum

<400> 11

Met Ala Asn Ser Asn Arg Val Glu His Ala Ser Glu Thr Trp Leu Thr

1 5 10 15

Asp Ala Thr Val Phe Glu Val Asn Arg Thr Pro Ala His Ser Asn His

20 25 30

Lys Cys Phe Thr His Asp Pro Gln Ser Gly Glu His Ser Asp Leu Thr

35 40 45

Gln Ser Leu Asp Gly Glu Trp Arg Val Glu Ile Val Gln Ala Ser Asp

50 55 60

Ile Asp Phe Asn Glu Glu Pro Phe Val Ala Glu Asn Phe Asp Asp Ser

65 70 75 80

Ser Phe Cys Arg Ala Gln Val Pro Gly His Leu Gln Met Ala Gly Leu

85 90 95

Leu Lys Asn Lys Tyr Val Asn Ile Gln Tyr Pro Trp Asp Gly His Glu

100 105 110

Asn Pro Leu Glu Pro Asn Val Pro Glu Asn Asn His Val Ala Leu Tyr

115 120 125

Arg Arg Lys Phe Val Val Ser Lys Arg Leu Ala Asp Thr Lys Glu Ser

130 135 140

Glu Gly Ser Val Ser Ile Val Phe His Gly Met Ala Thr Ala Ile Tyr

145 150 155 160

Val Trp Val Asn Gly Leu Phe Ala Gly Tyr Gly Glu Asp Gly Phe Thr

165 170 175

Pro Asn Glu Phe Asp Ile Thr Asp Leu Leu His Asp Gly Glu Asn Val

180 185 190

Val Ala Val Ala Cys Tyr Glu Tyr Ser Ser Ala Ser Trp Leu Glu Asp

195 200 205

Gln Asp Phe Trp Arg Leu His Gly Leu Phe Arg Ser Val Glu Leu Thr

210 215 220

Ala Gln Pro His Val His Val Glu Asn Met Gln Leu Glu Ala Asp Trp

225 230 235 240

Asp Ala Glu Ser Gly Thr Ala Ser Leu Asp Ala Ala Leu Ser Val Arg

245 250 255

Asn Ala Ser Asp Ala Ala Thr Ile Ser Ala Thr Leu Lys Asp Ser Glu

260 265 270

Gly Asn Val Val Trp Glu Ala Ser Thr Asn Ala Asp Ala Asn Thr Thr

275 280 285

Phe Ala Ser Gly Ser Leu Gln Gly Leu Glu Pro Trp Ser Ala Glu Ser

290 295 300

Pro Ser Leu Tyr Glu Leu Glu Val Asn Val Ile Asp Gln Ala Gly Asn

305 310 315 320

Ile Val Glu Ala Ala Val Gln Lys Val Gly Phe Arg Arg Phe Arg Ile

325 330 335

Glu Asn Gly Ile Met Thr Leu Asn Gly Lys Arg Ile Val Phe Lys Gly

340 345 350

Ala Asp Arg His Glu Phe Asp Ala Lys Arg Gly Arg Ser Ile Thr Glu

355 360 365

Gln Asp Met Ile Asp Asp Val Ile Phe Cys Lys Arg His Asn Ile Asn

370 375 380

Ala Ile Arg Thr Ser His Tyr Pro Asn Gln Glu Arg Trp Tyr Asp Leu

385 390 395 400

Cys Asp Glu Tyr Gly Ile Tyr Leu Ile Asp Glu Thr Asn Leu Glu Thr

405 410 415

His Gly Ser Trp Cys Leu Pro Gly Asp Val Val Thr Ala Glu Thr Ala

420 425 430

Val Pro Gly Ser Lys Ala His Trp Glu Gly Ala Cys Val Asp Arg Val

435 440 445

Asn Ser Met Val Arg Arg Asp Tyr Asn His Pro Ser Val Val Ile Trp

450 455 460

Ser Leu Gly Asn Glu Ser Tyr Thr Gly Asp Val Phe Arg Ala Met Tyr

465 470 475 480

Lys His Val His Asp Ile Asp Pro Asn Arg Pro Val His Tyr Glu Gly

485 490 495

Val Thr Lys Asn Arg Asp Tyr Asp Asp Val Thr Asp Ile Glu Thr Arg

500 505 510

Met Tyr Glu His Ala Asp Val Val Glu Glu Tyr Leu Lys Asn Asp Pro

515 520 525

Gln Lys Pro Tyr Ile Ser Cys Glu Tyr Met His Ala Met Gly Asn Ser

530 535 540

Val Gly Asn Leu Asp Glu Tyr Thr Ala Leu Glu Arg Tyr Pro His Tyr

545 550 555 560

Gln Gly Gly Phe Ile Trp Asp Phe Ile Asp Gln Ala Ile Tyr Ala Thr

565 570 575

Gln Pro Asp Gly Ser Thr Arg Leu Cys Tyr Gly Gly Asp Phe Gly Asp

580 585 590

Arg Pro Ser Asp Tyr Glu Phe Ser Gly Asn Gly Leu Val Phe Ala Asp

595 600 605

Arg Thr Pro Thr Pro Lys Ala Gln Glu Val Lys Gln Leu Tyr Ser Asn

610 615 620

Val His Ile Asp Val Thr Asp Arg Ser Val Ser Ile Lys Asn Asp Asn

625 630 635 640

Leu Phe Ile Ser Thr Gly Gly Tyr Gln Phe Val Leu Arg Ile Leu Ala

645 650 655

Asp Gly Glu Pro Val Trp Gln Ser Glu Arg Arg Phe Asp Val Pro Ala

660 665 670

Asp Ser Ala Cys Thr Phe Asp Val Glu Trp Pro Val Asp Leu Tyr Arg

675 680 685

Ala Asn Ala Asp Glu Leu Val Leu Glu Val Ser Gln Arg Leu Ala Glu

690 695 700

Ala Thr Asp Trp Ala Pro Ala Gly Tyr Glu Leu Ala Phe Gly Gln Thr

705 710 715 720

Ile Val Ala Gly Thr Lys Ala Ala Glu Asp Ala Ala Leu Pro Ala Asp

725 730 735

Gly Ile Val Thr Val Gly Arg Trp Asn Ala Gly Val Gln Gly Ser Gly

740 745 750

Arg Glu Ile Leu Leu Ser Arg Thr Gln Gly Gly Leu Val Ser Tyr Thr

755 760 765

Phe Asp Gly His Glu Phe Val Leu Arg Arg Pro Ala Ile Thr Thr Phe

770 775 780

Arg Ala Leu Thr Asp Asn Asp Arg Gly Ala Gly His Gly Phe Glu Arg

785 790 795 800

Ala Gln Trp Met Val Ala Gly Arg Tyr Ala Arg Cys Val Asp Asn Val

805 810 815

Ile Glu Gln Val Asp Glu Asp Thr Leu Lys Ala Val Tyr Thr Tyr Glu

820 825 830

Leu Ala Thr Pro Gln Cys Thr Lys Val Thr Val Gly Tyr Thr Ala Asp

835 840 845

Thr Thr Gly Arg Leu Asn Leu His Val Glu Tyr Pro Gly Glu Ser Gly

850 855 860

Glu Leu Pro Thr Ile Pro Ala Phe Gly Ile Glu Trp Thr Leu Pro Val

865 870 875 880

Gln Tyr Ser Asn Leu Arg Phe Phe Gly Ala Gly Pro Glu Glu Thr Tyr

885 890 895

Gln Asp Arg Lys His Ala Lys Leu Gly Val Trp Ser Thr Asp Ala Phe

900 905 910

Lys Asp His Ala Pro Tyr Leu Met Pro Gln Glu Thr Gly Asn His Glu

915 920 925

Glu Val Arg Trp Ala Glu Ile Thr Asp Glu Asn Gly His Gly Leu Arg

930 935 940

Val Ser Arg Ala Asn Gly Ala Ala Pro Phe Ala Val Ser Leu Gln Pro

945 950 955 960

Tyr Ser Ser Phe Met Ile Glu Glu Ala Gln His Gln Asp Glu Leu Pro

965 970 975

Ala Pro Lys His Met Phe Leu Arg Val Leu Ala Ala Gln Met Gly Val

980 985 990

Gly Gly Asp Asp Ser Trp Met Ser Pro Val His Ser Gln Tyr His Ile

995 1000 1005

Thr Ala Asp Gln Pro Ile Ser Leu Asp Val Asn Leu Glu Leu Ile

1010 1015 1020

<210> 12

<211> 1008

<212>PRT

<213> Lactobacillus delbrueckii subsp. bulgaricus

<400> 12

Met Ser Asn Lys Leu Val Lys Glu Lys Arg Val Asp Gln Ala Asp Leu

1 5 10 15

Ala Trp Leu Thr Asp Pro Glu Val Tyr Glu Val Asn Thr Ile Pro Pro

20 25 30

His Ser Asp His Glu Ser Phe Gln Ser Gln Glu Glu Leu Glu Glu Gly

35 40 45

Lys Ser Ser Leu Val Gln Ser Leu Asp Gly Asp Trp Leu Ile Asp Tyr

50 55 60

Ala Glu Asn Gly Gln Gly Pro Val Asn Phe Tyr Ala Glu Asp Phe Asp

65 70 75 80

Asp Ser Asn Phe Lys Ser Val Lys Val Pro Gly Asn Leu Glu Leu Gln

85 90 95

Gly Phe Gly Gln Pro Gln Tyr Val Asn Val Gln Tyr Pro Trp Asp Gly

100 105 110

Ser Glu Glu Ile Phe Pro Pro Gln Ile Pro Ser Lys Asn Pro Leu Ala

115 120 125

Ser Tyr Val Arg Tyr Phe Asp Leu Asp Glu Ala Phe Trp Asp Lys Glu

130 135 140

Val Ser Leu Lys Phe Asp Gly Ala Ala Thr Ala Ile Tyr Val Trp Leu

145 150 155 160

Asn Gly His Phe Val Gly Tyr Gly Glu Asp Ser Phe Thr Pro Ser Glu

165 170 175

Phe Met Val Thr Lys Phe Leu Lys Lys Glu Asn Asn Arg Leu Ala Val

180 185 190

Ala Leu Tyr Lys Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe

195 200 205

Trp Arg Met Ser Gly Leu Phe Arg Ser Val Thr Leu Gln Ala Lys Pro

210 215 220

Arg Leu His Leu Glu Asp Leu Lys Leu Thr Ala Ser Leu Thr Asp Asn

225 230 235 240

Tyr Gln Lys Gly Lys Leu Glu Val Glu Ala Asn Ile Ala Tyr Arg Leu

245 250 255

Pro Asn Ala Ser Phe Lys Leu Glu Val Arg Asp Ser Glu Gly Asp Leu

260 265 270

Val Ala Glu Lys Leu Gly Pro Ile Arg Ser Glu Gln Leu Glu Phe Thr

275 280 285

Leu Ala Asp Leu Pro Val Ala Ala Trp Ser Ala Glu Lys Pro Asn Leu

290 295 300

Tyr Gln Val Arg Leu Tyr Leu Tyr Gln Ala Gly Ser Leu Leu Glu Val

305 310 315 320

Ser Arg Gln Glu Val Gly Phe Arg Asn Phe Glu Leu Lys Asp Gly Ile

325 330 335

Met Tyr Leu Asn Gly Gln Arg Ile Val Phe Lys Gly Ala Asn Arg His

340 345 350

Glu Phe Asp Ser Lys Leu Gly Arg Ala Ile Thr Glu Glu Asp Met Ile

355 360 365

Trp Asp Ile Lys Thr Met Lys Arg Ser Asn Ile Asn Ala Val Arg Cys

370 375 380

Ser His Tyr Pro Asn Gln Ser Leu Phe Tyr Arg Leu Cys Asp Lys Tyr

385 390 395 400

Gly Leu Tyr Val Ile Asp Glu Ala Asn Leu Glu Ser His Gly Thr Trp

405 410 415

Glu Lys Val Gly Gly His Glu Asp Pro Ser Phe Asn Val Pro Gly Asp

420 425 430

Asp Gln His Trp Leu Gly Ala Ser Leu Ser Arg Val Lys Asn Met Met

435 440 445

Ala Arg Asp Lys Asn His Ala Ser Ile Leu Ile Trp Ser Leu Gly Asn

450 455 460

Glu Ser Tyr Ala Gly Thr Val Phe Ala Gln Met Ala Asp Tyr Val Arg

465 470 475 480

Lys Ala Asp Pro Thr Arg Val Gln His Tyr Glu Gly Val Thr His Asn

485 490 495

Arg Lys Phe Asp Asp Ala Thr Gln Ile Glu Ser Arg Met Tyr Ala Pro

500 505 510

Ala Lys Val Ile Glu Glu Tyr Leu Thr Asn Lys Pro Ala Lys Pro Phe

515 520 525

Ile Ser Val Glu Tyr Ala His Ala Met Gly Asn Ser Val Gly Asp Leu

530 535 540

Ala Ala Tyr Thr Ala Leu Glu Lys Tyr Pro His Tyr Gln Gly Gly Phe

545 550 555 560

Ile Trp Asp Trp Ile Asp Gln Gly Leu Glu Lys Asp Gly His Leu Leu

565 570 575

Tyr Gly Gly Asp Phe Asp Asp Arg Pro Thr Asp Tyr Glu Phe Cys Gly

580 585 590

Asn Gly Leu Val Phe Ala Asp Arg Thr Glu Ser Pro Lys Leu Ala Asn

595 600 605

Val Lys Ala Leu Tyr Ala Asn Leu Lys Leu Glu Val Lys Asp Gly Gln

610 615 620

Leu Phe Leu Lys Asn Asp Asn Leu Phe Thr Asn Ser Ser Ser Tyr Tyr

625 630 635 640

Phe Leu Thr Ser Leu Leu Val Asp Gly Lys Leu Thr Tyr Gln Ser Arg

645 650 655

Pro Leu Thr Phe Gly Leu Glu Pro Gly Glu Ser Gly Thr Phe Ala Leu

660 665 670

Pro Trp Pro Glu Val Ala Asp Glu Lys Gly Glu Val Val Tyr Arg Val

675 680 685

Thr Ala His Leu Lys Glu Asp Leu Pro Trp Ala Asp Glu Gly Phe Thr

690 695 700

Val Ala Glu Ala Glu Glu Val Ala Gln Lys Leu Pro Glu Phe Lys Pro

705 710 715 720

Glu Gly Arg Pro Asp Leu Val Asp Ser Asp Tyr Asn Leu Gly Leu Lys

725 730 735

Gly Asn Asn Phe Gln Ile Leu Phe Ser Lys Val Lys Gly Trp Pro Val

740 745 750

Ser Leu Lys Tyr Ala Gly Arg Glu Tyr Leu Lys Arg Leu Pro Glu Phe

755 760 765

Thr Phe Trp Arg Ala Leu Thr Asp Asn Asp Arg Gly Ala Gly Tyr Gly

770 775 780

Tyr Asp Leu Ala Arg Trp Glu Asn Ala Gly Lys Tyr Ala Arg Leu Lys

785 790 795 800

Asp Ile Ser Cys Glu Val Lys Glu Asp Ser Val Leu Val Lys Thr Ala

805 810 815

Phe Thr Leu Pro Val Ala Leu Lys Gly Asp Leu Thr Val Thr Tyr Glu

820 825 830

Val Asp Gly Arg Gly Lys Ile Ala Val Thr Ala Asp Phe Pro Gly Ala

835 840 845

Glu Glu Ala Gly Leu Leu Pro Ala Phe Gly Leu Asn Leu Ala Leu Pro

850 855 860

Lys Glu Leu Thr Asp Tyr Arg Tyr Tyr Gly Leu Gly Pro Asn Glu Ser

865 870 875 880

Tyr Pro Asp Arg Leu Glu Gly Asn Tyr Leu Gly Ile Tyr Gln Gly Ala

885 890 895

Val Lys Lys Asn Phe Ser Pro Tyr Leu Arg Pro Gln Glu Thr Gly Asn

900 905 910

Arg Ser Lys Val Arg Trp Tyr Gln Leu Phe Asp Glu Lys Gly Gly Leu

915 920 925

Glu Phe Thr Ala Asn Gly Ala Asp Leu Asn Leu Ser Ala Leu Pro Tyr

930 935 940

Ser Ala Ala Gln Ile Glu Ala Ala Asp His Ala Phe Asp Leu Thr Asn

945 950 955 960

Asn Tyr Thr Trp Val Arg Ala Leu Ser Ala Gln Met Gly Val Gly Gly

965 970 975

Asp Asp Ser Trp Gly Gln Lys Val His Pro Glu Phe Cys Leu Asp Ala

980 985 990

Gln Lys Ala Arg Gln Leu Arg Leu Val Ile Gln Pro Leu Leu Leu Lys

995 1000 1005

<210> 13

<211> 1007

<212>PRT

<213> Lactobacillus delbrueckii subsp. lactis

<400> 13

Met Ser Asn Lys Leu Val Lys Glu Lys Arg Val Asp Gln Ala Asp Leu

1 5 10 15

Ala Trp Leu Thr Asp Pro Glu Val Tyr Glu Val Asn Thr Ile Pro Pro

20 25 30

His Ser Asp His Glu Ser Phe Gln Ser Gln Glu Glu Leu Glu Glu Gly

35 40 45

Lys Ser Ser Leu Val Gln Ser Leu Asp Gly Asn Trp Leu Ile Asp Tyr

50 55 60

Ala Glu Asn Gly Gln Gly Pro Ile Asn Phe Tyr Ala Glu Asp Phe Asp

65 70 75 80

Asp Ser Asn Phe Lys Ser Val Lys Val Pro Gly Asn Leu Glu Leu Gln

85 90 95

Gly Phe Gly Gln Pro Gln Tyr Val Asn Ile Gln Tyr Pro Trp Asp Gly

100 105 110

Ser Glu Glu Ile Phe Pro Pro Gln Val Pro Ser Lys Asn Pro Leu Ala

115 120 125

Ser Tyr Val Arg Tyr Phe Asp Leu Asp Glu Ala Leu Trp Asp Lys Glu

130 135 140

Val Ser Leu Lys Phe Ala Gly Ala Ala Thr Ala Ile Tyr Val Trp Leu

145 150 155 160

Asn Gly His Phe Val Gly Tyr Gly Glu Asp Ser Phe Thr Pro Ser Glu

165 170 175

Phe Met Val Thr Lys Phe Leu Lys Lys Glu Gly Asn Arg Leu Ala Val

180 185 190

Ala Leu Tyr Lys Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe

195 200 205

Trp Arg Leu Ser Gly Leu Phe Arg Ser Val Thr Leu Glu Ala Lys Pro

210 215 220

Leu Leu His Leu Glu Asp Leu Lys Leu Thr Ala Ser Leu Thr Asp Asn

225 230 235 240

Tyr Gln Lys Gly Lys Leu Glu Val Glu Ala Asn Ile Ala Tyr Arg Leu

245 250 255

Pro Asn Ala Ser Phe Lys Leu Glu Val Arg Asp Ser Glu Gly Asp Leu

260 265 270

Val Ala Glu Lys Val Gly Pro Ile Arg Ser Glu Lys Leu Gly Phe Ser

275 280 285

Leu Ala Asp Leu Pro Val Ala Ala Trp Ser Ala Glu Lys Pro Asn Leu

290 295 300

Tyr Gln Val Arg Leu Tyr Leu Tyr Gln Ala Gly Ser Leu Leu Glu Val

305 310 315 320

Ser Arg Gln Glu Val Gly Phe Arg Asn Phe Glu Leu Lys Asp Gly Ile

325 330 335

Met Tyr Leu Asn Gly Gln Arg Ile Val Phe Lys Gly Val Asn Arg His

340 345 350

Glu Phe Asp Ser Lys Leu Gly Arg Ala Ile Thr Glu Ala Asp Met Ile

355 360 365

Trp Asp Ile Lys Thr Met Lys Gln Ser Asn Ile Asn Ala Val Arg Cys

370 375 380

Ser His Tyr Pro Asn Gln Ser Leu Phe Tyr Arg Leu Cys Asp Lys Tyr

385 390 395 400

Gly Leu Tyr Val Ile Asp Glu Ala Asn Leu Glu Ser His Gly Thr Trp

405 410 415

Glu Lys Val Gly His Glu Asp Pro Ser Phe Asn Val Pro Gly Asp Asp

420 425 430

Gln His Trp Leu Gly Ala Ser Leu Ser Arg Val Lys Asn Met Met Ala

435 440 445

Arg Asp Lys Asn His Ala Ser Ile Leu Ile Trp Ser Leu Gly Asn Glu

450 455 460

Ser Tyr Ala Gly Thr Val Phe Ala Gln Met Ala Asp Tyr Val Arg Lys

465 470 475 480

Ala Asp Pro Thr Arg Val Gln His Tyr Glu Gly Val Thr His Asn Arg

485 490 495

Lys Phe Asp Asp Ala Thr Gln Ile Glu Ser Arg Met Tyr Ala Pro Ala

500 505 510

Lys Glu Ile Glu Glu Tyr Leu Thr Lys Lys Pro Ala Lys Pro Phe Ile

515 520 525

Ser Val Glu Tyr Ala His Ala Met Gly Asn Ser Val Gly Asp Leu Ala

530 535 540

Ala Tyr Thr Ala Leu Glu Lys Tyr Pro His Tyr Gln Gly Gly Phe Ile

545 550 555 560

Trp Asp Trp Ile Asp Gln Gly Leu Glu Lys Asp Gly His Leu Leu Tyr

565 570 575

Gly Gly Asp Phe Asp Asp Arg Pro Thr Asp Tyr Glu Phe Cys Gly Asp

580 585 590

Gly Leu Val Phe Ala Asp Arg Thr Thr Ser Pro Lys Leu Ala Asn Val

595 600 605

Lys Ala Leu Tyr Ser Asn Leu Lys Leu Glu Val Lys Asp Gly Gln Leu

610 615 620

Phe Ile Lys Asn Asp Asn Leu Phe Thr Asn Ser Ser Ala Tyr Tyr Phe

625 630 635 640

Leu Ala Ser Leu Leu Val Asp Gly Lys Leu Thr Tyr Gln Ser Gln Pro

645 650 655

Leu Thr Phe Gly Leu Glu Pro Gly Glu Ser Gly Thr Phe Val Leu Pro

660 665 670

Trp Pro Glu Val Glu Asp Glu Lys Gly Glu Ile Val Tyr Gln Val Thr

675 680 685

Ala His Leu Lys Glu Asp Leu Pro Trp Ala Asp Glu Gly Phe Thr Val

690 695 700

Ala Glu Ala Glu Glu Ala Val Thr Lys Leu Pro Glu Phe Tyr Pro Ala

705 710 715 720

Gly Arg Pro Glu Leu Val Asp Ser Asp Phe Asn Leu Gly Leu Lys Gly

725 730 735

Asn Gly Phe Arg Ile Leu Phe Ser Lys Ala Lys Gly Trp Pro Val Ser

740 745 750

Ile Lys Tyr Ala Gly Arg Glu Tyr Leu Lys Arg Leu Pro Glu Phe Thr

755 760 765

Phe Trp Arg Ala Leu Thr Asp Asn Asp Arg Gly Ala Gly Tyr Gly Tyr

770 775 780

Asp Leu Ala Lys Trp Glu Asn Ala Gly Lys Tyr Ala Arg Leu Gln Asp

785 790 795 800

Ile Ser Tyr Glu Ile Lys Glu Asn Ser Ala Leu Val Lys Thr Thr Phe

805 810 815

Thr Leu Pro Val Ala Leu Lys Gly Asp Leu Thr Ile Thr Tyr Glu Val

820 825 830

Asp Ser Leu Gly Lys Ile Ala Val Thr Ala Asn Phe Pro Gly Ala Val

835 840 845

Glu Asn Gly Leu Leu Pro Ala Phe Gly Leu Asn Phe Ala Leu Pro Lys

850 855 860

Glu Leu Ser Asp Tyr Arg Tyr Tyr Gly Leu Gly Pro Asn Glu Ser Tyr

865 870 875 880

Ala Asp Arg Leu Glu Gly Ser Tyr Leu Gly Ile Tyr Gln Gly Ala Val

885 890 895

Glu Lys Asn Phe Thr Pro Tyr Leu Arg Pro Gln Glu Ala Gly Asn Arg

900 905 910

Ser Lys Val Arg Tyr Tyr Gln Leu Phe Asp Glu Glu Gly Gly Leu Glu

915 920 925

Phe Thr Ala Asn Gly Ala Asp Leu Asn Leu Ser Ala Leu Pro Tyr Ser

930 935 940

Ala Ala Gln Ile Glu Ala Ala Asp His Ala Phe Glu Leu Thr Asn Asn

945 950 955 960

Tyr Thr Trp Val Arg Ala Leu Ala Ala Gln Met Gly Val Gly Gly Asp

965 970 975

Asp Ser Trp Gly Gln Lys Val His Pro Glu Phe Cys Leu Asp Ala Gln

980 985 990

Glu Ala Arg Gln Leu Lys Leu Val Ile Gln Pro Leu Leu Leu Lys

995 1000 1005

<210> 14

<211> 1008

<212>PRT

<213> Lactobacillus delbrueckii subsp. bulgaricus

<400> 14

Met Ser Asn Lys Leu Val Lys Glu Lys Arg Val Asp Gln Ala Asp Leu

1 5 10 15

Ala Trp Leu Thr Asp Pro Glu Val Tyr Glu Val Asn Thr Ile Pro Pro

20 25 30

His Phe Asp His Glu Ser Phe Gln Ser Gln Glu Glu Leu Glu Glu Gly

35 40 45

Lys Ser Ser Leu Val Gln Ser Leu Asp Gly Asp Trp Leu Ile Asp Tyr

50 55 60

Ala Glu Asn Gly Gln Gly Pro Val Asn Phe Tyr Ala Glu Asp Phe Asp

65 70 75 80

Asp Ser Asn Phe Lys Ser Val Lys Val Pro Gly Asn Leu Glu Leu Gln

85 90 95

Gly Phe Gly Gln Pro Gln Tyr Val Asn Val Gln Tyr Pro Trp Asp Gly

100 105 110

Ser Glu Glu Ile Phe Pro Pro Gln Ile Pro Ser Lys Asn Pro Leu Ala

115 120 125

Ser Tyr Val Arg Tyr Phe Asp Leu Asp Glu Ala Phe Trp Asp Lys Glu

130 135 140

Val Ser Leu Lys Phe Asp Gly Ala Ala Thr Ala Ile Tyr Val Trp Leu

145 150 155 160

Asn Gly His Phe Val Gly Tyr Gly Glu Asp Ser Phe Thr Pro Ser Glu

165 170 175

Phe Met Val Thr Lys Phe Leu Lys Lys Glu Asn Asn Arg Leu Ala Val

180 185 190

Ala Leu Tyr Lys Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe

195 200 205

Trp Arg Met Ser Gly Leu Phe Arg Ser Val Thr Leu Gln Ala Lys Pro

210 215 220

Arg Leu His Leu Glu Asp Leu Lys Leu Thr Ala Ser Leu Thr Asp Asn

225 230 235 240

Tyr Gln Lys Gly Lys Leu Glu Val Glu Ala Asn Ile Ala Tyr Arg Leu

245 250 255

Pro Asn Ala Ser Phe Lys Leu Glu Val Arg Asp Ser Glu Gly Asp Leu

260 265 270

Val Ala Glu Lys Leu Gly Pro Ile Arg Ser Glu Gln Leu Glu Phe Thr

275 280 285

Leu Ala Asp Leu Pro Val Ala Ala Trp Ser Ala Glu Lys Pro Asn Leu

290 295 300

Tyr Gln Val Arg Leu Tyr Leu Tyr Gln Ala Gly Ser Leu Leu Glu Val

305 310 315 320

Ser Arg Gln Glu Val Gly Phe Arg Asn Phe Glu Leu Lys Asp Gly Ile

325 330 335

Met Tyr Leu Asn Gly Gln Arg Ile Val Phe Lys Gly Ala Asn Arg His

340 345 350

Glu Phe Asp Ser Lys Leu Gly Arg Ala Ile Thr Glu Glu Asp Met Ile

355 360 365

Trp Asp Ile Lys Thr Met Lys Arg Ser Asn Ile Asn Ala Val Arg Cys

370 375 380

Ser His Tyr Pro Asn Gln Ser Leu Phe Tyr Arg Leu Cys Asp Lys Tyr

385 390 395 400

Gly Leu Tyr Val Ile Asp Glu Ala Asn Leu Glu Ser His Gly Thr Trp

405 410 415

Glu Lys Val Gly Gly His Glu Asp Pro Ser Phe Asn Val Pro Gly Asp

420 425 430

Asp Gln His Trp Leu Gly Ala Ser Leu Ser Arg Val Lys Asn Met Met

435 440 445

Ala Arg Asp Lys Asn His Ala Ser Ile Leu Ile Trp Ser Leu Gly Asn

450 455 460

Glu Ser Tyr Ala Gly Thr Val Phe Ala Gln Met Ala Asp Tyr Val Arg

465 470 475 480

Lys Ala Asp Pro Thr Arg Val Gln His Tyr Glu Gly Val Thr His Asn

485 490 495

Arg Lys Phe Asp Asp Ala Thr Gln Ile Glu Ser Arg Met Tyr Ala Pro

500 505 510

Ala Lys Val Ile Glu Glu Tyr Leu Thr Asn Lys Pro Ala Lys Pro Phe

515 520 525

Ile Ser Val Glu Tyr Ala His Ala Met Gly Asn Ser Val Gly Asp Leu

530 535 540

Ala Ala Tyr Thr Ala Leu Glu Lys Tyr Pro His Tyr Gln Gly Gly Phe

545 550 555 560

Ile Trp Asp Trp Ile Asp Gln Gly Leu Glu Lys Asp Gly His Leu Leu

565 570 575

Tyr Gly Gly Asp Phe Asp Asp Arg Pro Thr Asp Tyr Glu Phe Cys Gly

580 585 590

Asn Gly Leu Val Phe Ala Asp Arg Thr Glu Ser Pro Lys Leu Ala Asn

595 600 605

Val Lys Ala Leu Tyr Ala Asn Leu Lys Leu Glu Val Lys Asp Gly Gln

610 615 620

Leu Phe Leu Lys Asn Asp Asn Leu Phe Thr Asn Ser Ser Ser Tyr Tyr

625 630 635 640

Phe Leu Thr Ser Leu Leu Val Asp Gly Lys Leu Thr Tyr Gln Ser Arg

645 650 655

Pro Leu Thr Phe Gly Leu Glu Pro Gly Glu Ser Gly Thr Phe Ala Leu

660 665 670

Pro Trp Pro Glu Val Ala Asp Glu Lys Gly Glu Val Val Tyr Arg Val

675 680 685

Thr Ala His Leu Lys Glu Asp Leu Pro Trp Ala Asp Glu Gly Phe Thr

690 695 700

Val Ala Glu Ala Glu Glu Val Ala Gln Lys Leu Pro Glu Phe Lys Pro

705 710 715 720

Glu Gly Arg Pro Asp Leu Val Asp Ser Asp Tyr Asn Leu Gly Leu Lys

725 730 735

Gly Asn Asn Phe Gln Ile Leu Phe Ser Lys Val Lys Gly Trp Pro Val

740 745 750

Ser Leu Lys Tyr Ala Gly Arg Glu Tyr Leu Lys Arg Leu Pro Glu Phe

755 760 765

Thr Phe Trp Arg Ala Leu Thr Asp Asn Asp Arg Gly Ala Gly Tyr Gly

770 775 780

Tyr Asp Leu Ala Arg Trp Glu Asn Ala Gly Lys Tyr Ala Arg Leu Lys

785 790 795 800

Asp Ile Ser Cys Glu Val Lys Glu Asp Ser Val Leu Val Lys Thr Ala

805 810 815

Phe Thr Leu Pro Val Ala Leu Lys Gly Asp Leu Thr Val Thr Tyr Glu

820 825 830

Val Asp Gly Arg Gly Lys Ile Ala Val Thr Ala Asp Phe Pro Gly Ala

835 840 845

Glu Glu Ala Gly Leu Leu Pro Ala Phe Gly Leu Asn Leu Ala Leu Pro

850 855 860

Lys Glu Leu Thr Asp Tyr Arg Tyr Tyr Gly Leu Gly Pro Asn Glu Ser

865 870 875 880

Tyr Pro Asp Arg Leu Glu Gly Asn Tyr Leu Gly Ile Tyr Gln Gly Ala

885 890 895

Val Lys Lys Asn Phe Ser Pro Tyr Leu Arg Pro Gln Glu Thr Gly Asn

900 905 910

Arg Ser Lys Val Arg Trp Tyr Gln Leu Phe Asp Glu Lys Gly Gly Leu

915 920 925

Glu Phe Thr Ala Asn Gly Ala Asp Leu Asn Leu Ser Ala Leu Pro Tyr

930 935 940

Ser Ala Ala Gln Ile Glu Ala Ala Asp His Ala Phe Glu Leu Thr Asn

945 950 955 960

Asn Tyr Thr Trp Val Arg Ala Leu Ser Ala Gln Met Gly Val Gly Gly

965 970 975

Asp Asp Ser Trp Gly Gln Lys Val His Pro Glu Phe Cys Leu Asp Ala

980 985 990

Gln Lys Ala Arg Gln Leu Arg Leu Val Ile Gln Pro Leu Leu Leu Lys

995 1000 1005

<210> 15

<211> 1008

<212>PRT

<213> Lactobacillus delbrueckii subsp. bulgaricus

<400> 15

Met Ser Asn Lys Leu Val Lys Glu Lys Arg Val Asp Gln Ala Asp Leu

1 5 10 15

Ala Trp Leu Thr Asp Pro Glu Val Tyr Glu Val Asn Thr Ile Pro Pro

20 25 30

His Ser Asp His Glu Ser Phe Gln Ser Gln Glu Glu Leu Glu Glu Gly

35 40 45

Lys Ser Ser Leu Val Gln Ser Leu Asp Gly Asp Trp Leu Ile Asp Tyr

50 55 60

Ala Glu Asn Gly Gln Gly Pro Val Asn Phe Tyr Ala Glu Asp Phe Asp

65 70 75 80

Asp Ser Asn Phe Lys Ser Val Lys Val Pro Gly Asn Leu Glu Leu Gln

85 90 95

Gly Phe Gly Gln Pro Gln Tyr Val Asn Val Gln Tyr Pro Trp Asp Gly

100 105 110

Ser Glu Glu Ile Phe Pro Pro Gln Ile Pro Ser Lys Asn Pro Leu Ala

115 120 125

Ser Tyr Val Arg Tyr Phe Asp Leu Asp Glu Ala Phe Trp Asp Lys Glu

130 135 140

Val Ser Leu Lys Phe Asp Gly Ala Ala Thr Ala Ile Tyr Val Trp Leu

145 150 155 160

Asn Gly His Phe Val Gly Tyr Gly Glu Asp Ser Phe Thr Pro Ser Glu

165 170 175

Phe Met Val Thr Lys Phe Leu Lys Lys Glu Asn Asn Arg Leu Ala Val

180 185 190

Ala Leu Tyr Lys Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe

195 200 205

Trp Arg Met Ser Gly Leu Phe Arg Ser Val Thr Leu Gln Ala Lys Pro

210 215 220

Arg Leu His Leu Glu Asp Leu Lys Leu Thr Ala Ser Leu Thr Asp Asn

225 230 235 240

Tyr Gln Lys Gly Lys Leu Glu Val Glu Ala Asn Ile Ala Tyr Arg Leu

245 250 255

Pro Asn Ala Ser Phe Lys Leu Glu Val Arg Asp Ser Glu Gly Asp Leu

260 265 270

Val Ala Glu Lys Leu Gly Pro Ile Arg Ser Glu Gln Leu Glu Phe Thr

275 280 285

Leu Ala Asp Leu Pro Val Ala Ala Trp Ser Ala Glu Lys Pro Asn Leu

290 295 300

Tyr Gln Val Arg Leu Tyr Leu Tyr Gln Ala Gly Ser Leu Leu Glu Val

305 310 315 320

Ser Arg Gln Glu Val Gly Phe Arg Asn Phe Glu Leu Lys Asp Gly Ile

325 330 335

Met Tyr Leu Asn Gly Gln Arg Ile Val Phe Lys Gly Ala Asn Arg His

340 345 350

Glu Phe Asp Ser Lys Leu Gly Arg Ala Ile Thr Glu Glu Asp Met Ile

355 360 365

Trp Asp Ile Lys Thr Met Lys Arg Ser Asn Ile Asn Ala Val Arg Cys

370 375 380

Ser His Tyr Pro Asn Gln Ser Leu Phe Tyr Arg Leu Cys Asp Lys Tyr

385 390 395 400

Gly Leu Tyr Val Ile Asp Glu Ala Asn Leu Glu Ser His Gly Thr Trp

405 410 415

Glu Lys Val Gly Gly His Glu Asp Pro Ser Phe Asn Val Pro Gly Asp

420 425 430

Asp Gln His Trp Leu Gly Ala Ser Leu Ser Arg Val Lys Asn Met Met

435 440 445

Ala Arg Asp Lys Asn His Ala Ser Ile Leu Ile Trp Ser Leu Gly Asn

450 455 460

Glu Ser Tyr Ala Gly Thr Val Phe Ala Gln Met Ala Asp Tyr Val Arg

465 470 475 480

Lys Ala Asp Pro Thr Arg Val Gln His Tyr Glu Gly Val Thr His Asn

485 490 495

Arg Lys Phe Asp Asp Ala Thr Gln Ile Glu Ser Arg Met Tyr Ala Pro

500 505 510

Ala Lys Val Ile Glu Glu Tyr Leu Thr Asn Lys Pro Ala Lys Pro Phe

515 520 525

Ile Ser Val Glu Tyr Ala His Ala Met Gly Asn Ser Val Gly Asp Leu

530 535 540

Ala Ala Tyr Thr Ala Leu Glu Lys Tyr Pro His Tyr Gln Gly Gly Phe

545 550 555 560

Ile Trp Asp Trp Ile Asp Gln Gly Leu Glu Lys Asp Gly His Leu Leu

565 570 575

Tyr Gly Gly Asp Phe Asp Asp Arg Pro Thr Asp Tyr Glu Phe Cys Gly

580 585 590

Asn Gly Leu Val Phe Ala Asp Arg Thr Glu Ser Pro Lys Leu Ala Asn

595 600 605

Val Lys Ala Leu Tyr Ala Asn Leu Lys Leu Glu Val Lys Asp Gly Gln

610 615 620

Leu Phe Leu Lys Asn Asp Asn Leu Phe Thr Asn Ser Ser Ser Tyr Tyr

625 630 635 640

Phe Leu Thr Ser Leu Leu Val Asp Gly Lys Leu Thr Tyr Gln Ser Arg

645 650 655

Pro Leu Thr Phe Gly Leu Glu Pro Gly Glu Ser Gly Thr Phe Ala Leu

660 665 670

Pro Trp Pro Glu Val Ala Asp Glu Lys Gly Glu Val Val Tyr Arg Val

675 680 685

Thr Ala His Leu Lys Glu Asp Leu Pro Trp Ala Asp Glu Gly Phe Thr

690 695 700

Val Ala Glu Ala Glu Glu Val Ala Gln Lys Leu Pro Glu Phe Lys Pro

705 710 715 720

Glu Gly Arg Pro Asp Leu Val Asp Ser Asp Tyr Asn Leu Gly Leu Lys

725 730 735

Gly Asn Asn Phe Gln Ile Leu Phe Ser Lys Val Lys Gly Trp Pro Val

740 745 750

Ser Leu Lys Tyr Ala Gly Arg Glu Tyr Leu Lys Arg Leu Pro Glu Phe

755 760 765

Thr Phe Trp Arg Ala Leu Thr Asp Asn Asp Arg Gly Ala Gly Tyr Gly

770 775 780

Tyr Asp Leu Ala Arg Trp Glu Asn Ala Gly Lys Tyr Ala Arg Leu Lys

785 790 795 800

Asp Ile Ser Cys Glu Val Lys Glu Asp Ser Val Leu Val Lys Thr Ala

805 810 815

Phe Thr Leu Pro Val Ala Leu Lys Gly Asp Leu Thr Val Thr Tyr Glu

820 825 830

Val Asp Gly Arg Gly Lys Ile Ala Val Thr Ala Asp Phe Pro Gly Ala

835 840 845

Glu Glu Ala Gly Leu Leu Pro Ala Phe Gly Leu Asn Leu Ala Leu Pro

850 855 860

Lys Glu Leu Thr Asp Tyr Arg Tyr Tyr Gly Leu Gly Pro Asn Glu Ser

865 870 875 880

Tyr Pro Asp Arg Leu Glu Gly Asn Tyr Leu Gly Ile Tyr Gln Gly Ala

885 890 895

Val Lys Lys Asn Phe Ser Pro Tyr Leu Arg Pro Gln Glu Thr Gly Asn

900 905 910

Arg Ser Lys Val Arg Trp Tyr Gln Leu Phe Asp Glu Lys Gly Gly Leu

915 920 925

Glu Phe Thr Ala Asn Gly Ala Asp Leu Asn Leu Ser Ala Leu Pro Tyr

930 935 940

Ser Ala Ala Gln Ile Glu Ala Ala Asp His Ala Phe Glu Leu Thr Asn

945 950 955 960

Asn Tyr Thr Trp Val Arg Ala Leu Ser Ala Gln Met Gly Val Gly Gly

965 970 975

Asp Asp Ser Trp Gly Gln Lys Val His Pro Glu Phe Cys Leu Asp Ala

980 985 990

Gln Lys Ala Arg Gln Leu Arg Leu Val Ile Gln Pro Leu Leu Leu Lys

995 1000 1005

<210> 16

<211> 1008

<212>PRT

<213> Lactobacillus delbrueckii subsp. lactis

<400> 16

Met Ser Asn Lys Leu Val Lys Glu Lys Arg Val Asp Gln Ala Asp Leu

1 5 10 15

Ala Trp Leu Thr Asp Pro Glu Val Tyr Glu Val Asn Thr Ile Pro Pro

20 25 30

His Ser Asp His Glu Ser Phe Gln Ser Gln Glu Glu Leu Glu Glu Gly

35 40 45

Lys Ser Ser Leu Val Gln Ser Leu Asp Gly Asp Trp Leu Ile Asp Tyr

50 55 60

Ala Glu Asn Gly Gln Gly Pro Val Asn Phe Tyr Ala Glu Asp Phe Asp

65 70 75 80

Asp Ser Asn Phe Lys Ser Val Lys Val Pro Gly Asn Leu Glu Leu Gln

85 90 95

Gly Phe Gly Gln Pro Gln Tyr Val Asn Ile Gln Tyr Pro Trp Asp Gly

100 105 110

Ser Glu Glu Ile Phe Pro Pro Gln Val Pro Ser Lys Asn Pro Leu Ala

115 120 125

Ser Tyr Val Arg Tyr Phe Asp Leu Asp Glu Ala Phe Trp Asp Lys Glu

130 135 140

Val Ser Leu Lys Phe Ala Gly Ala Ala Thr Ala Ile Tyr Val Trp Leu

145 150 155 160

Asn Gly His Phe Val Gly Tyr Gly Glu Asp Ser Phe Thr Pro Ser Glu

165 170 175

Phe Met Val Thr Lys Phe Leu Lys Lys Glu Asn Asn Arg Leu Ala Val

180 185 190

Ala Leu Tyr Lys Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe

195 200 205

Trp Arg Leu Ser Gly Leu Phe Arg Ser Val Thr Leu Gln Ala Lys Pro

210 215 220

Leu Leu His Leu Glu Asp Leu Lys Leu Thr Ala Ser Leu Thr Asp Asn

225 230 235 240

Tyr Gln Lys Gly Lys Leu Glu Val Glu Ala Asn Ile Ala Tyr Arg Leu

245 250 255

Pro Asn Ala Ser Phe Lys Leu Glu Val Arg Asp Ser Glu Gly Asp Leu

260 265 270

Val Ala Glu Lys Leu Gly Pro Ile Arg Ser Glu Gln Leu Glu Phe Thr

275 280 285

Leu Ala Asp Leu Pro Val Ala Ala Trp Ser Ala Glu Lys Pro Asn Leu

290 295 300

Tyr Gln Val Arg Leu Tyr Leu Tyr Gln Ala Gly Ser Leu Leu Glu Val

305 310 315 320

Ser Arg Gln Glu Val Gly Phe Arg Asn Phe Glu Leu Lys Asp Gly Ile

325 330 335

Met Tyr Leu Asn Gly Gln Arg Ile Val Phe Lys Gly Val Asn Arg His

340 345 350

Glu Phe Asp Ser Lys Leu Gly Arg Ala Ile Thr Glu Glu Asp Met Ile

355 360 365

Trp Asp Ile Lys Thr Met Lys Arg Ser Asn Ile Asn Ala Val Arg Cys

370 375 380

Ser His Tyr Pro Asn Gln Ser Leu Phe Tyr Arg Leu Cys Asp Lys Tyr

385 390 395 400

Gly Leu Tyr Val Ile Asp Glu Ala Asn Leu Glu Ser His Gly Thr Trp

405 410 415

Glu Lys Val Gly His Glu Asp Pro Ser Phe Asn Val Pro Gly Asp Asp

420 425 430

Gln His Trp Leu Gly Ala Ser Leu Ser Arg Val Lys Asn Met Met Ala

435 440 445

Arg Asp Lys Asn His Ala Ser Ile Leu Ile Trp Ser Leu Gly Asn Glu

450 455 460

Ser Tyr Ala Gly Thr Val Phe Ala Gln Met Ala Asp Tyr Val Arg Lys

465 470 475 480

Ala Asp Pro Thr Arg Val Gln His Tyr Glu Gly Val Thr His Asn Arg

485 490 495

Lys Phe Asp Asp Ala Thr Gln Ile Glu Ser Arg Met Tyr Ala Pro Ala

500 505 510

Lys Glu Ile Glu Glu Tyr Leu Thr Lys Lys Pro Ala Lys Pro Phe Ile

515 520 525

Ser Val Glu Tyr Ala His Ala Met Gly Asn Ser Val Gly Asp Leu Ala

530 535 540

Ala Tyr Thr Ala Leu Glu Lys Tyr Pro His Tyr Gln Gly Gly Phe Ile

545 550 555 560

Trp Asp Trp Ile Asp Gln Gly Leu Glu Lys Asp Gly His Leu Leu Tyr

565 570 575

Gly Gly Gly Asp Phe Asp Asp Arg Pro Thr Asp Tyr Glu Phe Cys Gly

580 585 590

Asn Gly Leu Val Phe Ala Asp Arg Thr Thr Ser Pro Lys Leu Ala Asn

595 600 605

Val Lys Ala Leu Tyr Ser Asn Leu Lys Leu Glu Val Lys Asp Gly Gln

610 615 620

Leu Phe Leu Lys Asn Asp Asn Leu Phe Thr Asn Ser Ser Ala Tyr Tyr

625 630 635 640

Phe Leu Thr Ser Leu Leu Val Asp Gly Lys Leu Thr Tyr Gln Ser Gln

645 650 655

Pro Leu Thr Phe Gly Leu Glu Pro Gly Glu Ser Gly Thr Phe Val Leu

660 665 670

Pro Trp Pro Glu Val Glu Asp Glu Lys Gly Glu Ile Val Tyr Gln Val

675 680 685

Thr Ala His Leu Lys Glu Asp Leu Pro Trp Ala Asp Glu Gly Phe Thr

690 695 700

Val Ala Glu Ala Glu Glu Ala Val Thr Lys Leu Pro Glu Phe Tyr Pro

705 710 715 720

Ala Gly Arg Pro Glu Leu Val Asp Ser Asp Phe Asn Leu Gly Leu Lys

725 730 735

Gly Asn Gly Phe Arg Ile Leu Phe Ser Lys Ala Lys Gly Trp Pro Val

740 745 750

Ser Ile Lys Tyr Ala Gly Arg Glu Tyr Leu Lys Arg Leu Pro Glu Phe

755 760 765

Thr Phe Trp Arg Ala Leu Thr Asp Asn Asp Arg Gly Ala Gly Tyr Gly

770 775 780

Tyr Asp Leu Ala Lys Trp Glu Asn Ala Gly Lys Tyr Ala Arg Leu Gln

785 790 795 800

Asp Ile Ser Tyr Glu Ile Lys Glu Asn Ser Val Leu Val Lys Thr Ala

805 810 815

Phe Thr Leu Pro Val Ala Leu Lys Gly Asp Leu Thr Ile Thr Tyr Glu

820 825 830

Val Asp Ser Leu Gly Lys Ile Ala Val Thr Ala Asn Phe Pro Gly Ala

835 840 845

Val Glu Asn Gly Leu Leu Pro Ala Phe Gly Leu Asn Phe Ala Leu Pro

850 855 860

Lys Glu Leu Ser Asp Tyr Arg Tyr Tyr Gly Leu Gly Pro Asn Glu Ser

865 870 875 880

Tyr Ala Asp Arg Leu Glu Gly Ser Tyr Leu Gly Ile Tyr Gln Gly Ala

885 890 895

Val Glu Lys Asn Phe Thr Pro Tyr Leu Arg Pro Gln Glu Ala Gly Asn

900 905 910

Arg Ser Lys Val Arg Tyr Tyr Gln Leu Phe Asp Glu Glu Ser Gly Leu

915 920 925

Glu Phe Thr Ala Asn Gly Ala Asp Leu Asn Leu Ser Ala Leu Pro Tyr

930 935 940

Ser Ala Ala Gln Ile Glu Ala Ala Asp His Ala Phe Glu Leu Ser Asn

945 950 955 960

Asn Tyr Thr Trp Val Arg Ala Leu Ala Ala Gln Met Gly Val Gly Gly

965 970 975

Asp Asp Ser Trp Gly Gln Lys Val His Pro Glu Phe Cys Leu Asp Ala

980 985 990

Gln Glu Ala Arg Gln Leu Lys Leu Val Ile Gln Pro Leu Leu Leu Lys

995 1000 1005

<210> 17

<211> 1008

<212>PRT

<213> Lactobacillus delbrueckii subsp. bulgaricus

<400> 17

Met Ser Asn Lys Leu Val Lys Glu Lys Arg Val Asp Gln Ala Asp Leu

1 5 10 15

Ala Trp Leu Thr Asp Pro Glu Val Tyr Glu Val Asn Thr Ile Pro Pro

20 25 30

His Ser Asp His Glu Ser Phe Gln Ser Gln Glu Glu Leu Glu Glu Gly

35 40 45

Lys Ser Ser Leu Val Gln Ser Leu Asp Gly Asp Trp Leu Ile Asp Tyr

50 55 60

Ala Glu Asn Gly Gln Gly Pro Val Asn Phe Tyr Ala Glu Asp Phe Asp

65 70 75 80

Asp Ser Asn Phe Lys Ser Val Lys Val Pro Gly Asn Leu Glu Leu Gln

85 90 95

Gly Phe Gly Gln Pro Gln Tyr Val Asn Val Gln Tyr Pro Trp Asp Gly

100 105 110

Ser Glu Glu Ile Phe Pro Pro Gln Ile Pro Ser Lys Asn Pro Leu Ala

115 120 125

Ser Tyr Val Arg Tyr Phe Asp Leu Asp Glu Ala Phe Trp Asp Lys Glu

130 135 140

Val Ser Leu Lys Phe Asp Gly Ala Ala Thr Ala Ile Tyr Val Trp Leu

145 150 155 160

Asn Gly His Phe Val Gly Tyr Gly Glu Asp Ser Phe Thr Pro Ser Glu

165 170 175

Phe Met Val Thr Lys Phe Leu Lys Lys Glu Asn Asn Arg Leu Ala Val

180 185 190

Ala Leu Tyr Lys Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe

195 200 205

Trp Arg Met Ser Gly Leu Phe Arg Ser Val Thr Leu Gln Ala Lys Pro

210 215 220

Arg Leu His Leu Glu Asp Leu Lys Leu Thr Ala Ser Leu Thr Asp Asn

225 230 235 240

Tyr Gln Lys Gly Lys Leu Glu Val Glu Ala Asn Ile Ala Tyr Arg Leu

245 250 255

Pro Asn Ala Ser Phe Lys Leu Glu Val Arg Asp Ser Glu Gly Asp Leu

260 265 270

Val Ala Glu Lys Leu Gly Pro Ile Arg Ser Glu Gln Leu Glu Phe Thr

275 280 285

Leu Ala Asp Leu Pro Val Ala Ala Trp Ser Ala Glu Lys Pro Asn Leu

290 295 300

Tyr Gln Val Arg Leu Tyr Leu Tyr Gln Ala Gly Ser Leu Leu Glu Val

305 310 315 320

Ser Arg Gln Glu Val Gly Phe Arg Asn Phe Glu Leu Lys Asp Gly Ile

325 330 335

Met Tyr Leu Asn Gly Gln Arg Ile Val Phe Lys Gly Val Asn Arg His

340 345 350

Glu Phe Asp Ser Lys Leu Gly Arg Ala Ile Thr Glu Glu Asp Met Ile

355 360 365

Trp Asp Ile Lys Thr Ile Lys Arg Ser Asn Ile Asn Ala Val Arg Cys

370 375 380

Ser His Tyr Pro Asn Gln Ser Leu Phe Tyr Arg Leu Cys Asp Lys Tyr

385 390 395 400

Gly Leu Tyr Val Ile Asp Glu Ala Asn Leu Glu Ser His Gly Thr Trp

405 410 415

Glu Lys Val Gly Gly His Glu Asp Pro Ser Phe Asn Val Pro Gly Asp

420 425 430

Asp Gln His Trp Leu Gly Ala Ser Leu Ser Arg Val Lys Asn Met Met

435 440 445

Ala Arg Asp Lys Asn His Ala Ser Ile Leu Ile Trp Ser Leu Gly Asn

450 455 460

Glu Ser Tyr Ala Gly Thr Val Phe Ala Gln Met Ala Asp Tyr Val Arg

465 470 475 480

Lys Ala Asp Pro Thr Arg Val Gln His Tyr Glu Gly Val Thr His Asn

485 490 495

Arg Lys Phe Asp Asp Ala Thr Gln Ile Glu Ser Arg Met Tyr Ala Pro

500 505 510

Ala Lys Val Ile Glu Glu Tyr Leu Thr Asn Lys Pro Ala Lys Pro Phe

515 520 525

Ile Ser Val Glu Tyr Ala His Ala Met Gly Asn Ser Val Gly Asp Leu

530 535 540

Ala Ala Tyr Thr Ala Leu Glu Lys Tyr Pro His Tyr Gln Gly Gly Phe

545 550 555 560

Ile Trp Asp Trp Ile Asp Gln Gly Leu Glu Lys Asp Gly His Leu Leu

565 570 575

Tyr Gly Gly Asp Phe Asp Asp Arg Pro Thr Asp Tyr Glu Phe Cys Gly

580 585 590

Asn Gly Leu Val Phe Ala Asp Arg Thr Glu Ser Pro Lys Leu Ala Asn

595 600 605

Val Lys Ala Leu Tyr Ala Asn Leu Lys Leu Glu Val Lys Asp Gly Gln

610 615 620

Leu Phe Leu Lys Asn Asp Asn Leu Phe Thr Asn Ser Ser Ser Tyr Tyr

625 630 635 640

Phe Leu Thr Ser Leu Leu Val Asp Gly Lys Leu Thr Tyr Gln Ser Arg

645 650 655

Pro Leu Thr Phe Gly Leu Glu Pro Gly Glu Ser Gly Thr Phe Ala Leu

660 665 670

Pro Trp Pro Glu Val Ala Asp Glu Lys Gly Glu Val Val Tyr Arg Val

675 680 685

Thr Ala His Leu Lys Glu Asp Leu Pro Trp Ala Asp Glu Gly Phe Thr

690 695 700

Val Ala Glu Ala Glu Glu Val Ala Gln Lys Leu Pro Glu Phe Lys Pro

705 710 715 720

Glu Gly Arg Pro Asp Leu Val Asp Ser Asp Tyr Asn Leu Gly Leu Lys

725 730 735

Gly Asn Asn Phe Gln Ile Leu Phe Ser Lys Val Lys Gly Trp Pro Val

740 745 750

Ser Leu Lys Tyr Ala Gly Arg Glu Tyr Leu Lys Arg Leu Pro Glu Phe

755 760 765

Thr Phe Trp Arg Ala Leu Thr Asp Asn Asp Arg Gly Ala Gly Tyr Gly

770 775 780

Tyr Asp Leu Ala Arg Trp Glu Asn Ala Gly Lys Tyr Ala Arg Leu Lys

785 790 795 800

Asp Ile Ser Cys Glu Val Lys Glu Asp Ser Val Leu Val Lys Thr Ala

805 810 815

Phe Thr Leu Pro Val Ala Leu Lys Gly Asp Leu Thr Val Thr Tyr Glu

820 825 830

Val Asp Gly Arg Gly Lys Ile Ala Val Thr Ala Asp Phe Pro Gly Ala

835 840 845

Glu Glu Ala Gly Leu Leu Pro Ala Phe Gly Leu Asn Leu Ala Leu Pro

850 855 860

Lys Glu Leu Thr Asp Tyr Arg Tyr Tyr Gly Leu Gly Pro Asn Glu Ser

865 870 875 880

Tyr Pro Asp Arg Leu Glu Gly Asn Tyr Leu Gly Ile Tyr Gln Gly Ala

885 890 895

Val Lys Lys Asn Phe Ser Pro Tyr Leu Arg Pro Gln Glu Thr Gly Asn

900 905 910

Arg Ser Lys Val Arg Trp Tyr Gln Leu Phe Asp Glu Lys Gly Gly Leu

915 920 925

Glu Phe Thr Ala Asn Gly Ala Asp Leu Asn Leu Ser Ala Leu Pro Tyr

930 935 940

Ser Ala Ala Gln Ile Glu Ala Ala Asp His Ala Phe Asp Leu Thr Asn

945 950 955 960

Asn Tyr Thr Trp Val Arg Ala Leu Ser Ala Gln Met Gly Val Gly Gly

965 970 975

Asp Asp Ser Trp Gly Gln Lys Val His Pro Glu Phe Cys Leu Asp Ala

980 985 990

Gln Lys Ala Arg Gln Leu Arg Leu Val Ile Gln Pro Leu Leu Leu Lys

995 1000 1005

<210> 18

<211> 1008

<212>PRT

<213> Lactobacillus delbrueckii subsp. bulgaricus

<400> 18

Met Ser Asn Lys Leu Val Lys Glu Lys Arg Val Asp Gln Ala Asp Leu

1 5 10 15

Ala Trp Leu Thr Asp Pro Glu Val Tyr Glu Val Asn Thr Ile Pro Pro

20 25 30

His Ser Asp His Glu Ser Phe Gln Ser Gln Glu Glu Leu Glu Glu Gly

35 40 45

Lys Ser Ser Leu Val Gln Ser Leu Asp Gly Asp Trp Leu Ile Asp Tyr

50 55 60

Ala Glu Asn Gly Gln Gly Pro Val Asn Phe Tyr Ala Glu Asp Phe Asp

65 70 75 80

Asp Ser Asn Phe Lys Ser Val Lys Val Pro Gly Asn Leu Glu Leu Gln

85 90 95

Gly Phe Gly Gln Pro Gln Tyr Val Asn Val Gln Tyr Pro Trp Asp Gly

100 105 110

Ser Glu Glu Ile Phe Pro Pro Gln Ile Pro Ser Lys Asn Pro Leu Ala

115 120 125

Ser Tyr Val Arg Tyr Phe Asp Leu Asp Glu Ala Phe Trp Asp Lys Glu

130 135 140

Val Ser Leu Lys Phe Asp Gly Ala Ala Thr Ala Ile Tyr Val Trp Leu

145 150 155 160

Asn Gly His Phe Val Gly Tyr Gly Glu Asp Ser Phe Thr Pro Ser Glu

165 170 175

Phe Met Val Thr Lys Phe Leu Lys Lys Glu Asn Asn Arg Leu Ala Val

180 185 190

Ala Leu Tyr Lys Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe

195 200 205

Trp Arg Met Ser Gly Leu Phe Arg Ser Val Thr Leu Gln Ala Lys Pro

210 215 220

Arg Leu His Leu Glu Asp Leu Lys Leu Thr Ala Ser Leu Thr Asp Asn

225 230 235 240

Tyr Gln Lys Gly Lys Leu Glu Val Glu Ala Asn Ile Ala Tyr Arg Leu

245 250 255

Pro Asn Ala Ser Phe Lys Leu Glu Val Arg Asp Ser Glu Gly Asp Leu

260 265 270

Val Ala Glu Lys Leu Gly Pro Ile Gly Ser Glu Gln Leu Glu Phe Thr

275 280 285

Leu Ala Asp Leu Pro Val Ala Ala Trp Ser Ala Glu Lys Pro Asn Leu

290 295 300

Tyr Gln Val Arg Leu Tyr Leu Tyr Gln Ala Gly Ser Leu Leu Glu Val

305 310 315 320

Ser Arg Gln Glu Val Gly Phe Arg Asn Phe Glu Leu Lys Asp Gly Ile

325 330 335

Met Tyr Leu Asn Gly Gln Arg Ile Val Phe Lys Gly Val Asn Arg His

340 345 350

Glu Phe Asp Ser Lys Leu Gly Arg Ala Ile Thr Glu Glu Asp Met Ile

355 360 365

Trp Asp Ile Lys Thr Ile Lys Arg Ser Asn Ile Asn Ala Val Arg Cys

370 375 380

Ser His Tyr Pro Asn Gln Ser Leu Phe Tyr Arg Leu Cys Asp Lys Tyr

385 390 395 400

Gly Leu Tyr Val Ile Asp Glu Ala Asn Leu Glu Ser His Gly Thr Trp

405 410 415

Glu Lys Val Gly Gly His Glu Asp Pro Ser Phe Asn Val Pro Gly Asp

420 425 430

Asp Gln His Trp Leu Gly Ala Ser Leu Ser Arg Val Lys Asn Met Met

435 440 445

Ala Arg Asp Lys Asn His Ala Ser Ile Leu Ile Trp Ser Leu Gly Asn

450 455 460

Glu Ser Tyr Ala Gly Thr Val Phe Ala Gln Met Ala Asp Tyr Val Arg

465 470 475 480

Lys Ala Asp Pro Thr Arg Val Gln His Tyr Glu Gly Val Thr His Asn

485 490 495

Arg Lys Phe Asp Asp Ala Thr Gln Ile Glu Ser Arg Met Tyr Ala Pro

500 505 510

Ala Lys Val Ile Glu Glu Tyr Leu Thr Asn Lys Pro Ala Lys Pro Phe

515 520 525

Ile Ser Val Glu Tyr Ala His Ala Met Gly Asn Ser Val Gly Asp Leu

530 535 540

Ala Ala Tyr Thr Ala Leu Glu Lys Tyr Pro His Tyr Gln Gly Gly Phe

545 550 555 560

Ile Trp Asp Trp Ile Asp Gln Gly Leu Glu Lys Asp Gly His Leu Leu

565 570 575

Tyr Gly Gly Asp Phe Asp Asp Arg Pro Thr Asp Tyr Glu Phe Cys Gly

580 585 590

Asn Gly Leu Val Phe Ala Asp Arg Thr Glu Ser Pro Lys Leu Ala Asn

595 600 605

Val Lys Ala Leu Tyr Ala Asn Leu Lys Leu Glu Val Lys Asp Gly Gln

610 615 620

Leu Phe Leu Lys Asn Asp Asn Leu Phe Thr Asn Ser Ser Ser Tyr Tyr

625 630 635 640

Phe Leu Thr Ser Leu Leu Val Asp Gly Lys Leu Thr Tyr Gln Ser Arg

645 650 655

Pro Leu Thr Phe Gly Leu Glu Pro Gly Glu Ser Gly Thr Phe Ala Leu

660 665 670

Pro Trp Pro Glu Val Ala Asp Glu Lys Gly Glu Val Val Tyr Arg Val

675 680 685

Thr Ala His Leu Lys Glu Asp Leu Pro Trp Ala Asp Glu Gly Phe Thr

690 695 700

Val Ala Glu Ala Glu Glu Val Ala Gln Lys Leu Pro Glu Phe Lys Pro

705 710 715 720

Glu Gly Arg Pro Asp Leu Val Asp Ser Asp Tyr Asn Leu Gly Leu Lys

725 730 735

Gly Asn Asn Phe Gln Ile Leu Phe Ser Lys Val Lys Gly Trp Pro Val

740 745 750

Ser Leu Lys Tyr Ala Gly Arg Glu Tyr Leu Lys Arg Leu Pro Glu Phe

755 760 765

Thr Phe Trp Arg Ala Leu Thr Asp Asn Asp Arg Gly Ala Gly Tyr Gly

770 775 780

Tyr Asp Leu Ala Arg Trp Glu Asn Ala Gly Lys Tyr Ala Arg Leu Lys

785 790 795 800

Asp Ile Ser Cys Glu Val Lys Glu Asp Ser Val Leu Val Lys Thr Ala

805 810 815

Phe Thr Leu Pro Val Ala Leu Lys Gly Asp Leu Thr Val Thr Tyr Glu

820 825 830

Val Asp Gly Arg Gly Lys Ile Ala Val Thr Ala Asp Phe Pro Gly Ala

835 840 845

Glu Glu Ala Gly Leu Leu Pro Ala Phe Gly Leu Asn Leu Ala Leu Pro

850 855 860

Lys Glu Leu Thr Asp Tyr Arg Tyr Tyr Gly Leu Gly Pro Asn Glu Ser

865 870 875 880

Tyr Pro Asp Arg Leu Glu Gly Asn Tyr Leu Gly Ile Tyr Gln Gly Ala

885 890 895

Val Lys Lys Asn Phe Ser Pro Tyr Leu Arg Pro Gln Glu Thr Gly Asn

900 905 910

Arg Ser Lys Val Arg Trp Tyr Gln Leu Phe Asp Glu Lys Gly Gly Leu

915 920 925

Glu Phe Thr Ala Asn Gly Ala Asp Leu Asn Leu Ser Ala Leu Pro Tyr

930 935 940

Ser Ala Ala Gln Ile Glu Ala Ala Asp His Ala Phe Glu Leu Thr Asn

945 950 955 960

Asn Tyr Thr Trp Val Arg Ala Leu Ser Ala Gln Met Gly Val Gly Gly

965 970 975

Asp Asp Ser Trp Gly Gln Lys Val His Pro Glu Phe Cys Leu Asp Ala

980 985 990

Gln Lys Ala Arg Gln Leu Arg Leu Val Ile Gln Pro Leu Leu Leu Lys

995 1000 1005

<210> 19

<211> 1007

<212>PRT

<213> Lactobacillus delbrueckii subsp. lactis

<400> 19

Met Ser Asn Lys Leu Val Lys Glu Lys Arg Val Asp Gln Ala Asp Leu

1 5 10 15

Ala Trp Leu Thr Asp Pro Glu Val Tyr Glu Val Asn Thr Ile Pro Pro

20 25 30

His Ser Asp His Glu Ser Phe Gln Ser Gln Glu Glu Leu Glu Glu Gly

35 40 45

Lys Ser Ser Leu Val Gln Ser Leu Asp Gly Asn Trp Leu Ile Asp Tyr

50 55 60

Ala Glu Asn Gly Gln Gly Pro Ile Asn Phe Tyr Ala Glu Asp Phe Asp

65 70 75 80

Asp Ser Asn Phe Lys Ser Val Lys Val Pro Gly Asn Leu Glu Leu Gln

85 90 95

Gly Phe Gly Gln Pro Gln Tyr Val Asn Ile Gln Tyr Pro Trp Asp Gly

100 105 110

Ser Glu Glu Ile Phe Pro Pro Gln Val Pro Ser Lys Asn Pro Leu Ala

115 120 125

Ser Tyr Val Arg Tyr Phe Asp Leu Asp Glu Ala Leu Trp Asp Lys Glu

130 135 140

Val Ser Leu Lys Phe Ala Gly Ala Ala Thr Ala Ile Tyr Val Trp Leu

145 150 155 160

Asn Gly His Phe Val Gly Tyr Gly Glu Asp Ser Phe Thr Pro Ser Glu

165 170 175

Phe Met Val Thr Lys Phe Leu Lys Lys Glu Gly Asn Arg Leu Ala Val

180 185 190

Ala Leu Tyr Lys Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe

195 200 205

Trp Arg Leu Ser Gly Leu Phe Arg Ser Val Thr Leu Glu Ala Lys Pro

210 215 220

Leu Leu His Leu Glu Asp Leu Lys Leu Thr Ala Ser Leu Thr Asp Asn

225 230 235 240

Tyr Gln Lys Gly Lys Leu Glu Val Glu Ala Asn Ile Ala Tyr Arg Leu

245 250 255

Pro Asn Ala Ser Phe Lys Leu Glu Val Arg Asp Ser Glu Gly Asp Leu

260 265 270

Val Ala Glu Lys Val Gly Pro Ile Arg Ser Glu Lys Leu Asp Phe Ser

275 280 285

Leu Ala Asp Leu Pro Val Ala Ala Trp Ser Ala Glu Lys Pro Asn Leu

290 295 300

Tyr Gln Val Arg Leu Tyr Leu Tyr Gln Ala Gly Ser Leu Leu Glu Val

305 310 315 320

Ser Arg Gln Glu Val Gly Phe Arg Asn Phe Glu Leu Lys Asp Gly Ile

325 330 335

Met Tyr Leu Asn Gly Gln Arg Ile Val Phe Lys Gly Val Asn Arg His

340 345 350

Glu Phe Asp Ser Lys Leu Gly Arg Ala Ile Thr Glu Ala Asp Met Ile

355 360 365

Trp Asp Ile Lys Thr Met Lys Gln Ser Asn Ile Asn Ala Val Arg Cys

370 375 380

Ser His Tyr Pro Asn Gln Ser Leu Phe Tyr Arg Leu Cys Asp Lys Tyr

385 390 395 400

Gly Leu Tyr Val Ile Asp Glu Ala Asn Leu Glu Ser His Gly Thr Trp

405 410 415

Glu Lys Val Gly His Glu Asp Pro Ser Phe Asn Val Pro Gly Asp Asp

420 425 430

Gln His Trp Leu Gly Ala Ser Leu Ser Arg Val Lys Asn Met Met Ala

435 440 445

Arg Asp Lys Asn His Ala Ser Ile Leu Ile Trp Ser Leu Gly Asn Glu

450 455 460

Ser Tyr Ala Gly Thr Val Phe Ala Gln Met Ala Asp Tyr Val Arg Lys

465 470 475 480

Ala Asp Pro Thr Arg Val Gln His Tyr Glu Gly Val Thr His Asn Arg

485 490 495

Lys Phe Asp Asp Ala Thr Gln Ile Glu Ser Arg Met Tyr Ala Pro Ala

500 505 510

Lys Glu Ile Glu Glu Tyr Leu Thr Lys Lys Pro Ala Lys Pro Phe Ile

515 520 525

Ser Val Glu Tyr Ala His Ala Met Gly Asn Ser Val Gly Asp Leu Ala

530 535 540

Ala Tyr Thr Ala Leu Glu Lys Tyr Pro His Tyr Gln Gly Gly Phe Ile

545 550 555 560

Trp Asp Trp Ile Asp Gln Gly Leu Glu Lys Asp Gly His Leu Leu Tyr

565 570 575

Gly Gly Asp Phe Asp Asp Arg Pro Thr Asp Tyr Glu Phe Cys Gly Asp

580 585 590

Gly Leu Val Phe Ala Asp Arg Thr Thr Ser Pro Lys Leu Ala Asn Val

595 600 605

Lys Ala Leu Tyr Ser Asn Leu Lys Leu Glu Val Lys Asp Gly Gln Leu

610 615 620

Phe Ile Lys Asn Asp Asn Leu Phe Thr Asn Ser Ser Ala Tyr Tyr Phe

625 630 635 640

Leu Thr Ser Leu Leu Val Asp Gly Lys Leu Thr Tyr Gln Ser Gln Pro

645 650 655

Leu Thr Phe Gly Leu Glu Pro Gly Glu Ser Gly Thr Phe Ala Leu Pro

660 665 670

Trp Pro Glu Val Glu Asp Glu Lys Gly Glu Ile Val Tyr Gln Val Thr

675 680 685

Ala His Leu Lys Glu Asp Leu Pro Trp Ala Asp Glu Gly Phe Thr Val

690 695 700

Ala Glu Ala Glu Glu Ala Val Thr Lys Leu Pro Glu Phe Tyr Pro Ala

705 710 715 720

Gly Arg Pro Glu Leu Val Asp Ser Asp Phe Asn Leu Gly Leu Lys Gly

725 730 735

Asn Gly Phe Arg Ile Leu Phe Ser Lys Ala Lys Gly Trp Pro Val Ser

740 745 750

Ile Lys Tyr Ala Gly Arg Glu Tyr Leu Lys Arg Leu Pro Glu Phe Thr

755 760 765

Phe Trp Arg Ala Leu Thr Asp Asn Asp Arg Gly Ala Gly Tyr Gly Tyr

770 775 780

Asp Leu Ala Lys Trp Glu Asn Ala Gly Lys Tyr Ala Arg Leu Gln Asp

785 790 795 800

Ile Ser Tyr Glu Ile Lys Glu Asn Ser Ala Leu Val Lys Thr Ala Phe

805 810 815

Thr Leu Pro Val Ala Leu Lys Gly Asp Leu Thr Ile Thr Tyr Glu Val

820 825 830

Asp Ser Leu Gly Lys Ile Ala Val Thr Ala Asn Phe Pro Gly Ala Val

835 840 845

Glu Asn Gly Leu Leu Pro Ala Phe Gly Leu Asn Phe Ala Leu Pro Lys

850 855 860

Glu Leu Ser Asp Tyr Arg Tyr Tyr Gly Leu Gly Pro Asn Glu Ser Tyr

865 870 875 880

Ala Asp Arg Leu Glu Gly Ser Tyr Leu Gly Ile Tyr Gln Gly Met Val

885 890 895

Glu Lys Asn Phe Thr Pro Tyr Leu Arg Pro Gln Glu Ala Gly Asn Arg

900 905 910

Ser Lys Val Arg Tyr Tyr Gln Leu Phe Asp Glu Glu Gly Gly Leu Glu

915 920 925

Phe Thr Ala Asn Gly Ala Asp Leu Asn Leu Ser Ala Leu Pro Tyr Ser

930 935 940

Ala Ala Gln Ile Glu Ala Ala Asp His Ala Phe Glu Leu Thr Asn Asn

945 950 955 960

Tyr Thr Trp Val Arg Ala Leu Ala Ala Gln Met Gly Val Gly Gly Asp

965 970 975

Asp Ser Trp Gly Gln Lys Val His Pro Glu Phe Cys Leu Asp Ala Gln

980 985 990

Glu Ala Arg Gln Leu Lys Leu Val Ile Gln Pro Leu Leu Leu Lys

995 1000 1005

<210> 20

<211> 628

<212>PRT

<213> Lactobacillus helvaticus

<400> 20

Met Gln Ala Asn Ile Asn Trp Leu Asp Asn Pro Glu Val Phe Arg Val

1 5 10 15

Asn Gln Leu Pro Ala His Ser Asp His Pro Phe Phe Arg Asp Tyr Arg

20 25 30

Glu Trp Gln Lys Gln His Ser Ser Tyr Gln Gln Ser Leu Asn Gly Lys

35 40 45

Trp Lys Phe His Phe Ser Ala Asn Pro Met Asp Arg Pro Gln Asp Phe

50 55 60

Tyr Gln Arg Asp Phe Asp Ser Ser Asn Phe Asp Ser Ile Pro Val Pro

65 70 75 80

Ser Glu Ile Glu Leu Ser Asn Tyr Thr Gln Asn Gln Tyr Ile Asn Val

85 90 95

Leu Phe Pro Trp Glu Gly Lys Ile Phe Arg Arg Pro Ala Tyr Ala Leu

100 105 110

Asp Pro Asn Asp His Glu Glu Gly Ser Phe Ser Lys Gly Ala Asp Asn

115 120 125

Thr Val Gly Ser Tyr Leu Lys Arg Phe Asp Leu Ser Ser Ala Leu Ile

130 135 140

Gly Lys Asp Val His Ile Lys Phe Glu Gly Val Glu Gln Ala Met Tyr

145 150 155 160

Val Trp Leu Asn Gly His Phe Val Gly Tyr Ala Glu Asp Ser Phe Thr

165 170 175

Pro Ser Glu Phe Asp Leu Thr Pro Tyr Ile Gln Asp Lys Asp Asn Leu

180 185 190

Leu Ala Val Glu Val Phe Lys His Ser Thr Ala Ser Trp Leu Glu Asp

195 200 205

Gln Asp Met Phe Arg Phe Ser Gly Ile Phe Arg Ser Val Glu Leu Leu

210 215 220

Gly Ile Pro Ala Thr His Leu Met Asp Met Asp Leu Lys Pro Arg Val

225 230 235 240

Ala Asp Asn Tyr Gln Asp Gly Ile Phe Asn Leu Lys Leu His Phe Ile

245 250 255

Gly Lys Lys Ala Gly Ser Phe His Leu Leu Val Lys Asp Ile Lys Gly

260 265 270

His Thr Leu Leu Glu Lys Asn Glu Asp Ile Lys Glu Asn Val Gln Ile

275 280 285

Asn Asn Glu Lys Phe Glu Asn Val His Leu Trp Asn Asn His Asp Pro

290 295 300

Tyr Leu Tyr Gln Leu Leu Ile Glu Val Tyr Asp Glu Gln Gln Asn Leu

305 310 315 320

Leu Glu Leu Ile Pro Phe Gln Phe Gly Phe Arg Arg Ile Glu Ile Ser

325 330 335

Pro Glu Lys Val Val Leu Leu Asn Gly Lys Arg Leu Ile Ile Asn Gly

340 345 350

Val Asn Arg His Glu Trp Asp Ala Lys Arg Gly Arg Ser Ile Thr Met

355 360 365

Ser Asp Met Thr Thr Asp Ile Asn Thr Phe Lys Glu Asn Asn Ile Asn

370 375 380

Ala Val Arg Thr Cys His Tyr Pro Asn Gln Ile Pro Trp Tyr Tyr Leu

385 390 395 400

Cys Asp Gln Asn Gly Ile Tyr Val Met Ala Glu Asn Asn Leu Glu Ser

405 410 415

His Gly Thr Trp Gln Lys Met Gly Glu Ile Glu Pro Ser Asp Asn Val

420 425 430

Pro Gly Ser Ile Pro Gln Trp Lys Glu Ala Val Ile Asp Arg Ala Arg

435 440 445

Asn Asn Tyr Glu Thr Phe Lys Asn His Thr Ser Ile Leu Phe Trp Ser

450 455 460

Leu Gly Asn Glu Ser Tyr Ala Gly Asp Asn Ile Ile Ala Met Asn Glu

465 470 475 480

Phe Tyr Lys Ser His Asp Asp Thr Arg Leu Val His Tyr Glu Gly Val

485 490 495

Val His Arg Pro Glu Leu Lys Asp Lys Ile Ser Asp Val Glu Ser Cys

500 505 510

Met Tyr Leu Pro Pro Lys Lys Val Glu Glu Tyr Leu Gln Asn Asp Pro

515 520 525

Pro Lys Pro Phe Met Glu Cys Glu Tyr Met His Asp Met Gly Asn Ser

530 535 540

Asp Gly Gly Met Gly Ser Tyr Ile Lys Leu Leu Asp Lys Tyr Pro Gln

545 550 555 560

Tyr Phe Gly Gly Phe Ile Trp Asp Phe Ile Asp Gln Ala Leu Leu Val

565 570 575

His Asp Glu Ile Ser Gly His Asp Val Leu Arg Tyr Gly Gly Asp Phe

580 585 590

Asp Asp Arg His Ser Asp Tyr Glu Phe Ser Gly Asp Gly Leu Met Phe

595 600 605

Ala Asp Arg Thr Pro Lys Pro Ala Met Gln Glu Val Arg Tyr Tyr Tyr

610 615 620

Gly Leu His Lys

625

<210> 21

<211> 318

<212>PRT

<213> Lactobacillus helvaticus

<400> 21

Met Asp Tyr Thr Asn Asn Gln Leu His Ile Ile Tyr Gly Asp Ala Thr

1 5 10 15

Phe Gly Val Asn Gly Lys Asp Phe Gln Tyr Ile Phe Ser Tyr Glu Arg

20 25 30

Gly Gly Leu Glu Ser Leu Lys Val His Gly Lys Glu Trp Leu Tyr Arg

35 40 45

Val Pro Thr Pro Thr Phe Trp Arg Ala Thr Thr Asp Asn Asp Arg Gly

50 55 60

Ser Gly Phe Asn Leu Lys Ala Ala Gln Trp Leu Gly Ala Asp Met Phe

65 70 75 80

Thr Lys Cys Thr Asp Ile His Leu Lys Val Asp Arg His Asp Phe Ala

85 90 95

Glu Leu Pro Ile Ala Pro Phe Asn Asn Lys Phe Ser Asn His Glu Tyr

100 105 110

Ala Lys Ser Ala Glu Ile Ser Phe Thr Tyr Gln Thr Leu Thr Thr Pro

115 120 125

Ala Thr Asn Ala Lys Ile Ile Tyr Asn Ile Asp Asp Val Gly His Ile

130 135 140

Lys Val Thr Met Arg Tyr Tyr Gly Lys Lys Gly Leu Pro Pro Leu Pro

145 150 155 160

Val Ile Gly Ile Arg Leu Ile Met Pro Thr Ala Ala Thr Gly Phe Asp

165 170 175

Tyr Glu Gly Leu Ser Gly Glu Thr Tyr Pro Asp Arg Met Ala Gly Ala

180 185 190

Lys Glu Gly Lys Phe His Ile Asp Gly Leu Pro Val Thr Glu Tyr Leu

195 200 205

Val Pro Gln Glu Asn Gly Met His Met Gln Thr Lys Lys Leu Thr Ile

210 215 220

Asn Arg Glu Thr Thr Gln Asn Asn Val Asp Arg Thr Asn Glu Lys Phe

225 230 235 240

Ser Leu Ser Ile Gln Gln Ala Glu Lys Pro Phe Asn Phe Ser Cys Leu

245 250 255

Pro Tyr Thr Ala Glu Glu Leu Glu Asn Ala Thr His Ile Glu Glu Leu

260 265 270

Pro Leu Val Arg Arg Thr Val Leu Val Ile Ala Gly Ala Val Arg Gly

275 280 285

Val Gly Gly Ile Asp Ser Trp Gly Thr Asp Val Glu Ser Ala Tyr His

290 295 300

Ile Asn Pro Glu Leu Asp His Glu Phe Ser Phe Ile Leu Asn

305 310 315

<210> 22

<211> 1023

<212>PRT

<213> Bifidobacterium longum

<400> 22

Met Thr Asp Val Thr His Val Asp Arg Ala Ser Gln Ala Trp Leu Thr

1 5 10 15

Asp Pro Thr Val Phe Glu Val Asn Arg Thr Pro Ala His Ser Ser His

20 25 30

Lys Trp Tyr Ala Arg Asp Pro Gln Ser Gly Gln Trp Ser Asp Leu Lys

35 40 45

Gln Ser Leu Asp Gly Glu Trp Arg Val Glu Val Val Gln Ala Ala Asp

50 55 60

Ile Asn Leu Glu Glu Glu Pro Ala Thr Ala Glu Ser Phe Asp Asp Ser

65 70 75 80

Ser Phe Glu Arg Ile Gln Val Pro Gly His Leu Gln Thr Ala Gly Leu

85 90 95

Met Asn His Lys Tyr Val Asn Val Gln Tyr Pro Trp Asp Gly His Glu

100 105 110

Asn Pro Leu Glu Pro Asn Ile Pro Glu Asn Asn His Val Ala Leu Tyr

115 120 125

Arg Arg Lys Phe Thr Val Ser Ala Pro Val Ala Asn Ala Lys Gln Ala

130 135 140

Gly Gly Ser Val Ser Ile Val Phe His Gly Met Ala Thr Ala Ile Tyr

145 150 155 160

Val Trp Val Asn Gly Ala Phe Val Gly Tyr Gly Glu Asp Gly Phe Thr

165 170 175

Pro Asn Glu Phe Asp Ile Thr Glu Leu Leu His Asp Gly Glu Asn Val

180 185 190

Val Ala Val Ala Cys Tyr Glu Tyr Ser Ser Ala Ser Trp Leu Glu Asp

195 200 205

Gln Asp Phe Trp Arg Leu His Gly Leu Phe Arg Ser Val Glu Leu Ala

210 215 220

Ala Arg Pro His Val His Ile Glu Asn Thr Gln Ile Glu Ala Asp Trp

225 230 235 240

Asp Pro Glu Ala Gly Thr Ala Ser Leu Asp Ala Ala Leu Thr Val Leu

245 250 255

Asn Ala Ala Asp Ala Ala Thr Val Arg Ala Thr Leu Lys Asp Ala Asp

260 265 270

Gly Asn Thr Val Trp Gln Thr Thr Gly Asp Ala Glu Ala Gln Thr Ala

275 280 285

Ile Ser Ser Gly Pro Leu Gln Gly Ile Ala Pro Trp Ser Ala Glu Ser

290 295 300

Pro Thr Leu Tyr Glu Leu Asp Val Asp Val Ile Asp Gln Ala Gly Asp

305 310 315 320

Val Ile Glu Cys Thr Ser Gln Lys Val Gly Phe Arg Arg Phe Arg Ile

325 330 335

Glu Asp Gly Ile Leu Thr Ile Asn Gly Lys Arg Ile Val Phe Lys Gly

340 345 350

Ala Asp Arg His Glu Phe Asp Ala Glu Gln Gly Arg Ala Ile Thr Glu

355 360 365

Gln Asp Met Ile Asp Asp Val Val Phe Cys Lys Arg His Asn Ile Asn

370 375 380

Ser Ile Arg Thr Ser His Tyr Pro Asn Gln Glu Arg Trp Tyr Glu Leu

385 390 395 400

Cys Asp Glu Tyr Gly Ile Tyr Leu Ile Asp Glu Ala Asn Leu Glu Ala

405 410 415

His Gly Ser Trp Ser Leu Pro Gly Asp Val Leu Thr Glu Asp Thr Ile

420 425 430

Val Pro Gly Ser Lys Arg Glu Trp Glu Gly Ala Cys Val Asp Arg Val

435 440 445

Asn Ser Met Met Arg Arg Asp Tyr Asn His Pro Ser Val Leu Ile Trp

450 455 460

Ser Leu Gly Asn Glu Ser Tyr Val Gly Asp Val Phe Arg Ala Met Tyr

465 470 475 480

Lys His Val His Asp Ile Asp Pro Asn Arg Pro Val His Tyr Glu Gly

485 490 495

Val Thr His Asn Arg Asp Tyr Asp Asp Val Thr Asp Ile Glu Thr Arg

500 505 510

Met Tyr Ser His Ala Asp Glu Ile Glu Lys Tyr Leu Lys Asp Asp Pro

515 520 525

Lys Lys Pro Tyr Leu Ser Cys Glu Tyr Met His Ala Met Gly Asn Ser

530 535 540

Val Gly Asn Met Asp Glu Tyr Thr Ala Leu Glu Arg Tyr Pro Lys Tyr

545 550 555 560

Gln Gly Gly Phe Ile Trp Asp Phe Ile Asp Gln Ala Ile Tyr Ala Thr

565 570 575

Gln Pro Asp Gly Thr Arg Ser Leu Arg Tyr Gly Gly Asp Phe Gly Asp

580 585 590

Arg Pro Ser Asp Tyr Glu Phe Ser Gly Asp Gly Leu Leu Phe Ala Asn

595 600 605

Arg Lys Pro Ser Pro Lys Ala Gln Glu Val Lys Gln Leu Tyr Ser Asn

610 615 620

Val His Ile Asp Val Thr Lys Asp Ser Val Ser Val Lys Asn Asp Asn

625 630 635 640

Leu Phe Thr Ala Thr Gly Asp Tyr Val Phe Val Leu Ser Val Leu Ala

645 650 655

Asp Gly Lys Pro Val Trp Gln Ser Thr Arg Arg Phe Asp Val Pro Ala

660 665 670

Gly Glu Thr Arg Thr Phe Asp Val Ala Trp Pro Val Ala Ala Tyr Arg

675 680 685

Ala Asp Ala Arg Glu Leu Val Leu Gln Val Ser Gln Arg Leu Ala Lys

690 695 700

Ala Thr Asp Trp Ala Glu Ser Gly Tyr Glu Leu Ala Phe Gly Gln Thr

705 710 715 720

Val Val Pro Ala Asp Ala Thr Ala Thr Pro Asp Thr Lys Pro Ala Asp

725 730 735

Gly Thr Ile Thr Val Gly Arg Trp Asn Ala Gly Val Arg Gly Ala Gly

740 745 750

Arg Glu Val Leu Leu Ser Arg Thr Gln Gly Gly Met Val Ser Tyr Thr

755 760 765

Phe Ala Gly Asn Glu Phe Val Leu Arg Arg Pro Ala Ile Thr Thr Phe

770 775 780

Arg Pro Leu Thr Asp Asn Asp Arg Gly Ala Gly His Gly Phe Glu Arg

785 790 795 800

Val Gln Trp Leu Gly Ala Gly Arg Tyr Ala Arg Cys Val Asp Asn Val

805 810 815

Leu Glu Gln Ile Asp Asp Ser Thr Leu Lys Gly Thr Tyr Thr Tyr Glu

820 825 830

Leu Ala Thr Ala Gln Arg Thr Lys Val Thr Val Ser Tyr Thr Ala His

835 840 845

Thr Asp Gly Arg Val Asn Leu His Val Glu Tyr Pro Gly Glu Gln Gly

850 855 860

Asp Leu Pro Thr Ile Pro Ala Phe Gly Ile Glu Trp Thr Leu Pro Val

865 870 875 880

Gln Tyr Thr Asn Leu Arg Phe Phe Gly Thr Gly Pro Ala Glu Thr Tyr

885 890 895

Leu Asp Arg Lys His Ala Lys Leu Gly Val Trp Ser Thr Asn Ala Phe

900 905 910

Ala Asp His Ala Pro Tyr Leu Met Pro Gln Glu Thr Gly Asn His Glu

915 920 925

Asp Val Arg Trp Ala Glu Ile Thr Asp Asp His Gly His Gly Met Arg

930 935 940

Val Ser Arg Ala Asp Gly Ala Ala Pro Phe Ala Val Ser Leu Leu Pro

945 950 955 960

Tyr Ser Ser Phe Met Leu Glu Glu Ala Gln His Gln Asp Glu Leu Pro

965 970 975

Lys Pro Lys His Met Phe Leu Arg Val Leu Ala Ala Gln Met Gly Val

980 985 990

Gly Gly Asp Asp Ser Trp Met Ser Pro Val His Pro Gln Tyr His Ile

995 1000 1005

Pro Ala Asp Lys Pro Ile Ser Leu Asp Val Asp Leu Glu Leu Ile

1010 1015 1020

<210> 23

<211> 628

<212>PRT

<213> Lactobacillus reuteri

<400> 23

Met Asp Ala Asp Ile Lys Trp Leu Asp Glu Pro Glu Thr Phe Arg Val

1 5 10 15

Asn Gln Leu Pro Ala His Ser Asp His Tyr Tyr Tyr Gly Asn Tyr Asp

20 25 30

Glu Trp Arg His Asn Asn Ser Arg Phe Ala Gln Asn Leu Asp Gly Gln

35 40 45

Trp Gln Phe Asn Phe Ala Glu Asn Leu Arg Glu Arg Glu Asn Asp Phe

50 55 60

Tyr Lys Met Asp Tyr Asp Ser Ser Ser Phe Gly Thr Ile Glu Val Pro

65 70 75 80

Ser Glu Ile Glu Leu Asn Asn Tyr Ala Gln Asn Asn Tyr Ile Asn Thr

85 90 95

Leu Ile Pro Trp Glu Gly Lys Ile Tyr Arg Arg Pro Ala Tyr Thr Leu

100 105 110

Ser Pro Asp Asp Ala Gln Glu Gly Ser Phe Ser Asp Gly Asp Asp Asn

115 120 125

Thr Ile Gly Glu Tyr Leu Lys His Phe Asp Leu Asp Pro Ser Leu Arg

130 135 140

Gly Lys Gln Val Arg Ile Arg Phe Asp Gly Val Glu Arg Ala Met Tyr

145 150 155 160

Val Trp Leu Asn Gly His Phe Ile Gly Tyr Ala Glu Asp Ser Phe Thr

165 170 175

Pro Ser Glu Phe Asp Leu Thr Pro Tyr Ile Gln Asp Glu Gly Asn Val

180 185 190

Leu Ala Val Glu Val Phe Lys His Ser Thr Ala Ser Trp Ile Glu Asp

195 200 205

Gln Asp Met Phe Arg Phe Ser Gly Ile Phe Arg Ser Val Asn Leu Leu

210 215 220

Ala Gln Pro Leu Val His Val Glu Asp Leu Asn Ile Arg Pro Ile Val

225 230 235 240

Thr Asp Asn Tyr Gln Asp Gly Ile Phe Asn Val Glu Leu Gln Leu His

245 250 255

Gly Glu Lys Thr Gly Asn Val Asn Val Arg Val Ile Asp Asn Asp Gly

260 265 270

Asn Thr Leu Val Asn Glu Thr His Pro Val Asp Ser Thr Val Lys Val

275 280 285

Gln Asp Gln Phe Leu Glu Asn Val His Leu Trp Asp Asn His Asp Pro

290 295 300

Tyr Leu Tyr Gln Leu Leu Ile Glu Ile Arg Asp Asp Glu Gly Asn Leu

305 310 315 320

Val Glu Leu Val Pro Tyr Arg Phe Gly Phe Arg Arg Ile Glu Ile Asn

325 330 335

Lys Asp His Val Val Leu Leu Asn Gly Gln Arg Leu Ile Ile Asn Gly

340 345 350

Val Asn Arg His Glu Trp Asp Ala Arg Arg Gly Arg Ala Ile Thr Met

355 360 365

Asp Asp Met Thr Ser Asp Ile His Thr Phe Lys Glu Asn Asn Ile Asn

370 375 380

Ala Val Arg Thr Cys His Tyr Pro Asp Gln Ile Pro Trp Tyr Tyr Leu

385 390 395 400

Cys Asp Asp Asn Gly Ile Tyr Met Met Ala Glu Asn Asn Leu Glu Ser

405 410 415

His Ala Thr Trp Gln Lys Met Gly Ala Ile Glu Pro Ser Tyr Asn Val

420 425 430

Pro Gly Ser Val Pro Gln Trp Arg Asp Val Val Val Asp Arg Ala Arg

435 440 445

Thr Asn Tyr Glu Thr Phe Lys Asn His Pro Ser Ile Leu Phe Trp Ser

450 455 460

Leu Gly Asn Glu Ser Tyr Ala Gly Asp Asn Ile Val Lys Met Asn Glu

465 470 475 480

Phe Tyr Lys Lys His Asp Asp Ser Arg Leu Val His Tyr Glu Gly Val

485 490 495

Cys His Thr Pro Glu Tyr Arg Asp Arg Ile Ser Asp Val Glu Ser Trp

500 505 510

Met Tyr Leu Pro Pro Lys Glu Val Glu Glu Tyr Leu Lys Asn Asn Pro

515 520 525

Asp Lys Pro Phe Met Glu Cys Glu Tyr Met His Asp Met Gly Asn Ser

530 535 540

Asp Gly Gly Met Gly Ser Tyr Ile Ser Leu Leu Asp Lys Tyr Pro Gln

545 550 555 560

Tyr Phe Gly Gly Phe Ile Trp Asp Phe Ile Asp Gln Ala Leu Leu Val

565 570 575

Lys Asp Pro Val Ser Gly Gln Glu Val Met Arg Tyr Gly Gly Asp Phe

580 585 590

Asp Asp Arg His Ser Asp Tyr Glu Phe Ser Gly Asp Gly Leu Met Phe

595 600 605

Ala Asp Arg Thr Pro Lys Pro Ala Met Gln Glu Val Arg Tyr Tyr Tyr

610 615 620

Gly Leu His Lys

625

<210> 24

<211> 319

<212>PRT

<213> Lactobacillus reuteri

<400> 24

Met Ala Tyr Thr Asn Lys Leu Arg Val Ile Tyr Gly Asp Ala Thr Leu

1 5 10 15

Gly Leu Ser Gly Asp Gly Phe His Tyr Ile Phe Ser Tyr Glu Arg Gly

20 25 30

Gly Leu Glu Ser Leu Lys Leu Asn Gly Lys Glu Trp Leu Tyr Arg Glu

35 40 45

Pro Met Pro Thr Phe Trp Arg Ala Thr Thr Asp Asn Asp Arg Gly Ser

50 55 60

Gly Phe Asn Ile Arg Ser Ala Gln Trp Leu Ala Ala Asp Thr Phe His

65 70 75 80

Lys Cys Val Gly Ile Asp Leu Thr Val Asp Asn Gln His Phe Ala Glu

85 90 95

Leu Pro Ile Ala Pro Ile Thr Asn Glu Phe Ser Asp Pro Val Ser Ala

100 105 110

Glu Ser Val Lys Ile Lys Tyr Thr Phe Ala Thr Leu Thr Val Pro Ala

115 120 125

Thr Gln Val Thr Val Ile Tyr Glu Val Asn Gly Gln Gly Glu Ile Lys

130 135 140

Val Thr Met His Tyr Tyr Gly His Glu Asp Leu Pro Gly Leu Pro Val

145 150 155 160

Val Gly Met Arg Phe Ile Met Pro Thr Val Ala Thr Gly Phe Asp Tyr

165 170 175

Gln Gly Leu Ser Gly Glu Thr Tyr Pro Asp Arg Met Ala Gly Ala Thr

180 185 190

Glu Gly Thr Phe His Val Asp Gly Leu Pro Val Thr Lys Tyr Leu Val

195 200 205

Pro Gln Glu Asn Gly Met His Met Ala Thr His Ala Leu Thr Ile Thr

210 215 220

Arg Asp Ser Thr Gln Asn Asn Ala Asp His Ser Arg Glu Pro Phe Ser

225 230 235 240

Leu Thr Val Lys Gln Asp Ala Gln Pro Phe Ala Phe Ser Cys Leu Pro

245 250 255

Tyr Thr Ala Glu Glu Leu Glu Asn Ala Thr His Ile Glu Glu Leu Pro

260 265 270

Leu Ala Arg Arg Thr Val Leu Val Val Ala Gly Ala Val Arg Gly Val

275 280 285

Gly Gly Ile Asp Ser Trp Gly Ala Asp Val Glu Glu Gln Tyr His Ile

290 295 300

Pro Ala Asp Arg Asp Val Glu Phe Ser Phe Val Leu Asn Ala Lys

305 310 315

<210> 25

<211> 1007

<212>PRT

<213> Lactobacillus delbrueckii subsp. lactis

<400> 25

Met Ser Asn Lys Leu Val Lys Glu Lys Arg Val Asp Gln Ala Asp Leu

1 5 10 15

Ala Trp Leu Thr Asp Pro Glu Val Tyr Glu Val Asn Thr Ile Pro Pro

20 25 30

His Ser Asp His Glu Ser Phe Gln Ser Gln Glu Glu Leu Glu Glu Gly

35 40 45

Lys Ser Ser Leu Val Gln Ser Leu Asp Gly Asn Trp Leu Ile Asp Tyr

50 55 60

Ala Glu Asn Gly Gln Gly Pro Ile Asn Phe Tyr Ala Glu Asp Phe Asp

65 70 75 80

Asp Ser Asn Phe Lys Ser Val Lys Val Pro Gly Asn Leu Glu Leu Gln

85 90 95

Gly Phe Gly Gln Pro Gln Tyr Val Asn Ile Gln Tyr Pro Trp Asp Gly

100 105 110

Ser Glu Glu Ile Phe Pro Pro Gln Val Pro Ser Lys Ile Pro Leu Ala

115 120 125

Ser Tyr Val Arg Tyr Phe Asp Leu Asp Glu Ala Leu Trp Asp Lys Glu

130 135 140

Val Ser Leu Lys Phe Ala Gly Ala Ala Thr Ala Ile Tyr Val Trp Leu

145 150 155 160

Asn Gly His Phe Val Gly Tyr Gly Glu Asp Ser Phe Thr Pro Ser Glu

165 170 175

Phe Met Val Thr Lys Phe Leu Lys Lys Glu Gly Asn Arg Leu Ala Val

180 185 190

Ala Leu Tyr Lys Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe

195 200 205

Trp Arg Leu Ser Gly Leu Phe Arg Ser Val Thr Leu Glu Ala Lys Pro

210 215 220

Leu Leu His Leu Glu Asp Leu Lys Leu Thr Ala Ser Leu Thr Asp Asn

225 230 235 240

Tyr Gln Lys Gly Lys Leu Glu Val Glu Ala Asn Ile Ala Tyr Arg Leu

245 250 255

Pro Asn Ala Ser Phe Lys Leu Glu Val Arg Asp Ser Glu Gly Asp Leu

260 265 270

Val Ala Glu Lys Val Gly Pro Ile Arg Ser Glu Lys Leu Asp Phe Ser

275 280 285

Leu Ala Asp Leu Pro Val Ala Ala Trp Ser Ala Glu Lys Pro Asn Leu

290 295 300

Tyr Gln Val Arg Leu Tyr Leu Tyr Gln Ala Gly Ser Leu Leu Glu Val

305 310 315 320

Ser Arg Gln Glu Val Gly Phe Arg Asn Phe Glu Leu Lys Asp Gly Ile

325 330 335

Met Tyr Leu Asn Gly Gln Arg Ile Val Phe Lys Gly Val Asn Arg His

340 345 350

Glu Phe Asp Ser Lys Leu Gly Arg Ala Ile Thr Glu Ala Asp Met Ile

355 360 365

Trp Asp Ile Lys Thr Met Lys Gln Ser Asn Ile Asn Ala Val Arg Cys

370 375 380

Ser His Tyr Pro Asn Gln Ser Leu Phe Tyr Arg Leu Cys Asp Lys Tyr

385 390 395 400

Gly Leu Tyr Val Ile Asp Glu Ala Asn Leu Glu Ser His Gly Thr Trp

405 410 415

Glu Lys Val Gly His Glu Asp Pro Ser Phe Asn Val Pro Gly Asp Asp

420 425 430

Gln His Trp Leu Gly Ala Ser Leu Ser Arg Val Lys Asn Met Met Ala

435 440 445

Arg Asp Lys Asn His Ala Ser Ile Leu Ile Trp Ser Leu Gly Asn Glu

450 455 460

Ser Tyr Ala Gly Thr Val Phe Ala Gln Met Ala Asp Tyr Val Arg Lys

465 470 475 480

Ala Asp Pro Thr Arg Val Gln His Tyr Glu Gly Val Thr His Asn Arg

485 490 495

Lys Phe Asp Asp Ala Thr Gln Ile Glu Ser Arg Met Tyr Ala Pro Ala

500 505 510

Lys Glu Ile Glu Glu Tyr Leu Thr Lys Lys Pro Ala Lys Pro Phe Ile

515 520 525

Ser Val Glu Tyr Ala His Ala Met Gly Asn Ser Val Gly Asp Leu Ala

530 535 540

Ala Tyr Thr Ala Leu Glu Lys Tyr Pro His Tyr Gln Gly Gly Phe Ile

545 550 555 560

Trp Asp Trp Ile Asp Gln Gly Leu Glu Lys Asp Gly His Leu Leu Tyr

565 570 575

Gly Gly Asp Phe Asp Asp Arg Pro Thr Asp Tyr Glu Phe Cys Gly Asp

580 585 590

Gly Leu Val Phe Ala Asp Arg Thr Thr Ser Pro Lys Leu Ala Asn Val

595 600 605

Lys Ala Leu Tyr Ser Asn Leu Lys Leu Glu Val Lys Asp Gly Gln Leu

610 615 620

Phe Ile Lys Asn Asp Asn Leu Phe Thr Asn Ser Ser Ala Tyr Tyr Phe

625 630 635 640

Leu Thr Ser Leu Leu Val Asp Gly Lys Leu Thr Tyr Gln Ser Gln Pro

645 650 655

Leu Thr Phe Gly Leu Glu Pro Gly Glu Ser Gly Thr Phe Ala Leu Pro

660 665 670

Trp Pro Glu Val Glu Asp Glu Lys Gly Glu Ile Val Tyr Gln Val Thr

675 680 685

Ala His Leu Lys Glu Asp Leu Pro Trp Ala Asp Glu Gly Phe Thr Val

690 695 700

Ala Glu Ala Glu Glu Ala Val Thr Lys Leu Pro Glu Phe Tyr Pro Ala

705 710 715 720

Gly Arg Pro Glu Leu Val Asp Ser Asp Phe Asn Leu Gly Leu Lys Gly

725 730 735

Asn Gly Phe Arg Ile Leu Phe Ser Lys Ala Lys Gly Trp Pro Val Ser

740 745 750

Ile Lys Tyr Ala Gly Arg Glu Tyr Leu Lys Arg Leu Pro Glu Phe Thr

755 760 765

Phe Trp Arg Ala Leu Thr Asp Asn Asp Arg Gly Ala Gly Tyr Gly Tyr

770 775 780

Asp Leu Ala Lys Trp Glu Asn Ala Gly Lys Tyr Ala Arg Leu Gln Asp

785 790 795 800

Ile Ser Tyr Glu Ile Lys Glu Asn Ser Ala Leu Val Lys Thr Thr Phe

805 810 815

Thr Leu Pro Val Ala Leu Lys Gly Asp Leu Thr Ile Thr Tyr Glu Val

820 825 830

Asp Ser Leu Gly Lys Ile Ala Val Thr Ala Asn Phe Pro Gly Ala Val

835 840 845

Glu Asn Gly Leu Leu Pro Ala Phe Gly Leu Asn Phe Ala Leu Pro Lys

850 855 860

Glu Leu Ser Asp Tyr Arg Tyr Tyr Gly Leu Gly Pro Asn Glu Ser Tyr

865 870 875 880

Ala Asp Arg Leu Glu Gly Ser Tyr Leu Gly Ile Tyr Gln Gly Met Val

885 890 895

Glu Lys Asn Phe Thr Pro Tyr Leu Arg Pro Gln Glu Ala Gly Asn Arg

900 905 910

Ser Lys Val Arg Tyr Tyr Gln Leu Phe Asp Glu Glu Gly Gly Leu Glu

915 920 925

Phe Thr Ala Asn Gly Ala Asp Leu Asn Leu Ser Ala Leu Pro Tyr Ser

930 935 940

Ala Ala Gln Ile Glu Ala Ala Asp His Ala Phe Glu Leu Thr Asn Asn

945 950 955 960

Tyr Thr Trp Val Arg Ala Leu Ala Ala Gln Met Gly Val Gly Gly Asp

965 970 975

Asp Ser Trp Gly Gln Lys Val His Pro Glu Phe Cys Leu Asp Ala Gln

980 985 990

Glu Ala Arg Gln Leu Lys Leu Val Ile Gln Pro Leu Leu Leu Lys

995 1000 1005

<210> 26

<211> 628

<212>PRT

<213> Lactobacillus helvaticus

<400> 26

Met Gln Ala Asn Ile Asn Trp Leu Asp Asn Pro Glu Val Phe Arg Val

1 5 10 15

Asn Gln Leu Pro Ala His Ser Asp His Pro Phe Phe Arg Asp Tyr Arg

20 25 30

Glu Trp Gln Lys Gln His Ser Ser Tyr Gln Gln Ser Leu Asn Gly Lys

35 40 45

Trp Lys Phe His Phe Ser Ala Asn Pro Met Asp Arg Pro Gln Asp Phe

50 55 60

Tyr Gln Arg Asp Phe Asp Ser Ser Asn Phe Asp Ser Ile Pro Val Pro

65 70 75 80

Ser Glu Ile Glu Leu Ser Asn Tyr Thr Gln Asn Gln Tyr Ile Asn Val

85 90 95

Leu Phe Pro Trp Glu Gly Lys Ile Phe Arg Arg Pro Ala Tyr Ala Leu

100 105 110

Asp Pro Asn Asp His Glu Glu Gly Ser Phe Ser Lys Gly Ala Asp Asn

115 120 125

Thr Val Gly Ser Tyr Leu Lys Arg Phe Asp Leu Ser Ser Ala Leu Ile

130 135 140

Gly Lys Asp Val His Ile Lys Phe Glu Gly Val Glu Gln Ala Met Tyr

145 150 155 160

Val Trp Leu Asn Gly His Phe Val Gly Tyr Ala Glu Asp Ser Phe Thr

165 170 175

Pro Ser Glu Phe Asp Leu Thr Pro Tyr Ile Gln Glu Lys Asp Asn Leu

180 185 190

Leu Ala Val Glu Val Phe Lys His Ser Thr Ala Ser Trp Leu Glu Asp

195 200 205

Gln Asp Met Phe Arg Phe Ser Gly Ile Phe Arg Ser Val Glu Leu Leu

210 215 220

Gly Ile Pro Ala Thr His Leu Met Asp Met Asp Leu Lys Pro Arg Val

225 230 235 240

Ala Asp Asn Tyr Gln Asp Gly Ile Phe Asn Leu Lys Leu His Phe Ile

245 250 255

Gly Lys Lys Ala Gly Ser Phe His Leu Leu Val Lys Asp Ile Lys Gly

260 265 270

His Thr Leu Leu Glu Lys Asn Glu Asp Ile Lys Glu Asn Val Gln Ile

275 280 285

Asn Asn Glu Lys Phe Glu Asn Val His Leu Trp Asn Asn His Asp Pro

290 295 300

Tyr Leu Tyr Gln Leu Leu Ile Glu Val Tyr Asp Glu Gln Gln Asn Leu

305 310 315 320

Leu Glu Leu Ile Pro Phe Gln Phe Gly Phe Arg Arg Ile Glu Ile Ser

325 330 335

Pro Glu Lys Val Val Leu Leu Asn Gly Lys Arg Leu Ile Ile Asn Gly

340 345 350

Val Asn Arg His Glu Trp Asp Ala Lys Arg Gly Arg Ser Ile Thr Met

355 360 365

Ser Asp Met Thr Thr Asp Ile Asn Thr Phe Lys Glu Asn Asn Ile Asn

370 375 380

Ala Val Arg Thr Cys His Tyr Pro Asn Gln Ile Pro Trp Tyr Tyr Leu

385 390 395 400

Cys Asp Gln Asn Gly Ile Tyr Val Met Ala Glu Asn Asn Leu Glu Ser

405 410 415

His Gly Thr Trp Gln Lys Met Gly Glu Ile Glu Pro Ser Asp Asn Val

420 425 430

Pro Gly Ser Ile Pro Gln Trp Lys Glu Ala Val Ile Asp Arg Ala Arg

435 440 445

Asn Asn Tyr Glu Thr Phe Lys Asn His Thr Ser Ile Leu Phe Trp Ser

450 455 460

Leu Gly Asn Glu Ser Tyr Ala Gly Asp Asn Ile Ile Ala Met Asn Glu

465 470 475 480

Phe Tyr Lys Ser His Asp Asp Thr Arg Leu Val His Tyr Glu Gly Val

485 490 495

Val His Arg Pro Glu Leu Lys Asp Lys Ile Ser Asp Val Glu Ser Cys

500 505 510

Met Tyr Leu Pro Pro Lys Lys Val Glu Glu Tyr Leu Gln Asn Asp Pro

515 520 525

Pro Lys Pro Phe Met Glu Cys Glu Tyr Met His Asp Met Gly Asn Ser

530 535 540

Asn Gly Gly Met Asp Ser Tyr Ile Lys Leu Leu Asp Lys Tyr Pro Gln

545 550 555 560

Tyr Phe Gly Gly Phe Ile Trp Asp Phe Ile Asp Gln Ala Leu Leu Val

565 570 575

His Asp Glu Ile Ser Gly His Asp Val Leu Arg Tyr Gly Gly Asp Phe

580 585 590

Asp Asp Arg His Ser Asp Tyr Glu Phe Ser Gly Asp Gly Leu Met Phe

595 600 605

Ala Asp Arg Lys Pro Lys Pro Ala Met Gln Glu Val Arg Tyr Tyr Tyr

610 615 620

Gly Leu His Lys

625

<210> 27

<211> 318

<212>PRT

<213> Lactobacillus helvaticus

<400> 27

Met Asp Tyr Thr Asn Asn Gln Leu His Ile Ile Tyr Gly Asp Ala Thr

1 5 10 15

Phe Gly Val Asn Gly Lys Asp Phe Gln Tyr Ile Phe Ser Tyr Glu Arg

20 25 30

Gly Gly Leu Glu Ser Leu Lys Val His Gly Lys Glu Trp Leu Tyr Arg

35 40 45

Val Pro Thr Pro Thr Phe Trp Arg Ala Thr Thr Asp Asn Asp Arg Gly

50 55 60

Ser Gly Phe Asn Leu Lys Ala Ala Gln Trp Leu Gly Ala Asp Met Phe

65 70 75 80

Thr Lys Cys Thr Asp Ile His Leu Lys Val Asp Arg His Asp Phe Ala

85 90 95

Glu Leu Pro Ile Ala Pro Phe Asn Asn Lys Phe Ser Asn His Glu Tyr

100 105 110

Ala Lys Ser Ala Glu Ile Ser Phe Thr Tyr Gln Thr Leu Thr Thr Pro

115 120 125

Ala Thr Asn Ala Lys Ile Ile Tyr Asn Ile Asp Asp Gly Gly His Ile

130 135 140

Lys Val Thr Met Arg Tyr Tyr Gly Lys Lys Gly Leu Pro Pro Leu Pro

145 150 155 160

Val Ile Gly Ile Arg Leu Ile Met Pro Thr Ala Ala Thr Gly Phe Asp

165 170 175

Tyr Glu Gly Leu Ser Gly Glu Thr Tyr Pro Asp Arg Met Ala Gly Ala

180 185 190

Lys Glu Gly Lys Phe His Ile Asp Gly Leu Pro Val Thr Glu Tyr Leu

195 200 205

Val Pro Gln Glu Asn Gly Met His Met Gln Thr Lys Lys Leu Thr Ile

210 215 220

Asn Arg Glu Thr Thr Gln Asn Asn Val Asp Arg Thr Asn Glu Lys Phe

225 230 235 240

Ser Leu Ser Ile Gln Gln Ala Glu Lys Pro Phe Asn Phe Ser Cys Leu

245 250 255

Pro Tyr Thr Ala Glu Glu Leu Glu Asn Ala Thr His Ile Glu Glu Leu

260 265 270

Pro Leu Val Arg Arg Thr Val Leu Val Ile Ala Gly Ala Val Arg Gly

275 280 285

Val Gly Gly Ile Asp Ser Trp Gly Thr Asp Val Glu Ser Ala Tyr His

290 295 300

Ile Asn Pro Asp Leu Asp His Glu Phe Ser Phe Ile Leu Asn

305 310 315

<210> 28

<211> 626

<212>PRT

<213> Lactobacillus crispatus

<400> 28

Met Lys Ala Asn Ile Lys Trp Leu Asp Asp Pro Glu Val Phe Arg Ile

1 5 10 15

Asn Gln Leu Pro Ala His Ser Asp His Pro Phe Tyr Lys Asp Tyr Arg

20 25 30

Glu Trp Gln Lys His Ser Ser Ser Phe Lys Gln Ser Leu Asn Gly Ala

35 40 45

Trp Gln Phe His Phe Ser Lys Asp Pro Gln Ser Arg Pro Ile Asp Phe

50 55 60

Tyr Lys Leu Ser Phe Asp Ser Ser Ser Phe Asp Thr Ile Pro Val Pro

65 70 75 80

Ser Glu Ile Glu Leu Asn Gly Tyr Ala Gln Asn Gln Tyr Thr Asn Ile

85 90 95

Leu Tyr Pro Trp Glu Ser Lys Ile Tyr Arg Lys Pro Ala Tyr Thr Leu

100 105 110

Gly Arg Gly Ile Lys Asp Gly Asp Phe Ser Gln Gly Lys Asp Asn Thr

115 120 125

Val Gly Ser Tyr Leu Lys His Phe Asp Leu Asn Pro Ala Leu Ala Gly

130 135 140

His Asp Ile His Ile Gln Phe Glu Gly Val Glu Arg Ala Met Tyr Val

145 150 155 160

Tyr Leu Asn Gly His Phe Ile Gly Tyr Ala Glu Asp Ser Phe Thr Pro

165 170 175

Ser Glu Phe Asp Leu Thr Pro Tyr Ile Gln Ala Lys Asp Asn Ile Leu

180 185 190

Ala Val Glu Val Phe Lys His Ser Thr Ala Ser Trp Leu Glu Asp Gln

195 200 205

Asp Met Phe Arg Phe Ser Gly Ile Phe Arg Ser Val Glu Leu Leu Ala

210 215 220

Leu Pro Arg Thr His Leu Met Asp Leu Asp Ile Lys Pro Thr Val Val

225 230 235 240

Asn Asp Tyr His Asp Gly Val Phe Asn Ala Lys Leu His Phe Met Gly

245 250 255

Lys Thr Ser Gly Asn Val His Val Leu Ile Glu Asp Ile Asp Gly Lys

260 265 270

Thr Leu Leu Asn Lys Lys Leu Pro Leu Lys Ser Thr Val Glu Ile Glu

275 280 285

Asn Glu Thr Phe Ala Asn Val His Leu Trp Asp Asn His Asp Pro Tyr

290 295 300

Leu Tyr Gln Leu Ile Ile Glu Val His Asp Gln Asp Gly Lys Leu Val

305 310 315 320

Glu Leu Ile Pro Tyr Gln Phe Gly Phe Arg Lys Ile Glu Ile Thr Lys

325 330 335

Asp His Val Val Leu Leu Asn Gly Lys Arg Leu Ile Ile Asn Gly Val

340 345 350

Asn Arg His Glu Trp Asp Ala Lys Arg Gly Arg Ser Ile Thr Leu Ala

355 360 365

Asp Met Lys Gln Asp Ile Ala Thr Phe Lys His Asn Asn Ile Asn Ala

370 375 380

Val Arg Thr Cys His Tyr Pro Asn Gln Ile Pro Trp Tyr Tyr Leu Cys

385 390 395 400

Asp Gln Asn Gly Ile Tyr Met Met Ala Glu Asn Asn Leu Glu Ser His

405 410 415

Gly Thr Trp Gln Lys Leu Gly Gln Val Glu Ala Thr Ser Asn Val Pro

420 425 430

Gly Ser Ile Pro Glu Trp Arg Glu Val Val Val Asp Arg Ala Arg Ser

435 440 445

Asn Tyr Glu Thr Phe Lys Asn His Thr Ala Ile Leu Phe Trp Ser Leu

450 455 460

Gly Asn Glu Ser Tyr Ala Gly Ser Asn Ile Ala Ala Met Asn Lys Leu

465 470 475 480

Tyr Lys Asp His Asp Ser Ser Arg Leu Thr His Tyr Glu Gly Val Phe

485 490 495

His Ala Pro Glu Phe Lys Lys Glu Ile Ser Asp Leu Glu Ser Cys Met

500 505 510

Tyr Leu Pro Pro Lys Glu Ala Glu Glu Tyr Leu Gln Asn Pro Lys Lys

515 520 525

Pro Leu Val Glu Cys Glu Tyr Met His Asp Met Gly Asn Ser Asp Gly

530 535 540

Gly Ile Gly Ser Tyr Ile Lys Leu Ile Asp Lys Tyr Pro Gln Tyr Met

545 550 555 560

Gly Gly Phe Ile Trp Asp Phe Ile Asp Gln Ala Leu Leu Val His Asp

565 570 575

Pro Val Ser Gly Gln Asp Val Leu Arg Tyr Gly Gly Asp Phe Asp Asp

580 585 590

Arg His Ser Asp Tyr Glu Phe Ser Gly Asp Gly Leu Met Phe Ala Asp

595 600 605

Arg Thr Pro Lys Pro Ala Met Gln Glu Val Arg Tyr Tyr Tyr Gly Leu

610 615 620

His Lys

625

<210> 29

<211> 316

<212>PRT

<213> Lactobacillus crispatus

<400> 29

Met Ala Tyr Thr Asn Asn Leu His Val Val Tyr Gly Glu Ala Ser Leu

1 5 10 15

Gly Val Asn Gly Gln Asp Phe Ala Tyr Leu Phe Ser Tyr Glu Arg Gly

20 25 30

Val Leu Glu Ser Leu Lys Ile Lys Asp Lys Glu Trp Leu Tyr Arg Thr

35 40 45

Pro Thr Pro Thr Phe Trp Arg Ala Thr Thr Asp Asn Asp Arg Gly Ser

50 55 60

Gly Phe Asn Gln Lys Ala Ala Gln Trp Leu Gly Ala Asp Met Phe Thr

65 70 75 80

Lys Cys Val Gly Ile His Val Gln Val Asp Asp His Gln Phe Asp Glu

85 90 95

Leu Pro Ile Ala Pro Ile Asn Asn Gln Phe Ser Asn Gln Glu Phe Ala

100 105 110

His Glu Val Lys Val Ala Phe Asp Tyr Glu Thr Leu Thr Thr Pro Ala

115 120 125

Thr Lys Val Lys Ile Ile Tyr Asn Ile Asn Asp Ala Gly His Met Thr

130 135 140

Ile Thr Met His Tyr Phe Gly Lys Lys Gly Leu Pro Pro Leu Pro Val

145 150 155 160

Ile Gly Met Arg Phe Ile Met Pro Thr Lys Ala Lys Ser Phe Asp Tyr

165 170 175

Thr Gly Leu Ser Gly Glu Thr Tyr Pro Asp Arg Met Ala Gly Ala Glu

180 185 190

Arg Gly Thr Phe His Ile Asp Gly Leu Pro Val Thr Lys Tyr Leu Val

195 200 205

Pro Gln Glu Asn Gly Met His Met Gln Thr Asn Glu Leu Val Ile Thr

210 215 220

Arg Asn Ser Thr Gln Asn Asn Ala Asp Lys Asp Gly Asp Phe Ser Leu

225 230 235 240

Lys Ile Thr Gln Thr Lys Gln Pro Phe Asn Phe Ser Leu Leu Pro Tyr

245 250 255

Thr Ala Glu Glu Leu Glu Asn Ala Thr His Ile Glu Glu Leu Pro Leu

260 265 270

Ala Arg Arg Ser Val Leu Val Ile Ala Gly Ala Val Arg Gly Val Gly

275 280 285

Gly Ile Asp Ser Trp Gly Ser Asp Val Glu Glu Gln Tyr His Ile Asp

290 295 300

Pro Glu Gln Asp His Glu Phe Ser Phe Thr Leu Asn

305 310 315

<210> 30

<211> 1025

<212>PRT

<213> Streptococcus thermophilus

<400> 30

Met Asn Met Thr Lys Ile Gln Thr Tyr Leu Asn Asp Pro Lys Ile Val

1 5 10 15

Ser Val Asn Thr Val Asp Ala His Ser Asp His Lys Tyr Phe Glu Ser

20 25 30

Leu Glu Glu Phe Ser Glu Gly Glu Met Lys Leu Arg Gln Ser Leu Asn

35 40 45

Gly Lys Trp Lys Ile His Tyr Ala Gln Asn Thr Asn Gln Val Leu Lys

50 55 60

Asp Phe Tyr Lys Thr Glu Phe Asp Glu Thr Asp Leu Asn Phe Ile Asn

65 70 75 80

Val Pro Gly His Leu Glu Leu Gln Gly Phe Gly Ser Pro Gln Tyr Val

85 90 95

Asn Thr Gln Tyr Pro Trp Asp Gly Lys Glu Phe Leu Arg Pro Pro Gln

100 105 110

Val Pro Gln Glu Ser Asn Ala Val Ala Ser Tyr Val Lys His Phe Thr

115 120 125

Leu Asn Asp Ala Leu Lys Asp Lys Lys Val Phe Ile Ser Phe Gln Gly

130 135 140

Val Ala Thr Ser Ile Phe Val Trp Val Asn Gly Asn Phe Val Gly Tyr

145 150 155 160

Ser Glu Asp Ser Phe Thr Pro Ser Glu Phe Glu Ile Ser Asp Tyr Leu

165 170 175

Val Glu Gly Asp Asn Lys Leu Ala Val Ala Val Tyr Arg Tyr Ser Thr

180 185 190

Ala Ser Trp Leu Glu Asp Gln Asp Phe Trp Arg Leu Tyr Gly Ile Phe

195 200 205

Arg Asp Val Tyr Leu Tyr Ala Ile Pro Lys Val His Val Gln Asp Leu

210 215 220

Phe Val Lys Gly Asp Tyr Asp Tyr Gln Thr Lys Ala Gly Gln Leu Asp

225 230 235 240

Ile Asp Leu Lys Thr Val Gly Asp Tyr Glu Asp Lys Lys Ile Lys Tyr

245 250 255

Val Leu Ser Asp Tyr Glu Gly Ile Val Thr Glu Gly Asp Ala Ser Val

260 265 270

Asn Gly Asp Gly Glu Leu Ser Val Ser Leu Glu Asn Leu Lys Ile Lys

275 280 285

Pro Trp Ser Ala Glu Ser Pro Lys Leu Tyr Asp Leu Ile Leu His Val

290 295 300

Leu Asp Asp Asp Gln Val Val Glu Val Val Pro Val Lys Val Gly Phe

305 310 315 320

Arg Arg Phe Glu Ile Lys Asp Lys Leu Met Leu Leu Asn Gly Lys Arg

325 330 335

Ile Val Phe Lys Gly Val Asn Arg His Glu Phe Asn Ala Arg Thr Gly

340 345 350

Arg Cys Ile Thr Glu Glu Asp Met Leu Trp Asp Ile Lys Val Met Lys

355 360 365

Gln His Asn Ile Asn Ala Val Arg Thr Ser His Tyr Pro Asn Gln Thr

370 375 380

Arg Trp Tyr Glu Leu Cys Asp Glu Tyr Gly Leu Tyr Val Ile Asp Glu

385 390 395 400

Ala Asn Leu Glu Thr His Gly Thr Trp Gln Lys Leu Gly Leu Cys Glu

405 410 415

Pro Ser Trp Asn Ile Pro Ala Ser Glu Pro Glu Trp Leu Pro Ala Cys

420 425 430

Leu Asp Arg Ala Asn Asn Met Phe Gln Arg Asp Lys Asn His Ala Ser

435 440 445

Val Ile Ile Trp Ser Cys Gly Asn Glu Ser Tyr Ala Gly Lys Asp Ile

450 455 460

Ala Asp Met Ala Asp Tyr Phe Arg Ser Val Asp Asn Thr Arg Pro Val

465 470 475 480

His Tyr Glu Gly Val Ala Trp Cys Arg Glu Phe Asp Tyr Ile Thr Asp

485 490 495

Ile Glu Ser Arg Met Tyr Ala Lys Pro Ala Asp Ile Glu Glu Tyr Leu

500 505 510

Thr Thr Gly Lys Leu Val Asp Leu Ser Ser Val Ser Asp Lys His Phe

515 520 525

Ala Ser Gly Asn Leu Thr Asn Lys Pro Gln Lys Pro Tyr Ile Ser Cys

530 535 540

Glu Tyr Met His Thr Met Gly Asn Ser Gly Gly Gly Leu Gln Leu Tyr

545 550 555 560

Thr Asp Leu Glu Lys Tyr Pro Glu Tyr Gln Gly Gly Phe Ile Trp Asp

565 570 575

Phe Ile Asp Gln Ala Ile Tyr Lys Thr Leu Pro Asn Gly Ser Glu Phe

580 585 590

Leu Ser Tyr Gly Gly Asp Trp His Asp Arg Pro Ser Asp Tyr Glu Phe

595 600 605

Cys Gly Asn Gly Ile Val Phe Ala Asp Arg Thr Leu Thr Pro Lys Leu

610 615 620

Gln Thr Val Lys His Leu Tyr Ser Asn Ile Lys Ile Ala Val Asp Glu

625 630 635 640

Lys Ser Val Thr Ile Lys Asn Asp Asn Leu Phe Glu Asp Leu Ser Ala

645 650 655

Tyr Thr Phe Leu Ala Arg Val Tyr Glu Asp Gly Arg Lys Val Ser Glu

660 665 670

Ser Glu Tyr His Phe Asp Val Lys Pro Gly Glu Glu Ala Thr Phe Pro

675 680 685

Val Asn Phe Val Val Glu Ala Ser Asn Ser Glu Gln Ile Tyr Glu Val

690 695 700

Ala Cys Val Leu Arg Glu Ala Thr Lys Trp Ala Pro Lys Gly His Glu

705 710 715 720

Ile Val Arg Gly Gln Tyr Val Val Glu Lys Ile Ser Thr Glu Thr Pro

725 730 735

Val Lys Ala Pro Leu Asn Val Val Glu Gly Asp Phe Asn Ile Gly Ile

740 745 750

Gln Gly Gln Asn Phe Ser Ile Leu Leu Ser Arg Ala Gln Asn Thr Leu

755 760 765

Val Ser Ala Lys Tyr Asn Gly Val Glu Phe Ile Glu Lys Gly Pro Lys

770 775 780

Leu Ser Phe Thr Arg Ala Tyr Thr Asp Asn Asp Arg Gly Ala Gly Tyr

785 790 795 800

Pro Phe Glu Met Ala Gly Trp Lys Val Ala Gly Asn Tyr Ser Lys Val

805 810 815

Thr Asp Thr Gln Ile Gln Ile Glu Asp Asp Ser Val Lys Val Thr Tyr

820 825 830

Val His Glu Leu Pro Gly Leu Ser Asp Val Glu Val Lys Val Thr Tyr

835 840 845

Gln Val Asp Tyr Lys Gly Arg Ile Phe Val Thr Ala Asn Tyr Asp Gly

850 855 860

Lys Ala Gly Leu Pro Asn Phe Pro Glu Phe Gly Leu Glu Phe Ala Ile

865 870 875 880

Gly Ser Gln Phe Thr Asn Leu Ser Tyr Tyr Gly Tyr Gly Ala Glu Glu

885 890 895

Ser Tyr Arg Asp Lys Leu Pro Gly Ala Tyr Leu Gly Arg Tyr Glu Thr

900 905 910

Ser Val Glu Lys Thr Phe Ala Pro Tyr Leu Met Pro Gln Glu Ser Gly

915 920 925

Asn His Tyr Gly Thr Arg Glu Phe Thr Val Ser Asp Asp Asn His Asn

930 935 940

Gly Leu Lys Phe Thr Ala Leu Asn Lys Ala Phe Glu Phe Ser Ala Leu

945 950 955 960

Arg Asn Ser Thr Glu Gln Ile Glu Asn Ala Arg His Gln Tyr Glu Leu

965 970 975

Gln Glu Ser Asp Ala Thr Trp Ile Lys Val Leu Ala Ala Gln Met Gly

980 985 990

Val Gly Gly Asp Asp Ser Trp Gly Ala Pro Val His Asp Glu Phe Leu

995 1000 1005

Leu Ser Ser Ala Asp Ser Tyr Gln Leu Ser Phe Met Ile Glu Pro

1010 1015 1020

Leu Asn

1025

<210> 31

<211> 1008

<212>PRT

<213> Lactobacillus delbrueckii subsp. indicus

<400> 31

Met Asn Asn Lys Leu Ala Gln Val Lys Arg Val Asp Gln Ala Asp Leu

1 5 10 15

Ala Trp Leu Thr Asp Pro Glu Ile Tyr Glu Val Asn Thr Ile Pro Pro

20 25 30

His Ser Asp His Glu Ser Phe Gln Ser Leu Glu Glu Leu Glu Glu Gly

35 40 45

Lys Ser Ser Leu Val Gln Ser Leu Asp Gly Asp Trp Leu Ile Asp Tyr

50 55 60

Ala Glu Asn Gly Glu Gly Pro Ala Asn Phe Tyr Glu Glu Asn Phe Asp

65 70 75 80

Asp Ser Ser Phe Lys Ser Val Lys Val Pro Gly Asn Leu Glu Leu Gln

85 90 95

Gly Phe Gly Gln Pro Gln Tyr Val Asn Val Gln Tyr Pro Trp Asp Gly

100 105 110

Ser Asp Glu Ile Phe Pro Pro Met Ile Pro Ser Lys Asn Pro Val Ala

115 120 125

Ser Tyr Val Arg Tyr Phe Asp Leu Glu Glu Ala Phe Trp Asp Lys Glu

130 135 140

Val Ser Leu Lys Phe Ala Gly Ala Ala Thr Ala Ile Tyr Val Trp Leu

145 150 155 160

Asn Gly His Phe Val Gly Tyr Gly Glu Asp Ser Phe Thr Pro Ser Glu

165 170 175

Phe Met Val Thr Lys Phe Leu Lys Lys Glu Gly Asn Arg Leu Ala Val

180 185 190

Ala Leu Tyr Lys Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe

195 200 205

Trp Arg Met Ser Gly Leu Phe Arg Ser Val Thr Leu Glu Ala Lys Pro

210 215 220

Leu Leu His Leu Gln Asp Leu Lys Leu Thr Ala Ser Leu Thr Asn Asp

225 230 235 240

Tyr Gln Lys Gly Ser Leu Gln Val Glu Ala Asp Ile Asp Tyr Arg Leu

245 250 255

Pro Asn Ser Ser Phe Lys Leu Glu Leu Arg Asp Ser Ala Gly Glu Leu

260 265 270

Val Ala Glu Lys Val Gly Pro Ile Arg Ser Glu Lys Leu Asp Phe Ser

275 280 285

Leu Ala Asp Leu Pro Val Ala Ala Trp Ser Ala Glu Glu Pro Asn Leu

290 295 300

Tyr Gln Val Arg Leu Ser Leu Tyr Gln Gln Gly Ser Leu Leu Glu Val

305 310 315 320

Ser Arg Gln Glu Val Gly Phe Arg Asn Phe Glu Leu Lys Asp Gly Ile

325 330 335

Met Tyr Leu Asn Gly Lys Arg Ile Val Phe Lys Gly Val Asn Arg His

340 345 350

Glu Phe Asp Ser Lys Leu Gly Arg Ala Ile Thr Glu Ala Asp Met Ile

355 360 365

Trp Asp Ile Lys Thr Met Lys Gln Ser Asn Ile Asn Ala Val Arg Cys

370 375 380

Ser His Tyr Pro Asn Gln Ser Ile Phe Tyr His Leu Cys Asp Lys Tyr

385 390 395 400

Gly Leu Tyr Val Ile Asp Glu Ala Asn Leu Glu Ser His Gly Thr Trp

405 410 415

Glu Lys Val Gly Gly His Glu Asp Pro Ser Phe Asn Val Pro Gly Asp

420 425 430

Asp Gln Arg Trp Leu Gly Ala Ser Leu Ser Arg Val Lys Asn Met Met

435 440 445

Ala Arg Asp Lys Asn His Ala Ser Ile Leu Ile Trp Ser Leu Gly Asn

450 455 460

Glu Ser Tyr Ala Gly Lys Val Phe Ala Gln Met Ala Asp Tyr Val Arg

465 470 475 480

Gln Ala Asp Pro Thr Arg Val Gln His Tyr Glu Gly Val Thr His Asn

485 490 495

Arg Lys Phe Asp Asp Ala Thr Gln Ile Glu Ser Arg Met Tyr Ala Pro

500 505 510

Ala Lys Glu Ile Glu Glu Tyr Leu Thr Lys Lys Pro Ala Lys Pro Phe

515 520 525

Val Ser Cys Glu Tyr Ala His Ala Met Gly Asn Ser Val Gly Asp Leu

530 535 540

Ala Ala Tyr Thr Ala Leu Glu Lys Tyr Pro His Tyr Gln Gly Gly Phe

545 550 555 560

Ile Trp Asp Trp Ile Asp Gln Gly Leu Glu Lys Glu Gly His Leu Leu

565 570 575

Tyr Gly Gly Asp Phe Asp Asp Arg Pro Ser Asp Tyr Glu Phe Cys Gly

580 585 590

Asp Gly Leu Val Phe Ala Asp Arg Thr Thr Ser Pro Lys Leu Ala Asn

595 600 605

Val Lys Ala Leu Tyr Ser Asn Leu Lys Leu Glu Leu Lys Asp Gly Gln

610 615 620

Leu Phe Leu Lys Asn Asp Asn Leu Phe Thr Asn Ser Ser Ala Tyr Tyr

625 630 635 640

Phe Leu Thr Ser Leu Leu Val Asp Gly Lys Leu Thr Tyr Gln Ser Gln

645 650 655

Pro Leu Thr Phe Ala Leu Glu Pro Gly Glu Ser Gly Thr Phe Ala Leu

660 665 670

Pro Trp Pro Glu Val Glu Asp Glu Lys Gly Glu Ile Val Tyr Gln Val

675 680 685

Thr Ala His Leu Lys Glu Asp Leu Pro Trp Ala Asp Glu Gly Phe Thr

690 695 700

Val Ala Glu Ala Glu Glu Ala Val Thr Lys Leu Pro Glu Phe Tyr Pro

705 710 715 720

Ala Gly Arg Pro Glu Leu Val Asp Ser Asp Tyr Asn Leu Gly Ile Lys

725 730 735

Gly Asn Gly Phe Arg Ile Leu Phe Ser Lys Ala Lys Gly Trp Pro Val

740 745 750

Ser Ile Lys Tyr Ala Gly Arg Glu Tyr Leu Lys Arg Leu Pro Glu Phe

755 760 765

Thr Phe Trp Arg Ala Leu Thr Asp Asn Asp Arg Gly Ala Gly Tyr Gly

770 775 780

Tyr Asp Leu Ala Lys Trp Glu Asn Ala Gly Lys Tyr Ala Arg Leu Gln

785 790 795 800

Asp Ile Ser Tyr Glu Ile Lys Glu Asn Ser Val Leu Val Lys Thr Ala

805 810 815

Phe Thr Leu Pro Val Ala Leu Lys Gly Asp Leu Thr Ile Thr Tyr Glu

820 825 830

Val Asp Ser Leu Gly Lys Ile Ala Val Thr Ala Asn Phe Pro Gly Ala

835 840 845

Val Glu Asn Gly Leu Leu Pro Ala Phe Gly Leu Asn Phe Ala Leu Pro

850 855 860

Lys Glu Leu Ser Asp Tyr Arg Tyr Tyr Gly Leu Gly Pro Asn Glu Ser

865 870 875 880

Tyr Ala Asp Arg Leu Glu Gly Ser Tyr Leu Gly Ile Tyr Gln Gly Ala

885 890 895

Val Glu Lys Asn Phe Thr Pro Tyr Leu Arg Pro Gln Glu Val Gly Asn

900 905 910

Arg Ser Lys Val Arg Tyr Tyr Gln Leu Phe Asp Glu Glu Gly Gly Leu

915 920 925

Glu Phe Thr Ala Asn Gly Ala Asn Leu Asn Leu Ser Ala Leu Pro Tyr

930 935 940

Ser Ala Ala Gln Ile Glu Ala Ala Asp His Ala Phe Glu Leu Thr Asn

945 950 955 960

Asn Tyr Thr Trp Val Arg Ala Leu Ala Ala Gln Met Gly Val Gly Gly

965 970 975

Asp Asp Ser Trp Gly Gln Lys Val His Pro Glu Phe Cys Leu Asp Ala

980 985 990

Gln Glu Ala Arg Gln Leu Lys Leu Val Ile Gln Pro Leu Phe Thr Glu

995 1000 1005

<210> 32

<211> 1049

<212>PRT

<213> Bifidobacterium adolescentis

<400> 32

Met Ala Asp Thr Ala Glu Leu Ala Ile Val His Ala Thr Thr Ala Ser

1 5 10 15

Ala Ser Trp Leu Thr Asp Pro Thr Val Phe Ala Ala Asn Arg Lys Pro

20 25 30

Ala His Ser Ser His Arg Tyr Val Ile Gly Glu Thr Ser Glu Pro Lys

35 40 45

Gln Ser Leu Asp Gly Glu Trp Lys Val Arg Ile Glu Gln Ala Arg Asn

50 55 60

Val Asp Val Glu Ser Ala Pro Phe Ala Ala Val Asp Phe Glu Asp Gly

65 70 75 80

Asp Phe Gly Ala Ile Glu Val Pro Gly His Leu Gln Met Ala Gly Tyr

85 90 95

Leu Lys Asn Lys Tyr Val Asn Ile Gln Tyr Pro Trp Asp Gly His Glu

100 105 110

Asp Pro Gln Ala Pro Asn Ile Pro Glu Asn Asn His Val Ala Ile Tyr

115 120 125

Arg Arg Arg Phe Ala Leu Asp Ala Gln Leu Ala Arg Thr Leu Glu Asn

130 135 140

Asp Gly Thr Val Ser Leu Thr Phe His Gly Ala Ala Thr Ala Ile Tyr

145 150 155 160

Val Trp Leu Asp Gly Thr Phe Val Gly Tyr Gly Glu Asp Gly Phe Thr

165 170 175

Pro Ser Glu Phe Asp Val Thr Glu Ala Leu Arg Asn Gly Asn Gly Asn

180 185 190

Ala Ala Asp Ser Pro Glu Ala Glu His Thr Leu Thr Val Ala Cys Tyr

195 200 205

Glu Tyr Ser Ser Ala Ser Trp Leu Glu Asp Gln Asp Phe Trp Arg Leu

210 215 220

His Gly Leu Phe Arg Thr Val Glu Leu Ala Ala Gln Pro His Thr His

225 230 235 240

Val Glu Thr Val Gln Leu Glu Ala Asp Tyr Thr Ala Ala Asp Thr Ala

245 250 255

Gly Thr Ala Asp Thr Ala Glu Leu Asn Ala Ala Leu Thr Leu Arg Asn

260 265 270

Pro Ala Asp Ala Met Thr Ile Glu Ser Thr Leu Arg Asp Gly Asp Gly

275 280 285

Asn Val Val Trp Glu Ser Thr Gln Ala Cys Asn Gly Glu Ile Ala Leu

290 295 300

Asn Ser Gly Lys Met Thr Asn Ile Ala Pro Trp Ser Ala Glu Ser Pro

305 310 315 320

Thr Leu Tyr Thr Leu Thr Val Arg Val Val Gly His Asp Gly Ala Ile

325 330 335

Ile Glu Thr Val Thr Gln Lys Ile Gly Phe Arg Thr Phe Arg Ile Glu

340 345 350

Asn Gly Ile Met Thr Leu Asn Gly Lys Arg Ile Val Phe Lys Gly Ala

355 360 365

Asp Arg His Glu Phe Asp Ala Lys Arg Gly Arg Ala Ile Thr Arg Glu

370 375 380

Asp Met Leu Ser Asp Val Val Phe Cys Lys Arg His Asn Ile Asn Ala

385 390 395 400

Ile Arg Thr Ser His Tyr Pro Asn Gln Glu Tyr Trp Tyr Asp Leu Cys

405 410 415

Asp Glu Tyr Gly Leu Tyr Leu Ile Asp Glu Thr Asn Met Glu Thr His

420 425 430

Gly Thr Trp Val Ala Asn Asn Val Glu Arg Pro Glu Asp Gly Ile Pro

435 440 445

Gly Ser Arg Pro Glu Trp Glu Asp Ala Cys Val Asp Arg Ile Asn Ser

450 455 460

Met Met Arg Arg Asp Tyr Asn His Pro Ser Val Leu Ile Trp Ser Leu

465 470 475 480

Gly Asn Glu Ser Ser Ala Gly Glu Val Phe Arg Ala Met Tyr Arg His

485 490 495

Ala His Thr Ile Asp Pro Asn Arg Pro Val His Tyr Glu Gly Ser Val

500 505 510

His Met Arg Glu Phe Glu Asp Val Thr Asp Ile Glu Ser Arg Met Tyr

515 520 525

Ala His Ala Asp Glu Ile Glu Arg Tyr Leu Asn Asp Gly Ser Pro Ala

530 535 540

His Thr Asp Gly Pro Lys Lys Pro Tyr Ile Ser Cys Glu Tyr Met His

545 550 555 560

Ala Met Gly Asn Ser Cys Gly Asn Met Asp Glu Tyr Thr Ala Leu Glu

565 570 575

Arg Tyr Pro Met Tyr Gln Gly Gly Phe Ile Trp Asp Phe Ile Asp Gln

580 585 590

Ala Ile Glu Thr Lys Leu Pro Asp Gly Thr Thr Arg Met Cys Tyr Gly

595 600 605

Gly Asp Phe Gly Asp Arg Pro Ser Asp Tyr Glu Phe Ser Gly Asp Gly

610 615 620

Leu Leu Phe Ala Asp Arg Thr Pro Ser Pro Lys Ala Gln Glu Val Lys

625 630 635 640

Gln Leu Tyr Ala Asn Val Lys Ile Ala Val Ser Val Asp Glu Ala Arg

645 650 655

Ile Thr Asn Asp Asn Leu Phe Val Ser Thr Gly Asp Tyr Arg Phe Val

660 665 670

Leu Arg Ile Leu Ala Asp Gly Lys Pro Val Trp Ser Thr Thr Arg Arg

675 680 685

Phe Asp Val Ala Ala Gly Glu Ser Ala Ser Phe Glu Val Asp Trp Pro

690 695 700

Val Asp Asp Tyr Arg Ser Asn Ala Glu Glu Leu Val Leu Glu Val Ser

705 710 715 720

Gln Gln Leu Gly Asn Ala Cys Asp Trp Ala Pro Ala Gly Tyr Glu Leu

725 730 735

Ala Phe Gly Gln Cys Val Val Ala Gly Ala Lys Thr Thr Ala Asp Ala

740 745 750

Val Asp Ala Ala Gly Ala Pro Ala Asp Gly Thr Val Thr Leu Gly Arg

755 760 765

Trp Asn Ala Gly Val Arg Gly Gln Gly Arg Glu Ala Leu Phe Ser Arg

770 775 780

Thr Gln Gly Gly Met Val Ser Tyr Thr Phe Gly Glu Arg Glu Phe Val

785 790 795 800

Leu Arg Arg Pro Ser Ile Thr Thr Phe Arg Pro Leu Thr Asp Asn Asp

805 810 815

Arg Gly Ala Gly His Ala Phe Glu Arg Ala Ala Trp Ala Val Ala Gly

820 825 830

Lys Tyr Ala Arg Cys Val Asp Cys Ala Ile Ala Asn Arg Gly Glu Asn

835 840 845

Ala Val Glu Ala Thr Tyr Thr Tyr Glu Leu Ala Ile Pro Gln Arg Thr

850 855 860

Lys Val Thr Val Arg Tyr Val Ala Asp Thr Ala Gly Leu Val Ser Leu

865 870 875 880

Asp Val Glu Tyr Pro Gly Glu Lys Asn Gly Asp Leu Pro Thr Ile Pro

885 890 895

Ala Phe Gly Ile Glu Trp Ala Leu Pro Val Glu Tyr Ala Asn Leu Arg

900 905 910

Phe Tyr Gly Ala Gly Pro Glu Glu Thr Tyr Ala Asp Arg Arg His Ala

915 920 925

Lys Leu Gly Val Trp Ser Thr Thr Ala Gly Asp Asp Cys Ala Pro Tyr

930 935 940

Leu Leu Pro Gln Glu Thr Gly Asn His Glu Asp Val Arg Trp Ala Glu

945 950 955 960

Ile Thr Asp Asp Ser Gly His Gly Val Arg Val Lys Arg Gly Ala Gly

965 970 975

Ala Lys Pro Phe Ala Met Ser Leu Leu Pro Tyr Ser Ser Thr Met Leu

980 985 990

Glu Glu Ala Leu His Gln Asp Glu Leu Pro Lys Pro Arg His Met Phe

995 1000 1005

Leu Arg Leu Leu Ala Ala Gln Met Gly Val Gly Gly Asp Asp Ser

1010 1015 1020

Trp Met Ser Pro Val His Glu Gln Tyr Gln Leu Pro Ala Asp Gln

1025 1030 1035

Pro Leu Ser Leu Asn Val Gln Leu Lys Leu Phe

1040 1045

<210> 33

<211> 1023

<212>PRT

<213> Bifidobacterium adolescentis

<400> 33

Met Ala Asn Glu Thr Arg Ile Glu His Ala Ser Glu Thr Trp Leu Ala

1 5 10 15

Asp Ser Thr Val Phe Glu Val Asn Arg Val Pro Ala His Ser Asp His

20 25 30

Lys Cys Tyr Ala His Asp Ser Gln Thr Asn Glu Trp Ser Asp Leu Arg

35 40 45

Gln Ser Leu Asp Gly Glu Trp Arg Val Glu Val Val Gln Ala Ser Asp

50 55 60

Ile Glu Phe Asn Glu Glu Pro Phe Val Arg Glu Asn Phe Asp Asp Ser

65 70 75 80

Ala Phe Glu Arg Ile Gln Val Pro Gly His Leu Gln Met Ala Gly Leu

85 90 95

Met Asn Asn Lys Tyr Val Asn Ile Gln Tyr Pro Trp Asp Gly His Glu

100 105 110

Asn Pro Ala Glu Pro Asn Ile Pro Glu Asn Asn His Val Ala Leu Tyr

115 120 125

Arg Lys Thr Phe Thr Met Ala Asn Arg Leu Ala Asp Thr Lys Asn Ala

130 135 140

Gly Gly Thr Val Ser Ile Val Phe His Gly Met Ala Thr Ala Ile Tyr

145 150 155 160

Val Trp Val Asn Gly Met Phe Val Gly Tyr Gly Glu Asp Gly Phe Thr

165 170 175

Pro Asn Glu Phe Asp Ile Thr Glu Met Leu His Asp Gly Glu Asn Val

180 185 190

Val Ala Val Ala Cys Tyr Glu Tyr Ser Ser Ala Ser Trp Leu Glu Asp

195 200 205

Gln Asp Phe Trp Arg Leu His Gly Leu Phe Arg Ser Val Glu Leu Ala

210 215 220

Ala Gln Pro His Val His Ile Glu Asn Met Gln Ile Glu Ser Asp Trp

225 230 235 240

Asp Pro Glu Ser Gly Ser Ala Ser Leu Asp Ala Ala Leu Thr Val Arg

245 250 255

Asn Ala Ala Asp Ala Ala Thr Ile Ser Ala Thr Leu Lys Asp Ser Asp

260 265 270

Gly Asn Val Val Trp Glu Thr Ala Asn Cys Ala Asp Pro Asp Thr Ser

275 280 285

Ile Ser Thr Gly Ser Leu Asn Gly Ile Arg Pro Trp Ser Ala Glu Asp

290 295 300

Pro Val Leu Tyr Glu Phe Glu Val Thr Val Ile Asp His Ala Gly Asn

305 310 315 320

Ile Ala Glu Val Ala Val Gln Lys Val Gly Phe Arg Arg Phe Arg Ile

325 330 335

Glu Asp Gly Ile Met Thr Ile Asn Gly Lys Arg Ile Val Phe Lys Gly

340 345 350

Ala Asp Arg His Glu Phe Asp Pro Lys Arg Gly Arg Ala Ile Thr Glu

355 360 365

Gln Asp Met Ile Asp Asp Val Val Phe Cys Lys Arg His Asn Leu Asn

370 375 380

Ala Ile Arg Thr Ser His Tyr Pro Asn Gln Glu Arg Trp Tyr Glu Leu

385 390 395 400

Cys Asp Glu Tyr Gly Ile Tyr Leu Ile Asp Glu Thr Asn Leu Glu Thr

405 410 415

His Gly Ser Trp Cys Leu Pro Gly Asp Val Leu Thr Glu Glu Thr Ala

420 425 430

Val Pro Gly Ser Lys Ala His Trp Glu Gly Ala Cys Val Asp Arg Val

435 440 445

Asn Ser Met Val Arg Arg Asp Tyr Asn His Pro Ser Val Leu Ile Trp

450 455 460

Ser Leu Gly Asn Glu Ser Tyr Thr Gly Asp Val Phe Arg Ala Met Tyr

465 470 475 480

Lys Arg Val His Asp Ile Asp Pro Asn Arg Pro Val His Tyr Glu Gly

485 490 495

Val Thr His Asn Arg Asp Tyr Asn Asp Val Thr Asp Ile Glu Thr Arg

500 505 510

Met Tyr Ala His Ala Asp Ala Ile Glu Glu Tyr Leu Lys Asn Asp Pro

515 520 525

Gln Lys Pro Tyr Ile Ser Cys Glu Tyr Met His Ala Met Gly Asn Ser

530 535 540

Cys Gly Asn Met Asp Glu Tyr Thr Ala Leu Glu Arg Tyr Pro Lys Tyr

545 550 555 560

Gln Gly Gly Phe Ile Trp Asp Phe Ile Asp Gln Ala Ile Tyr Ala Thr

565 570 575

Gln Pro Asp Gly Thr Thr Ser Leu Arg Tyr Gly Gly Asp Phe Gly Asp

580 585 590

Arg Pro Ser Asp Tyr Glu Phe Ser Gly Asn Gly Leu Val Phe Ala Asp

595 600 605

Arg Lys Pro Thr Pro Lys Ala Gln Glu Val Lys Gln Leu Tyr Ser Asn

610 615 620

Val His Ile Asp Val Ala Glu Asp Ser Val Thr Ile Lys Asn Asp Asn

625 630 635 640

Leu Phe Thr Ser Thr Gly Glu Tyr Thr Phe Val Leu Arg Val Leu Ala

645 650 655

Asp Gly Glu Pro Val Trp Gln Ser Glu Arg Arg Phe Asp Val Pro Ala

660 665 670

Gly Ser Thr Glu Lys Leu Asp Val Asp Trp Pro Leu Asp Leu Tyr Arg

675 680 685

Asp Gly Ala Ser Glu Leu Val Leu Glu Val Ser Gln Arg Leu Ala Lys

690 695 700

Ala Thr Asn Trp Ala Val Ala Gly Tyr Glu Leu Ala Phe Gly Gln Thr

705 710 715 720

Val Val Ala Gly Ser Lys Lys Ala Ser Ala Pro Val Lys Pro Val Asp

725 730 735

Gly Ile Val Thr Val Gly Arg Trp Asn Val Gly Val Gln Gly Ser Gly

740 745 750

Arg Glu Val Leu Leu Ser Arg Thr Gln Gly Gly Leu Val Ser Tyr Thr

755 760 765

Phe Asn Asn Arg Glu Phe Val Leu Arg Arg Pro Ala Val Thr Thr Phe

770 775 780

Arg Ala Leu Thr Asp Asn Asp Arg Gly Ala Gly His Gly Phe Glu Arg

785 790 795 800

Ala Gln Trp Leu Gly Ala Gly Arg Tyr Ala Arg Cys Ile Gly Asn Glu

805 810 815

Ile Glu Gln Ile Asp Glu Asn Thr Val Lys Ala Ser Tyr Thr Tyr Glu

820 825 830

Leu Ala Thr Pro Gln Arg Thr Lys Val Thr Val Ser Tyr Thr Ala Asp

835 840 845

Thr Thr Gly Arg Val Asn Leu His Val Glu Tyr Pro Gly Glu Pro Gly

850 855 860

Asp Leu Pro Thr Ile Pro Ala Phe Gly Ile Glu Trp Thr Leu Pro Val

865 870 875 880

Gln Tyr Ser Asn Leu Arg Phe Phe Gly Ala Gly Pro Glu Glu Thr Tyr

885 890 895

Gln Asp Arg Lys His Ala Lys Leu Gly Val Trp Ser Thr Asp Ala Phe

900 905 910

Lys Asp His Ala Pro Tyr Leu Met Pro Gln Glu Thr Gly Asn His Glu

915 920 925

Asp Val Arg Trp Ala Glu Ile Thr Asp Glu Lys Gly His Gly Leu Arg

930 935 940

Ile Ser Arg Ala Glu Gly Ala Glu Pro Phe Ala Met Ser Leu Gln Pro

945 950 955 960

Tyr Ser Ser Phe Met Leu Glu Glu Ala Gln His Gln Asp Glu Leu Pro

965 970 975

Ala Pro Lys His Met Phe Leu Arg Val Leu Ala Glu Gln Met Gly Val

980 985 990

Gly Gly Asp Asp Ser Trp Met Ser Pro Val His Pro Gln Tyr His Ile

995 1000 1005

Pro Ala Asp Gln Pro Ile Ser Leu Asp Val Asp Leu Asp Leu Ile

1010 1015 1020

<210> 34

<211> 1305

<212>PRT

<213> Bifidobacterium bifidum

<400> 34

Met Val Glu Asp Ala Thr Arg Ser Asp Ser Thr Thr Gln Met Ser Ser

1 5 10 15

Thr Pro Glu Val Val Tyr Ser Ser Ala Val Asp Ser Lys Gln Asn Arg

20 25 30

Thr Ser Asp Phe Asp Ala Asn Trp Lys Phe Met Leu Ser Asp Ser Val

35 40 45

Gln Ala Gln Asp Pro Ala Phe Asp Asp Ser Ala Trp Gln Gln Val Asp

50 55 60

Leu Pro His Asp Tyr Ser Ile Thr Gln Lys Tyr Ser Gln Ser Asn Glu

65 70 75 80

Ala Glu Ser Ala Tyr Leu Pro Gly Gly Thr Gly Trp Tyr Arg Lys Ser

85 90 95

Phe Thr Ile Asp Arg Asp Leu Ala Gly Lys Arg Ile Ala Ile Asn Phe

100 105 110

Asp Gly Val Tyr Met Asn Ala Thr Val Trp Phe Asn Gly Val Lys Leu

115 120 125

Gly Thr His Pro Tyr Gly Tyr Ser Pro Phe Ser Phe Asp Leu Thr Gly

130 135 140

Asn Ala Lys Phe Gly Gly Glu Asn Thr Ile Val Val Lys Val Glu Asn

145 150 155 160

Arg Leu Pro Ser Ser Arg Trp Tyr Ser Gly Ser Gly Ile Tyr Arg Asp

165 170 175

Val Thr Leu Thr Val Thr Asp Gly Val His Val Gly Asn Asn Gly Val

180 185 190

Ala Ile Lys Thr Pro Ser Leu Ala Thr Gln Asn Gly Gly Asn Val Thr

195 200 205

Met Asn Leu Thr Thr Lys Val Ala Asn Asp Thr Glu Ala Ala Ala Asn

210 215 220

Ile Thr Leu Lys Gln Thr Val Phe Pro Lys Gly Gly Lys Thr Asp Ala

225 230 235 240

Ala Ile Gly Thr Val Thr Thr Ala Ser Lys Ser Ile Ala Ala Gly Ala

245 250 255

Ser Ala Asp Val Thr Ser Thr Ile Thr Ala Ala Ser Pro Lys Leu Trp

260 265 270

Ser Ile Lys Asn Pro Asn Leu Tyr Thr Val Arg Thr Glu Val Leu Asn

275 280 285

Gly Asp Thr Val Leu Asp Thr Tyr Asp Thr Glu Tyr Gly Phe Arg Trp

290 295 300

Thr Gly Phe Asp Ala Thr Ser Gly Phe Ser Leu Asn Gly Glu Lys Val

305 310 315 320

Lys Leu Lys Gly Val Ser Met His His Asp Gln Gly Ser Leu Gly Ala

325 330 335

Val Ala Asn Arg Arg Ala Ile Glu Arg Gln Val Glu Ile Leu Gln Lys

340 345 350

Met Gly Val Asn Ser Ile Arg Thr Thr His Asn Pro Ala Ala Lys Ala

355 360 365

Leu Ile Asp Val Cys Asn Glu Lys Gly Val Leu Val Val Glu Glu Val

370 375 380

Phe Asp Met Trp Asn Arg Ser Lys Asn Gly Asn Thr Glu Asp Tyr Gly

385 390 395 400

Lys Trp Phe Gly Gln Thr Ile Ala Gly Asp Asn Ala Val Leu Gly Gly

405 410 415

Asp Lys Asp Glu Thr Trp Ala Lys Phe Asp Leu Thr Ser Thr Ile Asn

420 425 430

Arg Asp Arg Asn Ala Pro Ser Val Ile Met Trp Ser Leu Gly Asn Glu

435 440 445

Met Met Glu Gly Ile Ser Gly Ser Val Ser Asp Phe Pro Ala Thr Ser

450 455 460

Ala Lys Leu Val Ala Trp Thr Lys Ala Ala Asp Ser Thr Arg Pro Met

465 470 475 480

Thr Tyr Gly Asp Asn Lys Ile Lys Ala Asn Trp Asn Glu Ser Asn Thr

485 490 495

Met Gly Asp Asn Leu Thr Ala Asn Gly Gly Val Val Gly Thr Asn Tyr

500 505 510

Ser Asp Gly Ala Asn Tyr Asp Lys Ile Arg Thr Thr His Pro Ser Trp

515 520 525

Ala Ile Tyr Gly Ser Glu Thr Ala Ser Ala Ile Asn Ser Arg Gly Ile

530 535 540

Tyr Asn Arg Thr Thr Gly Gly Ala Gln Ser Ser Asp Lys Gln Leu Thr

545 550 555 560

Ser Tyr Asp Asn Ser Ala Val Gly Trp Gly Ala Val Ala Ser Ser Ala

565 570 575

Trp Tyr Asp Val Val Gln Arg Asp Phe Val Ala Gly Thr Tyr Val Trp

580 585 590

Thr Gly Phe Asp Tyr Leu Gly Glu Pro Thr Pro Trp Asn Gly Thr Gly

595 600 605

Ser Gly Ala Val Gly Ser Trp Pro Ser Pro Lys Asn Ser Tyr Phe Gly

610 615 620

Ile Val Asp Thr Ala Gly Phe Pro Lys Asp Thr Tyr Tyr Phe Tyr Gln

625 630 635 640

Ser Gln Trp Asn Asp Asp Val His Thr Leu His Ile Leu Pro Ala Trp

645 650 655

Asn Glu Asn Val Val Ala Lys Gly Ser Gly Asn Lys Val Pro Val Val

660 665 670

Val Tyr Thr Asp Ala Ala Lys Val Lys Leu Tyr Phe Thr Pro Lys Gly

675 680 685

Ser Thr Glu Lys Arg Leu Ile Gly Glu Lys Ser Phe Thr Lys Lys Thr

690 695 700

Thr Ala Ala Gly Tyr Thr Tyr Gln Val Tyr Glu Gly Thr Asp Lys Asp

705 710 715 720

Ser Thr Ala His Lys Asn Met Tyr Leu Thr Trp Asn Val Pro Trp Ala

725 730 735

Glu Gly Thr Ile Ser Ala Glu Ala Tyr Asp Glu Asn Asn Arg Leu Ile

740 745 750

Pro Glu Gly Ser Thr Glu Gly Asn Ala Ser Val Thr Thr Thr Gly Lys

755 760 765

Ala Ala Lys Leu Lys Ala Asp Ala Asp Arg Lys Thr Ile Thr Ala Asp

770 775 780

Gly Lys Asp Leu Ser Tyr Ile Glu Val Asp Val Thr Asp Ala Asn Gly

785 790 795 800

His Ile Val Pro Asp Ala Ala Asn Arg Val Thr Phe Asp Val Lys Gly

805 810 815

Ala Gly Lys Leu Val Gly Val Asp Asn Gly Ser Ser Pro Asp His Asp

820 825 830

Ser Tyr Gln Ala Asp Asn Arg Lys Ala Phe Ser Gly Lys Val Leu Ala

835 840 845

Ile Val Gln Ser Thr Lys Glu Ala Gly Glu Ile Thr Val Thr Ala Lys

850 855 860

Ala Asp Gly Leu Gln Ser Ser Thr Val Lys Ile Ala Thr Thr Ala Val

865 870 875 880

Pro Gly Thr Ser Thr Glu Lys Thr Val Arg Ser Phe Tyr Tyr Ser Arg

885 890 895

Asn Tyr Tyr Val Lys Thr Gly Asn Lys Pro Ile Leu Pro Ser Asp Val

900 905 910

Glu Val Arg Tyr Ser Asp Gly Thr Ser Asp Arg Gln Asn Val Thr Trp

915 920 925

Asp Ala Val Ser Asp Asp Gln Ile Ala Lys Ala Gly Ser Phe Ser Val

930 935 940

Ala Gly Thr Val Ala Gly Gln Lys Ile Ser Val Arg Val Thr Met Ile

945 950 955 960

Asp Glu Ile Gly Ala Leu Leu Asn Tyr Ser Ala Ser Thr Pro Val Gly

965 970 975

Thr Pro Ala Val Leu Pro Gly Ser Arg Pro Ala Val Leu Pro Asp Gly

980 985 990

Thr Val Thr Ser Ala Asn Phe Ala Val His Trp Thr Lys Pro Ala Asp

995 1000 1005

Thr Val Tyr Asn Thr Ala Gly Thr Val Lys Val Pro Gly Thr Ala

1010 1015 1020

Thr Val Phe Gly Lys Glu Phe Lys Val Thr Ala Thr Ile Arg Val

1025 1030 1035

Gln Arg Ser Gln Val Thr Ile Gly Ser Ser Val Ser Gly Asn Ala

1040 1045 1050

Leu Arg Leu Thr Gln Asn Ile Pro Ala Asp Lys Gln Ser Asp Thr

1055 1060 1065

Leu Asp Ala Ile Lys Asp Gly Ser Thr Thr Val Asp Ala Asn Thr

1070 1075 1080

Gly Gly Gly Ala Asn Pro Ser Ala Trp Thr Asn Trp Ala Tyr Ser

1085 1090 1095

Lys Ala Gly His Asn Thr Ala Glu Ile Thr Phe Glu Tyr Ala Thr

1100 1105 1110

Glu Gln Gln Leu Gly Gln Ile Val Met Tyr Phe Phe Arg Asp Ser

1115 1120 1125

Asn Ala Val Arg Phe Pro Asp Ala Gly Lys Thr Lys Ile Gln Ile

1130 1135 1140

Ser Ala Asp Gly Lys Asn Trp Thr Asp Leu Ala Ala Thr Glu Thr

1145 1150 1155

Ile Ala Ala Gln Glu Ser Ser Asp Arg Val Lys Pro Tyr Thr Tyr

1160 1165 1170

Asp Phe Ala Pro Val Gly Ala Thr Phe Val Lys Val Thr Val Thr

1175 1180 1185

Asn Ala Asp Thr Thr Thr Pro Ser Gly Val Val Cys Ala Gly Leu

1190 1195 1200

Thr Glu Ile Glu Leu Lys Thr Ala Thr Ser Lys Phe Val Thr Asn

1205 1210 1215

Thr Ser Ala Ala Leu Ser Ser Leu Thr Val Asn Gly Thr Lys Val

1220 1225 1230

Ser Asp Ser Val Leu Ala Ala Gly Ser Tyr Asn Thr Pro Ala Ile

1235 1240 1245

Ile Ala Asp Val Lys Ala Glu Gly Glu Gly Asn Ala Ser Val Thr

1250 1255 1260

Val Leu Pro Ala His Asp Asn Val Ile Arg Val Ile Thr Glu Ser

1265 1270 1275

Glu Asp His Val Thr Arg Lys Thr Phe Thr Ile Asn Leu Gly Thr

1280 1285 1290

Glu Gln Glu Phe Pro Ala Asp Ser Asp Glu Arg Asp

1295 1300 1305

<210> 35

<211> 1025

<212>PRT

<213> Kluyveromyces lactis

<400> 35

Met Ser Cys Leu Ile Pro Glu Asn Leu Arg Asn Pro Lys Lys Val His

1 5 10 15

Glu Asn Arg Leu Pro Thr Arg Ala Tyr Tyr Tyr Asp Gln Asp Ile Phe

20 25 30

Glu Ser Leu Asn Gly Pro Trp Ala Phe Ala Leu Phe Asp Ala Pro Leu

35 40 45

Asp Ala Pro Asp Ala Lys Asn Leu Asp Trp Glu Thr Ala Lys Lys Trp

50 55 60

Ser Thr Ile Ser Val Pro Ser His Trp Glu Leu Gln Glu Asp Trp Lys

65 70 75 80

Tyr Gly Lys Pro Ile Tyr Thr Asn Val Gln Tyr Pro Ile Pro Ile Asp

85 90 95

Ile Pro Asn Pro Pro Thr Val Asn Pro Thr Gly Val Tyr Ala Arg Thr

100 105 110

Phe Glu Leu Asp Ser Lys Ser Ile Glu Ser Phe Glu His Arg Leu Arg

115 120 125

Phe Glu Gly Val Asp Asn Cys Tyr Glu Leu Tyr Val Asn Gly Gln Tyr

130 135 140

Val Gly Phe Asn Lys Gly Ser Arg Asn Gly Ala Glu Phe Asp Ile Gln

145 150 155 160

Lys Tyr Val Ser Glu Gly Glu Asn Leu Val Val Val Lys Val Phe Lys

165 170 175

Trp Ser Asp Ser Thr Tyr Ile Glu Asp Gln Asp Gln Trp Trp Leu Ser

180 185 190

Gly Ile Tyr Arg Asp Val Ser Leu Leu Lys Leu Pro Lys Lys Ala His

195 200 205

Ile Glu Asp Val Arg Val Thr Thr Thr Phe Val Asp Ser Gln Tyr Gln

210 215 220

Asp Ala Glu Leu Ser Val Lys Val Asp Val Gln Gly Ser Ser Tyr Asp

225 230 235 240

His Ile Asn Phe Thr Leu Tyr Glu Pro Glu Asp Gly Ser Lys Val Tyr

245 250 255

Asp Ala Ser Ser Leu Leu Asn Glu Glu Asn Gly Asn Thr Thr Phe Ser

260 265 270

Thr Lys Glu Phe Ile Ser Phe Ser Thr Lys Lys Asn Glu Glu Thr Ala

275 280 285

Phe Lys Ile Asn Val Lys Ala Pro Glu His Trp Thr Ala Glu Asn Pro

290 295 300

Thr Leu Tyr Lys Tyr Gln Leu Asp Leu Ile Gly Ser Asp Gly Ser Val

305 310 315 320

Ile Gln Ser Ile Lys His His Val Gly Phe Arg Gln Val Glu Leu Lys

325 330 335

Asp Gly Asn Ile Thr Val Asn Gly Lys Asp Ile Leu Phe Arg Gly Val

340 345 350

Asn Arg His Asp His His Pro Arg Phe Gly Arg Ala Val Pro Leu Asp

355 360 365

Phe Val Val Arg Asp Leu Ile Leu Met Lys Lys Phe Asn Ile Asn Ala

370 375 380

Val Arg Asn Ser His Tyr Pro Asn His Pro Lys Val Tyr Asp Leu Phe

385 390 395 400

Asp Lys Leu Gly Phe Trp Val Ile Asp Glu Ala Asp Leu Glu Thr His

405 410 415

Gly Val Gln Glu Pro Phe Asn Arg His Thr Asn Leu Glu Ala Glu Tyr

420 425 430

Pro Asp Thr Lys Asn Lys Leu Tyr Asp Val Asn Ala His Tyr Leu Ser

435 440 445

Asp Asn Pro Glu Tyr Glu Val Ala Tyr Leu Asp Arg Ala Ser Gln Leu

450 455 460

Val Leu Arg Asp Val Asn His Pro Ser Ile Ile Ile Trp Ser Leu Gly

465 470 475 480

Asn Glu Ala Cys Tyr Gly Arg Asn His Lys Ala Met Tyr Lys Leu Ile

485 490 495

Lys Gln Leu Asp Pro Thr Arg Leu Val His Tyr Glu Gly Asp Leu Asn

500 505 510

Ala Leu Ser Ala Asp Ile Phe Ser Phe Met Tyr Pro Thr Phe Glu Ile

515 520 525

Met Glu Arg Trp Arg Lys Asn His Thr Asp Glu Asn Gly Lys Phe Glu

530 535 540

Lys Pro Leu Ile Leu Cys Glu Tyr Gly His Ala Met Gly Asn Gly Pro

545 550 555 560

Gly Ser Leu Lys Glu Tyr Gln Glu Leu Phe Tyr Lys Glu Lys Phe Tyr

565 570 575

Gln Gly Gly Phe Ile Trp Glu Trp Ala Asn His Gly Ile Glu Phe Glu

580 585 590

Asp Val Ser Thr Ala Asp Gly Lys Leu His Lys Ala Tyr Ala Tyr Gly

595 600 605

Gly Asp Phe Lys Glu Glu Val His Asp Gly Val Phe Ile Met Asp Gly

610 615 620

Leu Cys Asn Ser Glu His Asn Pro Thr Pro Gly Leu Val Glu Tyr Lys

625 630 635 640

Lys Val Ile Glu Pro Val His Ile Lys Ile Ala His Gly Ser Val Thr

645 650 655

Ile Thr Asn Lys His Asp Phe Ile Thr Thr Asp His Leu Leu Phe Ile

660 665 670

Asp Lys Asp Thr Gly Lys Thr Ile Asp Val Pro Ser Leu Lys Pro Glu

675 680 685

Glu Ser Val Thr Ile Pro Ser Asp Thr Thr Tyr Val Val Ala Val Leu

690 695 700

Lys Asp Asp Ala Gly Val Leu Lys Ala Gly His Glu Ile Ala Trp Gly

705 710 715 720

Gln Ala Glu Leu Pro Leu Lys Val Pro Asp Phe Val Thr Glu Thr Ala

725 730 735

Glu Lys Ala Ala Lys Ile Asn Asp Gly Lys Arg Tyr Val Ser Val Glu

740 745 750

Ser Ser Gly Leu His Phe Ile Leu Asp Lys Leu Leu Gly Lys Ile Glu

755 760 765

Ser Leu Lys Val Lys Gly Lys Glu Ile Ser Ser Lys Phe Glu Gly Ser

770 775 780

Ser Ile Thr Phe Trp Arg Pro Pro Thr Asn Asn Asp Glu Pro Arg Asp

785 790 795 800

Phe Lys Asn Trp Lys Lys Tyr Asn Ile Asp Leu Met Lys Gln Asn Ile

805 810 815

His Gly Val Ser Val Glu Lys Gly Ser Asn Gly Ser Leu Ala Val Val

820 825 830

Thr Val Asn Ser Arg Ile Ser Pro Val Val Phe Tyr Tyr Gly Phe Glu

835 840 845

Thr Val Gln Lys Tyr Thr Ile Phe Ala Asn Lys Ile Asn Leu Asn Thr

850 855 860

Ser Met Lys Leu Thr Gly Glu Tyr Gln Pro Pro Asp Phe Pro Arg Val

865 870 875 880

Gly Tyr Glu Phe Trp Leu Gly Asp Ser Tyr Glu Ser Phe Glu Trp Leu

885 890 895

Gly Arg Gly Pro Gly Glu Ser Tyr Pro Asp Lys Lys Glu Ser Gln Arg

900 905 910

Phe Gly Leu Tyr Asp Ser Lys Asp Val Glu Glu Phe Val Tyr Asp Tyr

915 920 925

Pro Gln Glu Asn Gly Asn His Thr Asp Thr His Phe Leu Asn Ile Lys

930 935 940

Phe Glu Gly Ala Gly Lys Leu Ser Ile Phe Gln Lys Glu Lys Pro Phe

945 950 955 960

Asn Phe Lys Ile Ser Asp Glu Tyr Gly Val Asp Glu Ala Ala His Ala

965 970 975

Cys Asp Val Lys Arg Tyr Gly Arg His Tyr Leu Arg Leu Asp His Ala

980 985 990

Ile His Gly Val Gly Ser Glu Ala Cys Gly Pro Ala Val Leu Asp Gln

995 1000 1005

Tyr Arg Leu Lys Ala Gln Asp Phe Asn Phe Glu Phe Asp Leu Ala

1010 1015 1020

PheGlu

1025

<210> 36

<211> 30

<212> DNA

<213> Primer

<220>

<221> other_sign

<222> (9)..(9)

<223> n represents a deoxyuracil residue at 9 positions

<400> 36

attaaccang cgacgcaact tcgaatggcc

thirty

<210> 37

<211> 31

<212> DNA

<213> Primer

<220>

<221> other_sign

<222> (9)..(9)

<223> n represents a deoxyuracil residue at 9 positions

<400> 37

atcttctcnt taccgcctta ccacgagcac g

31

<210> 38

<211> 33

<212> DNA

<213> Primer

<220>

<221> other_sign

<222> (9)..(9)

<223> n represents a deoxyuracil residue at 9 positions

<400> 38

agagaagant ttcagcctga tacagattaa atc

33

<210> 39

<211> 33

<212> DNA

<213> Primer

<220>

<221> other_sign

<222> (9)..(9)

<223> n represents a deoxyuracil residue at 9 positions

<400> 39

atggttaant cctcctgtta gcccaaaaaa cgg

33

<210> 40

<211> 22

<212> DNA

<213> Primer

<400> 40

cggcgtcaca ctttgctatg cc

22

<210> 41

<211> 20

<212> DNA

<213> Primer

<400> 41

ccgcgctact gccgccaggc

20

<---

Claims (18)

1. A peptide exhibiting beta-galactosidase enzymatic activity, which has the amino acid sequence represented by SEQ ID NO: 1 or 32. 2. A method of reducing the lactose content in a composition containing lactose, such as dairy products, comprising the step of contacting said composition with a peptide exhibiting beta-galactosidase enzymatic activity, wherein the peptide has the amino acid sequence represented by SEQ ID NO: 1, 4, 7 -8, 10, 12-19, 22, 25-27, 32 or 33; or with a host cell expressing any of these peptides, at a pH in the range of 3-10 and at a temperature in the range of 0-140°C. 3. The use of a peptide exhibiting beta-galactosidase enzymatic activity, wherein the peptide has the amino acid sequence represented by SEQ ID NO: 1, 4, 7-8, 10, 12-19, 22, 25-27, 32 or 33; or a host cell expressing any of these peptides to produce a lactose-reduced dairy product. 4. A method for producing a dairy product, including the stages: a) providing a lactose-containing milk-based substrate; b) adding to said lactose-containing milk-based substrate a peptide exhibiting beta-galactosidase activity, where the peptide has the amino acid sequence represented by SEQ ID NO: 1, 4, 7-8, 10, 12-19, 22, 25-27, 32 or 33; And c) treating said milk-based substrate with said peptide exhibiting beta-galactosidase activity. 5. The method according to claim 4, where step c) is carried out at a pH in the range of 3–10, for example in the range of 3–9, for example in the range of 3–8, for example in the range of 3–7, for example in the range of 3–6, for example in the range 3–5, e.g. range 3–4, e.g. range 4–10, e.g. range 4–9, e.g. range 4–8, e.g. range 4–7, e.g. range 4–6, e.g. range 4 –5, for example in the range 5–10, for example in the range 5–9, for example in the range 5–8, for example in the range 5–7, for example in the range 5–6, for example in the range 6–10, for example in the range 6– 9, for example in the range 6–8, for example in the range 6–7. 6. The method according to any one of paragraphs. 4, 5, where step c) or part of step c) is carried out at a temperature of no more than about 25°C, for example no more than about 20°C, for example no more than about 18°C, for example no more than about 16°C , for example not more than about 14°C, for example not more than about 12°C, for example not more than about 10°C, for example not more than about 8°C, for example not more than about 7°C, for example not more than about 6°C, eg no more than about 5°C, eg no more than about 4°C, eg no more than about 3°C, eg no more than about 2°C. 7. Method according to any one of paragraphs. 4, 5, where stage c) or part of stage c) carried out at a temperature of at least about 25°C, for example at least about 30°C, for example at least about 35°C, for example at least about 40°C, for example at least about 45°C, for example at least at least about 50°C, for example at least about 55°C, for example at least about 60°C, for example at least about 65°C, for example at least about 70°C, for example at least about 75°C , for example at least about 80°C, for example at least about 85°C, for example at least about 90°C, for example at least about 95°C, for example at least about 100°C, for example at least about 110°C, for example at least about 120°C, for example at least about 130°C, for example at least about 135°C, for example at least about 140°C. 8. The method according to any one of paragraphs. 4-7, wherein the galactose inhibition of said peptide is less than 60%, such as less than 55%, such as less than 50%, such as less than about 45%, such as less than about 40%. 9. The method according to any one of paragraphs. 4-8, wherein said dairy product is a fermented milk product and said step b) is carried out during or before fermentation. 10. The method according to claim 9, which does not require the addition of additional enzyme after fermentation. eleven. The method according to any one of paragraphs. 4-9, where the specified dairy product is a fermented milk product and the specified stage b) is carried out immediately after fermentation. 12. The method according to any one of paragraphs. 4-9, wherein said dairy product is fresh milk and said step b) is carried out before, in combination with or immediately after the pasteurization step. 13. The method according to any one of paragraphs. 4-9, wherein said dairy product is ultra-pasteurized (UHT) milk and said step b) is carried out before, in combination with, or immediately after the UHT step. 14. The method according to any one of paragraphs. 4–13, where step c) is started at a temperature from 40 to 100°C, for example at a temperature from 50 to 100°C, for example at a temperature from 60 to 100°C, for example at a temperature from 70 to 100°C, for example at temperature from 80 to 100°C, for example at a temperature from 40 to 90°C, for example at a temperature from 40 to 80°C, for example at a temperature from 40 to 70°C, for example at a temperature from 40 to 60°C, for example at temperature from 40 to 50°C.