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EP0382738A1 - Lipase immobilisee, positionnellement non specifique, sa production et son utilisation - Google Patents

Lipase immobilisee, positionnellement non specifique, sa production et son utilisation

Info

Publication number
EP0382738A1
EP0382738A1 EP88907259A EP88907259A EP0382738A1 EP 0382738 A1 EP0382738 A1 EP 0382738A1 EP 88907259 A EP88907259 A EP 88907259A EP 88907259 A EP88907259 A EP 88907259A EP 0382738 A1 EP0382738 A1 EP 0382738A1
Authority
EP
European Patent Office
Prior art keywords
lipase
use according
immobilized
preparation
specific
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
EP88907259A
Other languages
German (de)
English (en)
Inventor
Tom Busk Room No. 501 Nielsen
Miyoko Room No. 201 Hasida
Hidesato Room No. 105 Shimoto
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Novo Nordisk AS
Original Assignee
Novo Nordisk AS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Novo Nordisk AS filed Critical Novo Nordisk AS
Publication of EP0382738A1 publication Critical patent/EP0382738A1/fr
Ceased legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6409—Fatty acids
    • C12P7/6418—Fatty acids by hydrolysis of fatty acid esters
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
    • C12N11/089—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
    • C12N11/091—Phenol resins; Amino resins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)
    • C12N9/20—Triglyceride splitting, e.g. by means of lipase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/62—Carboxylic acid esters
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6436—Fatty acid esters
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6436—Fatty acid esters
    • C12P7/6445—Glycerides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6436—Fatty acid esters
    • C12P7/6445—Glycerides
    • C12P7/6458—Glycerides by transesterification, e.g. interesterification, ester interchange, alcoholysis or acidolysis

Definitions

  • Lipase is taken to mean an enzyme that catalyzes reactions involving ester bonds (such as hydrolysis, synthesis and exhange of ester bonds) in water-insoluble carboxylic acid esters e.g. at an interface between aqueous and an organic phase.
  • Lipase may be in soluble, derivatized or immobilized form.
  • Immobilized lipase denotes lipase in the form of immobilized enzyme or immobilized cells, as defined in "Guidelines for the characterization of immobilized biocatalysts” (1983), Enzyme Microb. Technol., 5_, 304-307.
  • Derivatized lipase denotes lipase that has been chemically modified without immobiliz.ing " it T__5oluble lipase denotes un-modified lipase. T *-- ⁇ ⁇ " "y ⁇ - - ⁇ - ⁇ ⁇ ⁇
  • Lipases may be divided according to their positional specificity. As used in this specification, a positionally specific lipase (or specific lipase for short) is one that reacts only with the fatty acyl groups in the 1- and 3- positions of a triglyceride molecule, and a positionally non-specific lipase (or non-specific lipase for short) is one that reacts with all three fatty acyl groups of a triglyceride.
  • Non-specific lipase activity can be determined in an interesteri-fication reaction with a triglyceride by measuring the reaction rate in the middle position, e.g. by reacting pure triolein or cocoa butter stearin with a pure fatty acid.
  • Lipase-catalyzed processes are ester hydrolysis, ester synthesis and interesterification.
  • the esters involved may be triglycerides or other carboxylic acid esters.
  • Random interesterification of fats refers to interesterification reactions in which all three acyl groups in the triglycerides react, whereby a nearly random distribution of fatty acids in all three positions can be obtained. This may be achieved by use of a chemical catalyst or a non-specific lipase.
  • This,-invention relates to an immobilized, non ⁇ specific lipase preparation in particulate form, to a method for producing it, and to its use in a lipase- catalyzed process, i.e. interesterification, ester synthesis and ester hydrolysis.
  • a lipase- catalyzed process i.e. interesterification, ester synthesis and ester hydrolysis.
  • it relates to such an immobilized lipase with improved heat
  • the lipase is derived from Candida cylindracea, and the data in- the article show that the immobilized lipase has optimum temperature about 40°C, and that there is significant deactivation at 50°C.
  • thermostable, immobilized, non-specific lipase for
  • the object of the invention to provide immobilized, non-specific lipase that is thermostable enough for long-term use at 60°C or higher.
  • the lipase should be microbial, as these can be produced
  • thermostability data have been published for the following: Staphylococcus aureus (Vadehra, D.V. (1974). Lipids, 9_, 158), Penicillium cyclopium (Okumura, S., et al. (1976). Agricultural and Biological Chemistry, 40, 655 and Renshaw E.C. and San Clemente CL. (1966) Developments in Industrial Microbiology, , 214), Corynebacterium acnes (Hassing, G.S. (1971). Biochimica et Biophysica Acta, 242, 381 and Pablo
  • positionally non-specific lipase can be obtained from Pseudomonas cepacia. And it has surprisingly been found that this lipase in immobilized form is heatstable enough for long- term use at 60°C, and is thus more heat-stable than previously known immobilized, non-specific lipases.
  • the immobilized lipase -i-s " -particularly suited for randomization of fat at temperatures as high as 60-80°C. Most fats are liquid at such temperatures and can be randomized without the presence of a solvent.
  • Lipases are known from Pseudomonas cepacia and from other species of Pseudomonas, but none of these are known to be positionally non-specific towards triglycerides.
  • P. Eigtved et al. in a paper presented at the AOCS/JOCS Meeting in Honolulu on 1986-05-17 describe a Pseudomonas cepacia lipase that can esterify secondary alcohols (in contrast to most specific lipases), but is positionally specific ir its action on triglycerides.
  • the first aspect of the invention provides an immobilized, positionally non-specific lipase preparation in particulate form, characterized in that the non-specific lipase is producible from Pseudomonas cepacia and/ or that it has immunological properties identical to those of the non-specific lipase from the P. cepacia strain DSM 3959.
  • This preparation has a half-life at 60°C above 1,000 hours when measured in continuous, fixed-bed interesterification.
  • Another aspect of the invention provides use of the above-mentioned immobilized preparation in a lipase- catalyzed process.
  • non-specific lipases that can be used in the practice of the invention are those that are producible by a strain of Pseudomonas cepacia and/or have immunological properties identical to those of the non ⁇ specific lipase from P. cepacia strain DSM 3959. Immunological identity may be determined according to N.H. Axelsen et al. (ed.): Quantitative Immunoelectrophoresis, -(Blackwell Scientific Publications, 1973), especially • . __ chapter 10 and to Ivan Roitt: Essential Immunology, 5th ed. (Blackwell Scientific Publications, 1984), especially Chapter 6.
  • Non-specific lipase for use in the invention may contain specific lipase as well, e.g.
  • Non-specific lipase for use in the invention may be produced by cultivating the strain DSM 3959 under aerobic conditions in a nutrient medium containing assimilable carbon and nitrogen together with other essential nutrients, the medium being composed in accordance with principles known in the art.
  • the strain DSM 3959 was deposited at Deutsche Sammlung von Mikroorganismen (DSM) in West Germany on Jan. 30, 1987 under the terms of the Budapest Treaty. It has been identified as Pseudomonas cepacia. The strain also produces specific lipase.
  • Non-specific lipase for use in the invention may also be obtained according to Japanese published application 57-63,087 from the P. cepacia strain with deposit number 5494 at the Fermentation Research Institute, Japan. Specificity of the lipase preparation -..isclosed in said application has not previously been described, but we have found that the preparation contains non-specific lipase.
  • lipase may be immobilized by any method known in the art, e.g. in . Mosbach (ed. ): Methods in Enzymology, _4_4, "Immobilized Enzymes", (Academic Press, New York, 1976).
  • Available methods for enzyme immobilization include: cross-linking of cell homogenates, covalent coupling to insoluble inorganic or organic carriers, entrapment in gels and adsorption on ion-exchange resins or other adsorbent materials.
  • coating on a particulate support may be used, as described in Macrae A.R. and Hammond R.C. (1985), Biotechnology and Genetic Engineering Reviews, 3, 193.
  • a preferred immobilization method uses a particulate, macroporous resin.
  • the lipase may be simply adsorbed on the resin, or it may be attached to the resin by cross-linking with glutaraldehyde or other cross-linking agent known in the art.
  • a preferred resin type is weakly basic anion exchange resin, e.g. of acrylic, polystyrene or phenolic type.
  • An example of a commercial product is Lewatit® E 1999/85 (product of Bayer, West Germany).
  • the immobilization on this type of resin is preferably according to EP 0 140 542, incorporated herein by reference.
  • Another preferred resin type is an adsorbent resin of the phenol-formaldehyde type.
  • the immobilization on this resin is preferably done according to DK 85/878, incorporated herein by reference.
  • Another preferred immobilization method uses an inorganic support material, and the lipase is preferably attached to the support by adsorption or covalent coupling.
  • Such support materials and immobilization techniques are describe ' d in K. Mosbach (ed.): Methods in Enzymol ⁇ gy, _44, "Immobilized Enzymes” (Academic Press, 1976).
  • the lipase-catalyzed process of this invention may be any of the following types. In each case reactant types are listed in parentheses:
  • the alcohol may be any mono- or polyvalent primary and/or secondary alcohol or a mixture of these.
  • the acid may be any carboxylic acid or a mixture of these.
  • the ester may be any ester derived from the mentioned alcohol and acid, or a mixture of these. Use of the immobilized lipase of the invention is particularly advantageous with triglycerides, where reaction in all three positions is desired.
  • the temperature in the process of this invention is preferably above 60°C, where most substrates and products of interest are liquid. Higher temperatures are generally preferred as reaction rate increases, and the diffusion resistance to mass transfer into and out of the immobilized lipase decreases. Also, in the case of column operation higher temperatures may be preferred to reduce the pressure drop over the column. On the other hand, in many cases the substrates and products will be degraded at the higher temperatures. Thus, a preferred range is 60- 90°C, more preferably 60-80°C.
  • Preferred embodiments of this process are fat splitting and hydrolysis of cholesterol esters. This may be performed either batch-wise or continuously.
  • the fat and water are mixed mechanically together with the necessary amount of immobilized lipase.
  • the water content will usually be kept below 40% w/w.
  • the temperature should be above the melting point of the fat, and may be as high as 80°C. Reaction time depends on enzyme dosage and desired conversion, but may be up to several days.
  • the immobilized lipase may be recovered and reused, thereby improving process economy.
  • fat above its melting point is passed through a reactor in which the immobilized lipase is retained.
  • Water may be added to the system in several ways, e.g. by dispersing water in the fat or by intermittently absorbing water in the immobilized lipase.
  • the process of this invention is particularly advantageous for the synthesis of esters of secondary alcohols that are otherwise difficult to produce, including those where the acid or alcohol is high-melting.
  • the process may be performed batch-wise or continuously.
  • the immobilized lipase may be recovered and reused to improve economy.
  • water is removed during reaction, e.g. by vacuum distillation or by absorption on molecular sieves.
  • the temperature should be such that the reaction mixture is liquid, preferably 60-90°C, preferably 60-80°C.
  • the reactants comprise a triglyceride fat and a fatty acid.
  • a preferred embodiment of this process is random interesterification of fat, where the reactant mixture comprises triglyceride fat, and reaction occurs by exchange of acyl groups between triglyceride molecules.
  • the reactant mixture may consist of a single fat fraction, whereby exchange between acyl groups in the three different positions occurs.
  • the reactant mixture may also consist of two or more types of fat, especially one being liquid at ambient temperature and one being a high-melting fat. The latter may be obtained by fractionation from natural sources or by hydrogenation. The product obtained by randomization of such mixtures is useful in margarine production.
  • the reactants comprise a triglyceride fat and a carboxyl acid ester, especially a methyl or ethyl ester.
  • the reactant mixture may also include a small amount of water, in order to maintain the activity of the enzyme. Water content up to saturation may be used, but a high water content leads to an undesired high degree of by-product formation by hydrolysis.
  • purification may be needed prior to carrying out the reaction in order to achieve the highest productivity of the immobilized lipase.
  • Conventional purification methods may be used, such as treatment with bleaching-earth or activated carbon.
  • reaction temperature may be as high as 80°C.
  • the lower limit for reaction temperature is determined by the melting point and viscosity of the reactant mixture. Preferred temperatures are from 60 to 90°C, most preferably from 60 to 80°C.
  • the reaction may be performed batch-wise or con- tinuously.
  • the substrate and if convenient solvent is mixed in a batch reactor whi>_-h is heated to the preferred temperature together with the immobilized lipase.
  • the substrate can be partly or fully saturated with water.
  • the enzyme dosage can be up to 10% depending on the desired conversion and reaction time.
  • the reaction time can be from a few hours to several days. After reaction the enzyme can be filtered off and reused, if convenient after a solvent wash.
  • the substrate is passed through a column containing the immobilized lipase.
  • the substrate can if convenient be dissolved in hexane or similar inert solvents.
  • the substrate can be partly or fu ' i y saturated with water before entering the enzyme column. This can e.g. be done by a precolumn containing water saturated resin or by saturating the substrate in the substrate container.
  • the desired conversion can be achieved by adjusting the flow rate through the column, i.e. changing the contact time.
  • the operation time in such a system can be up to several thousand hours.
  • the slow loss of activity occuring can be compensated for by decreasing the flow rate, i.e. increasing the residence time of the reactant mixture.
  • a typical initial residence time will depend. on desired conversion and can be from 5 min up to 2 hours.
  • the products may be further processed. ⁇ By-products such as free fatty acids may be removed afterwards by conventional methods such as caustic refining.
  • the product itself can be fractionated, blended with other oils or similar, depending on the specific application.
  • the method is based on hydrolysis of tributyrin in a pH-stat.
  • 1 LU Lipase Unit
  • 1 LU is the amount of enzyme which liberates 1 ⁇ mol titratable butyric acid per minute at 30°C, pH 7.0 with gum arabic as an emulsifier. Further details are given in Novo Analytical Method AF 95/5, available on request.
  • FAME fatty acid methyl esters
  • LPC gas chromatography
  • the ⁇ decrease-in oleic acid content expresses the non ⁇ specific lipase activity, and conveniently a Non- Specificity Index (NSI) may be calculated from the decrease of oleic acid and the total incorporation of lauric acid.
  • NBI Non- Specificity Index
  • the immobilized lipase (generally 250 mg as dry matter) is hydrated as required for activation usually to about 10% of water.
  • the following mixture is used: - 345 mg of cocoa butter stearin, (supplied by Aarhus Olie A/S, Denmark, and containing about 95% of SOS, POS and
  • NSI 3 x 33,3 % ° % La where % 0 is mole % oleic acid, and % La is mole % lauric acid
  • a culture of Pseudomonas cepacia strain DSM 3959 was transferred to a 500 ml shakeflask with 100 ml PSC-3 medium and shaken at 30°C for 1 day.
  • the composition of PSC-3 was as follows:
  • the resulting broth was used as seed culture for a 5
  • the medium was inoculated with 30 ml seed culture and fermented 5 days with good agitation and aeration at 30 C with a continuous feed of 1.5 ml/hour soybea oil and 4.2 ml/hour 18.6% (NH 4 )_S0 4 .
  • the pH was controlled at 6.3 with carbonate.
  • the culture broth was concentrated to 800 ml by an Amicon Diaflow hollow fibre cartridge. One volume 96% ethanol was added to the concentrate, the mixture was stirred for 60 minutes at 4°C and centrifugated at 4200 g. The supernatant was freezedried, .-.',> ⁇ the powder was dissolved in water to giv 8000 LU/ml.
  • lipase preparation is non ⁇ specific.
  • Cocoa " Butter Stearin (CBS) (27.2 mole-% palmitic acid, 39.6 mole-% stearic acid, 31.3 mole-% oleic acid, 0.9 mole-% linoleic acid and 1.1 mole-% arachidic acid) was used as test substrate as it has the unsaturated fatty acid (oleic and linoleic) concentrated in the 2-position of the triglycerides.
  • CBS Butter Stearin
  • the chemical randomization was carried out as follows: 3.0 g CBS was dried at 95°C for 30 minutes under vacuum by a rotary evaporator. Before addition of 30 mg sodium methoxylate (NaOCH 3 ) the temperature was reduced to 85°C and the vacuum was replaced with nitrogen at atmospheric pressure. The reaction was carried out for 1 1/2 hour under rotation before it was stopped by addition of 0.6 ml 1M HCl and washed 3 times with 5 ml deionized water at 60°C. The sample was dried at 95°C for 1 hour in the rotary evaporator before analysis. The enzymatic interesterifications were carried out with immobilized P. cepacia lipase (prepared similarly to
  • Example 2 but with a lipase load of 32,000 LU/g resin) and with Lipozyme TM IM20 (product of Novo Industri A/S), an immobilized, positionally specific lipase from Mucor miehei prepared according to EP 0 140 542. Both by the following procedure: 250 mg dry weight immobilized lipase was hydrated to 10% and 1.7 g CBS was added. The mixture was placed in a
  • the fatty acid composition in the 2-position of the triglycerides was analysed as follows: 100 mg CBS or interest -xified CBS, 3 ml pancreatic lipase solution (250 mg porcine pancreas lipase grade II from Sigma cat. no. L3126 dissolved in 10 ml 1M trisbuffer pH 8), 300 ⁇ l 2M CaCl 2 , and 0.75 ml 0.2% w/v taurocholate were mixed.
  • the emulsion was heated in a water bath at 40°C for 2 minutes and mixed on a Whirley mixer for 1 1/2 minute before the reaction was stoppe by addition of 4 ml 96% ethanol.
  • the sample was transferred t a separation funnel and extracted with 4 x 20 ml diethyl
  • the monoglyceride band was identified by iodine vapour, scraped off and extracted by 3 times 10 ml diethyl ether.
  • the ether fase was filtered, evaporated and the sample was methylated and analysed on GLC (procedure as described in AF 206/2 available from Novo Industri). The results are presented below:
  • Unsaturated fatty acid oleic and linoleic acid
  • Example 2 4.5 g of the immobilized lipase of Example 2 was filled into a water jacketed column, having an internal diameter of 1.5 cm.
  • the column was heated by use of hot circulating water, i.e. 60°C.
  • a precolumn containing water-saturated resin, (Duolite® ES561) was placed before the enzyme column ⁇ d also heated to 60 C.
  • a substrate consisting of 71% highly refined bleached and deoinized soy bean oil with a peroxide value less than 3 and 29% analytical grade lauric acid was pumped through the columns.
  • At the outlet from the enzyme column samples were taken for analysis, and the incorporation of lauric acid measured by GLC. An incorporation of 14% w/w lauric acid was attempted and the flow rate was adjusted in order to keep the conversion at that value. Whenever the precolumn was dry it was replaced by a fresh one.
  • the samples were analysed by removing the free fatty acid and mono- and diglyceride by Al 2 0 3 -column chromatography thereafter methylation of the triglyceride by NaOCH, and finally analysis of the methylester on a GLC.

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Abstract

La lipase immobilisée est positionnellement non spécifique et plus thermostable que celles précédemment connues. Elle est utile pour les processus d'hydrolyse d'esters, de synthèse d'esters et d'interestérification. Elle peut être produite par immobilisation d'une lipase à partir de Pseudomonas cepacia.
EP88907259A 1987-07-31 1988-07-22 Lipase immobilisee, positionnellement non specifique, sa production et son utilisation Ceased EP0382738A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DK399387A DK399387D0 (da) 1987-07-31 1987-07-31 Immobiliseret lipase og dennes anvendelse
DK3993/87 1987-07-31

Publications (1)

Publication Number Publication Date
EP0382738A1 true EP0382738A1 (fr) 1990-08-22

Family

ID=8129070

Family Applications (1)

Application Number Title Priority Date Filing Date
EP88907259A Ceased EP0382738A1 (fr) 1987-07-31 1988-07-22 Lipase immobilisee, positionnellement non specifique, sa production et son utilisation

Country Status (4)

Country Link
EP (1) EP0382738A1 (fr)
JP (1) JPH02504342A (fr)
DK (1) DK399387D0 (fr)
WO (1) WO1989001032A1 (fr)

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* Cited by examiner, † Cited by third party
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ATE91151T1 (de) * 1989-02-17 1993-07-15 Unichema Chemie Bv Herstellung von estern.
DK336889D0 (da) * 1989-07-07 1989-07-07 Novo Nordisk As Polypeptider
DK95490D0 (da) * 1990-04-18 1990-04-18 Novo Nordisk As Fremgangsmaade til fremstilling af triglycerid og triglyceridsammensaetning
ES2102995T3 (es) 1990-09-14 1997-08-16 Clorox Co Complejo superficie-lipasa y procedimientos de formacion y uso.
ATE141950T1 (de) * 1990-12-24 1996-09-15 Hoechst Ag Verfahren zur acylierung von alkoholen mit einem immobilisierten pseudomonas-lipase
JPH04240286A (ja) * 1991-01-25 1992-08-27 Novo Nordisk As 耐熱性リパーゼによるピッチトラブル防止法
DE69227508D1 (de) * 1991-04-10 1998-12-10 Novo Nordisk As Lipasekatalysierte esterhydrolyse
JP2986595B2 (ja) * 1991-11-08 1999-12-06 ダイセル化学工業株式会社 新規リパーゼ
JPH07504561A (ja) 1991-12-20 1995-05-25 ノボ　ノルディスク　アクティーゼルスカブ リパーゼの製造のための方法
EP0675944B1 (fr) * 1992-12-22 2003-06-25 Novozymes A/S Lipase alcaline
JPH08238088A (ja) * 1995-03-06 1996-09-17 Showa Denko Kk 高温下で高活性を有するリパーゼ
US6936289B2 (en) 1995-06-07 2005-08-30 Danisco A/S Method of improving the properties of a flour dough, a flour dough improving composition and improved food products
CA2224203C (fr) 1995-06-07 2003-08-12 Jorn Borch Soe Procede d'amelioration des proprietes d'une pate a base de farine, composition d'amelioration de la pate et produit alimentaire ameliore
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DK399387D0 (da) 1987-07-31
JPH02504342A (ja) 1990-12-13

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