CN1662264A - Deodorant composition - Google Patents

Abstract

The present invention relates to methods and compositions for reducing odor from a locus by exposing the locus to an enzymatic agent, an oxidizing agent, and chopped, sliced or comminuted plant material that naturally contains one or more phenolic compounds.

Description

Deodorant composition

field of invention

The present invention relates to enzymatic methods and compositions capable of reducing odors from human, animal and industrial sources.

Background of the invention

A wide variety of odors in the environment are part of our daily lives, such as refrigerator odors, pet and livestock odors. Since some of these odors are objectionable, various methods have been proposed for their elimination. One such conventional deodorization method is to adsorb substances that cause bad odors. Activated carbon is an example of such an adsorbent. However, since activated carbon is not easy to handle and dispose of in large quantities, natural-based, environmentally friendly alternatives are desired. The present invention provides such an alternative.

Summary of the invention

The present invention provides a deodorant composition comprising an enzymatic agent, an oxidizing agent and chopped, sliced or pulverized plant material which naturally contains one or more phenolic compounds.

In another aspect, the invention provides a method of reducing odor from a locus comprising contacting the locus with a composition of the invention.

definition

The term "odour" in the present invention is intended to mean an unpleasant smell that can be smelled by ordinary people.

The term "odorous substance" is intended to mean a substance that produces an unpleasant odour. Odor precursors refer to substances that can be converted, for example, into odorants by biochemical reactions.

The term "deodorant" refers to a substance that reduces odor.

Detailed description of the invention

Enzyme Reagent

Enzymatic reagents in the present invention include phenol oxidase and oxidizing agents.

In a preferred embodiment, the enzymatic reagent also contains one or more other enzymatic activities, such as: protease, lipase, cutinase, amylase, carbohydrase, cellulase, pectinase, mannanase , arabinanase, galactanase and/or xylanase.

phenol oxidase

In the present invention, phenol oxidase is an enzyme or a compound exhibiting phenol oxidation activity. Phenoloxidase may be peroxidase or a compound exhibiting peroxidase activity, laccase or a compound exhibiting laccase activity, catechol oxidase (EC 1.10.3.1), o-aminophenol oxidase (EC 1.10.3.4), or bilirubin oxidase (EC 1.3.3.5).

In the present invention, phenoloxidase can be typically present in an amount of 1 to 10000 μg enzyme protein per liter of aqueous solution, preferably 5 to 2000 μg enzyme protein per liter of aqueous solution, more preferably 5 to 1000 μg of enzyme protein per liter of aqueous solution, most preferably per liter of aqueous solution Concentrations of 1 to 500 μg enzyme protein are present.

Assays for detecting the activity of these enzymes are well known to those skilled in the art.

It is to be understood that phenoloxidase variants such as enzymes produced by recombinant techniques are also included within the meaning of the term "phenoloxidase".

Laccases and compounds exhibiting laccase activity

The compound exhibiting laccase activity may be any laccase in the enzyme class of EC 1.10.3.2, or any fragment thereof exhibiting laccase activity.

Preferred laccases and/or compounds exhibiting laccase activity are those enzymes derived from microorganisms. These enzymes may also be of plant, bacterial or fungal origin (including filamentous fungi and yeast).

Suitable examples from fungi can be derived from strains of Aspergillus, Neurospora, such as: N. crassa, Podospora, Botrytis ), Collybia, Fomes, Lentinus, Pleurotus, Trametes, eg, T.villosa and T.versicolor, Rhizoctonia Genus (Rhizoctonia), such as Rhizoctonia solani (R.solani), Coprinus, such as C. cinereus, C. comatus, Coprinus ferris (C. friesii) and C. plicatilis, Psathyrella, such as P. condelleana, Panaeolus, such as Butterfly P.papilionaceus, Myceliophthora, such as M.thermophila, Schytalidium, such as S.thermophilum, Polyporus, such as P.pinsitus, Phytophthora Laccases of Phlebia, such as P. radita (WO 92/01046), or Coriolus, such as C. hirsutus (JP 2-238885).

Suitable examples of bacterial origin include laccases which may be derived from strains of Bacillus.

Laccases preferably derived from Coprinus, Myceliophthora, Polyporus, Scytalidium or Rhizoctonia, in particular derived from Coprinus cinerea, Myceliophthora thermophila, Polyporus pinsitus, Scytalidiumthermophilum or Rhizoctonia solani bacterial laccase.

Laccase or laccase-related enzymes may also be produced by culturing host cells transformed with a recombinant DNA vector carrying a DNA sequence encoding said laccase and a DNA sequence encoding a function allowing expression of the laccase-encoding DNA sequence , wherein said culturing is carried out in a culture medium under conditions allowing expression of the laccase, and the laccase is recovered from the culture.

Determination of Laccase Activity (LACU)

Laccase activity can be determined by oxidation of syringaldazine under aerobic conditions (particularly applicable to Polyporus laccases). The resulting purple color was measured photometrically at 530 nm. The analysis conditions are: 19 mM syringaldazine, 23 mM acetate buffer, pH 5.5, 30° C., and the reaction time is 1 minute.

One laccase unit (LACU) is the amount of enzyme that catalyzes the conversion of 1.0 mmole of syringaldazine per minute under these conditions.

Determination of Laccase Activity (LAMU)

Laccase activity can be determined by oxidation of syringaldazine under aerobic conditions. The resulting purple color was measured at 530nm. The analysis conditions are: 19mM syringaldazine, 23mM Tris/maleate buffer, pH 7.5, 30°C, 1 minute reaction time.

One laccase unit (LAMU) is the amount of enzyme that catalyzes the conversion of 1.0 mmole of syringaldazine per minute under these conditions.

Peroxidases and compounds exhibiting peroxidase activity

The compound exhibiting peroxidase activity may be any peroxidase in the enzyme class (EC 1.11.1.7), or any fragment derived therefrom having peroxidase activity. In the present invention, compounds exhibiting peroxidase activity include peroxidase and peroxidase active fragments derived from cytochrome, hemoglobin or peroxidase.

The peroxidase used in the composition of the present invention can preferably be produced by plants (eg horseradish peroxidase or soybean peroxidase) or microorganisms such as fungi or bacteria.

Some preferred fungi include strains belonging to the subphylum Deuteromycotina, Hyphomycetes, such as: Fusarium, Humicola, Trichoderma, Lacquer Myrothecium, Verticillum, Arthromyces, Caldariomyces, Ulocladium, Embellisia, Cladosporium or Dreschlera, especially Fusarium oxysporum (DSM 2672 ), Humicola insolens, Trichoderma resii, Myrothecium verrucaria (IFO 6113), Verticillum alboatrum, Verticillum dahlie, Arthromyces ramosus (FERM P-7754) , Caldariomyces fumago, Ulocladium chartarum, Embellisia alli or Dreschlerahalodes.

Other preferred fungi include strains belonging to the subdivision Basidiomycotina, class Basidiomycetes, such as: Coprinus, Phanerochaete, Coriolus or Trametes, especially Coprinus cinereus f. microsporus (IFO 8371) , Coprinus macrorhizus, Phanerochaete chrysosporium (eg NA-12) or Trametes (formerly known as Polyporus), eg T.versicolor (eg PR4 28-A).

Other preferred fungi include strains of the subdivision Zygomycotina, class Mycoraceae, eg Rhizopus or Mucor, especially Mucorhiemalis.

Some preferred bacteria include strains of the order Actinomycetales, such as: Streptomyces spheroides (ATTC 23965), Streptomycesthermoviolaceus (IFO 12382) or Streptoverticillum verticillium ssp. verticillium.

Other preferred bacteria include Bacillus pumilus (ATCC 12905), Bacillus stearothermophilus, Rhodobacter sphaeroides, Rhodomonas palustri, Streptococcus lactis, Pseudomonas purrocinia (ATCC 15958) or Pseudomonas fluorescens Pseudomonas fluorescens (NRRL B-11).

Other preferred bacteria include strains belonging to the genus Myxococcus, such as M. virescens.

Peroxidase may also be produced by culturing host cells transformed with a recombinant DNA vector carrying a DNA sequence encoding said peroxidase and an encoding function allowing expression of the peroxidase-encoding DNA sequence wherein the culturing is carried out in a culture medium under conditions allowing expression of the peroxidase, and the peroxidase is recovered from the culture.

In particular, the recombinantly produced peroxidase is a peroxidase derived from Coprinus species, in particular C. macrorhizus or C. cinerea according to WO 92/16634.

Detection of peroxidase activity (POXU)

One peroxidase unit (POXU) is the amount of enzyme that catalyzes the conversion of 1 micromole of hydrogen peroxide per minute under the following conditions:

0.1M phosphate buffer pH7.0, 0.88mM hydrogen peroxide, 1.67mM 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 30°C.

The reaction was followed for 60 seconds (15 seconds after mixing) by the change in absorbance at 418 nm, which should be in the range of 0.15-0.30.

For the calculation of activity, an absorption coefficient of ^oxidized ABTS of 36 mM ^cm and _a stoichiometry of conversion of 1 micromole of _H2O2 per two micromole of ABTS oxidized were used.

oxidizing agent

If the phenol oxidase requires a source of hydrogen peroxide as an oxidizing agent in order to exhibit phenol oxidation activity (eg, peroxidase activity), such source can be hydrogen peroxide or a hydrogen peroxide precursor that generates hydrogen peroxide in situ. Any solid entity that upon dissolution releases peroxide that can be utilized by phenoloxidase can be used as a source of hydrogen peroxide. Compounds that can generate hydrogen peroxide upon dissolution in water or an appropriate aqueous medium include, but are not limited to: metal peroxides, percarbonates, persulfates, perphosphates, peroxyacids, alkyl peroxides , acyl peroxides, peroxyesters, carbamide peroxides, perborates and peroxycarboxylic acids or peroxycarboxylates.

Any compound that produces a peroxide that can be used by phenol oxidase to exhibit phenol oxidation activity (e.g., peroxidase activity) is an acceptable source of hydrogen peroxide in the present invention; this includes those skilled in the art. Realized many compounds. Mixtures of two or more of these substances may also be used.

Another source of hydrogen peroxide is the enzymatic systems that produce hydrogen peroxide, such as oxidases and substrates for the oxidases. Examples of combinations of oxidase and substrate include, but are not limited to: amino acid oxidase (see, e.g., US 6,248,575) with appropriate amino acids, glucose oxidase (see, e.g., WO 95/29996) and glucose, lactate oxidase, and Lactic acid, galactose oxidase (see eg WO 00/50606) and galactose, and aldose oxidase (see eg WO 99/31990) and suitable aldoses.

From studies of enzymes EC 1.1.3._, EC 1.2.3._, EC 1.4.3._, and EC 1.5.3._ or similar enzyme classes (according to the International Union of Biochemistry), such oxidases Other examples of combinations with substrates will be readily apparent to those skilled in the art.

The source of hydrogen peroxide may be added at the beginning of the process or during the process, as typically in an amount corresponding to a level of 0.001 mM to 25 mM, preferably 0.005 mM to 5 mM, and especially 0.01 to 1 mM.

If the phenol oxidase requires a source of molecular oxygen as an oxidizing agent to exhibit phenol oxidation activity (eg, laccase activity), then this source of oxygen can be atmospheric oxygen or an oxygen precursor that generates oxygen in situ. Oxygen is normally present in sufficient amounts in the atmosphere. If more _O2 is required, additional oxygen can be added, eg compressed atmospheric air or pure compressed _O2 .

plant material

Plant material in the context of the present invention is any chopped, sliced, cut, comminuted plant matter.

These plant materials may be provided on the basis of frozen material, heat-treated material (such as tea or coffee grounds), fresh material, fermented material or in any other form known in the art. An extract may contain plant material, for example, the plant material may be mashed, pulped or homogenized.

According to the invention, plant material can be obtained, for example, from fruits, vegetables, cereals, leaves, wood or any other plant objects.

Usable fruits include, but are not limited to, pome and pome fruits (apple, pear, etc.), grapes, tomatoes, citrus (orange, lemon, lime, tangerine), plum, cherry, black currant, red currant, raspberry, Strawberries, cranberries, pineapples, and any other tropical fruit.

Usable vegetables include, but are not limited to, potatoes, carrots, celery, and onions.

Useful grains include, but are not limited to, wheat and corn.

Useful leaves include, but are not limited to, tea leaves (eg, green tea leaves). Tea leaves may be residues from the tea brewing process.

Plant material may also include mixtures of various plant objects from the same plant, or more than one plant material from different plants.

Plant material may have been extracted with hot water, such as tea leaves. Hot water refers to water at a temperature of 60-120°C, preferably 70-110°C, more preferably 80-100°C. Water at a temperature above 100°C must be kept under pressure (above 1 atmosphere).

The plant material of the invention naturally contains one or more phenolic compounds. Examples of such natural phenolic compounds include, but are not limited to, lignin, lignosulfonic acid, humic acid, nitrohumic acid, tannin, catechin, gallic acid, urishiol, hesperidin, guaiac guaiachol, 4-methyl-guaiacol, 4-ethyl-guaiacol, 4-isopropylcycloheptadienolone.

combination

The present invention provides a deodorant composition comprising:

- enzymatic reagents containing phenoloxidase and oxidizing agents; and

- Chopped, sliced or pulverized plant material which naturally contains one or more phenolic compounds.

The deodorant composition can be formulated as a slurry, suspension, paste, solid or dry product formulation. The dry product formulation can then be rehydrated to form an active composition that can be used in the methods of the invention.

When formulating the composition as a dry product, the ingredients may be blended, arranged in separate layers or packaged individually.

When formulated as a solid, all ingredients may be mixed together, for example, in powder form.

When using the composition other than in dry form, and even then, it is preferred to use a two-part formulation system that separates the enzyme from the rest of the composition.

The compositions of the present invention may also contain adjuvants such as wetting agents, thickeners, buffers, stabilizers, fragrances, colorants, fillers and the like.

Useful wetting agents are surfactants, ie, nonionic, anionic, amphoteric or zwitterionic surfactants.

The compositions of the invention may be concentrated products or ready-to-use products. In use, the concentrated product is typically diluted with water to provide a medium with effective odor reducing activity, applied to the locus to be deodorized and allowed to react with odorants or odorant precursors present.

Method and use

The present invention provides a method of reducing odor from a locus comprising exposing the locus to:

- enzymatic reagents containing phenoloxidase and oxidizing agents; and

- Chopped, sliced or comminuted plant material which naturally contains one or more phenolic compounds.

A locus can be brought into contact with a composition of the invention by soaking the locus with an aqueous formulation (e.g., a suspension) of the composition of the invention, by spraying the locus with the composition of the invention, by applying the composition of the invention, e.g. Material coating place. The composition can also be mixed with objects containing odorous substances. The loci are brought into contact with the composition, thereby allowing the odorants in the loci to react with the composition. Any method of applying the composition to a locus whereby odors from the locus can be reduced is an acceptable method of practice. Those skilled in the art will readily recognize such methods.

In another embodiment, the locus is contacted with the composition at a temperature of 5 to 80°C, preferably 10 to 50°C, more preferably 15 to 40°C, most preferably room temperature.

In another embodiment, the locus is contacted with the composition between pH 2 and pH 10, preferably between pH 3.5 and pH 9, more preferably between pH 5 and pH 9.

In yet another embodiment, the phenoloxidase and oxidizing agent are laccase and a source of oxygen, or peroxidase and a source of hydrogen peroxide.

After 2 days at 20° C. and a relative humidity of 60-90%, compared to those same surfaces that do not implement the method of the present invention, the method of the present invention can reduce the odor from the place to below 80% (preferably below 50%, more preferably 30% or less). % or less, most preferably less than 20%, especially less than 10%).

Odor Evaluation

Odor can be assessed by a sensory panel of at least 5 trained individuals. The intensity of the odor is indicated on a scale of 0 to 10, with 0 being "no odor" and 10 being "very strong odor". All evaluations were performed twice and averages were calculated.

Alternatively, since compounds emitting bad taste (odor substances) are often volatile compounds such as methyl mercaptan, conventional analytical methods such as gas chromatography and gas detection tubes can be used to detect and quantify these compounds.

The following examples describe the present invention in more detail, and these examples should not be construed as limiting the scope of the present invention.

Example

Chemical reagents used as buffers and substrates were commercial products of at least reagent grade.

Example 1

Deodorant composition with laccase and tea leaves

Enzyme: Myceliophthora thermophila laccase with an activity of 276 LAMU/g (disclosed in patent application WO9533836, available from Novozymes A/S, Denmark)

Tea: Green tea (dried form), trade name "maccha iri ryokucha" (ITO EN, Ltd., Japan).

odor detector

Gas sampling pump kit: GV-100S (Gastec Corporation)

Gas detection tube: No.71 H, methyl mercaptan (Gastec Corporation)

Preparation of green tea residue

400ml of water (80°C) was mixed with 20g of green tea, and after waiting for 40 seconds, the water was removed with a cotton fiber filter.

Preparation of standard odor solutions

Methyl mercaptan, sodium salt, about 15% dissolved in water (Tokyo Kasei Kogyo Co., Ltd.), diluted with water at a ratio of 1:100.

experimental method

In four 50ml glass test tubes, green tea residue was mixed with standard odor solution and enzyme in the amounts shown in Tables 1-4, and then thoroughly mixed. The detection tube was sealed with plastic film and incubated at 25°C for 1 hour. Then use a gas detection tube (No.71 H, with a gas sampling pump: GV-100S) to sample 50 microliters from each detection tube, and read the color scale (ppm value of methyl mercaptan) on the detection tube for analysis. This experiment was repeated 4 times.

result

The "Deodorant Efficacy" values in Tables 1-4 clearly show that the combination of laccase and plant material (green tea grounds) is very effective in removing odorants (methyl mercaptan).

Table 1 Test tube number Standard odor solution (microliter) Green tea residue (g) Laccase solution (microliter) Gas detector reading Deodorant efficacy (%) 1 100 0 100 140 0 2 100 0.5 0 140 0 3 100 0.5 50 25 82 4 100 0.5 100 5 96

Table 2 Test tube number Standard odor solution (microliter) Green tea residue (g) Laccase solution (microliter) Gas detector reading Deodorant efficacy (%) 1 100 0 100 150 0 2 100 0.5 0 150 0 3 100 0.5 50 25 83 4 100 0.5 100 8 95

table 3 Test tube number Standard odor solution (microliter) Green tea residue (g) Laccase solution (microliter) Gas detector reading Deodorant efficacy (%) 1 100 0 100 150 0 2 100 0.5 0 150 0 3 100 0.5 50 30 80 4 100 0.5 100 7 95

Table 4 Test tube number Standard odor solution (microliter) Green tea residue (g) Laccase solution (microliter) Gas detector reading Deodorant efficacy (%) 1 100 0 100 140 0 2 100 0.5 0 140 0 3 100 0.5 50 25 82 4 100 0.5 100 5 96

Claims (5)

1, a kind of deodorant compositions, it comprises:

-contain the zymetology reagent of phenol oxidase and oxidant, reach

-that shred, section or comminuted plants material, natural one or more phenolic compounds that contains of this vegetable material.

2, according to the compositions of claim 1, wherein vegetable material is timber, tea, coffee or Semen Fagopyri Esculenti.

3, according to the compositions of claim 1 or 2, vegetable material wherein extracts with hot water.

4, remove method, comprise that each the compositions that makes place and claim 1-3 contacts from the abnormal smells from the patient in place.

5, the natural vegetable material that contains one or more phenolic compounds laccase and chopping, section or that pulverize is reducing from the purposes in the abnormal smells from the patient in place.