CN1130483C - Fabric treated with cellulose and oxidoreductase - Google Patents

Abstract

The present invention relates to a method for imparting a worn appearance to dyed fabric with reduced strength loss, the method comprising: (a) contacting the dyed fabric with cellulase enzymes in an aqueous medium at a concentration of 0.01-250 μg enzyme protein; (b) Simultaneously or subsequently treating the above fabric with a phenol oxidase system and an enhancer.

Description

Fabric treated with cellulase and oxidoreductase

field of invention

The present invention relates to a method of imparting a worn appearance to dyed fabrics, especially cellulosic fabrics such as denim.

Background technique

Over the past few years, a new industry has emerged, the so-called "jeans laundering" branch, which responds to the popular need for stylish, casual and comfortable clothing.

Initially, all indigo jeans on the market are stiff and uncomfortable when initially purchased due to the finishing system used in the denim fabric.

The first step in the development of the finishing method was to sell the manufacturer's starched jeans. But these "-prewashed" jeans faded only slightly and had a softer, more comfortable feel, as if they had been washed several times. This is a popular trend today, and consumers are willing to pay extra for this additional processing.

Shortly after the introduction of prewashed jeans, the idea of using whetstones to speed up the aging process was developed, and "stonewashing" became a second step in process improvement. Include volcanic stones in the wash, or tumble volcanic stones with wet clothing to soften the stiffest parts like trouser waists, cuffs, and pockets.

However, the use of stones to wear jeans can be extremely damaging to equipment and fabrics, so cellulase treatment is now often used instead of stone washing, or a combination of the two methods is applied to achieve the desired worn appearance. See "AATCC: Handbook of Apparel Wet Processing Techniques", 1994, published by the American Association of Textile Chemists and Colorists, pp. 19-21.

Application of the stonewashing process described above reduces fabric strength, and non-stonewashing cellulase treatments alone do not achieve the desired worn appearance, so there is a need in the industry for a more gentle treatment.

Summary of the invention

Surprisingly, it has been found that by combining a cellulase treatment process with a treatment using a phenol oxidase system and a strengthening agent, it is possible to achieve the desired fabric worn appearance with minimal loss of strength. Accordingly, the present invention relates to a method of imparting a worn appearance to dyed fabrics with reduced strength loss, the method comprising:

(a) bringing the dyed fabric into contact with cellulase at a concentration corresponding to 0.01 to 250 μg of enzyme protein per g of fabric in an aqueous medium;

在该式中，A表示诸如-D、-CH＝CH-D、-CH＝CH-CH＝CH-D、-CH＝N-D、-N＝N-D或者-N＝CH-D之类的基团，其中D选自由-CO-E、-SO₂-E、-N-XY和-N⁺-XYZ组成的组，其中E可以是-H、-OH，-R或者-OR，并且X、Y和Z可以是相同的或者不同的且选自-H和-R；R是C₁-C₁₆烷基，优选的是C₁-C₈烷基，其中烷基可以是饱和或不饱和的，有支链或无支链的，同时可以可选择地由羧基、磺基或者氨基基团取代；并且B和C可以相同或不同，且可选自C_mH_2m+1；1≤m≤5；或由式II表示：

(b) Simultaneously or subsequently treating the aforementioned fabric with a phenol oxidase system and a reinforcing agent, wherein the reinforcing agent may be described by formula I:

In this formula, A represents a group such as -D, -CH=CH-D, -CH=CH-CH=CH-D, -CH=ND, -N=ND or -N=CH-D , where D is selected from the group consisting of -CO-E, -SO ₂ -E, -N-XY and -N ⁺ -XYZ, where E can be -H, -OH, -R or -OR, and X, Y and Z may be the same or different and selected from -H and -R; R is a C ₁ -C ₁₆ alkyl group, preferably a C ₁ -C ₈ alkyl group, wherein the alkyl group may be saturated or unsaturated, Branched or unbranched, and optionally substituted by carboxyl, sulfo or amino groups; and B and C may be the same or different, and may be selected from C _m H _2m+1 ; 1≤m≤5 ; or represented by formula II:

In this formula, X represents -O- or -S-, and the substituent groups R ¹ -R ⁹ may be the same or different, and independently represent any of the following groups: hydrogen, halogen, hydroxyl, formyl, Carboxyl and esters and their salts, carbamoyl, sulfo and esters and their salts, sulfamoyl, nitro, amino, phenyl, C ₁ -C ₁₄ -alkyl, C ₁ -C ₅ - Alkoxy, carbonyl-C ₁ -C ₅ -alkyl, aryl-C ₁ -C ₅ -alkyl; wherein carbamoyl, sulfamoyl and amino groups can also be unsubstituted or substituted by the substituent R ¹⁰ One or two times; where phenyl may also be unsubstituted or substituted by one or more substituents R ¹⁰ ; where C ₁ -C ₁₄ -alkyl, C ₁ -C ₅ -alkoxy, carbonyl-C ₁ - C ₅ -Alkyl and aryl-C ₁ -C ₅ -Alkyl groups can be saturated or unsaturated, branched or unbranched and can also be unsubstituted or replaced by one or more substituents R ¹⁰ replaced;

Wherein the substituent ^R represents any of the following groups: halogen, hydroxyl, formyl, carboxyl and esters and their salts, carbamoyl, sulfo and esters and their salts, sulfamoyl, nitro, amino , phenyl, aminoalkyl, piperidino, piperazinyl, pyrrolidino (pyrrolidino), C ₁ -C ₅ -alkyl, C ₁ -C ₅ -alkoxy; wherein carbamoyl, sulfamoyl Acyl and amino groups can also be unsubstituted or substituted once or twice by hydroxyl, C ₁ -C ₅ -alkyl, C ₁ -C ₅ -alkoxy; wherein phenyl can also be substituted by one or more of the following groups Substituted by: halogen, hydroxy, amino, formyl, carboxyl and esters and their salts, carbamoyl, sulfo and esters and their salts, and sulfamoyl; where C ₁ -C ₅ -alkyl and C ₁ -C ₅ -Alkoxy can also be saturated or unsaturated, branched or unbranched, and can also be substituted one or two times by any of the following groups: halogen, hydroxyl, amino, methyl Acyl, carboxyl and esters and their salts, carbamoyl, sulfo and esters and their salts, and sulfamoyl;

Or in the general formula, two of the substituents R ¹ -R ⁹ may jointly form a group -B-, wherein B represents any of the following groups: (-CHR ¹⁰ -N=N-), (- CH=CH-) _n , (-CH=N-) _n or (-N=CR ¹⁰ -NR ¹¹ -), n in the group is an integer between 1-3, and R ¹⁰ is the substitution defined above group, R ¹¹ is defined as R ¹⁰ . DETAILED DESCRIPTION OF THE INVENTION Comparison between Bleached Appearance and Worn Appearance

Those skilled in the art of evaluating denim finishing processes will be able to differentiate the bleached appearance of denim from the worn appearance.

The former is the result of dye removal (bleaching) from dyed warp yarns. Since bleaching occurs over the entire surface of each dyed warp yarn, the result is an overall reduction in color depth. Thus, the bleached look of a pair of indigo jeans is characterized by a lighter bluish hue than the corresponding reference standard.

The latter - a scuffed appearance, is the result of treating denim with cellulase and/or pumice stones. This method has the characteristic of removing the dye from the fabric non-uniformly, so it creates a sharp contrast between the dyed and bleached areas (areas where the dye has been removed).

Typically, a worn appearance is obtained from a method comprising cellulase and/or pumice, while a bleached appearance may be obtained from a method comprising a non-enzymatic bleach such as hypochlorite or a method comprising an oxidoreductase and a booster.

The present invention relates to a method of providing a worn look rather than a bleached look comprising a mild treatment with a cellulase followed by a mild treatment with an oxidoreductase and a strengthening agent. dyed fabric

The invention can be applied to any dyed fabric known in the art, especially synthetic or natural fabrics such as polyester.

The present invention is most suitable for use on cellulose-containing fabrics, such as cotton, viscose, rayon, ramie, flax, Tencel or mixtures thereof, or mixtures of any of the above fibers, or combinations of any of the above fibers and synthetic fibers. mixture. Especially denim fabric.

Fabrics can be dyed with vat dyes such as indigo, or dyes related to indigo such as thioindigo. Fabrics can also be dyed with more than one dye, for example: first with sulfur dyes and then with indigo dyes, or vice versa.

In the most preferred embodiment of the invention, the fabric is a denim with a sulfur base colored indigo (i.e., the denim is first dyed with a sulfur dye and then indigo); including fabrics made therefrom. clothing. cellulase

As used herein, the term "cellulase" refers to an enzyme that catalyzes the degradation of cellulose into glucose, cellobiose, triose and other cellulosic oligosaccharides.

In this context, the term "cellulase" is understood to include the mature protein or its precursor form or a functional fragment thereof (which fragment must have full-length enzymatic activity). Furthermore, the term "cellulase" is meant to include homologues or analogs of the above enzymes. Such homologues contain an amino acid sequence that exhibits at least 60% identity to the amino acid sequence of the parent enzyme (ie, the parent cellulase). The degree of identity can be determined by conventional methods, see for example: Altshul et al., Advances in Mathematical Biology (Bull. Math. Bio.), 48, 1986, pp. 603-616, and Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA.89, 1992, pp.10915-10919.

The cellulases used in the present invention are preferably monocomponent (recombinant) cellulases, ie must not include other proteins or cellulase proteins. Recombinant cellulase components can be cloned and expressed according to standard routine techniques well known to those skilled in the art.

In a preferred embodiment of the invention, the cellulase used in the above method is an endoglucanase (EC 3.2.1.4), preferably a monocomponent (recombinant) endoglucanase.

The cellulase is preferably a microbial cellulase, more preferably a bacterial or fungal cellulase.

Examples of bacterial cellulases are cellulases derived from or produced by bacteria from the genus group consisting of Pseudomonas or Bacillus, especially Bacillus candidiasis.

The cellulase or endoglucanase may be an acidic, neutral or alkaline cellulase or endoglucanase, i.e. showing maximum cellulolysis in the acidic, neutral or alkaline range respectively Active cellulase or endoglucanase.

Thus, useful cellulases are acid cellulases, preferably fungal acid cellulases, derived from or consisting of species from the genera Trichoderma, Actinomyces, Myromyces, Aspergillus and Botrytis. Produced by fungi in the genus group.

Preferred useful acid cellulases are derived from or produced by fungi from the group of genus consisting of Trichoderma viride, Trichoderma reesei, Trichoderma longibrachiatum, Myrothecium verrucaria, Aspergillus niger, Aspergillus oryzae and Botrytis cinerea.

Another useful cellulase or endoglucanase is a neutral or alkaline cellulase, preferably a fungal neutral or alkaline cellulase derived from or derived from Aspergillus, Penicillium, Myceliophthora, Humicola, Rakemycetes, Fusarium, Botrytis, Pleurotus, Chaetomium, Verrucospora, Verticillium, Lactophylla, Produced by fungi in the group of genera consisting of Papulospora, Gliocladium, Cephalosporium and Acremonium.

Preferred alkaline cellulases are derived from or produced by fungi from the group of genus consisting of Humicola insolens, Fusarium oxysporum, Myceliopthora thermophila or Cephalosporium species, preferably derived from or produced by Humicola insolens (DSM 1800 ), Fusarium oxysporum (DSM2672), Myceliopthora thermophila (CBS117.65) or Cephalosporium species (RYM-202) by fungi in the group of genera.

A preferred example of a native or parent cellulase is an alkaline dextranase that is synthesized against a highly purified ~43kD endoglucanase derived from Humicola insolens (DSM1800) The antibodies produced were immunoreactive, or the alkaline glucanase was a derivative of a ~43kD endoglucanase that exhibited cellulase activity.

Other examples of useful cellulases are variants of the parent cellulase having fungal origin (eg, cellulases derived from fungal strains of Humicola, Trichoderma, or Fusarium).

According to the invention, the cellulase concentration in the aqueous medium may be 0.01-250 μg enzyme protein per g fabric, preferably 0.1-250 μg enzyme protein per g fabric, especially 0.5-50 μg enzyme protein per g fabric. Determination of cellulase activity (ECU)

In the context of the present invention cellulase activity may be expressed by the ECU. Cellulolytic enzymes hydrolyze CMC, thereby increasing the viscosity of the cultured mixture. The resulting decrease in viscosity can be measured by an oscillating viscometer (eg MIVI 3000 from Sofraser, France).

Cellulolytic activity according to the ECU assay can be determined according to the following analytical method: The ECU assay quantifies the amount of catalytic activity present in a sample by measuring the ability of the sample to reduce the viscosity of a carboxymethylcellulose (CMC) solution. The conditions under which the assay was carried out were: temperature 40°C, pH 7.5, 0.1M phosphate buffer, time 30 minutes, use of a relevant enzyme standard to reduce the viscosity of the CMC (carboxymethylcellulose Hercules 7LFD) substrate, enzyme The concentration is about 0.15ECU/ml. The main standard is set at 8200 ECU/g. phenoloxidase system

The term "phenol oxidase system" refers to a system in which an enzyme is able to oxidize organic compounds containing phenolic groups by using hydrogen peroxide or molecular oxygen. Examples of such enzymes are peroxidases and oxidases.

If the phenol oxidase system requires a source of hydrogen peroxide, then the source may be hydrogen peroxide or an in situ generated hydrogen peroxide precursor for hydrogen peroxide, such as percarbonate or perborate, or Enzyme systems that generate hydrogen peroxide, such as oxidases and oxidase substrates, are either amino acid oxidases and suitable amino acids, or peroxycarboxylic acids or salts thereof. Hydrogen peroxide may be added at the beginning or during the process, for example at _a concentration comparable to 0.001-25 mM _H2O2 .

If the phenoloxidase system requires molecular oxygen, then molecular oxygen from the atmosphere is usually sufficient.

The enzyme in the phenoloxidase system may be an enzyme exhibiting peroxidase activity, or a laccase or a laccase-related enzyme as described below.

According to the present invention, the concentration of phenoloxidase in the aqueous medium may be 0.01-250 μg enzyme protein per g fabric, preferably 0.1-250 μg enzyme protein per g fabric, especially 0.5-50 μg enzyme protein per g fabric. Peroxidases and compounds that act as peroxidases

The enzyme exhibiting peroxidase activity may be any peroxidase included in the enzyme classification (EC 1.11.1.7), or any fragment derived therefrom exhibiting peroxidase activity, or Synthetic or semi-synthetic derivatives of peroxidases (for example, porphyrin ring systems or microperoxidases, see for example: US 4,077,768, EP 537 381, WO 91/05858 and WO 92/16634). These enzymes are well known from microorganisms, plants and animals.

The peroxidase used in the method of the invention is preferably produced in plants (eg horseradish or soybean peroxidase) or microorganisms such as fungi or bacteria. Some preferred fungi include those belonging to the subphylum Deuteromycotina, class Hyphomycetes, for example: Fusarium, Humicola, Trichoderma, Myromyces, Verticillium, Arthromyces, Caldariomyces, Ulocladium, Strains of Embellisia, Cladosporium, or Dreschlera, especially Fusarium oxysporum (DSM 2672), Humicola insolens, Trichoderma resii, Myrothecium verrucana (IFO6113), Verticillum alboatrum, Verticillum dahlie, Arthromycesramosus (FERM P-7754), Caldariomyces lofumago, art , Embellisia alli or Dreschlera halodes.

Other preferred fungi include strains belonging to the subdivision Basidiomycotina, class Basidiomycetes, e.g. Coprinus, Spreadtodenta, Coriolus or Trametes, especially Coprinus cinerea f. microsporus (IFO8371), Coprinus longifolia, Phanerochaete chrysosPorium (eg NA-12), or Trametes (formerly known as Polyporus), such as Trametes versicolor (eg PR428-A).

More preferred fungi include strains belonging to the subdivision Zygomycetes, class Mycoraceae, for example the genus Rhizopus or the genus Mucor, especially Mucor subtilis.

Some preferred bacteria include strains of the order Actinomycetes, such as Streptomyces sphaeroides (ATCC 23965), Streptomyces thermopurpurinas (IFO 12382) or Verticillium sp. Verticillium subsp.

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

More preferred bacteria include strains belonging to the genus Myxococcus, for example: Myxococcus viridans.

The peroxidase may also be a peroxidase produced by a method comprising: culturing a recombinant DNA vector (carrying the above-mentioned The DNA sequence of the oxidase and the DNA sequence encoding the function of enabling the expression of the DNA sequence encoding the above-mentioned peroxidase) transformed host cells and recovery of the peroxidase from the culture.

In particular, recombinantly produced peroxidases are produced from Coprinus species, especially Coprinus longifolia or Coprinus cinereus according to WO 92/16634, or derived from variants thereof, as described in WO 94/12621 variant of .

In the context of the present invention, compounds acting as peroxidases comprise active fragments of peroxidases derived from cytochromes, hemoglobin or peroxidases, and synthetic or semisynthetic derivatives thereof (e.g. iron porphyrin, iron porphyrin and iron phthalocyanine and their derivatives). Determination of peroxidase activity

1 peroxidase unit (PODU) refers to the amount of enzyme required to catalyze the conversion of 1 μmole hydrogen peroxide per minute under the following analytical conditions: 0.88 mM hydrogen peroxide, 1.67 mM 2,2'-azinobis(3- Ethylbenzothiazoline-6-sulfonate), 0.1M phosphate buffer, pH 7.0, culture temperature 30°C, wavelength measured by photometer 418nm. Laccase and laccase-related enzymes

In the context of the present invention, laccases and laccase-related enzymes mean any laccase included in the enzyme classification (EC 1.10.3.2), any chatechol included in the enzyme classification (EC 1.10.3.1) Oxidase, any bilirubin oxidase included in the enzyme class (EC 1.3.3.5), or any monohydric phenol mono(add)oxygenase included in the enzyme class (EC 1.14.99.1).

It is well known that laccases are derived from microbial and plant sources. Microbial laccases may be of bacterial or fungal origin (including filamentous fungi and yeasts), suitable examples include those obtained from Aspergillus, Neurospora (e.g. Neurospora crassa), Podospora, Botrytis, Phytophthora, Laminaria, Lentinus, Pleurotus, Trametes (e.g. T.villosa and Trametes versicolor), Rhizoctonia (e.g. Rhizoctonia solani), Coprinus (e.g. Coprinus and Coprinus cinerea), Psatyrella, Myceliophthora (e.g. M.thermophila), Schytalidium, Polyporus (e.g. P. pinsitus), Phytophthora (e.g. P.radita) (WO 92/01046)) or a laccase of a strain of Versicolor versicolor (eg Versicolor versicolor (JP 2-238885)).

In addition, the laccase or an enzyme related to laccase may be a laccase produced by a method comprising: culturing a recombinant DNA vector (carrying the The DNA sequence of the above-mentioned laccase and the DNA sequence encoding the function of enabling the expression of the DNA sequence encoding the above-mentioned laccase) transformed host cells and recovery of the laccase from the culture. Determination of Laccase Activity (LACU)

Laccase activity can be determined by oxidation of syringaldazine under aerobic conditions. The resulting violet color was analyzed photometrically at a wavelength of 530 nm. The analysis conditions were 19 μM syringaldazine, 23.2 mM acetate buffer, pH 5.5, 30° C., and a reaction time of 1 minute.

1 laccase unit (LACU) refers to the amount of enzyme required to catalyze the conversion of 1.0 μmole of syringaldazine per minute under the above conditions. enhancer

According to the present invention, an enhancer is any compound which enhances the bleaching process. A typical enhancer is an organic compound such as that described by one of the following formulas:

在式中，A是一个基团，如：-D、-CH＝CH-D、-CH＝CH-CH＝CH-D、-CH＝N-D、-N＝N-D或者-N＝CH-D，其中D选自由-CO-E、-SO₂-E、-N-XY和-N⁺-XYZ组成的组，其中E可以是-H、-OH、-R或者-OR，并且X、Y和Z可以是相同或者不同的且选自-H和-R；R是C₁-C₁₆烷基，优选的是C₁-C₈烷基，其中烷基可以是饱和或者不饱和的，是有支链或者无支链的，且可以可选择地用羧基、磺基或者氨基取代；同时B和C可以是相同或者不同的，并且可选自C_mH_2m+1；1≤m≤5。The enhancer can be represented by the following formula I:

In the formula, A is a group, such as: -D, -CH=CH-D, -CH=CH-CH=CH-D, -CH=ND, -N=ND or -N=CH-D, wherein D is selected from the group consisting of -CO-E, -SO ₂ -E, -N-XY and -N ⁺ -XYZ, wherein E can be -H, -OH, -R or -OR, and X, Y and Z can be the same or different and selected from -H and -R; R is C ₁ -C ₁₆ alkyl, preferably C ₁ -C ₈ alkyl, wherein the alkyl can be saturated or unsaturated, is Branched or unbranched, and may be optionally substituted with carboxy, sulfo or amino; while B and C may be the same or different, and may be selected from C _m H _2m+1 ; 1≤m≤5.

In a preferred embodiment, A in the above formula is -CO-E, wherein E can be -H, -OH, -R or -OR; R is C ₁ -C ₁₆ alkyl, preferably C ₁ - C ₈ Alkyl, where the alkyl can be saturated or unsaturated, branched or unbranched, optionally substituted with carboxyl, sulfo or amino; while B and C can be the same or different , and may be selected from C _m H _2m+1 ; 1≤m≤5.

In the above formula, A may be placed at the meta position of the hydroxyl group instead of at the para position shown in the above formula.

In particular embodiments, the enhancer is acetosyringone, methyl syringate, ethyl syringate, propyl syringate, butyl syringate, hexyl syringate, or octyl syringate.

The above enhancers can be prepared by methods well known to those skilled in the art; some enhancers are also commercially available, for example: syringone acetate. Methyl syringate, ethyl syringate, propyl syringate, butyl syringate, hexyl syringate and octyl syringate can be produced as disclosed in Chem. Ber. 67 , 1934, p.67.

The enhancer used in the present invention can also be represented by the following formula II:

In this formula, X represents (-O-) or (-S-); the substituents R ¹ -R ⁹ may be the same or different, and independently represent any of the following groups: hydrogen, halogen, hydroxyl, Formyl, carboxyl and esters and their salts, carbamoyl, sulfo and esters and their salts, sulfamoyl, nitro, amino, phenyl, C ₁ -C ₁₄ -alkyl, C ₁ - C ₅ -alkoxy, carbonyl-C ₁ -C ₅ -alkyl, aryl-C ₁ -C ₅ -alkyl; where carbamoyl, sulfamoyl and amino can also be unsubstituted or substituted by R ¹⁰ is substituted once or twice; where phenyl can also be unsubstituted or substituted by one or more substituents R ¹⁰ ; where C ₁ -C ₁₄ -alkyl, C ₁ -C ₅ -alkoxy, carbonyl-C ₁ -C ₅ -Alkyl, aryl-C ₁ -C ₅ -Alkyl can be saturated or unsaturated, branched or unbranched, unsubstituted or replaced by one or more Substituent R ¹⁰ is substituted;

Wherein the substituent R ¹⁰ represents any of the following groups: halogen, hydroxyl, formyl, carboxyl and esters and their salts, carbamoyl, sulfo, and esters and their salts, sulfamoyl, nitro , amino, phenyl, aminoalkyl, piperidino, piperazinyl, pyrrolidinyl, C ₁ -C ₅ -alkyl, C ₁ -C ₅ -alkoxy; wherein carbamoyl, sulfamoyl And the amino group can also be unsubstituted or substituted once or twice by hydroxyl, C ₁ -C ₅ -alkyl, C ₁ -C ₅ -alkoxy; at the same time, the phenyl can also be substituted by one or more of the following groups Substituted by: halogen, hydroxyl, amino, formyl, carboxyl and esters and their salts, carbamoyl, sulfo and esters and their salts, and sulfamoyl; where C ₁ -C ₅ -alkyl , C ₁ -C ₅ -alkoxy can be saturated or unsaturated, branched or unbranched, and can be substituted once or twice by any of the following groups: halogen, hydroxyl, amino, Formyl, carboxyl and esters and their salts, carbamoyl, sulfo and esters and their salts, and sulfamoyl;

Alternatively, in the general formula, two of the substituents R ¹ -R ⁹ may together form a group -B-, wherein B represents any of the following groups: (-CHR ¹⁰ -N=N-), (-CH =CH-) _n , (-CH=N-) _n or (-N=CR ¹⁰ -NR ¹¹ -), in the above groups n represents an integer from 1 to 3, and R ¹⁰ is a substituent as defined above , R ¹¹ is defined as R ¹⁰ .

In particular embodiments, the enhancer is 10-methylphenothiazine, phenothiazine-10-propionic acid, N-hydroxysuccinamide, phenothiazine-10-propionic acid, 10-ethylphenothiazine -4-carboxylic acid, 10-ethylphenothiazine, 10-propylphenothiazine, 10-isopropylphenothiazine, phenothiazine-10-methyl propionate, 10-phenylphenothiazine, 10-allylphenothiazine, 10-(3-(4-methylphenothiazin-1-yl)propyl)phenothiazine, 10-(2-pyrrolidin-1-yl-ethyl)phen Thiazine, 2-methoxy-10-methyl-phenothiazine, 1-methoxy-10-methylphenothiazine, 3-methoxy-10-methylphenothiazine, 3,10- Dimethylphenothiazine, 3,7,10-trimethylphenothiazine, 10-(2-hydroxyethyl)phenothiazine, 10-(3-hydroxypropyl)phenothiazine, 3-(2 -Hydroxyethyl)-10-methylphenothiazine, 3-hydroxymethyl-10-methylphenothiazine, 3,7-dibromophenothiazine-10-propionic acid, phenothiazine-10-propane Amide, chlorpromazine, 2-chloro-10-methylphenothiazine, 2-acetyl-10-methylphenoxazine, 10-methylphenoxazine, 10-ethylphenoxazine, phenoxazine Oxazine-10-propionic acid, 10-(2-hydroxyethyl)phenoxazine or 4-carboxyphenoxazine-10-propionic acid.

Enhancers can be obtained from Sigma-Aldrich, Janssen Chimica, Kodak, Tokyo Kasai Organic Chemicals, Daiichi Pure Chemicals, or Boehringer Mannheim; N-methylated derivatives of phenothiazines and phenoxazines can be obtained by base iodides (as described by Cornel Bodea and Ioan Silberg in "Recent Advances in Phenothiazine Chemistry" (Advances in Heterocyclic Chemistry, 1968, Vol.9, pp.321-460); B. Cardillo and G. Casnati in four body, 1967, Vol.23, p.3771 described) methylation preparation; phenothiazine and phenoxazine propionic acid can be used as described in Journal of Organic Chemistry, 15, 1950, pp.1125-1130 method of preparation. The hydroxyethyl and hydroxypropyl derivatives of phenothiazines and phenoxazines can be prepared as described by G. Cauquil, Bulletin de la Society Chemique de France, 1960, p.1049.

The strengthening agent of the present invention may be present in a concentration of 0.05-500 μmole strengthening agent per g denim, preferably from 0.05-100 μmole strengthening agent per g denim, especially from 0.05-20 μmole strengthening agent per g denim. industrial application

Typically, the invention is applied to industrial machine production for the cellulase treatment of fabrics.

The fabric is typically loaded into the machine at the machine capacity stated by each manufacturer. The fabric can be added to the machine before or after it is in the water.

Typically, cellulase treatment is performed first, followed by treatment with the phenol oxidase system and enhancer, but the two methods can be performed simultaneously or vice versa.

Cellulase enzymes can be placed in the water before being placed on the fabric, or can be added after the fabric has been soaked. In general, buffers can be utilized in order to approximate the pH optimum of the enzyme in question. After contacting the fabric with the cellulase enzyme, the fabric should be agitated in the machine long enough to ensure complete wetting of the fabric and activity of the enzyme. Typically, a reaction time of 5-60 minutes and a reaction temperature of 20°C-90°C (preferably 30°C-80°C, most preferably 40°C-70°C) will be suitable.

The phenoloxidase system and enhancer of the present invention can be added to the water prior to incorporation into the fabric, or can be added after the fabric has been soaked. The phenol oxidase system can be added at the same time as the enhancer, or it can be added separately. Typically, a buffer is utilized in order to approximate the pH optimum of the enzyme in question. After contacting the fabric with the phenoloxidase system and enhancer of the present invention, the fabric should be agitated in the machine for a sufficient time to ensure complete wetting of the fabric and activation of the enzyme system and enhancer. Typically, a reaction time of 5-60 minutes and a reaction temperature of 20°C-90°C (preferably 30°C-80°C, most preferably 40°C-70°C) will be suitable.

The process steps described above can be carried out once or can be repeated two or three times, depending on what kind of worn look the dyed fabric is to achieve.

Additionally, the invention will be illustrated by the following examples, which are in no way intended to limit the scope of the claimed invention.

Example 1

Treatment of denim with cellulase and laccase/enhancer

Treatment of denim was carried out on industrial scale equipment (450 liters) using 50 kg of denim.

Five different types of denim (all produced by the Levi Strauss Company) were used in the present invention. All five denims are based on sulfur dyes, but the ratio of indigo to sulfur dye varies and the structure of the fabric varies.

Step 1 : Abrasion with Cellulase/Pumice Stone

Denim is broken down into two distinct wear methods:

1) The standard abrasion method is to use neutral cellulase + pumice or

2) Abrasion method without pumice

        1: 750g MTF12EB (neutral cellulase, available from UK

Acquired by T.S. Chemicals]

64kg pumice stone

50 minutes, pH6.5, 60℃

                50kg each time denim denim

2: 750g MTF12EB (neutral cellulase, available from T.S.Chemicals, UK)

  50 minutes, pH6.5, 60°C

              50kg each time denim

Step 2 : Treatment with Laccase and Enhancer

The jeans treated in the first step (excluding one pair of jeans of each type kept as a control) were then treated with laccase and enhancer at the following doses and conditions:

      270g Monosodium Phosphate

68g disodium phosphate

        40.5g PPT (Phenothiazine 10-propionate)

40000 LACU (=625mg) Trametes villosa laccase (available from Novo Nordisk A/S)

     12 minutes, pH6-6.5, 60°C

              50kg each time denim

As a result of the treatment: each denim produced four different looks (+/- pumice in cellulase treatment and +/- laccase treatment method). The above treated jeans were then subjected to an appearance review by 6 experts in the field (proficient in evaluating denim finishing methods). The main conclusion from their evaluation was that the cellulase treatment without pumice produced significantly less abrasive appearance than the corresponding pumice treatment.

The process consisting of a cellulase treatment step without pumice followed by a treatment with laccase and a strengthening agent gave jeans a strong worn look without a bleached look. There is equally keen interest in the evaluation of this appearance and methods of producing this appearance which would otherwise require greater amounts of cellulase and large amounts of pumice. Additionally, this method produced a strong worn look without the damage to the fabric that would be the result of a pumice or cellulase/pumice treatment (if the same worn look was to be achieved).

Example 2:

Wear enhancement using Myceliophthora thermophila laccase and methyl syringate as enhancers powerful

Fabric :

Abrasion with Swift denim fabric (Dakota type): Denim was abraded using a 12 kg Wascator FL 120 wash extractor.

Denim load: 2.6 kg

Water: 401

Buffer: 30g KH ₂ PO ₄

10g Na ₂ HPO ₄

pH: 6.8 Enzyme: 70g Denimax ^™ T (a commercial product available from NovoNordisk A/S, Bagsvaerd, Denmark)

Time: 2 hours

Temperature: 55°C Abrasion Enhancement A Wascator FOM 71 wash extractor was used for abrasion enhancement of denim.

Denim load: 0.8kg

Water: 20l

Buffer: 4.2g sodium acetate, _3H2O

4.0g succinic acid

pH: 5.1

Enzyme: 670 LACU Myceliophthora thermophila laccase (available from NovoNordisk A/S)

Enhancer: 0.5g methyl syringate

Time: 20 minutes

Temperature: 60°C Evaluation:

Results were assessed visually in a lightbox and by measuring reflections. For the latter, Texflash 2000 (provided by Datacolor) was used to evaluate the degree of bleaching and lightness using the variation of the color space coordinates L ^* a ^* b ^* :

L gives a variation of black (-L ^* )/white (+L ^* ), a gives a variation of green (-a ^* )/red (+a ^* ), b gives a variation of blue (-b ^* )/yellow (+b ^* ) change.

Decreasing L ^* value means increasing black (decreasing white), increasing L ^* value means increasing white (decreasing black), decreasing a ^* value means increasing green (decreasing red), increasing a ^* means increasing red (decreasing green) ), a decrease in b ^* value means an increase in blue (decrease in yellow), and an increase in b ^* value means an increase in yellow (decrease in blue).

Texflash 2000 operates in the L ^* a ^* b ^* coordinate system. The light source utilized was CIE Light Standard C. Each measurement is the average of 10 measurements. The instrument is calibrated using calibration plates (black and white). result:

The result is shown in the following table (Table 1): Table 1 processing steps L ^* a ^* b ^* ΔL ^* wear and tear 31.60 -1.45 -17.41 Wear and Enhancement 34.88 -1.69 -16.64 3.28

In terms of visual evaluation, the denim produced by the abrasion enhancement method had a highly worn appearance without a bleached appearance, similar to the results obtained in Example 1. Thus, the Myceliophthora thermophila laccase and methyl syringate enhancer functioned similarly to the laccase and enhancer used in Example 1.

Example 3 Wear Reinforced Fabrics with Different Levels of Cellulase and Different Dosages of Laccase and Strengthening Agent : Swift denim fabric (Dakota type) was utilized. Abrasion: A 12kg Wascator FL 120 wash extractor was used for denim abrasion. Three different doses of cellulase were applied.

Weight of loaded denim: 2.6kg

Water: 40l

Buffer _: 30g _KH2PO4

10g Na ₂ HPO ₄

Optimum pH: 6.8

Enzyme: Denimax ^TM Ultra MG (a commercial single-component cellulase product available from

Obtained by Novo Nordisk A/S)

1: 8g (=3.7μg cellulase/g fabric)

2: 28g (=12.9μg cellulase/g fabric)

3: 54g (=24.9μg cellulase/g fabric)

Time: 2 hours

Temperature: 55°C Wear Enhancement:

Use the Wascator FOM 71 Wash Extractor for Denim Wear Enhancement. The amounts of laccase and mediator were varied in the three experiments.

The weight of the denim of the load: 0.8kg

Water: 20l

Buffer: 4.2g sodium acetate, 3H ₂ O

4.0g succinic acid

Optimum pH: 5.1

Enzyme: Trametes villosa laccase (available from Novo Nordisk A/S)

A: 300 LACU (=5.9 μg laccase/g fabric)

B: 600 LACU (=11.7μg laccase/g fabric)

C: 900 LACU (=17.6 μg laccase/g fabric)

Enhancer: phenothiazine 10-propionate

A: 0.15g (=0.7μmole/g fabric)

B: 0.30g (=1.4μmole/g fabric)

C: 0.45g (=2.1μmole/g fabric)

Time: 20 minutes

Temperature 60°C evaluation:

As described in Example 2. Result: the result is shown in the table below (table 2):

Table 2 Cellulase dosage (g) Laccase dosage (LACU) Enhancer dosage (g) L ^* ΔL ^* visual evaluation effect 8 - - 28.69 - no wear appearance 8 300 0.15 32.47 3.78 Wear Enhancement (Worn Appearance) 8 600 0.30 34.23 5.54 Wear Enhancement (Worn Appearance) 8 900 0.45 35.93 7.24 Bleach (bleached appearance) 28 - - 31.58 - no wear appearance 28 300 0.15 33.90 2.32 Wear Enhancement (Worn Appearance) 28 600 0.30 35.74 4.16 Wear Enhancement (Worn Appearance) 28 900 0.45 38.50 6.92 Bleach (bleached appearance) 54 - - 32.91 - no wear appearance 54 300 0.15 35.32 2.41 Wear Enhancement (Worn Appearance) 54 600 0.30 37.90 4.99 Bleach (bleached appearance) 54 900 0.45 40.46 7.55 Bleach (bleached appearance) As can be seen, wear enhancement is only obtained when the dosage of laccase and enhancer is below a certain limit (otherwise the effect would be a bleached appearance). It can also be seen that this limit depends on the dose of cellulase in the attrition step - the higher the dose of cellulase, the lower the limit, i.e. the following approximate law: at 4 μg monocomponent cellulase/g fabric conditions, obtaining wear-enhancing fabrics requires:

<15μg laccase/g fabric

<2 μmole enhancer/g fabric; under the condition of 13 μg monocomponent cellulase/g fabric, to obtain wear-enhanced fabric requires:

＜12μg laccase/g fabric

<1.5 μmole enhancer/g fabric; under the condition of 25 μg single-component cellulase/g fabric, to obtain wear-enhanced fabric requires:

<10μg laccase/g fabric

<1 μmole enhancer/g fabric.

Claims (18)

1. method that makes institute's dyeing and weaving thing tool worn appearance and have the loss of intensity of minimizing, this method comprises:

(a) in water-bearing media, institute's dyeing and weaving thing is contacted with cellulase, corresponding concentration is every gram fabric 0.1-250 μ g zymoprotein;

(b) simultaneously or subsequently handle above-mentioned fabrics with the phenoloxidase cascade reinforcing agent of unifying, wherein the concentration of phenol oxidase contains 0.5-50 μ g zymoprotein corresponding to every gram fabric, and the concentration of reinforcing agent in water-bearing media is every gram fabric 0.05-500 μ mole, and described reinforcing agent can be represented by formula I: In the formula, A be such as-D ,-CH=CH-D ,-CH=CH-CH=CH-D ,-CH=N-D ,-N=N-D or-group the N=CH-D, wherein D be selected from by-CO-E ,-SO ₂-E ,-N-XY and-N ⁺The group that-XYZ forms, wherein E can be-H ,-OH ,-R or-OR, X, Y and Z can be identical or different and be selected from-H and-R; R is C ₁-C ₁₆Alkyl, preferably C ₁-C ₈Alkyl, wherein alkyl can be saturated or undersaturated, and side chain or unbranched is arranged, and also can use carboxyl, sulfo group or amino group to replace; B and C can be identical or different and be selected from C _mH _2m+11≤m≤5; Perhaps represent by formula II: In the formula, X represents (O-) or (S-), substituent R ¹-R ⁹Can be identical or different, represent following arbitrary group independently: hydrogen, halogen, hydroxyl, formoxyl, carboxyl and ester class and its esters, carbamoyl, sulfo group and ester class and its esters, sulfamoyl, nitro, amino, phenyl, C ₁-C ₁₄-alkyl, C ₁-C ₅-alkoxyl, carbonyl-C ₁-C ₅-alkyl, aryl-C ₁-C ₅-alkyl; Carbamoyl wherein, sulfamoyl and amino group can also not be substituted or be substituted basic R ¹⁰Replace once or twice; Wherein phenyl can also not be substituted or by one or more substituent R ¹⁰Replace; C wherein ₁-C ₁₄-alkyl, C ₁-C ₅-alkoxyl, carbonyl-C ₁-C ₅-alkyl and aryl-C ₁-C ₅-alkyl group can be saturated or undersaturated, and side chain or unbranched is arranged, and can also not be substituted or by one or more substituent R ¹⁰Replace;

Substituent R wherein ¹⁰Represent following arbitrary group: halogen, hydroxyl, formoxyl, carboxyl and ester class and its esters; carbamoyl, sulfo group and ester class and its esters, sulfamoyl, nitro, amino, phenyl, aminoalkyl, piperidino, piperazinyl, pyrrolidinyl (pyrrolidino), C ₁-C ₅-alkyl, C ₁-C ₅-alkoxyl; Wherein carbamoyl, sulfamoyl and amino group can also be not by or by hydroxyl, C ₁-C ₅-alkyl, C ₁-C ₅-alkoxyl replaces once or twice; Wherein phenyl can also be replaced by one or more following groups: halogen, hydroxyl, amino, formoxyl, carboxyl and ester class and its esters, carbamoyl, sulfo group and ester class and its esters, and sulfamoyl; C wherein ₁-C ₅-alkyl and C ₁-C ₅-alkoxy base can be saturated or undersaturated, side chain or unbranched is arranged, can also be replaced once or twice by following arbitrary group: halogen, hydroxyl, amino, formoxyl, carboxyl and ester class and its esters, carbamoyl, sulfo group and ester class and its esters, and sulfamoyl;

Perhaps, substituent R in general formula ¹-R ⁹Two common form a group-B-, wherein B represents following arbitrary group: (CHR ¹⁰-N=N-), (CH=CH-) _n, (CH=N-) _nOr (N=CR ¹⁰-NR ¹¹-), n represents the integer of 1-3, R in the group ¹⁰Be the substituting group of above-mentioned definition, R ¹¹Definition is as R ¹⁰

2. according to the process of claim 1 wherein that fabric is dyeed by vat dyestuffs.

3. according to the method for claim 2, wherein vat dyestuffs is indigo or thioindigo.

4. according to arbitrary method of claim 1-3, wherein fabric is the mixture of cellulosic fabric or cellulose fibre or the mixture of cellulose fiber peacekeeping synthetic fiber.

5. according to the process of claim 1 wherein that fabric is a denim goods, preferably with denim goods indigo or thioindigo dyeing.

According to the process of claim 1 wherein cellulase available from the mould genus of humic (for example: Humicola insolens), Fusarium (for example: sharp sickle spore), myceliophthora (for example: Myceliophthora thermophila) or the kind of Cephalosporium.

7. according to the process of claim 1 wherein that the concentration of cellulase is corresponding to every gram fabric 0.5-50 μ g zymoprotein.

8. according to the process of claim 1 wherein that the phenol oxidase system is peroxidase and hydrogen peroxide source.

9. according to the method for claim 7, wherein peroxidase is a horseradish peroxidase, soybean peroxidase or available from Coprinus, for example Coprinus cinereus or long root ghost umbrella, bacillus, for example bacillus pumilus, myxobacter, for example peroxidase of myxococcus virescens.

10. according to the method for claim 8 or 9, wherein hydrogen peroxide source is hydrogen peroxide or a kind of hydrogen peroxide precursor, for example perborate or percarbonate or a hydrogen peroxide produce enzyme system, for example oxidizing ferment and its substrate or a kind of peroxycarboxylic acid or its salt.

11. according to the process of claim 1 wherein that water-bearing media contains H ₂O ₂Perhaps a kind of and 0.001-25mM H ₂O ₂The H that concentration is suitable ₂O ₂Precursor.

12. according to the process of claim 1 wherein that the phenol oxidase system is a kind of laccase or a kind of enzyme relevant with laccase and oxygen.

13. according to the method for claim 12, wherein laccase is available from Trametes, for example Trametesvillosa, Coprinus, for example Coprinus cinereus or myceliophthora, for example Myceliophthorathermophila.

14. according to the process of claim 1 wherein that the concentration of phenol oxidase contains 0.5-50 μ g zymoprotein corresponding to every gram fabric.

15. according to the process of claim 1 wherein that reinforcing agent belongs to the group of being made up of acetosyringone, syringaldehyde, methyl syringate and syringic acid.

16. according to the process of claim 1 wherein that reinforcing agent belongs to by 10-methyl-phenothiazine, phenothiazine-10-propionic acid, phenoxazine-10-propionic acid, phenoxazine-10-ethoxy, phenthazine-10-ethyl-4-carboxyl, 10-(3-dimethylamino-propyl) promazine hydrochloride and group that phenthazine-the 10-ethyl alcohol is formed.

17. according to the process of claim 1 wherein in water-bearing media, the concentration of reinforcing agent is every gram fabric 0.05-100 μ mole.

18. fabric that obtains according to the method for claim 1.