+

WO1997040229A1 - Tissu traite par cellulase et enzyme hybride comprenant une enzyme oxydant le phenol - Google Patents

Tissu traite par cellulase et enzyme hybride comprenant une enzyme oxydant le phenol Download PDF

Info

Publication number
WO1997040229A1
WO1997040229A1 PCT/DK1997/000176 DK9700176W WO9740229A1 WO 1997040229 A1 WO1997040229 A1 WO 1997040229A1 DK 9700176 W DK9700176 W DK 9700176W WO 9740229 A1 WO9740229 A1 WO 9740229A1
Authority
WO
WIPO (PCT)
Prior art keywords
enzyme
fabric
process according
cellulase
alkyl
Prior art date
Application number
PCT/DK1997/000176
Other languages
English (en)
Inventor
Thomas Vollmond
Original Assignee
Novo Nordisk A/S
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 A/S filed Critical Novo Nordisk A/S
Priority to AU23821/97A priority Critical patent/AU2382197A/en
Publication of WO1997040229A1 publication Critical patent/WO1997040229A1/fr

Links

Classifications

    • D—TEXTILES; PAPER
    • D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06P—DYEING OR PRINTING TEXTILES; DYEING LEATHER, FURS OR SOLID MACROMOLECULAR SUBSTANCES IN ANY FORM
    • D06P5/00—Other features in dyeing or printing textiles, or dyeing leather, furs, or solid macromolecular substances in any form
    • D06P5/02—After-treatment
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/38—Products with no well-defined composition, e.g. natural products
    • C11D3/386—Preparations containing enzymes, e.g. protease or amylase
    • C11D3/38645—Preparations containing enzymes, e.g. protease or amylase containing cellulase
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/38—Products with no well-defined composition, e.g. natural products
    • C11D3/386—Preparations containing enzymes, e.g. protease or amylase
    • C11D3/38654—Preparations containing enzymes, e.g. protease or amylase containing oxidase or reductase
    • D—TEXTILES; PAPER
    • D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
    • D06M16/00—Biochemical treatment of fibres, threads, yarns, fabrics, or fibrous goods made from such materials, e.g. enzymatic
    • D06M16/003—Biochemical treatment of fibres, threads, yarns, fabrics, or fibrous goods made from such materials, e.g. enzymatic with enzymes or microorganisms
    • D—TEXTILES; PAPER
    • D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06P—DYEING OR PRINTING TEXTILES; DYEING LEATHER, FURS OR SOLID MACROMOLECULAR SUBSTANCES IN ANY FORM
    • D06P5/00—Other features in dyeing or printing textiles, or dyeing leather, furs, or solid macromolecular substances in any form
    • D06P5/15—Locally discharging the dyes
    • D06P5/158—Locally discharging the dyes with other compounds

Definitions

  • the present invention relates to a process for providing a worn look in dyed fabric, especially cellulosic fabric such as denim.
  • the first step in the processing evolution was to sell jeans that had been laundered by the manufacturer.
  • pre-washed jeans had a slightly faded appearance and a softer hand that felt comfortable, as though they had been laundered several times. This trend became fashionable as well, and consumers were willing to pay the extra cost involved for this additional processing.
  • the fabric looses strength by using the stone- process described above, and the stone-free cellulase treatment does not alone give the desired worn look, so there is a need in industry for a more gentle process.
  • the present invention relates to a process for providing an abraded look with a reduced strength loss in dyed fabric comprising
  • an enhancing agent to the treatment with the hybrid enzyme mentioned above, said enhancing agent having a formula I or formula II:
  • formula X represents (-0-) or (-S-)
  • the substituent groups R ⁇ R 9 which may be identical or different, independently represents any of the following radicals: hydrogen, halogen, hydroxy, formyl, carboxy, and esters and salts hereof, carbamoyl, sulfo, and esters and salts hereof, sulfamoyl, nitro, amino, phenyl, C ⁇ -C ⁇ -alkyl, C ⁇ C ⁇ alkoxy, carbonyl-C ⁇ -C j -alkyl, aryl-C ⁇ C j -alkyl; which carbamoyl, sulfa- moyl, and amino groups may furthermore be unsubstituted or substituted once or twice with a substituent group R 10 ; and which phenyl may furthermore be unsubstituted or substituted with one or more substituent groups R 10 ; and which C x -C 14 -alkyl, and aryl-
  • Bleached versus worn look Persons skilled in the art of evaluating denim finishing processes, are capable of differentiating between a bleached look and a worn (or abraded) look of denim.
  • the former is a result of removal (bleaching) of dye from the dyed warp yarn. Since the bleaching takes place on the whole surface of every dyed yarn, the result is a general reduction in colour intensity. Thus, the bleached look of a pair of indigo-dyed jeans is characterised by a lighter blue shade than the corresponding reference.
  • the latter - the worn look - is a result of a treatment of denim with cellulase and/or pumice stone.
  • This process is characterised by an uneven removal of dye from the fabric, hence it results in a high level of contrast between dyed areas and areas from which dye has been removed.
  • the worn look is obtained by a process involving cellulase and/or pumice stone
  • the bleached look can be obtained by a process involving non-enzymatic bleaching agents such as hypochlorite or by a process involving oxidoreductase and an enhancing agent.
  • the present invention relates to a process of providing a worn but not bleached look, comprising a mild treatment with a cellulase and a subsequent mild treatment with a hybrid enzyme comprising a phenol oxidizing enzyme, or a mild treatment with a hybrid enzyme comprising a phenol oxidizing enzyme followed by a mild treatment with a cellulase, or a simultaneously mild treatment with a cellulase and a hybrid enzyme comprising a phenol oxidizing enzyme.
  • a cellulase it is preferred to use a cellulase, but instead of using a cellulase pumice stones or periite may be used, or a combination of cellulase and pumice stones or a combination of cellulase and periite as known in the art.
  • the invention may be applied to any dyed fabric known in the art, in particular to synthetic fabrics such as polyester or to natural fabrics.
  • the invention is most beneficially applied to cel ⁇ lulose-containing fabrics, such as cotton, viscose, rayon, ramie, linen, Tencel, or mixtures thereof, or mixtures of any of these fibres, or mixtures of any of these fibres together with synthetic fibres.
  • the fabric is denim.
  • the fabric may be dyed with vat dyes such as indigo, or indigo-related dyes such as thioindigo.
  • the fabric may also be dyed with more than one dye, e.g., first with a sulphur dye and then with an indigo dye, or vice versa.
  • the fabric is an indigo-dyed denim with a sulphur-bottorn, (i.e. the denim is first dyed with a sulphur dye and then with an indigo dye) ; including clothing items manufactured therefrom.
  • cellulase refers to an enzyme which catalyses the degradation of cellulose to glucose, cellobiose, triose and other cello-oligosaccharides.
  • cellulase is understood to include a mature protein or a precursor form thereof or a functional fragment thereof which essentially has the activity of the full-length enzyme.
  • cellulase is intended to include homologues or analogues of said enzyme.
  • homologues comprise an amino acid sequence exhibiting a degree of identity of at least 60% with the amino acid sequence of the parent enzyme, i.e. the parent cellulase.
  • the degree of identity may be determined by conventional methods, see for instance, Altshul et al. , Bull. Math. Bio. 4J . , 1986, pp. 603-616, and Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89, 1992, pp. 10915-10919.
  • the cellulase to be used in the present invention is a monocomponent (recombinant) cellulase, i.e. a cellulase essentially free from other proteins or cellulase proteins.
  • a recombinant cellulase component may be cloned and expressed according to standard techniques conventional to the skilled person.
  • the cellulase to be used in the method is an endoglucanase (EC 3.2.1.4), preferably a monocomponent (recombinant) endogluc ⁇ anase.
  • the cellulase is a microbial cellulase, more preferably a bacterial or fungal cellulase.
  • bacterial cellulases are cellulases der- ived from or producible by bacteria from the group of genera consisting of Pseudomonas or Bacillus, in particular Bacillus lautus.
  • the cellulase or endoglucanase may be an acid, a neutral or an alkaline cellulase or endoglucanase, i.e. exhibiting maximum cellulolytic activity in the acid, neutral or alkaline range, respectively.
  • a useful cellulase is an acid cellulase, preferably a fungal acid cellulase, which is der ⁇ ived from or producible by fungi from the group of genera con- sisting of Trichoderma, Actinomvces. Myrothecium, Aspergillus, and Botrytis.
  • a preferred useful acid cellulase is derived from or producible by fungi from the group of species consisting of Trichoderma viride. Trichoderma reesei, Trichoderma longibra- chiatum. Myrothecium verrucaria. Aspergillus niger, Aspergil ⁇ lus orvzae. and Botrytis cinerea.
  • Another useful cellulase or endoglucanase is a neutral or alkaline cellulase, preferably a fungal neutral or alkaline cellulase, which is derived from or producible by fungi from the group of genera consisting of Aspergillus, Penicillium. Myceliophthora, Humicola, Irpex, Fusarium, Sta- chvbotrys. Scopulariopsis. Chaetomium, Mycogone, Verticillium, Myrothecium. Papulospora, Gliocladium, Cephalosporium and Acremonium.
  • a preferred alkaline cellulase is derived from or producible by fungi from the group of species consisting of Humicola insolens, Fusarium oxysporum, Myceliopthora ther ⁇ mophila. or Cephalosporium sp. , preferably from the group of species consisting of Humicola insolens. DSM 1800, Fusarium oxysporum. DSM 2672, Myceliopthora thermophila, CBS 117.65, or Cephalosporium sp. , RYM-202.
  • useful cellulases are variants having, as a parent cellulase, a cellulase of fungal origin, e.g. a cellulase derivable from a strain of the fungal genus Humicola. Trichoderma or Fusarium.
  • the concentration of the cellulase enzyme in the aqueous medium may be 0.01-250 ⁇ g of enzyme protein per g of fabric, in particular 0.1-50 ⁇ g of enzyme protein per g of fabric.
  • cellulase activity can be expressed in ECU.
  • Cellulolytic enzymes hydrolyse CMC, thereby increasing the viscosity of the incubation mixture.
  • the resulting reduction in viscosity may be determined by a vibration viscosimeter (e.g. MIVI 3000 from Sofraser, France ) .
  • MIVI 3000 from Sofraser, France
  • Determination of the cellulolytic activity, measured in terms of ECU, may be determined according to the following analysis method (assay) :
  • the ECU assay quantifies the amount of catalytic activity present in the sample by measuring the ability of the sample to reduce the viscosity of a solution of carboxy-methylcellulose (CMC) .
  • the assay is carried out at 40°C; pH 7.5; 0. IM phosphate buffer; time 30 min; using a relative enzyme standard for reducing the viscosity of the CMC (carboxymethylcellulose Hercules 7 LFD) substrate; enzyme concentration approx. 0.15 ECU/ml .
  • the arch standard is defined to 8200 ECU/g .
  • an enzyme hybrid comprises a phenol oxidizing enzyme fused to an amino acid sequence having a cellulose-binding domain.
  • a phenol oxidizing enzyme is meant an enzyme, which by using hydrogen peroxide or molecular oxygen, is capable of oxidizing organic compounds containing phenolic groups . Examples of such enzymes are peroxidases and oxidases .
  • Enzyme hybrids are known in the art, (for reference see e.g. WO 90/00609 and WO 95/16782) : They may be prepared by transforming into a host cell a DNA construct comprising at least a fragment of DNA encoding the cellulose-binding domain ligated, with or without a linker, to a DNA sequence encoding the enzyme of interest and growing the host cell to express the fused gene.
  • the enzyme hybrids may be described by the following formula:
  • CBD can be either the N-terminal or the C-terminal region of an amino acid sequence corresponding to at least the cellulose-binding domain;
  • MR is the middle region (the linker) , and may be a bond, or a short linking group of from about 2 to about 100 carbon atoms, in particular of from 2 to 40 carbon atoms, or typically from about 2 to about 100 amino acids, in particular of from 2 to 40 amino acids, and
  • X can be either the N-terminal or the C-terminal region and is a phenol oxidizing enzyme.
  • cellulose-binding domain is intended to be understood as defined by Peter Tomme et al . "Cellulose- Binding Domains: Classification and Properties” in “Enzymatic Degradation of Insoluble Carbohydrates", John N. Saddler and Michael H. Penner (Eds.) , ACS Symposium Series, No. 618, 1996.
  • CBDs cellulose- binding domains
  • l-X cellulose- binding domains
  • CBDs are found in various enzymes such as cel- lulases, xylanases, mannanases, arabinofuranosidases, acetyl esterases and chitinases.
  • CBDs have also been found in algae, e.g., the red alga Porphyra purpurea as a non-hydrolytic polysaccharide-binding protein, for reference see Peter Tomme et al . , supra.
  • most of the CBDs are from cellulases and xylanases. CBDs are found at the N or C termini of proteins or are internal .
  • Cellulases useful for preparation of Cellulose-Binding Domains The techniques used in isolating a cellulase gene are well known in the art.
  • the term "cellulase” refers to an enzyme which catalyses the degradation of cellulose to glucose, cellobiose, triose and other cello-oligosaccharides.
  • the cellulase to be used in the method is an endoglucanase (EC 3.2.1.4), preferably a monocomponent (recombinant) endogluc ⁇ anase.
  • the cellulase is a microbial cellulase, more preferably a bacterial or fungal cellulase.
  • bacterial cellulases are cellul ⁇ ases derived from or producible by bacteria from the group consisting of Pseudomonas, Bacillus, Cellulomonas. Clostridium. Microspora, Thermotoga, Caldocellum and Actinomvcets such as Streptomyces, Termomonospora and Acidothemus. in particular from the group consisting of Pseudomonas cellulolvticus.
  • the cellulase may be an acid, a neutral or an alkaline cellulase, i.e. exhibiting maximum cellulolytic activity in the acid, neutral or alkaline range, respectively.
  • a useful cellulase is an acid cellulase, preferably a fungal acid cellulase, which is derived from or producible by fungi from the group of genera consisting of Trichoderma. Myrothecium. Aspergillus, Phanaerochaete, Neurospora, Neo- callimastix and Botrytis.
  • a preferred useful acid cellulase is derived from or producible by fungi from the group of species consisting of Trichoderma viride, Trichoderma reesei. Trichoderma longibra- chiatum, Myrothecium verrucaria, Aspergi1lus niger. Aspergil ⁇ lus oryzae. Phanaerochaete chrysosporium, Neurospora crassa. Neocallimastix partriciarum and Botrytis cinerea.
  • Another useful cellulase is a neutral or alkaline cellulase, preferably a fungal neutral or fungal alkaline cel ⁇ lulase, which is derived from or producible by fungi from the group of genera consisting of Aspergillus, Penicillium. Mvceliophthora. Humicola. Irpex, Fusarium, Stachvbotrys. Scopulariopsis. Chaetomium, Mycogone, Verticillium, Myrothe- cium, Papulospora, Gliocladium. Cephalosporium and Acremonium.
  • a preferred alkaline cellulase is derived from or producible by fungi from the group of species consisting of Humicola insolens, Fusarium oxysporum, Myceliopthora ther ⁇ mophila. Penicillium ianthinellum and Cephalosporium sp., preferably from the group of species consisting of Humicola insolens. DSM 1800, Fusarium oxysporum, DSM 2672, Myceliopt ⁇ hora thermophila, CBS 117.65, and Cephalosporium sp. , RYM-202.
  • a preferred example of a native or parent cellulase is an alkaline endoglucanase which is immunologically reactive with an antibody raised against a highly purified " 43kD endo ⁇ glucanase derived from Humicola insolens, DSM 1800, or which is a derivative of the " 43kD endoglucanase exhibiting cellulase activity.
  • useful cellulases are variants having, as a parent cellulase, a cellulase of fungal or bacterial origin, e.g. a cellulase derivable from a strain of the fungal genus Humicola. Trichoderma or Fusarium.
  • a cellulase In order to isolate the cellulose binding domain of e.g. a cellulase, several genetic approaches may be used.
  • One method uses restriction enzymes to remove a portion of the gene and then to fuse the remaining gene-vector fragment in frame to obtain a mutated gene that encodes a protein truncated for a particular gene fragment.
  • Another method involves the use of exonucleases such as Bal31 to systematically delete nucleotides either externally from the 5 ' and the 3 ' ends of the DNA or internally from a restricted gap within the gene. These gene deletion methods result in a mutated gene encoding a shortened gene molecule which may then be evaluated for substrate binding ability.
  • Appropriate substrates for evaluating the binding activity include compounds such as Avicel and cotton fibres.
  • nucleotide sequence encoding the substrate binding region may then be manipulated in a variety of ways to fuse it to a DNA sequence encoding the enzyme of interest.
  • the cellulose binding encoding fragment and the DNA encoding the enzyme of interest are then ligated with or without a linker.
  • the resulting ligated DNA may then be manipulated in a variety of ways to provide for expression.
  • Microbial hosts such as Aspergillus, e.g., A. niger and A. oryzae. Bacillus. E. coli or S.cerevisiae are preferred.
  • Peroxidases Suitable peroxidases to be fused with the sequence encoding the cellulose-binding domain may be any peroxidase enzyme comprised by the enzyme classification (EC 1.11.1.7) , or any fragment derived therefrom, exhibiting peroxidase activity.
  • the peroxidase employed in the method of the invention is producible by plants (e.g. horseradish or soybean peroxidase) or microorganisms such as fungi or bacteria.
  • Some preferred fungi include strains belonging to the subdivision Deuteromycotina, class Hvphomvcetes, e.g., Fu- sarium. Humicola, Tricoderma, Myrothecium, VerticilIum, Arthromvces.
  • fungi include strains belonging to the subdivision Basidiomvcotina, class Basidiomvcetes, e.g. Coprinus, Phanerochaete, Coriolus or Trametes, in particular
  • Coprinus cinereus f. microsporus (IFO 8371), Coprinus macror- hizus, Phanerochaete chrysosporium (e.g. NA-12) or Trametes
  • T. versicolor e.g. PR4
  • fungi include strains belonging to the subdivision Zygomvcotina, class Mvcoraceae, e.g. Rhizopus or Mucor, in particular Mucor hiemalis.
  • Some preferred bacteria include strains of the order Actinomvcetales, e.g., Streptomyces spheroides (ATTC 23965) , Streptomyces thermoviolaceus (IFO 12382) or Streptoverticillum verticillium ssp. verticillium.
  • Actinomvcetales e.g., Streptomyces spheroides (ATTC 23965) , Streptomyces thermoviolaceus (IFO 12382) or Streptoverticillum verticillium ssp. verticillium.
  • Bacillus pumilus Bacillus pumilus
  • Further preferred bacteria include strains belonging to Mvxococcus. e.g., M. virescens.
  • a recombinantly produced peroxidase is preferred, e.g., a peroxidase derived from a Coprinus sp. , in particular C. macrorhizus or C. cinereus according to WO 92/16634, or a variant thereof, e.g., a variant as described in WO 94/12621.
  • Suitable laccases to be fused with the sequence encoding the cellulose-binding domain include laccases and laccase related enzymes, i.e., any laccase enzyme comprised by the enzyme classification (EC 1.10.3.2) , any chatechol oxidase enzyme comprised by the enzyme classification (EC 1.10.3.1), any bilirubin oxidase enzyme comprised by the enzyme classifi ⁇ cation (EC 1.3.3.5) or any monophenol monooxygenase enzyme comprised by the enzyme classification (EC 1.14.18.1) .
  • laccase enzymes are known from microbial and plant origin.
  • the microbial laccase enzyme may be derived from bacteria or fungi (including filamentous fungi and yeasts) and suitable examples include a laccase derivable from a strain of Aspergillus. Neurospora. e.g., N. crassa, Podos p ora, Botrvtis, Collvbia, Fomes, Lentinus, Pleurotus, Trametes, e.g., T. vii- losa and T. versicolor, Rhizoctonia. e.g. , R. solani, Copri ⁇ nus, e.g. C. plicatilis and C. cinereus, Psatyrella. Mvceliophthora, e.g. M.
  • thermophila Scytalidium, Polyporus. e.g. , P. pinsitus, Phlebia, e.g. , P . radita (WO 92/01046) , or Coriolus. e.g. , C. hirsutus (JP 2-238885) , in particular laccases obtainable from Trametes . Mvceliophthora, Scytalidium or Polyporus.
  • Plasmids Preparation of plasmids capable of expressing fusion proteins having the amino acid sequences derived from fragments of more than one polypeptide are well known in the art, for reference see e.g. WO 90/00609.
  • the expression cassette may be included within a replication system for episomal maintenance in an appropiate cellular host or may be provided without a replication system, where it may become integrated into the host genome .
  • the DNA may be introduced into the host in accordance with known techniques such as transformation, microinjection or the like.
  • the host Once the fused gene has been introduced into the appropiate host, the host may be grown to express the fused gene. Normally it is desirable additionally to add a signal sequence which provides for secretion of the fused gene.
  • Typical examples of useful fused genes are:
  • Linker Cellulose-Binding Domain, in which the pro-peptide normally contains 5-25 amino acids.
  • the recombinant product may be glycosylated or non- glycosylated.
  • the source may be hydrogen peroxide or a hydrogen peroxide precursor for in situ production of hydrogen peroxide, e.g. , percarbonate or perborate, or a hydrogen peroxide generating enzyme system, e.g. an oxidase and a substrate for the oxidase, or an amino acid oxidase and a suitable amino acid, or a peroxycarboxylic acid or a salt thereof.
  • Hydrogen peroxide may be added at the beginning of or during the process, e.g. in a concentration corresponding to 0.001-25 mM H 2 0 2 .
  • the concentration of the hybrid enzyme in the aqueous medium where the localized variation in the colour density of the surface of the dyed fabric is taking place may be 0.01-250 ⁇ g of hybrid enzyme protein per g of fabric, preferably 0.1-50 ⁇ g of hybrid enzyme protein per g of fabric.
  • the peroxidase activity may be determined in the following way:
  • 1 peroxidase unit is the amount of enzyme that catalyzes the conversion of 1 ⁇ mole hydrogen peroxide per minute at the following analytical conditions: 0.88 mM hydrogen peroxide, 1.67 mM 2, 2 ' -azinobis (3- ethylbenzothiazoline-6-sulfonate) , 0.1 M phosphate buffer, pH 7.0, incubated at 30°C, photometrically followed at 418 nm. If the phenol oxidizing enzyme is a laccase, the laccase activity may be determined in the following way:
  • Laccase activity is determined from the oxidation of syringaldazin under aerobic conditions.
  • the violet colour produced is photometered at 530 nm.
  • the analytical conditions are 19 ⁇ M syringaldazin, 23.2 mM acetate buffer, pH 5.5, 30°C, 1 min. reaction time.
  • 1 laccase unit (LACU) is the amount of enzyme that catalyses the conversion of 1.0 ⁇ mole syringaldazin per minute at these conditions.
  • an enhancing agent is any compound that enhances the bleaching process .
  • the enhancing agent will typically be an organic compound, e.g., an organic compound described by one of the following formulas :
  • the enhancing agent may be described by the following formula I:
  • a in the above mentioned formula is -CO-E, in which E may be -H, -OH, -R, or -OR; R being a C-C 16 alkyl, preferably a C ⁇ C j , alkyl, which alkyl may be saturated or unsaturated, branched or unbranched and optionally substituted with a carboxy, sulfo or amino group; and B and C may be the same or different and selected from C m H 2m+1 ; 1 ⁇ m ⁇ 5.
  • the enhancing agent is acetosyringone, methylsyringate, ethylsyringate, propyl- syringate, butylsyringate, hexylsyringate, or octylsyringate .
  • the enhancing agents described above may be prepared using methods well known to those skilled in the art; some of the enhancing agents are also commercially available, e.g., acetosyringone. Methylsyringate, ethylsyringate, propyl- syringate, butylsyringate, hexylsyringate and octylsyringate may be produced as disclosed in Chem. Ber. 67, 1934, p. 67.
  • the enhancing agent used in the present invention may also be described by the following formula II:
  • formula X represents (-0-) or (-S-)
  • the substituent groups R ⁇ R 9 which may be identical or different, independently represents any of the following radicals: hydrogen, halogen, hydroxy, formyl, carboxy, and esters and salts hereof, carbamoyl, sulfo, and esters and salts hereof, sulfamoyl, nitro, amino, phenyl, C ⁇ C ⁇ -alkyl, C l - C 5 -alkoxy, carbonyl-C ⁇ G ; -alkyl, aryl-C ⁇ C j -alkyl; which car ⁇ bamoyl, sulfamoyl, and amino groups may furthermore be un ⁇ substituted or substituted once or twice with a substituent group R 10 ; and which phenyl may furthermore be unsubstituted or substituted with one or more substituent groups R 10 ; and which C.-C ⁇ -alkyl, C ⁇ -C 8
  • the enhancing agent is 10-methylphenothiazine, phenothiazine-10-propionic acid, N-hydroxysuccinimide phenothiazine-10-propionate, 10-ethyl- phenothiazine-4-carboxylic acid, 10-ethylphenothiazine, 10- propylphenothiazine, 10-isopropylphenothiazine, methyl pheno- thiazine-10-propionate, 10-phenylphenothiazine, 10-allylpheno- thiazine, 10- (3- (4-methylpiperazin-l-yl)propyl)phenothiazine, 10- (2-pyrrolidin-1-yl-ethyl)phenothiazine, 2-methoxy-10- methyl-phenothiazine, 1-methoxy-10-methylphenothiazine, 3- methoxy-10-methylphenothiazine, 3, 10-dimethylphenothiazin
  • the enhancing agents may be obtained from Sigma- Aldrich, Janssen Chimica, Kodak, Tokyo Kasai Organic Chemicals, Daiichi Pure Chemicals Co. or Boehringer Mannheim; N-methylated derivatives of phenothiazine and phenoxazine may be prepared by methylation with methyliodide as described by Cornel Bodea and loan Silberg in "Recent Advances in the Chemistry of Phenothiazines" (Advances in heterocyclic chemistry, 1968, Vol. 9, pp. 321-460) ; B. Cardillo & G. Casnati in Tetrahedron, 1967, Vol. 23, p. 3771.
  • Phenothiazine and phenoxazine propionic acids may be prepared as described in J. Org. Chem. 15, 1950, pp. 1125-1130. Hydroxyethyl and hydroxypropyl derivatives of phenothiazine and phenoxazine may be prepared as described by G. Cauquil in Bulletin de la Society Chemique de France. 1960, p.1049.
  • the enhancing agent of the invention may be present in concentrations of from 0.05 to 500 ⁇ mole per g of fabric, preferably 0.05 to 100 ⁇ mole per g of fabric, more preferably 0.05 to 10 ⁇ mole per g of fabric.
  • the present invention is typically used in industrial machines for cellulase treatment of fabric.
  • the fabric is normally added to the machine accord ⁇ ing to the machine capacity per the manufacturer's instruc ⁇ tions.
  • the fabric may be added to the machine prior to introducing water or the fabric may be added after water is introduced.
  • the cellulase treatment will be performed first, followed by the treatment with the enzyme hybrid and optionally the enhancing agent, but the two processes may be performed simultaneously, or vice versa.
  • the cellulase may be present in the water prior to adding the fabric or it may be added after the fabric has been wetted. Normally a buffer will be used in order to be close to the pH optimum of the enzyme in question. After the fabric has been contacted with the cellulase it should be agitated in the machine for a sufficient period of time to ensure that the fabric is fully wetted and to ensure the action of the enzyme.
  • a reaction time between 5 and 60 minutes and a reaction temperature between 20°C and 90°C, preferably between 30°C and 80°C, more preferably between 40°C and 70°C, will be suitable.
  • the enzyme hybrid, the hydrogen peroxide source if the enzyme hybrid comprises a peroxidase, and optionally the enhancing agent of the invention may be present in the water prior to adding the fabric or they may be added after the fabric has been wetted.
  • the enzyme hybrid may be added simultaneously with the enhancing agent or they may be added separately. Normally a buffer will be used in order to be close to the pH optimum of the enzyme in question.
  • a hydrogen peroxide source, and optionally the enhancing agent of the invention it should be agitated in the machine for a sufficient period of time to ensure that the fabric is fully wetted and to ensure the action of the enzyme and the enhancing agent.
  • a reaction time between 5 and 60 minutes and a reaction temperature between 20°C and 90°C, preferably between 30°C and 80°C, more preferably between 40°C and 70°C, will be suitable.
  • a person skilled in the art knows whether, e.g., a pair of jeans looks bleached or worn, so when testing the new process according to the invention it is best to let skilled persons do a visual evaluation comparing fabric treated with one of the traditional methods with fabric treated according to the present invention.
  • the evaluation of the worn look may be quantified by using a Minolta Chroma Meter CR200 or a Minolta Chroma Meter CR300.
  • a Minolta Chroma Meter CR200 or CR300 (available frcm Minolta Corp.) is used according to Manufacturer's instructions to evaluate the degree of abrasion as well as to estimate any discoloration using the change in the colour space coordinates L * aV (CIELAB-system) : L * gives the change in white/black at a scale of from 0 to 100, a * gives the change in green (-a * ) /red (+a * ) , and b' gives the change in blue
  • a decrease in L * means an increase in black colour (decrease of white colour)
  • an increase in L * means an increase in white colour (a decrease in black colour )
  • a decrease in a * means an increase in green colour (decrease in red colour)
  • an increase in a * means an increase in red colour
  • a decrease in b * means an increase in blue colour (a decrease in yellow colour)
  • an increase in b * means an increase in yellow colour (a decrease in blue colour) .
  • the fabric swatches treated according to the invention are compared to non-treated fabric swatches.
  • Meter CR300 is operated in the L * a * b * colour space (coordinate system) .
  • the light source used is a CIE light standard C.
  • Each measurement is an average of 3 measurements.
  • the instrument is calibrated using a Minolta calibration plate (white) .
  • 10 non- treated denim swatches are measured 2 times each and the average of the coordinates L * a * b * are calculated and entered as a reference.
  • the coordinates of the samples are then calculated as the difference ( ⁇ ) of the average of 3 measurements on each swatch from the reference value of the coordinates L * a * b * .
  • CiP-CBD and mCiP842-CBD This example concerns fusion proteins comprising a CBD linked to Coprinus cinereus peroxidase (CiP) or to a mutant thereof (mCiP842) (see, e.g., WO 95/10602) .
  • Yeast expression system The pJC106/YNG344 host/vector system was chosen as the standard expression system for all CiP experiments utilizing yeast expression.
  • Mutant mCiP842 contains the following amino acid substitutions relative to the parent CiP: V53A, E239G, M242I, Y272F.
  • the CBD-CiP fusion was constructed by amplifying four separate gene fragments using PCR.
  • H. insol ens family 45 cellulase The sequence of the H. insol ens family 45 cellulase is disclosed in WO 91/17244.
  • CiPpcrdwn CCCCCTTCCCTGGCGAATTCCGCATGAGG
  • JC20.1 ACCTTGGGGTAGAGCGAGGGCACCGATG Amplification
  • B 3.
  • JC20.2 TGCACTGCTGAGAGGTGGGC
  • Amplified products of reactions A) and B) were purified and phosphorylated using T4 polynucleotide kinase, ligated to one another for 15 min. at room temperature, and amplified with primers 1 and 4 to generate product AB.
  • Amplified products of reactions C) and D) were purified and mixed, then PCR-amplified to generate product CD.
  • Reaction products AB and CD were purified and phosphorylated using T4 polynucleotide kinase, ligated to one another for 15 min. at room temperature, and amplified with primers 1 and 8 to generate the final product.
  • Plasmid JC20A contains the wild type CiP gene, whereas plasmid JC20D contains the peroxide-stable mutant mCiP842. Transformants were selected on minimal media lacking uridine .
  • Yeast transformants were grown on minimal media plates containing 2% galactose (to induce the GAL1 yeast promoter driving CBD-CiP expression) that had been covered with a double filter layer consisting of cellulose acetate on top of nitrocellulose. After overnight growth, both filters were washed twice with 100 ml of 20 mM phosphate buffer, pH 7.0 for 5 minutes, after which no colony debris could be detected. Filters were then assayed for bound peroxidase activity by coating them with a 100 mM phosphate buffer, pH 7.0, containing 50 ⁇ g/ml of diamino-benzidine and 1 mM hydrogen peroxide. Bound peroxidase activity appears as a brown precipitate on the filter.
  • Liquid Assay Liquid cultures of mutants demonstrating cellulose binding in the filter assay were grown overnight in minimal media containing 2% galactose. 20 ⁇ l samples of culture broth were mixed with Avicel crystalline cellulose (20 g/L) in 0.1 M phosphate buffer, pH 7, 0.01% Tween 20 in a total volume of 100 ⁇ l and incubated at 22°C for 10 minutes. The mixture was then centrifuged to pellet the insoluble cellulose fraction, and the supernatants were assayed for peroxidase activity. Binding was scored as the % activity bound to the insoluble cellulose fraction based on the decrease in soluble activity.
  • This screening process utilizes broth samples from yeast cultures grown in microtiter plates.
  • the 96-well plate screen is performed by first growing yeast transformants of a pool of mutants in 50 ⁇ L volumes of URA(-) medium, pH 6.0 in 96- well microtiter plates. Cultures are inoculated by dilution into medium and pipetting (robotic or manual autopipettor) into 96-well plates. These are placed in an incubator set at
  • CiP and mCiP842 and the related fusion proteins were subjected to a combined pH - temperature - HO stress test: After an initial activity assay, cultures are diluted to ca. 0.06 PODU/ml, and incubated in 200 ⁇ M hydrogen peroxide, 100 mM phosphate/borate buffer, pH 10.5 at 50°C. After 0, 10, 20 and 30 minutes, samples are removed and residual activity is measured using the standard ABTS assay, pH 7.0. Improved mutants are those showing higher residual activity than CiP and are expressed as percent residual activity relative to the time 0 assay result.
  • Yeast expression plasmids designed to make H. insolens family 45 cellulase CBD-CiP fusions were constructed and sequenced.
  • JC20-series plasmids were transformed into S. cerevisae for expression and testing. After transformation, yeast colonies were grown on selective plates covered with a double filter layer: cellulose acetate filters on top of nitrocellulose. Wild type CiP secreted from yeast JC106 and the stable mutant mCiP842 pass through the cellulose acetate, then binds to the nitrocellulose where it can be visualized using diaminobenzidine (DAB) and H 2 0 2 . The cellulose acetate filter does not bind any wild-type or mCiP842 peroxidase.
  • DAB diaminobenzidine
  • the N-terminal CBD-CiP fusions encoded by plasmids JC20A, and JC20D are all detectable on both filters using the DAB assay, indicating that the fusion proteins have both peroxidase and cellulose-binding activities .
  • the peroxidase activity bound to the cellulose acetate filter remains bound even after washing extensively with buffer at pH 7.
  • the activity bound to the lower nitrocellulose filter suggests that binding of the CBD-CiP may be incomplete, or the cellulose filter gets saturated, allowing some of the fusion protein to pass through to the lower filter, or that some percentage of the fusion protein gets truncated to include only the peroxidase domain.
  • San Francisco denim fabric (standard sulphur bottom denim fabric from Swift, France) was desized with Aquazym (available from Novo Nordisk A/S) and abraded with DeniMax T (available from Novo Nordisk A/S) to obtain a mildly abraded look.
  • the resulting L*a*b* values of the San Francisco denim after abrasion was: L* : 30.87; a*: -0.16; b* : -15.36 Dakota denim fabric (standard pure indigo denim fabric from
  • CiP-CBD produced according to Example 1 : 3 ⁇ g enzyme protein per g denim,
  • Abrasion enhancement bv treating with cellulase and CBD- oxidoreductase (mCiP842-CBD)

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Textile Engineering (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Detergent Compositions (AREA)
  • Chemical Or Physical Treatment Of Fibers (AREA)

Abstract

Cette invention concerne un procédé permettant de conférer à des tissus un aspect usé mais non délavé. Ce procédé consiste à effectuer un traitement doux à l'aide d'une cellulase, ainsi qu'un autre traitement doux à l'aide d'une enzyme hybride qui comprend une enzyme oxydant le phénol.
PCT/DK1997/000176 1996-04-19 1997-04-18 Tissu traite par cellulase et enzyme hybride comprenant une enzyme oxydant le phenol WO1997040229A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU23821/97A AU2382197A (en) 1996-04-19 1997-04-18 Fabric treated with a cellulase and a hybrid enzyme comprising a phenol oxidizing enzyme

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DK46596 1996-04-19
DK0465/96 1996-04-19

Publications (1)

Publication Number Publication Date
WO1997040229A1 true WO1997040229A1 (fr) 1997-10-30

Family

ID=8093859

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DK1997/000176 WO1997040229A1 (fr) 1996-04-19 1997-04-18 Tissu traite par cellulase et enzyme hybride comprenant une enzyme oxydant le phenol

Country Status (2)

Country Link
AU (1) AU2382197A (fr)
WO (1) WO1997040229A1 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5912405A (en) * 1994-09-27 1999-06-15 Novo Nordisk A/S Enhancers such as acetosyringone
WO1999057155A1 (fr) * 1998-05-01 1999-11-11 The Procter & Gamble Company Detergent de lavage et/ou compositions respectant les tissus comprenant une proteine antimicrobienne modifiee
US7129069B2 (en) 2003-10-28 2006-10-31 Novo Zymes Als Hybrid enzymes
WO2007115723A3 (fr) * 2006-04-06 2007-11-29 Inst Francais Du Petrole Proteines resultant de la fusion entre des enzymes degradant la paroi cellulaire de plantes et une swollenine et leurs utilisations
US7319112B2 (en) 2000-07-14 2008-01-15 The Procter & Gamble Co. Non-halogenated antibacterial agents and processes for making same
CN102505532A (zh) * 2011-09-26 2012-06-20 青岛大学 一种利用纤维素酶促进涂料染色的方法
US8652455B2 (en) 2010-12-20 2014-02-18 E I Du Pont De Nemours And Company Targeted perhydrolases
CN113123144A (zh) * 2020-01-14 2021-07-16 尚科纺织企业工业及贸易公司 对纺织品进行染色的方法及用于其中的酶

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990000609A1 (fr) * 1988-07-08 1990-01-25 The University Of British Columbia Proteines de fusion se liant a la cellulose
WO1992018683A1 (fr) * 1991-04-12 1992-10-29 Novo Nordisk A/S Procede de blanchiment de textiles colores
WO1994007998A1 (fr) * 1992-10-06 1994-04-14 Novo Nordisk A/S Variantes de cellulase
WO1994012621A1 (fr) * 1992-12-01 1994-06-09 Novo Nordisk Amelioration de reactions enzymatiques
WO1994012578A1 (fr) * 1992-11-30 1994-06-09 Novo Nordisk A/S Procede de traitement de tissus en cellulose au moyen de cellulases

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990000609A1 (fr) * 1988-07-08 1990-01-25 The University Of British Columbia Proteines de fusion se liant a la cellulose
WO1992018683A1 (fr) * 1991-04-12 1992-10-29 Novo Nordisk A/S Procede de blanchiment de textiles colores
WO1994007998A1 (fr) * 1992-10-06 1994-04-14 Novo Nordisk A/S Variantes de cellulase
WO1994012578A1 (fr) * 1992-11-30 1994-06-09 Novo Nordisk A/S Procede de traitement de tissus en cellulose au moyen de cellulases
WO1994012621A1 (fr) * 1992-12-01 1994-06-09 Novo Nordisk Amelioration de reactions enzymatiques

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5912405A (en) * 1994-09-27 1999-06-15 Novo Nordisk A/S Enhancers such as acetosyringone
WO1999057155A1 (fr) * 1998-05-01 1999-11-11 The Procter & Gamble Company Detergent de lavage et/ou compositions respectant les tissus comprenant une proteine antimicrobienne modifiee
US7319112B2 (en) 2000-07-14 2008-01-15 The Procter & Gamble Co. Non-halogenated antibacterial agents and processes for making same
US7749744B2 (en) 2003-10-28 2010-07-06 Novozymes A/S Hybrid enzymes
US7129069B2 (en) 2003-10-28 2006-10-31 Novo Zymes Als Hybrid enzymes
US7312055B2 (en) 2003-10-28 2007-12-25 Novozymes A/S Hybrid enzymes
WO2007115723A3 (fr) * 2006-04-06 2007-11-29 Inst Francais Du Petrole Proteines resultant de la fusion entre des enzymes degradant la paroi cellulaire de plantes et une swollenine et leurs utilisations
US8652455B2 (en) 2010-12-20 2014-02-18 E I Du Pont De Nemours And Company Targeted perhydrolases
US8815550B2 (en) 2010-12-20 2014-08-26 E I Du Pont De Nemours And Company Targeted perhydrolases
CN102505532A (zh) * 2011-09-26 2012-06-20 青岛大学 一种利用纤维素酶促进涂料染色的方法
CN102505532B (zh) * 2011-09-26 2013-05-01 青岛大学 一种利用纤维素酶促进涂料染色的方法
CN113123144A (zh) * 2020-01-14 2021-07-16 尚科纺织企业工业及贸易公司 对纺织品进行染色的方法及用于其中的酶
EP3851574A1 (fr) 2020-01-14 2021-07-21 Sanko Tekstil Isletmeleri San. Ve Tic. A.S. Procédé de teinture de textiles et enzymes utilisés selon ce procédé
WO2021144356A1 (fr) 2020-01-14 2021-07-22 Sanko Tekstil Isletmeleri San. Ve Tic. A.S. Procédé de coloration de textiles et enzymes utilisées dans ce procédé
US11920290B2 (en) 2020-01-14 2024-03-05 Sanko Tekstil Isletmeleri San. Ve Tic A.S. Process for dyeing textiles and enzymes used therein

Also Published As

Publication number Publication date
AU2382197A (en) 1997-11-12

Similar Documents

Publication Publication Date Title
EP0787229B1 (fr) Procede de blanchiment consistant a utiliser un systeme enzyme oxydant le phenol et un agent facilitant
EP0857216B1 (fr) Cellulases, genes les codant et utilisation de ces cellulases
DE69730821T2 (de) Verfahren zum entfernen oder bleichen von verschmutzungen oder flecken aus zellulosekaltigem gewebe
EP0787230B1 (fr) Procede de blanchiment consistant a utiliser une enzyme oxydant le phenol, une source de peroxyde d'hydrogene et un agent de renforcement
KR100303622B1 (ko) 셀룰라제 변이체
FI119820B (fi) Menetelmä ja koostumuksia selluloosapitoisten kankaiden käsittelemiseksi käyttämällä typistettyjä sellulaasientsyymejä käsittäviä koostumuksia
EP0935692B1 (fr) Tissu traite avec de la cellulase et de l'oxydoreductase
US5908472A (en) Fabric treated with cellulase and oxidoreductase
EP1045934B1 (fr) Procede d'elimination de l'excedent de teinture des tissus imprimes ou teintes
EP0540784B1 (fr) Compositions empêchant le transfert de colorant
CA2164849C (fr) Compositions enzymatiques et methodes pour donner un aspect delave a du tissu denim colore a l'indigo
US7273748B2 (en) Cellulases, the genes encoding them and uses thereof
WO1997040229A1 (fr) Tissu traite par cellulase et enzyme hybride comprenant une enzyme oxydant le phenol
US6048367A (en) Process for removal of excess dye from printed or dyed fabric or yarn
US6184019B1 (en) Cellulases, the genes encoding them and uses thereof
WO1997040127A1 (fr) Blanchissage de tissus a l'aide d'une enzyme hybride contenant une enzyme oxydant un phenol qui est fusionnee a un domaine de liaison de la cellulose
US6248134B1 (en) Process for removal of excess dye from printed or dyed fabric or yarn
WO1997025469A1 (fr) Blanchiment/eclaircissement de textiles

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GE GH HU IL IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK TJ TM TR TT UA UG US UZ VN YU AM AZ BY KG KZ MD RU TJ TM

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH KE LS MW SD SZ UG AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

NENP Non-entry into the national phase

Ref country code: JP

Ref document number: 97537616

Format of ref document f/p: F

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载