WO2021259099A1

WO2021259099A1 - Use of cellulases for removing dust mite from textile

Info

Publication number: WO2021259099A1
Application number: PCT/CN2021/100308
Authority: WO
Inventors: Nan GAO; Yitong Liu; Ye Tao
Original assignee: Novozymes A/S
Priority date: 2020-06-24
Filing date: 2021-06-16
Publication date: 2021-12-30
Also published as: EP4172298A1; CN115836125A

Abstract

The present invention concerns a detergent composition comprising enzyme capable for degrading cellulosic materials and its use for removing dust mite from the surface of textile. In one aspect, said enzyme capable for degrading cellulosic materials is GH45 cellulase. In a further aspect, said GH45 cellulase is Thermothielavioides terrestris endoglucanase.

Description

USE OF CELLULASES FOR REMOVING DUST MITE FROM TEXTILE

FIELD OF THE INVENTION

The present invention concerns the use of enzymes capable for degrading cellulosic matrial, in particular the use of cellulases, for removing dust mite from textile surface.

Reference to a Sequence Listing

This application contains a Sequence Listing in computer readable form. The computer readable form is incorporated herein by reference.

BACKGROUND OF THE INVENTION

Dust mites are acarids (atype of arachnid, such as spiders and scorpions) . They range in size between 0.1 mm and 0.5 mm, live in all types of fibers, including carpets, clothing, fabric sofas, and bed sheets. In the past few decades it has become clear that dust mites are extremely allergenic and may trigger asthma and other allergy attacks. Attempts have been made to prevent contact with allergens by employing organic or inorganic miticides, cleaner, or ultraviolet rays. Specifically, US5672362A used disodium octaborate tetrahydrate (DOT) solution to kill dust mites. WO9836640 disclosed a compositions containing benzyl benzoate and alcohol to control dust mites. KR200287251 disclosed a protable UV sterilizer to remove dust mite. But so far the results have been disappointing because miticides or cleaner may bring in new allergy attack. US2004255983 disclosed a method for removing dust mites by washing the article repeatly to a desired level. However, too many wash cycles may break the intergrity and safety of the textile and thereby reduce the lifetime.

There is a need for a new method to remove dust mite from textile surface meanwhile not bring in new allergens or reduce the lifetime of the textile.

SUMMARY OF THE INVENTION

In one aspect, the present invention relates to the use of enzymes capable for degrading cellulosic material for reducing dust mite from a textile by adding said enzyme or enzymes in wash cycle. In another aspect, said enzyme is cellulase or hemicellulase. In a further aspect, said cellulase can be GH45 cellulase.

In another aspect, the present invention also relates to an enzyme composition for dust mites removing. Said enzyme composition comprises enzymes capable for degrading cellulosic materials and can be used in any washing, cleaning and/or fabric care methods including soaking methods, pre-treatment methods, methods with rinsing steps for which a separate rinse and said composition may be added and post-treatment methods.

In another aspect, said enzyme composition comprises one or more cellulases or hemicellulase. Further, said cellulase can be GH45 cellulase. In one aspect, said GH45 cellulase is Thermothielavioides terrestris (T. t ) endoglucanase.

In a further aspect, said T. t endoglucanase is selected from the group consisting of :

(1) T. t endoglucanase comprising or consisting of the mature polypeptide of SEQ ID NO: 2 or fragments thereof;

(2) T. t endoglucanase comprising or consisting of an amino acid sequence having at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 2;

(3) T. t endoglucanase encoded by a polynucleotide comprising or consisting of a nucleotide sequence having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1;

(4) T. t endoglucanase encoded by a polynucleotide that hybridizes under at least high stringency conditions, very high stringency conditions, with the mature polypeptide coding sequence of SEQ ID NO: 1 or the full-length complement thereof. In another aspect, said cellulase composition further comprises one or more detergent component or fabric soft agent.

In another aspect, the present application further comprises a detergent composition comprising the enzyme capable for degrading cellulosic material or comprising enzyme composition as mentioned above.

In still another aspect, the present application comprises a method for reducing or removing dust mite comprising:

(1) Washing or exposing the textile with a detergent composition comprising an enzyme capable of degrading cellulosic material or an enzyme composition comprising the enzyme capable of degrading cellulosic material, wherein, said enzyme capable of degrading cellulosic material comprises GH45 cellulase. Wherein, said GH45 cellulase is GH45 endoglucanase. In a further aspect, said endogluanase is T. t endoglucanase;

(2) Optionally rinsing the textile.

BRIEF DESCRIPTION OF FIGURES

Figure 1 shows the examplied anti-pilling experiments.

Figure 2 shows the anti-dust mite result of T. t. endoglucanase on E-252PA fabic.

Figure 3 shows the anti-dust mite result of T. t. endoglucanase on CN-42 fabic.

Definitions

Anti-pilling: The term "anti-pilling" denotes removal of pills from the textile surface and/or prevention of formation of pills on the textile surface.

Cellulolytic enzyme or cellulase: The term “cellulolytic enzyme” or “cellulase” means one or more (e.g., several) enzymes that hydrolyze a cellulosic material. Such enzymes include endoglucanase (s) , cellobiohydrolase (s) , beta-glucosidase (s) , or combinations thereof. The two basic approaches for measuring cellulolytic activity include: (1) measuring the total cellulolytic activity, and (2) measuring the individual cellulolytic activities (endoglucanases, cellobiohydrolases, and beta-glucosidases) as reviewed in Zhang et al., Outlook for cellulase improvement: Screening and selection strategies, 2006, Biotechnology Advances 24: 452-481. Total cellulolytic activity is usually measured using insoluble substrates, including Whatman №1 filter paper, microcrystalline cellulose, bacterial cellulose, algal cellulose, cotton, pretreated lignocellulose, etc. The most common total cellulolytic activity assay is the filter paper assay using Whatman №1 filter paper as the substrate. The assay was established by the International Union of Pure and Applied Chemistry (IUPAC) (Ghose, 1987, Measurement of cellulase activities, Pure Appl. Chem. 59: 257-68) .

For purposes of the present invention, cellulolytic enzyme activity is determined by measuring the increase in hydrolysis of a cellulosic material by cellulolytic enzyme (s) under the following conditions: add 1-50 mg of cellulolytic enzyme protein/g of cellulose in pretreated corn stover (PCS) (or other pretreated cellulosic material) for 3-7 days at a suitable temperature, e.g., 50℃, 55℃, or 60℃, compared to a control hydrolysis without addition of cellulolytic enzyme protein. Typical conditions are 1 ml reactions, washed or unwashed PCS, 5%insoluble solids, 50 mM sodium acetate pH 5, 1 mM MnSO ₄, 50℃, 55℃, or 60℃, 72 hours, sugar analysis by

HPX-87H column (Bio-Rad Laboratories, Inc., Hercules, CA, USA) . Other methods for measuring cellulolytic enzyme activity known in the art may also be applicable.

Cellulosic material: The term “cellulosic material” means any material containing cellulose. The predominant polysaccharide in the primary cell wall of biomass is cellulose, the second most abundant is hemicellulose, and the third is pectin. The secondary cell wall, produced after the cell has stopped growing, also contains polysaccharides and is strengthened by polymeric lignin covalently cross-linked to hemicellulose. Cellulose is a homopolymer of anhydrocellobiose and thus a linear beta- (1-4) -D-glucan, while hemicelluloses include a variety of compounds, such as xylans, xyloglucans, arabinoxylans, and mannans in complex branched structures with a spectrum of substituents. Although generally polymorphous, cellulose is found in plant tissue primarily as an insoluble crystalline matrix of parallel glucan chains. Hemicelluloses usually hydrogen bond to cellulose, as well as to other hemicelluloses, which help stabilize the cell wall matrix.

Cellulose is generally found, for example, in vegetable food products, such as salad, tomatoes, spinach, cabbage, grain or the like.

Detergent component: The term “detergent component” is defined herein to mean the types of chemicals which can be used in detergent compositions for e.g. laundry. Examples of detergent components are surfactants, builders, chelators or chelating agents, bleach system or bleach components, polymers, fabric conditioners, foam boosters, suds suppressors, dyes, perfume, tannish inhibitors, bactericides, fungicides, soil suspending agents, anti-corrosion agents, enzyme inhibitors or stabilizers, enzyme activators, transferase (s) , hydrolytic enzymes, oxido reductases, blueing agents and fluorescent dyes, antioxidants, and solubilizers.

Detergent Composition: The term “detergent composition” refers to compositions that find use in the removal of undesired compounds from surfaces to be cleaned, such as textile surfaces. The detergent composition may be used to e.g. clean textiles for both household cleaning and industrial cleaning. The terms encompass any materials/compounds selected for the particular type of cleaning composition desired and the form of the product (e.g., liquid, gel, powder, granulate, paste, or spray compositions) and includes, but is not limited to, detergent compositions (e.g., liquid and/or solid laundry detergents and fine fabric detergents; fabric fresheners; fabric softeners; and textile and laundry pre-spotters/pretreatment) . The detergent composition may contain one or more enzymes such as hemicellulases, peroxidases, proteases, cellulases, xylanases, lipases, phospholipases, esterases, cutinases, pectinases, mannanases, pectate lyases, keratinases, reductases, oxidases, phenoloxidases, lipoxygenases, ligninases, pullulanases, tannases, pentosanases, malanases, beta-glucanases, arabinosidases, hyaluronidase, chondroitinase, laccase, DNase, chlorophyllases, amylases, perhydrolases, peroxidases, xanthanase and mixtures thereof. The detergent composition may further comprise detergent component such as surfactants, builders, chelators or chelating agents, bleach system or bleach components, polymers, fabric conditioners, foam boosters, suds suppressors, dyes, perfume, tannish inhibitors, bactericides, fungicides, soil suspending agents, anti-corrosion agents, enzyme inhibitors or stabilizers, enzyme activators, transferase (s) , hydrolytic enzymes, oxido reductases, blueing agents and fluorescent dyes, antioxidants, and solubilizers.

Dust mite:

Dust mite (Dermatophagoides spp. ) is a kind of tiny arachnid arthropod, grow in a warm (22-26℃) and humid (75-80%humidity) environment. It lives on the dender and hair of humans or animals (cats or dogs) . There are 16 species of dust mites in home environment, including European house dust mites, American house dust mites, Maynella dust mites, and the sweet-eating mites with tropical claw mites and acarid mites. Tymite, etc., wherein, three species, i.e., European house dust mites (Dermatophagoides pteronyssinus) , American house dust mites (Dermatophagoides farinae) and tropical claw mites (Blomia tropicalis) are allergens. Specifically, European house dust mites (Dermatophagoides pteronyssinus) is the major allergen for asthma, sneezing, allergic conjunctivitis and atopic dermatitis.

Endoglucanase: The term “endoglucanase” means an endo-1, 4- (1, 3; 1, 4) -beta-D-glucan 4-glucanohydrolase (E.C. 3.2.1.4) that catalyzes endohydrolysis of 1, 4-beta-D-glycosidic linkages in cellulose, cellulose derivatives (such as carboxymethyl cellulose and hydroxyethyl cellulose) , lichenin, beta-1, 4 bonds in mixed beta-1, 3 glucans such as cereal beta-D-glucans or xyloglucans, and other plant material containing cellulosic components. Endoglucanase activity can be determined by measuring reduction in substrate viscosity or increase in reducing ends determined by a reducing sugar assay (Zhang et al., 2006, Biotechnology Advances 24: 452-481) . For purposes of the present invention, endoglucanase activity is determined using carboxymethyl cellulose (CMC) as substrate according to the procedure of Ghose, 1987, Pure and Appl. Chem. 59: 257-268, at pH 5, 40℃.

Fragment: The term “fragment” means a polypeptide having one or more (e.g., several) amino acids absent from the amino and/or carboxyl terminus of a mature polypeptide main; wherein the fragment has enzyme activity. In one aspect, a fragment contains at least 85%, e.g., at least 90%or at least 95%of the amino acid residues of the mature polypeptide of an enzyme.

Hemicellulolytic enzyme or hemicellulase: The term “hemicellulolytic enzyme” or “hemicellulase” means one or more (e.g., several) enzymes that hydrolyze a hemicellulosic material. See, for example, Shallom, D. and Shoham, Y. Microbial hemicellulases. Current Opinion In Microbiology, 2003, 6 (3) : 219-228) . Hemicellulases are key components in the degradation of plant biomass. Examples of hemicellulases include, but are not limited to, an acetylmannan esterase, an acetylxylan esterase, an arabinanase, an arabinofuranosidase, a coumaric acid esterase, a feruloyl esterase, a galactosidase, a glucuronidase, a glucuronoyl esterase, a mannanase, a mannosidase, a xylanase, and a xylosidase. The substrates of these enzymes, the hemicelluloses, are a heterogeneous group of branched and linear polysaccharides that are bound via hydrogen bonds to the cellulose microfibrils in the plant cell wall, crosslinking them into a robust network. Hemicelluloses are also covalently attached to lignin, forming together with cellulose a highly complex structure. The variable structure and organization of hemicelluloses require the concerted action of many enzymes for its complete degradation. The catalytic modules of hemicellulases are either glycoside hydrolases (GHs) that hydrolyze glycosidic bonds, or carbohydrate esterases (CEs) , which hydrolyze ester linkages of acetate or ferulic acid side groups. These catalytic modules, based on homology of their primary sequence, can be assigned into GH and CE families. Some families, with an overall similar fold, can be further grouped into clans, marked alphabetically (e.g., GH-A) . A most informative and updated classification of these and other carbohydrate active enzymes is available in the Carbohydrate-Active Enzymes (CAZy) database. Hemicellulolytic enzyme activities can be measured according to Ghose and Bisaria, 1987, Pure &AppI. Chem. 59: 1739-1752, at a suitable temperature, e.g., 50℃, 55℃, or 60℃, and pH, e.g., 5.0 or 5.5.

Improved wash performance: The term “improved wash performance” is defined herein as an enzyme displaying an increased wash performance relative to the wash performance of a similar wash without the enzyme e.g. by increased dust mites removal or increased dust mites reduction.

Low stringency conditions: The term “low stringency conditions” means for probes of at least 100 nucleotides in length, prehybridization and hybridization at 42℃ in 5X SSPE, 0.3%SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 25%formamide, following standard Southern blotting procedures for 12 to 24 hours. The carrier material is finally washed three times each for 15 minutes using 2X SSC, 0.2%SDS at 50℃.

Mature polypeptide: The term “mature polypeptide” means a polypeptide in its final form following translation and any post-translational modifications, such as N-terminal processing, C-terminal truncation, glycosylation, phosphorylation, etc. In the present invention, the mature polypeptide of SEQ ID NO: 2 is amino acid 22 to 300 of SEQ ID NO: 2 based on the SignalP 3.0 program (Bendtsen et al., 2004, J. Mol. Biol. 340: 783-795) that predicts amino acids 1 to 21 of SEQ ID NO: 2 are a signal peptide.

Mature polypeptide coding sequence: The term “mature polypeptide coding sequence” means a polynucleotide that encodes a mature polypeptide having enzymatic activity. In one aspect, the mature polypeptide coding sequence is nucleotides 64 to 858 of SEQ ID NO: 1 and nucleotides 1 to 63 of SEQ ID NO: 1 encode a signal peptide based on the program SignalP (Nielsen et al., 1997, Protein Engineering 10: 1-6) .

Medium stringency conditions: The term “medium stringency conditions” means for probes of at least 100 nucleotides in length, prehybridization and hybridization at 42℃ in 5X SSPE, 0.3%SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 35%formamide, following standard Southern blotting procedures for 12 to 24 hours. The carrier material is finally washed three times each for 15 minutes using 2X SSC, 0.2%SDS at 55℃.

Medium-high stringency conditions: The term “medium-high stringency conditions” means for probes of at least 100 nucleotides in length, prehybridization and hybridization at 42℃ in 5X SSPE, 0.3%SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 35%formamide, following standard Southern blotting procedures for 12 to 24 hours. The carrier material is finally washed three times each for 15 minutes using 2X SSC, 0.2%SDS at 60℃.

Sequence identity: The relatedness between two amino acid sequences or between two nucleotide sequences is described by the parameter “sequence identity” .

For purposes of the present invention, the sequence identity between two amino acid sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16: 276-277) , preferably version 5.0.0 or later. The parameters used are gap open penalty of 10, gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. The output of Needle labeled “longest identity” (obtained using the –nobrief option) is used as the percent identity and is calculated as follows:

For purposes of the present invention, the sequence identity between two deoxyribonucleotide sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, supra) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, supra) , preferably version 5.0.0 or later. The parameters used are gap open penalty of 10, gap extension penalty of 0.5, and the EDNAFULL (EMBOSS version of NCBI NUC4.4) substitution matrix. The output of Needle labeled “longest identity” (obtained using the –nobrief option) is used as the percent identity and is calculated as follows:

Textile: The term “textile” means any textile material including yarns, yarn intermediates, fibers, non-woven materials, natural materials, synthetic materials, and any other textile material, fabrics made of these materials and products made from fabrics (e.g., garments and other articles) . The textile or fabric may be in the form of knits, wovens, denims, non-wovens, felts, yarns, and towelling. The textile may be cellulose based such as natural cellulosics, including cotton, flax/linen, jute, ramie, sisal or coir or manmade cellulosics (e.g. originating from wood pulp) including viscose/rayon, cellulose acetate fibers (tricell) , lyocell or blends thereof. The textile or fabric may also be non-cellulose based such as natural polyamides including wool, camel, cashmere, mohair, rabbit and silk or synthetic polymers such as nylon, aramid, polyester, acrylic, polypropylene and spandex/elastane, or blends thereof as well as blends of cellulose based and non-cellulose based fibers. Examples of blends are blends of cotton and/or rayon/viscose with one or more companion material such as wool, synthetic fiber (e.g. polyamide fiber, acrylic fiber, polyester fiber, polyvinyl chloride fiber, polyurethane fiber, polyurea fiber, aramid fiber) , and/or cellulose-containing fiber (e.g. rayon/viscose, ramie, flax/linen, jute, cellulose acetate fiber, lyocell) . Fabric may be conventional washable laundry, for example stained household laundry. When the term fabric or garment is used it is intended to include the broader term textiles as well.

Wash cycle: The term “wash cycle” is defined herein as a washing operation wherein textile is exposed to the wash liquor for a period of time by circulating the wash liquor and optionally mechanically treat the textile in order to clean the textile and finally the superfluous wash liquor is removed. A wash cycle may be repeated one, two, three, four, five or even six times at the same or at different temperatures. Hereafter the textile is generally rinsed and dried. One of the wash cycles can be a soaking step, where the textile is left soaking in the wash liquor for a period.

Wash liquor: The term “wash liquor” is intended to mean the solution or mixture of water and a detergent component optionally including enzymes used for e.g. laundry.

Warewashing detergent composition: The term “warewashing detergent composition” refers to compositions that find use in the removal and/or reduction of undesired compounds from surfaces to be cleaned, such as surfaces of ware or the surfaces present in the interior of a warewashing machine.

The warewashing detergent composition can be any compositions intended for reducing and/or removing soil from dishes, table ware, pots, pans, cutlery and all forms of compositions for reducing and/or removing soil from the inner surfaces in dishwashing machines. The present invention is not restricted to any particular type of warewashing detergent composition. The terms encompass any detergent component selected for the particular type of cleaning composition desired and the form of the product (e.g., liquid, gel, powder, bar, granulate, paste, or spray compositions) . The detergent composition can contain enzymes in addition to the amylase and/or a protease comprised in the composition, e.g. one or more additional enzymes such as proteases, amylases, lipases, cutinases, cellulases, endoglucanases, xyloglucanases, pectinases, pectin lyases, xanthanases, peroxidaes, haloperoxygenases, catalases and mannanases, or any mixture thereof and a detergent component.

Warewashing machine: The term “warewashing machine” means any kind of washing machine that can be used for industrial or institutional warewashing. The term includes but is not limited to a door warewashing machine, a hood warewashing machine, a conveyor warewashing machine, an undercounter warewashing machine, a glasswasher, a flight warewashing machine, a pot and pan warewashing machine and a utensil washer.

Ware: The term “ware” is intended to mean any form of dishes, kitchen utensil, dinner set or tableware such as but not limited to pans, plates, drinking glasses, cups, knives, forks, spoons, porcelain etc. The ware can be made of any suitable material such as metal, glass, rubber, plastic, PVC, acrylics, ceramics, china or porcelain.

Wash performance: The term “wash performance” is used as an enzyme’s ability to remove or reduce dust mites present on a surface or textile to be cleaned during e.g. a cleaning cycle.

Wash time: The term “wash time” is defined herein as the time it takes for the entire washing process; i.e. the time for the wash cycle (s) and rinse cycle (s) together.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to the use of an enzyme/enzymes capable for degrading cellulosic material for reducing dust mite from a textile by adding said enzyme to detergent in wash cycle. The present inventors have found that by adding an enzyme capable for degrading cellulosic material to a detergent, the number of dust mites adhering on textile can be reduced significantly. In one aspect, said enzyme is an enzyme composition comprising cellulase or hemicellulase.

In another aspect, the present invention relates to an enzyme composition used for dust mite removal. Wherein, said enzyme composition used for dust mite removal comprises one or more cellulase or hemicellulase. Wherein, said cellulase can be GH45 cellulase. Further, said GH45 cellulase can be GH45 endoglucanase. In a further aspect, said cellulase is Thermothielavioides terrestris endoglucanase of SEQ ID NO: 2 or a cellulase comprises or consists of an amino acid sequence having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%sequence identity to the mature polypeptide of SEQ ID NO: 2.

In still another aspect, the present application comprises a method for removing dust mite comprising:

(2) Optionally rinsing the textile.

Enzymes

The enzymes or enzyme composition comprising cellulase or hemicellulase may further comprise one or more enzymes su

ch as a protease, lipase, cutinase, an amylase, carbohydrase, cellulase, pectinase, mannanase, arabinase, galactanase, xylanase, oxidase, e.g., a laccase, and/or peroxidase.

In general, the properties of the selected enzyme (s) should be compatible with the selected detergent, (i.e., pH-optimum, compatibility with other enzymatic and non-enzymatic ingredients, etc. ) , and the enzyme (s) should be present in effective amounts.

Cellulases

Suitable cellulases include those of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Suitable cellulases include cellulases from the genera Bacillus, Pseudomonas, Humicola, Fusarium, Thielavia, Acremonium, e.g., the fungal cellulases produced from Humicola insolens, Myceliophthora thermophila and Fusarium oxysporum disclosed in US 4,435,307, US 5,648,263, US 5,691,178, US 5,776,757 and WO 89/09259.

Especially suitable cellulases are the alkaline or neutral cellulases having colour care benefits. Examples of such cellulases are cellulases described in EP 0 495 257, EP 0 531 372, WO 96/11262, WO 96/29397, WO 98/08940. Other examples are cellulase variants such as those described in WO 94/07998, EP 0 531 315, US 5, 457, 046, US 5, 686, 593, US 5, 763, 254, WO 95/24471, WO 98/12307 and WO99/001544.

Other cellulases are endo-beta-1, 4-glucanase enzyme having a sequence of at least 97%identity to the amino acid sequence of position 1 to position 773 of SEQ ID NO: 2 of WO 2002/099091 or a family 44 xyloglucanase, which a xyloglucanase enzyme having a sequence of at least 60%identity to positions 40-559 of SEQ ID NO: 2 of WO 2001/062903.

Commercially available cellulases include Celluzyme ^TM, and Carezyme ^TM (Novozymes A/S) Carezyme Premium ^TM (Novozymes A/S) , Celluclean ^TM (Novozymes A/S) , Celluclean Classic ^TM (Novozymes A/S) , Cellusoft ^TM (Novozymes A/S) , Whitezyme ^TM (Novozymes A/S) , Clazinase ^TM, and Puradax HA ^TM (Genencor International Inc. ) , and KAC-500 (B) ^TM (Kao Corporation) .

GH45 cellulase

The GH Family 45 cellulase enzymes (formerly Family K) act with inversion of anomeric configuration to generate the alpha-D anomer of the oligosaccharide as a product. It has been elucidated that, in the active site, one aspartic acid amino acid acts as a general acid and another as a general base.

The three dimensional structure of Family 45 enzymes has been elucidated (see, for example, the structure of Humicolainsolens in Davies et al, 1996, ActaCrystallographica Section D-Biological Crystallography 52: 7-17 Part 1) . The enzymes contain a six-stranded beta-barrel to which a seventh strand is appended. The structure contains both parallel and anti-parallel beta-strands. The active center is located in an open substrate-binding groove.

As used herein, the term “GH45 cellulase” , “Family 45 cellulase” or “Cel45” means a carbohydrate active cellulase enzyme that contains a glycoside hydrolase Family 45 catalytic domain that is classified under EC 3.2.1.4. The term encompasses a carbohydrate active enzyme that hydrolyzes cellulose and cello-oligosaccharides using an inverting mechanism, and has either of the following two signature sequences in the vicinity of the catalytic aspartic acid amino acids: (i) both a first conserved signature sequence of A/S/T -T -R/N/T -Y/F/T -X -D -X -X -X -X -X -C/A-A/G/S-W/C and a second conserved signature sequence of H/Q/D/N -F/L -D -I/L/F; or (ii) has the second conserved signature sequence of H/Q/D/N -F/L -D -I/L/F but lacks said first conserved sequence. In one embodiment, the second conserved signature sequence is H-F-D-I.

Table 1: Family 45 cellulase subfamily B members:

Organism	Abreviated Name	GenBank Accession Number
Trichoderma reesei	TrCel45A	CAA83846.1
Trichomderma viride	TvEGV	AAQ21385.1
Penicillium decumbens	PdCel 45A	ACF33814.1
Aspergillus nidulans	AnAN6786.2	EAA58604.1
Hadiotis discus discus	HddEG1	ABO26608.1
Ampullaria crossean	AcEG27I	ABR92637.1
Ampullaria crossean	AcEG27II	ABR92638.1
Mytilus edulis	MeEG	CAC59695.1
Phanerochaete chrysosporium	PcCel45A	BAG68300.1

Table 2: Family 45 cellulase subfamily A members:

In a preferred embodiment, GH45 cellulase is Thermothielavioides terrestris endoglucanase.

(1) T. t endoglucanase comprising or consisting of the mature polypeptide of SEQ ID NO: 1 or fragments thereof;

(2) T. t endoglucanase comprising or consisting of an amino acid sequence having at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 1;

(3) T. t endoglucanase encoded by a polynucleotide comprising or consisting of a nucleotide sequence having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 2;

(4) T. t endoglucanase encoded by a polynucleotide that hybridizes under at least high stringency conditions, very high stringency conditions, with the mature polypeptide coding sequence of SEQ ID NO: 2 or the full-length complement thereof.

Detergent composition

The enzymes capable of degrading cellulosic material can be present in a laundry detergent composition comprising at least one or more additional enzyme (s) . The additional enzyme can be selected from the group consisting of hemicellulases, peroxidases, proteases, cellulases, xylanases, lipases, phospholipases, esterases, cutinases, pectinases, mannanases, pectate lyases, keratinases, reductases, oxidases, phenoloxidases, lipoxygenases, ligninases, pullulanases, tannases, pentosanases, malanases, β-glucanases, arabinosidases, hyaluronidase, chondroitinase, laccase, DNase, chlorophyllases, amylases, perhydrolases, peroxidases, xanthanase and mixtures thereof. In one embodiment of the invention the at least one or more additional enzyme is selected from proteases, lipases, mannanases, pectate lyases and amylases.

The detergent composition comprises a surfactant, which can be selected from the group consisting of anionic, cationic, non-ionic, semi-polar and zwitterionic surfactants. In addition other detergent components such as builders and polymers can be comprised in the detergent composition. When improving the water absorption of a textile, it has the benefit that items such as towels, can absorb more water when used for drying a skin or surfaces.

The invention further comprises a method for removing dust mite comprising subjecting a textile to an enzyme capable of degrading cellulosic material, or to an enzyme composition comprising the enzyme capable of degrading cellulosic material.

In one aspect, the method for removing dust mite comprising:

(3) Washing or exposing the textile with a detergent composition comprising an enzyme capable of degrading cellulosic material or an enzyme composition comprising the enzyme capable of degrading cellulosic material, wherein, said enzyme capable of degrading cellulosic material comprises GH45 cellulase. Wherein, said GH45 cellulase is GH45 endoglucanase. In a further aspect, said endogluanase is T. t endoglucanase.

(4) Optionally rinsing the textile.

The detergent composition may be e.g. a laundry detergent composition and may be in the form of a powder, a bar, a homogenous tablet, a tablet having two or more layers, a pouch having one or more compartments, a regular or compact powder, a granule, a paste, a gel, or a regular, compact or concentrated liquid. The composition can be a powder or a granule where the acidic material is coated on the powder or granule as an outer layer.

Mannanases

Suitable mannanases include those of bacterial or fungal origin. Chemically or genetically modified mutants are included. The mannanase may be an alkaline mannanase of Family 5 or 26. It may be a wild-type from Bacillus or Humicola, particularly B. agaradhaerens, B. licheniformis, B. halodurans, B. clausii, or H. insolens. Suitable mannanases are described in WO 1999/064619. A commercially available mannanase is Mannaway (Novozymes A/S) .

Peroxidases/Oxidases

Suitable peroxidases/oxidases include those of plant, bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Examples of useful peroxidases include peroxidases from Coprinus, e.g., from C. cinereus, and variants thereof as those described in WO 93/24618, WO 95/10602, and WO 98/15257. Commercially available peroxidases include Guardzyme ^TM (Novozymes A/S) . A peroxidase according to the invention also includes a haloperoxidase enzyme, such as chloroperoxidase, bromoperoxidase and compounds exhibiting chloroperoxidase or bromoperoxidase activity. Haloperoxidases are classified according to their specificity for halide ions. Chloroperoxidases (E. C. 1.11.1.10) catalyze formation of hypochlorite from chloride ions.

In an embodiment, the haloperoxidase of the invention is a chloroperoxidase. Preferably, the haloperoxidase is a vanadium haloperoxidase, i.e., a vanadate-containing haloperoxidase. In a preferred method of the present invention the vanadate-containing haloperoxidase is combined with a source of chloride ion.

Haloperoxidases have been isolated from many different fungi, in particular from the fungus group dematiaceous hyphomycetes, such as Caldariomyces, e.g., C. fumago, Alternaria, Curvularia, e.g., C. verruculosa and C. inaequalis, Drechslera, Ulocladium and Botrytis.

Haloperoxidases have also been isolated from bacteria such as Pseudomonas, e.g., P. pyrrocinia and Streptomyces, e.g., S. aureofaciens.

In a preferred embodiment, the haloperoxidase is derivable from Curvularia sp., in particular Curvularia verruculosa or Curvularia inaequalis, such as C. inaequalis CBS 102.42 as described in WO 95/27046; or C. verruculosa CBS 147.63 or C. verruculosa CBS 444.70 as described in WO 97/04102; or from Drechslera hartlebii as described in WO 01/79459, Dendryphiella salina as described in WO 01/79458, Phaeotrichoconis crotalarie as described in WO 01/79461, or Geniculosporium sp. as described in WO 01/79460.

An oxidase according to the invention include, in particular, any laccase enzyme comprised by the enzyme classification EC 1.10.3.2, or any fragment derived therefrom exhibiting laccase activity, or a compound exhibiting a similar activity, such as a catechol oxidase (EC 1.10.3.1) , an o-aminophenol oxidase (EC 1.10.3.4) , or a bilirubin oxidase (EC 1.3.3.5) .

Proteases

Suitable proteases include those of bacterial, fungal, plant, viral or animal origin e.g. vegetable or microbial origin. Microbial origin is preferred. Chemically modified or protein engineered mutants are included. It may be an alkaline protease, such as a serine protease or a metalloprotease. A serine protease may for example be of the S1 family, such as trypsin, or the S8 family such as subtilisin. A metalloproteases protease may for example be a thermolysin from e.g. family M4 or other metalloprotease such as those from M5, M7 or M8 families.

The term "subtilases" refers to a sub-group of serine protease according to Siezen et al., Protein Engng. 4 (1991) 719-737 and Siezen et al. Protein Science 6 (1997) 501-523. Serine proteases are a subgroup of proteases characterized by having a serine in the active site, which forms a covalent adduct with the substrate. The subtilases may be divided into 6 sub-divisions, i.e. the Subtilisin family, the Thermitase family, the Proteinase K family, the Lantibiotic peptidase family, the Kexin family and the Pyrolysin family.

Examples of subtilases are those derived from Bacillus such as Bacillus lentus, B. alkalophilus, B. subtilis, B. amyloliquefaciens, Bacillus pumilus and Bacillus gibsonii described in;US7262042 and WO09/021867, and subtilisin lentus, subtilisin Novo, subtilisin Carlsberg, Bacillus licheniformis, subtilisin BPN’, subtilisin 309, subtilisin 147 and subtilisin 168 described in WO89/06279 and protease PD138 described in (WO93/18140) . Other useful proteases may be those described in WO92/175177, WO01/016285, WO02/026024 and WO02/016547. Examples of trypsin-like proteases are trypsin (e.g. of porcine or bovine origin) and the Fusarium protease described in WO89/06270, WO94/25583 and WO05/040372, and the chymotrypsin proteases derived from Cellumonas described in WO05/052161 and WO05/052146.

A further preferred protease is the alkaline protease from Bacillus lentus DSM 5483, as described for example in WO95/23221, and variants thereof which are described in WO92/21760, WO95/23221, EP1921147 and EP1921148.

Examples of metalloproteases are the neutral metalloprotease as described in WO07/044993 (Genencor Int. ) such as those derived from Bacillus amyloliquefaciens.

Examples of useful proteases are the variants described in: WO92/19729, WO96/034946, WO98/20115, WO98/20116, WO99/011768, WO01/44452, WO03/006602, WO04/03186, WO04/041979, WO07/006305, WO11/036263, WO11/036264, especially the variants with substitutions in one or more of the following positions: 3, 4, 9, 15, 27, 36, 57, 68, 76, 87, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 106, 118, 120, 123, 128, 129, 130, 160, 167, 170, 194, 195, 199, 205, 206, 217, 218, 222, 224, 232, 235, 236, 245, 248, 252 and 274 using the BPN’ numbering. More preferred the subtilase variants may comprise the mutations: S3T, V4I, S9R, A15T, K27R, *36D, V68A, N76D, N87S, R, *97E, A98S, S99G, D, A, S99AD, S101G, M, R S103A, V104I, Y, N, S106A, G118V, R, H120D, N, N123S, S128L, P129Q, S130A, G160D, Y167A, R170S, A194P, G195E, V199M, V205I, L217D, N218D, M222S, A232V, K235L, Q236H, Q245R, N252K, T274A (using BPN’ numbering) .

Suitable commercially available protease enzymes include those sold under the trade names

Duralase ^Tm, Durazym ^Tm,

Ultra,

and

(Novozymes A/S) , those sold under the tradename

Purafect

and

(Danisco/DuPont) , Axapem ^TM (Gist-Brocases N.V. ) , BLAP (sequence shown in Figure 29 of US5352604) and variants hereof (Henkel AG) and KAP (Bacillus alkalophilus subtilisin) from Kao.

Lipases and Cutinases

Suitable lipases and cutinases include those of bacterial or fungal origin. Chemically modified or protein engineered mutant enzymes are included. Examples include lipase from Thermomyces, e.g. from T. lanuginosus (previously named Humicola lanuginosa) as described in EP258068 and EP305216, cutinase from Humicola, e.g. H. insolens (WO96/13580) , lipase from strains of Pseudomonas (some of these now renamed to Burkholderia) , e.g. P. alcaligenes or P. pseudoalcaligenes (EP218272) , P. cepacia (EP331376) , P. sp. strain SD705 (WO95/06720 &WO96/27002) , P. wisconsinensis (WO96/12012) , GDSL-type Streptomyces lipases (WO10/065455) , cutinase from Magnaporthe grisea (WO10/107560) , cutinase from Pseudomonas mendocina (US5, 389, 536) , lipase from Thermobifida fusca (WO11/084412) , Geobacillus stearothermophilus lipase (WO11/084417) , lipase from Bacillus subtilis (WO11/084599) , and lipase from Streptomyces griseus (WO11/150157) and S. pristinaespiralis (WO12/137147) .

Other examples are lipase variants such as those described in EP407225, WO92/05249, WO94/01541, WO94/25578, WO95/14783, WO95/30744, WO95/35381, WO95/22615, WO96/00292, WO97/04079, WO97/07202, WO00/34450, WO00/60063, WO01/92502, WO07/87508 and WO09/109500.

Preferred commercial lipase products include include Lipolase ^TM, LipexTM; Lipolex ^TM and Lipoclean ^TM (Novozymes A/S) , Lumafast (originally from Genencor) and Lipomax (originally from Gist-Brocades) .

Still other examples are lipases sometimes referred to as acyltransferases or perhydrolases, e.g. acyltransferases with homology to Candida antarctica lipase A (WO10/111143) , acyltransferase from Mycobacterium smegmatis (WO05/56782) , perhydrolases from the CE 7 family (WO09/67279) , and variants of the M. smegmatis perhydrolase in particular the S54V variant used in the commercial product Gentle Power Bleach from Huntsman Textile Effects Pte Ltd (WO10/100028) .

Amylases

Suitable amylases which can be used together with the enzyme preparation of the invention may be an alpha-amylase or a glucoamylase and may be of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Amylases include, for example, alpha-amylases obtained from Bacillus, e.g., a special strain of Bacillus licheniformis, described in more detail in GB 1,296,839.

Suitable amylases include amylases having SEQ ID NO: 2 in WO 95/10603 or variants having 90%sequence identity to SEQ ID NO: 3 thereof. Preferred variants are described in WO 94/02597, WO 94/18314, WO 97/43424 and SEQ ID NO: 4 of WO 99/019467, such as variants with substitutions in one or more of the following positions: 15, 23, 105, 106, 124, 128, 133, 154, 156, 178, 179, 181, 188, 190, 197, 201, 202, 207, 208, 209, 211, 243, 264, 304, 305, 391, 408, and 444.

Different suitable amylases include amylases having SEQ ID NO: 6 in WO 02/010355 or variants thereof having 90%sequence identity to SEQ ID NO: 6. Preferred variants of SEQ ID NO: 6 are those having a deletion in positions 181 and 182 and a substitution in position 193.

Other amylases which are suitable are hybrid alpha-amylase comprising residues 1-33 of the alpha-amylase derived from B. amyloliquefaciens shown in SEQ ID NO: 6 of WO 2006/066594 and residues 36-483 of the B. licheniformis alpha-amylase shown in SEQ ID NO: 4 of WO 2006/066594 or variants having 90%sequence identity thereof. Preferred variants of this hybrid alpha-amylase are those having a substitution, a deletion or an insertion in one of more of the following positions: G48, T49, G107, H156, A181, N190, M197, I201, A209 and Q264. Most preferred variants of the hybrid alpha-amylase comprising residues 1-33 of the alpha-amylase derived from B. amyloliquefaciens shown in SEQ ID NO: 6 of WO 2006/066594 and residues 36-483 of SEQ ID NO: 4 are those having the substitutions:

M197T;

H156Y+A181T+N190F+A209V+Q264S; or

G48A+T49I+G107A+H156Y+A181T+N190F+I201F+A209V+Q264S.

Further amylases which are suitable are amylases having SEQ ID NO: 6 in WO 99/019467 or variants thereof having 90%sequence identity to SEQ ID NO: 6. Preferred variants of SEQ ID NO: 6 are those having a substitution, a deletion or an insertion in one or more of the following positions: R181, G182, H183, G184, N195, I206, E212, E216 and K269. Particularly preferred amylases are those having deletion in positions R181 and G182, or positions H183 and G184.

Additional amylases which can be used are those having SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 2 or SEQ ID NO: 7 of WO 96/023873 or variants thereof having 90%sequence identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 7. Preferred variants of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 7 are those having a substitution, a deletion or an insertion in one or more of the following positions: 140, 181, 182, 183, 184, 195, 206, 212, 243, 260, 269, 304 and 476, using SEQ ID 2 of WO 96/023873 for numbering. More preferred variants are those having a deletion in two positions selected from 181, 182, 183 and 184, such as 181 and 182, 182 and 183, or positions 183 and 184. Most preferred amylase variants of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 7 are those having a deletion in positions 183 and 184 and a substitution in one or more of positions 140, 195, 206, 243, 260, 304 and 476.

Other amylases which can be used are amylases having SEQ ID NO: 2 of WO 08/153815, SEQ ID NO: 10 in WO 01/66712 or variants thereof having 90%sequence identity to SEQ ID NO: 2 of WO 08/153815 or 90%sequence identity to SEQ ID NO: 10 in WO 01/66712. Preferred variants of SEQ ID NO: 10 in WO 01/66712 are those having a substitution, a deletion or an insertion in one of more of the following positions: 176, 177, 178, 179, 190, 201, 207, 211 and 264.

Further suitable amylases are amylases having SEQ ID NO: 2 of WO 09/061380 or variants having 90%sequence identity to SEQ ID NO: 2 thereof. Preferred variants of SEQ ID NO: 2 are those having a truncation of the C-terminus and/or a substitution, a deletion or an insertion in one of more of the following positions: Q87, Q98, S125, N128, T131, T165, K178, R180, S181, T182, G183, M201, F202, N225, S243, N272, N282, Y305, R309, D319, Q320, Q359, K444 and G475. More preferred variants of SEQ ID NO: 2 are those having the substitution in one of more of the following positions: Q87E, R, Q98R, S125A, N128C, T131I, T165I, K178L, T182G, M201L, F202Y, N225E, R, N272E, R, S243Q, A, E, D, Y305R, R309A, Q320R, Q359E, K444E and G475K and/or deletion in position R180 and/or S181 or of T182 and/or G183. Most preferred amylase variants of SEQ ID NO: 2 are those having the substitutions:

N128C+K178L+T182G+Y305R+G475K;

N128C+K178L+T182G+F202Y+Y305R+D319T+G475K;

S125A+N128C+K178L+T182G+Y305R+G475K; or

S125A+N128C+T131I+T165I+K178L+T182G+Y305R+G475K wherein the variants are C-terminally truncated and optionally further comprises a substitution at position 243 and/or a deletion at position 180 and/or position 181.

Further suitable amylases are amylases having SEQ ID NO: 1 of WO13184577 or variants having 90%sequence identity to SEQ ID NO: 1 thereof. Preferred variants of SEQ ID NO: 1 are those having a substitution, a deletion or an insertion in one of more of the following positions: K176, R178, G179, T180, G181, E187, N192, M199, I203, S241, R458, T459, D460, G476 and G477. More preferred variants of SEQ ID NO: 1 are those having the substitution in one of more of the following positions: K176L, E187P, N192FYH, M199L, I203YF, S241QADN, R458N, T459S, D460T, G476K and G477K and/or deletion in position R178 and/or S179 or of T180 and/or G181. Most preferred amylase variants of SEQ ID NO: 1 are those having the substitutions:

E187P+I203Y+G476K

E187P+I203Y+R458N+T459S+D460T+G476K

wherein the variants optionally further comprises a substitution at position 241 and/or a deletion at position 178 and/or position 179.

Further suitable amylases are amylases having SEQ ID NO: 1 of WO10104675 or variants having 90%sequence identity to SEQ ID NO: 1 thereof. Preferred variants of SEQ ID NO: 1 are those having a substitution, a deletion or an insertion in one of more of the following positions: N21, D97, V128 K177, R179, S180, I181, G182, M200, L204, E242, G477 and G478. More preferred variants of SEQ ID NO: 1 are those having the substitution in one of more of the following positions: N21D, D97N, V128I K177L, M200L, L204YF, E242QA, G477K and G478K and/or deletion in position R179 and/or S180 or of I181 and/or G182. Most preferred amylase variants of SEQ ID NO: 1 are those having the substitutions:

N21D+D97N+V128I wherein the variants optionally further comprises a substitution at position 200 and/or a deletion at position 180 and/or position 181.

Other suitable amylases are the alpha-amylase having SEQ ID NO: 12 in WO01/66712 or a variant having at least 90%sequence identity to SEQ ID NO: 12. Preferred amylase variants are those having a substitution, a deletion or an insertion in one of more of the following positions of SEQ ID NO: 12 in WO01/66712: R28, R118, N174; R181, G182, D183, G184, G186, W189, N195, M202, Y298, N299, K302, S303, N306, R310, N314; R320, H324, E345, Y396, R400, W439, R444, N445, K446, Q449, R458, N471, N484. Particular preferred amylases include variants having a deletion of D183 and G184 and having the substitutions R118K, N195F, R320K and R458K, and a variant additionally having substitutions in one or more position selected from the group: M9, G149, G182, G186, M202, T257, Y295, N299, M323, E345 and A339, most preferred a variant that additionally has substitutions in all these positions.

Other examples are amylase variants such as those described in WO2011/098531, WO2013/001078 and WO2013/001087.

Commercially available amylases are Duramyl ^TM, Termamyl ^TM, Fungamyl ^TM, Stainzyme ^TM, Stainzyme Plus ^TM, Natalase ^TM, Liquozyme X and BAN ^TM (from Novozymes A/S) , and Rapidase ^TM, Purastar ^TM/Effectenz ^TM, Powerase, Preferenz S1000, Preferenz S100 and Preferenz S110 (from Genencor International Inc. /DuPont) .

Concentration of the enzyme

In one embodiment of the present invention, the enzyme capable for degrading cellulosic material of the present invention may be used in the detergent composition in an amount corresponding to 0.001-200 mg of protein, such as 0.005-100 mg of protein, preferably 0.01-50 mg of protein, more preferably 0.05-20 mg of protein, even more preferably 0.1-10 mg of protein per liter of wash liquor.

The enzyme (s) of the detergent composition of the invention may be stabilized using conventional stabilizing agents, e.g. a polyol such as propylene glycol or glycerol, a sugar or sugar alcohol, lactic acid, boric acid, or a boric acid derivative, e.g. an aromatic borate ester, or a phenyl boronic acid derivative such as 4-formylphenyl boronic acid, and the composition may be formulated as described in, for example, WO92/19709 and WO92/19708.

Surfactants

The laundry detergent composition may comprise one or more surfactants, which may be anionic and/or cationic and/or non-ionic and/or semi-polar and/or zwitterionic, or a mixture thereof.

In a particular embodiment, the detergent composition includes a mixture of one or more nonionic surfactants and one or more anionic surfactants. The surfactant (s) is typically present at a level of from about 0.1%to 60%by weight, such as about 1%to about 40%, or about 3%to about 20%, or about 3%to about 10%. The surfactant (s) is chosen based on the desired cleaning application, and may include any conventional surfactant (s) known in the art.

When included therein, the detergent will usually contain from about 1%to about 40%by weight of an anionic surfactant, such as from about 5%to about 30%, including from about 5%to about 15%, or from about 15%to about 20%, or from about 20%to about 25%of an anionic surfactant. Non-limiting examples of anionic surfactants include sulfates and sulfonates, in particular, linear alkylbenzenesulfonates (LAS) , isomers of LAS, branched alkylbenzenesulfonates (BABS) , phenylalkanesulfonates, alpha-olefinsulfonates (AOS) , olefin sulfonates, alkene sulfonates, alkane-2, 3-diylbis (sulfates) , hydroxyalkanesulfonates and disulfonates, alkyl sulfates (AS) such as sodium dodecyl sulfate (SDS) , fatty alcohol sulfates (FAS) , primary alcohol sulfates (PAS) , alcohol ethersulfates (AES or AEOS or FES, also known as alcohol ethoxysulfates or fatty alcohol ether sulfates) , secondary alkanesulfonates (SAS) , paraffin sulfonates (PS) , ester sulfonates, sulfonated fatty acid glycerol esters, alpha-sulfo fatty acid methyl esters (alpha-SFMe or SES) including methyl ester sulfonate (MES) , alkyl-or alkenylsuccinic acid, dodecenyl/tetradecenyl succinic acid (DTSA) , fatty acid derivatives of amino acids, diesters and monoesters of sulfo-succinic acid or salt of fatty acids (soap) , and combinations thereof.

When included therein, the detergent will usually contain from about from about 1%to about 40%by weigh of a cationic surfactant, for example from about 0.5%to about 30%, in particular from about 1%to about 20%, from about 3%to about 10%, such as from about 3%to about 5%, from about 8%to about 12%or from about 10%to about 12%. Non-limiting examples of cationic surfactants include alkyldimethylethanolamine quat (ADMEAQ) , cetyltrimethylammonium bromide (CTAB) , dimethyldistearylammonium chloride (DSDMAC) , and alkylbenzyldimethylammonium, alkyl quaternary ammonium compounds, alkoxylated quaternary ammonium (AQA) compounds, ester quats, and combinations thereof.

When included therein, the detergent will usually contain from about 0.2%to about 40%by weight of a nonionic surfactant, for example from about 0.5%to about 30%, in particular from about 1%to about 20%, from about 3%to about 10%, such as from about 3%to about 5%, from about 8%to about 12%, or from about 10%to about 12%. Non-limiting examples of nonionic surfactants include alcohol ethoxylates (AE or AEO) , alcohol propoxylates, propoxylated fatty alcohols (PFA) , alkoxylated fatty acid alkyl esters, such as ethoxylated and/or propoxylated fatty acid alkyl esters, alkylphenol ethoxylates (APE) , nonylphenol ethoxylates (NPE) , alkylpolyglycosides (APG) , alkoxylated amines, fatty acid monoethanolamides (FAM) , fatty acid diethanolamides (FADA) , ethoxylated fatty acid monoethanolamides (EFAM) , propoxylated fatty acid monoethanolamides (PFAM) , polyhydroxyalkyl fatty acid amides, or N-acyl N-alkyl derivatives of glucosamine (glucamides, GA, or fatty acid glucamides, FAGA) , as well as products available under the trade names SPAN and TWEEN, and combinations thereof.

When included therein, the detergent will usually contain from about 0%to about 20%by weight of a semipolar surfactant. Non-limiting examples of semipolar surfactants include amine oxides (AO) such as alkyldimethylamineoxide, N- (coco alkyl) -N, N-dimethylamine oxide and N-(tallow-alkyl) -N, N-bis (2-hydroxyethyl) amine oxide, and combinations thereof.

When included therein, the detergent will usually contain from about 0%to about 20%by weight of a zwitterionic surfactant. Non-limiting examples of zwitterionic surfactants include betaines such as alkyldimethylbetaines, sulfobetaines, and combinations thereof.

A polypeptide of the present invention may also be incorporated in the detergent formulations disclosed in WO97/07202, which is hereby incorporated by reference.

Builders and Co-Builders

The detergent composition may comprise about 0-10%by weight, such as about 0.1%to about 5%of a builder or co-builder, or a mixture thereof. In a softener, the level of builder is typically0-1%, particularly 0-0, 5%. The builder and/or co-builder may particularly be a chelating agent that forms water-soluble complexes with Ca and Mg. Any builder and/or co-builder known in the art for use in softener may be utilized. Non-limiting examples of builders include zeolites, diphosphates (pyrophosphates) , triphosphates such as sodium triphosphate (STP or STPP) , carbonates such as sodium carbonate, soluble silicates such as sodium metasilicate, layered silicates (e.g., SKS-6 from Hoechst) , ethanolamines such as 2-aminoethan-1-ol (MEA) , diethanolamine (DEA, also known as 2, 2’-iminodiethan-1-ol) , triethanolamine (TEA, also known as 2, 2’, 2”-nitrilotriethan-1-ol) , and (carboxymethyl) inulin (CMI) , and combinations thereof.

The detergent composition may also comprise 0-5%by weight, such as about 0%to about 2%, of a detergent co-builder. The detergent composition may include include a co-builder alone, or in combination with a builder, for example a zeolite builder. Non-limiting examples of co-builders include homopolymers of polyacrylates or copolymers thereof, such as poly (acrylic acid) (PAA) or copoly (acrylic acid/maleic acid) (PAA/PMA) . Further non-limiting examples include citrate, chelators such as aminocarboxylates, aminopolycarboxylates and phosphonates, and alkyl-or alkenylsuccinic acid. Additional specific examples include 2, 2’, 2”-nitrilotriacetic acid (NTA) , ethylenediaminetetraacetic acid (EDTA) , diethylenetriaminepentaacetic acid (DTPA) , iminodisuccinic acid (IDS) , ethylenediamine-N, N’-disuccinic acid (EDDS) , methylglycinediacetic acid (MGDA) , glutamic acid-N, N-diacetic acid (GLDA) , 1-hydroxyethane-1, 1-diphosphonic acid (HEDP) , ethylenediaminetetra (methylenephosphonic acid) (EDTMPA) , diethylenetriaminepentakis (methylenephosphonic acid) (DTMPA or DTPMPA) , N- (2-hydroxyethyl) iminodiacetic acid (EDG) , aspartic acid-N-monoacetic acid (ASMA) , aspartic acid-N, N-diacetic acid (ASDA) , aspartic acid-N-monopropionic acid (ASMP) , iminodisuccinic acid (IDA) , N- (2-sulfomethyl) -aspartic acid (SMAS) , N- (2-sulfoethyl) -aspartic acid (SEAS) , N- (2-sulfomethyl) -glutamic acid (SMGL) , N- (2-sulfoethyl) -glutamic acid (SEGL) , N-methyliminodiacetic acid (MIDA) , α-alanine-N, N-diacetic acid (α-ALDA) , serine-N, N-diacetic acid (SEDA) , isoserine-N, N-diacetic acid (ISDA) , phenylalanine-N, N-diacetic acid (PHDA) , anthranilic acid-N, N-diacetic acid (ANDA) , sulfanilic acid-N, N-diacetic acid (SLDA) , taurine-N, N-diacetic acid (TUDA) and sulfomethyl-N, N-diacetic acid (SMDA) , N- (2-hydroxyethyl) ethylenediamine-N, N’, N”-triacetic acid (HEDTA) , diethanolglycine (DEG) , diethylenetriamine penta (methylenephosphonic acid) (DTPMP) , aminotris (methylenephosphonic acid) (ATMP) , and combinations and salts thereof. Further exemplary builders and/or co-builders are described in, e.g., WO 09/102854, US 5977053

Polymers

The detergent may comprise 0-10%by weight, such as 0.5-5%, 2-5%, 0.5-2%or 0.2-1%of a polymer. Any polymer known in the art for use in softeners may be utilized. The polymer may function as a co-builder as mentioned above, or may provide antiredeposition, fiber protection, soil release, dye transfer inhibition, anti-foaming properties, perfume encapsulation and lubricity. Some polymers may have more than one of the above-mentioned properties and/or more than one of the below-mentioned motifs. Exemplary polymers include polyquaterniums, melamine polymers, siloxanes, silicones, carboxymethyl) cellulose (CMC) , poly (vinyl alcohol) (PVA) , poly (vinylpyrrolidone) (PVP) , poly (ethyleneglycol) or poly (ethylene oxide) (PEG) , ethoxylated poly (ethyleneimine) , carboxymethyl inulin (CMI) , and polycarboxylates such as PAA, PAA/PMA, poly-aspartic acid, and lauryl methacrylate/acrylic acid copolymers , hydrophobically modified CMC (HM-CMC) , copolymers of terephthalic acid and oligomeric glycols, copolymers of poly (ethylene terephthalate) and poly (oxyethene terephthalate) (PET-POET) , PVP, poly (vinylimidazole) (PVI) , poly (vinylpyridine-N-oxide) (PVPO or PVPNO) and polyvinylpyrrolidone-vinylimidazole (PVPVI) . Further exemplary polymers include sulfonated polycarboxylates, polyethylene oxide and polypropylene oxide (PEO-PPO) and diquaternium ethoxy sulfate. Other exemplary polymers are disclosed in, e.g., WO 2006/130575. Salts of the above-mentioned polymers are also contemplated.

Adjunct materials

Any detergent component known in the art for use in softeners may also be utilized. Other optional softener components include solvents (including isopropyl alcohol, propylene glycol, alkane/cycloalkane) , anti-shrink agents, anti-soil redeposition agents, anti-wrinkling agents, bactericides, preservatives (including benzisothiazolinone, methylisothiazolinone and/or lactic acid) , binders, dyes, enzyme stabilizers (including boric acid, borates, CMC, and/or polyols such as propylene glycol) , emulsion stabilizers, antifoam agents (including dimethicone) , skin conditioning agents (including caprylic/capric glycerides, ethylhexyl stearate, or cocos oil , either alone or in combination. Any ingredient known in the art for use in softeners may be utilized. The choice of such ingredients is well within the skill of the artisan.

The present invention is further described by the following examples that should not be construed as limiting the scope of the invention.

Anti-redeposition agents

The detergent compositions of the present invention may also include one or more anti-redeposition agents such as carboxymethylcellulose (CMC) , polyvinyl alcohol (PVA) , polyvinylpyrrolidone (PVP) , polyoxyethylene and/or polyethyleneglycol (PEG) , homopolymers of acrylic acid, copolymers of acrylic acid and maleic acid, and ethoxylated polyethyleneimines. The cellulose based polymers described under soil release polymers above may also function as anti-redeposition agents.

Formulation of detergent products

The detergent composition of the invention may be in any convenient form, e.g., a bar, a homogenous tablet, a tablet having two or more layers, a pouch having one or more compartments, a regular or compact powder, a granule, a paste, a gel, or a regular, compact or concentrated liquid.

Pouches can be configured as single or multicompartments. It can be of any form, shape and material which is suitable for hold the composition, e.g. without allowing the release of the composition to release of the composition from the pouch prior to water contact. The pouch is made from water soluble film which encloses an inner volume. Said inner volume can be divided into compartments of the pouch. Preferred films are polymeric materials preferably polymers which are formed into a film or sheet. Preferred polymers, copolymers or derivates thereof are selected polyacrylates, and water soluble acrylate copolymers, methyl cellulose, carboxy methyl cellulose, sodium dextrin, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, malto dextrin, poly methacrylates, most preferably polyvinyl alcohol copolymers and, hydroxypropyl methyl cellulose (HPMC) . Preferably the level of polymer in the film for example PVA is at least about 60%. Preferred average molecular weight will typically be about 20,000 to about 150,000. Films can also be of blended compositions comprising hydrolytically degradable and water soluble polymer blends such as polylactide and polyvinyl alcohol (known under the Trade reference M8630 as sold by MonoSol LLC, Indiana, USA) plus plasticisers like glycerol, ethylene glycerol, propylene glycol, sorbitol and mixtures thereof. The pouches can comprise a solid laundry cleaning composition or part components and/or a liquid cleaning composition or part components separated by the water soluble film. The compartment for liquid components can be different in composition than compartments containing solids: US2009/0011970 A1.

Detergent ingredients can be separated physically from each other by compartments in water dissolvable pouches or in different layers of tablets. Thereby negative storage interaction between components can be avoided. Different dissolution profiles of each of the compartments can also give rise to delayed dissolution of selected components in the wash solution.

A liquid or gel detergent , which is not unit dosed, may be aqueous, typically containing at least 20%by weight and up to 95%water, such as up to about 70%water, up to about 65%water, up to about 55%water, up to about 45%water, up to about 35%water. Other types of liquids, including without limitation, alkanols, amines, diols, ethers and polyols may be included in an aqueous liquid or gel. An aqueous liquid or gel detergent may contain from 0-30%organic solvent.

A liquid or gel detergent may be non-aqueous.

Formulation of enzyme in co-granule

The enzyme of the invention may be formulated as a granule for example as a co-granule that combines one or more enzymes. Each enzyme will then be present in more granules securing a more uniform distribution of enzymes in the detergent. This also reduces the physical segregation of different enzymes due to different particle sizes. Methods for producing multi-enzyme co-granulates for the detergent industry are disclosed in the IP. com disclosure IPCOM000200739D.

Another example of formulation of enzymes by the use of co-granulates are disclosed in WO 2013/188331, which relates to a detergent composition comprising (a) a multi-enzyme co-granule; (b) less than 10 wt zeolite (anhydrous basis) ; and (c) less than 10 wt phosphate salt (anhydrous basis) , wherein said enzyme co-granule comprises from 10 to 98 wt%moisture sink component and the composition additionally comprises from 20 to 80 wt%detergent moisture sink component.

WO 2013/188331 also relates to a method of treating and/or cleaning a surface, preferably a fabric surface comprising the steps of (i) contacting said surface with the detergent composition as claimed and described herein in an aqueous wash liquor, (ii) rinsing and/or drying the surface.

The multi-enzyme co-granule may comprise an enzyme of the invention and (a) one or more enzymes selected from the group consisting of first-wash lipases, cleaning cellulases, xyloglucanases, perhydrolases, peroxidases, lipoxygenases, laccases and mixtures thereof; and (b) one or more enzymes selected from the group consisting of hemicellulases, proteases, care cellulases, cellobiose dehydrogenases, xylanases, phospho lipases, esterases, cutinases, pectinases, mannanases, pectate lyases, keratinases, reductases, oxidases, phenoloxidases, ligninases, pullulanases, tannases, pentosanases, lichenases glucanases, arabinosidases, hyaluronidase, chondroitinase, amylases, and mixtures thereof.

EXAMPLES

Washing method

The compositions of the invention were used in essentially any washing, cleaning and/or fabric care methods, including but not limited to soaking methods, pre-treatment methods, methods with rinsing steps for which a separate rinse aid composition was added and post-treatment methods.

The process described herein contacted fabrics with a laundering solution in the usual manner and exemplified here under. A conventional laundry method treated soiled fabric with an aqueous liquid dissolved or dispensed therein an effective amount of the laundry detergent and/or fabric care composition. The process of the invention was conveniently carried out in the course of the cleaning process. The method of cleaning was preferably carried out at 5℃ to 95℃, especially between 10℃ and 60℃. The pH of the treatment solution is 6 -12, preferably from 7 to 10.

The following examples meant to exemplify compositions of the present invention but not necessarily meant to limit or otherwise define the scope of the invention. In the detergent compositions, the enzymes levels were expressed by pure enzyme of the total composition by weight and unless otherwise specified, the detergent ingredients levels were expressed by detergent ingredient of the total compositions by weight.

Materials and Method

Test enzyme:

Thermothielavioides terrestris endoglucanase of SEQ ID NO: 2 obtained according to WO 2012/089024;

Fabrics:

CN-42, knitted cotton, commercially purchased from Center for Testmaterials B. V (CFT) , Netherlands;

E-252, striped colored cotton, commercially purchased from Center for Testmaterials B.V, Netherlands;

E-252PA, striped colored cotton, derived from E-252 mentioned above, which was prepared by the following process: preaged E-252 in Wascator auto washing machine for 12 hours at temperature 40℃ to generate visible pilling on the surface;

Model Detergent A:

Model detergent A was prepared with the following ingredients:

Method:

Washing method

Terg-O-tometer (TOM) wash assay

The Terg-O-tometer (TOM) was a medium scale model wash system that could be applied to test 16 different wash conditions simultaneously. TOM was basically a large temperature-controlled water bath with up to 16 open metal beakers submerged into it. Each beaker constituted one small top loader style washing machine and during an experiment, each of them contained a solution of a specific detergent/enzyme system and the performance of the cotton fabrics were tested. A rotating stirring arm was used to achieve desired mechanical stress. Said arm stirred the liquid within each beaker. Because the TOM beakers had no lid, it was possible to withdraw samples during a TOM experiment and assay for information on-line.

Equipment: The water bath with 16 steel beakers and 1 rotating stirrer per beaker was used with capacity of 1000mL of detergent solution. Temperature was set at 40℃. The water bath was filled with water. Rotational speed was set up to 200rpm/min.

1. Temperature in the Terg-O-Tometer was set to 40℃ and the rotation was started in the water bath. The temperature was kept at 40 +/-0.5℃.

2. Wash solution was prepared by adding desired amount of detergent into said solution and magnet stirred for 10 min under 40+/-0.5℃ and 14dH were set up in a bucket. Wash solution should be used within 30 to 60 min after preparation.

3. 1000ml wash solution was added into a TOM beaker.

4. Agitation was started at 200rpm and optionally enzymes were added to the beaker.

5. Sprinkled the fabrics into the beaker and then the ballast load.

6. Time measurement was started when the swatches and ballast were added to the beaker.

7. Washed for 70 minutes.

8. Agitation was stopped.

9. Transferred the wash load from TOM beaker to a sieve and rinsed with cold tap water.

10. Separated the test materials from the ballast load. The test materials were transferred to a 5L beaker with cold tap water under running water.

11. Set the timer to 5 minutes.

12. Pressed gently the water out by hand and placed the test swatches on a tray covered with a paper. Another paper was added on top of the swatches.

13. Dried the swatches in tumble dryer and then measured at the Color Eye as described below.

Anti-pilling effect determination

The E-252PA was measured by Color Eye equipment (CE7000/CE7000A, Largo AB) . L value was used to quantify the antipilling benefit. L indicated the change in white/black on a scale from 0 to 100, and a decrease in L meant an increase in black color (decrease in white color) and an increase in L meant an increase in white color (decrease in black color) . So, lower L value meant better antipilling effect.

The CN-42 was evaluated by sensory panel, 8-12 panelists were invited to give a value on the antipilling effect. The higher the score, the better effect the CN-42 had.

Anti-dust mite experiment:

The test procedure as well as materials are as following:

Dust mite: Dermatophagoides farina Hughhes

Steps for testing dust-mite reduction effect:

1. Sample preparation: CN-42 was cut in to 6*6cm size and washed according to previous washing method. Each sample was cut into a circle with diameter of 58mm.

2. Prevention test protocol: A sponge with side length 200mm, height 10mm was put into a plastic container with lid (side length 200-300mm, height 50-100mm, with venthole in the middle of diameter of 50±10mm, covered by PTFE film which was fixed on the plastic container lid by tape) , and then saturated NaCl solution was injected into said sponge (the sponge was immersed by satureated NaCl solution) .

3. 7 petri dishes were used and one of them was put in the center, and the other 6 petri dishes were put in the periphery area. Every 2 adjacent petri dishes were bridged by scotch tape. Then all of the 7 dishes were fixed on a plate.

4. 3 test samples and 3 benchmark samples were put into 6 petri dishes in periphery. Every test sample was adjacent to one benchmark sample. 0.05g dust mite feed was placed in each petri dish (see figure 1) .

5. 2000±200 alive dust mites were put in the petri dish in the center.

6. The plate with petri dishes on top was transferred onto the sponge in container. The lid was closed. And the container was put in the temperature and humidity chamber with temperature of 25±2℃, and humidity of 75±5%.

7. After 24h incubation, samples were taken out and counted for the number of dust mites with anatomical lens.

Example 1. Fabric treatment in presence of cellulase

2 gram model detergent A was added in Terg-o-tometer washing assay. Thermothielavioides terrestris of SEQ ID NO: 2 was used herein. The enzyme protein concentration in said detergent was 0.005%. Each detergent was diluted with water to 1L. Ballast was needed to fill up the wash items to 40g.

The fabric E-252PA in size of 8*8cm2 was used to evaluate the antipilling effect of the test enzyme. Such fabric was mixed with the diluted mode detergent A in Terg-o-tometer washing assay and washed for 3 cycles at 40℃ for 70min. L*value is used to show antipilling effect. Result was shown in Figure 2. Figure 2 shown that better anti-pilling effect was achieved in presence of cellulases in comparison with those without cellulase treatment (blank) . L value was reduced from more than 34.1 to about 27.9

The fabric CN-42 in size of 10*10 cm2 was used to evaluate the anti-dust mite effect. Such fabric was mixed with the diluted model detergent A in Terg-o-tometer washing assay and washed for 3 cycles at 40℃ for 70min. Figure 3 shown the result of anti-dust mite experiment in comparison with those without cellulase treatment (blank) . Seen from Fig. 3, the number of dust mite had been reduced to one third of the blank (from157 to 48) .

Claims

Use of an enzyme capable of degrading cellulosic material in reducing/removing dust mite on textile.
The use of claim 1, wherein, said enzyme capable of degrading cellulosic material is GH45 cellulase.
The use of claim 1, wherein, said enzyme capable of degrading cellulosic material are any one or more GH45 cellulases selected from table below:
The use of claim 3, wherein, said GH45 cellulase is Thermothielavioides terrestris endoglucanase.
The use of claim 4, wherein, said Thermothielavioides terrestris endoglucanase is selected from the group consisting of :

(1) T. t endoglucanase comprising or consisting of the mature polypeptide of SEQ ID NO: 2 or fragments thereof;

(2) T. t endoglucanase comprising or consisting of an amino acid sequence having at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%sequence identity to the mature polypeptide of SEQ ID NO: 2;

(3) T. t endoglucanase encoded by a polynucleotide comprising or consisting of a nucleotide sequence having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1;

(4) T. t endoglucanase encoded by a polynucleotide that hybridizes under at least high stringency conditions, very high stringency conditions, with the mature polypeptide coding sequence of SEQ ID NO: 1 or the full-length complement thereof, In another aspect, said cellulase composition further comprises one or more detergent component or fabric soft agent.
An enzyme composition for reducing or removing dust mite from textile comprising enzyme capable of degrading cellulosic material.
The enzyme composition of claim 6, wherein, said enzyme capable of degrading cellulosic material is GH45 cellulase.
The enzyme composition of claim 6, wherein, said enzyme capable of degrading cellulosic material is GH45 cellulase selected from table below:
The enzyme composition of claim 8, wherein, said GH45 cellulase is Thermothielavioides terrestris endoglucanase.
The enzyme composition of claim 9, wherein, said Thermothielavioides terrestris endoglucanase is selected from the group consisting of :

(1) T. t endoglucanase comprising or consisting of the mature polypeptide of SEQ ID NO: 2 or fragments thereof;

(2) T. t endoglucanase comprising or consisting of an amino acid sequence having at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%sequence identity to the mature polypeptide of SEQ ID NO: 2;

(3) T. t endoglucanase encoded by a polynucleotide comprising or consisting of a nucleotide sequence having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1;

(4) T. t endoglucanase encoded by a polynucleotide that hybridizes under at least high stringency conditions, very high stringency conditions, with the mature polypeptide coding sequence of SEQ ID NO: 1 or the full-length complement thereof; In another aspect, said cellulase composition further comprises one or more detergent component or fabric soft agent.
The enzyme composition of claim 6, further comprising at least one or more additional enzyme selected from the group consisting of hemicellulases, peroxidases, proteases, cellulases, xylanases, lipases, phospholipases, esterases, cutinases, pectinases, mannanases, pectate lyases, keratinases, reductases, oxidases, phenoloxidases, lipoxygenases, ligninases, pullulanases, tannases, pentosanases, malanases, β-glucanases, arabinosidases, hyaluronidase, chondroitinase, laccase, DNase, chlorophyllases, amylases, perhydrolases, peroxidases, xanthanase and mixtures thereof.
The enzyme composition of claim 11, wherein said at least one or more additional enzyme is selected from proteases, lipases, mannanases, pectate lyases and amylases.
A detergent composition for reducing or removing dust mite from textile comprising the enzyme composition of any of claims 6-12, a surfanctant, a builder and a polymer.
The detergent composition of claim 13, wherein, said surfactant is selected from the group consisting of anionic, cationic, non-ionic, semi-polar and zwitterionic surfactants.
The detergent composition of claim 13 or 14, further comprises one or more components selected from the group consisting of bleaching systems, bleach activators, bleach catalysts, silicates and dyestuff.
The detergent composition of any of the preceding claims, wherein the composition is in the form of a powder, a bar, a homogenous tablet, a tablet having two or more layers, a pouch having one or more compartments, a regular or compact powder, a granule, a paste, a gel, or a regular, compact or concentrated liquid.
The detergent composition of any of the preceding claims, wherein the enzyme composition used in the detergent composition is in an amount corresponding to 0.001-200 mg of protein, such as 0.005-100 mg of protein, preferably 0.01-50 mg of protein, more preferably 0.05-20 mg of protein, even more preferably 0.1-10 mg of protein per liter of wash liquor.
A method for reducing or removing dust mite from textile comprising:

(1) Washing or exposing the textile with any of detergent composition of claims 13-17 or with any of the enzyme composition of claims 6-12;

(2) Optionally rinsing the textile.
Method of claim 18, wherein the method is carried out by hand or machine.
The method according to any of claims 18-19, wherein the method is a laundry or wash method or a warewashing method.