Cleaning compositions containing an alginate lyase enzyme

Abstract

A detergent composition comprising an alginate lyase enzyme and a cleaning adjunct. A method of treating a surface by contacting the surface with an aqueous wash solution having the detergent composition. The compositions and methods are, among others, for improving the whiteness of fabrics, for improving stain removal from fabrics, for removing malodors from fabrics, for anti-wrinkle benefits, for anti-redeposition benefits, and / or for improving the drying of surfaces.

Description

[Technical Field]

[0001] (Reference to sequence listing) The present specification contains a Sequence Listing in computer readable form, which is incorporated herein by reference.

[0002] The present invention relates to laundry detergent compositions and cleaning methods containing specific alginate lyase enzymes. A particular problem can be the buildup of soil over time. This is problematic for both colored and white fabrics, but can be particularly noticeable on white or light-colored fabrics, for example, around incompletely cleaned collars and cuffs. This can be problematic because it can also result in unpleasant odors. The compositions and methods of the present invention are suitable for use in hand-wash and automatic laundry detergent compositions. The present invention also relates to cleaning methods and methods for making the laundry detergent compositions. [Background technology]

[0003] In cleaning applications, the loss of whiteness over time and the removal of soil or stains are continuing problems. Many cleaning technologies exist that aim to alleviate these problems, but providing improved effectiveness, especially in an environmentally friendly manner, is a constant challenge. In automatic washing machines, these problems are compounded by the increasing use of lower wash temperatures (e.g., cold water) and shorter wash cycles, which reduces the stain / soil removal efficacy of detergent compositions and exacerbates the problems of soil redeposition on surfaces during the cleaning process and whiteness loss over multiple washes. Summary of the Invention [Problem to be solved by the invention]

[0004] It is therefore an object of the present invention to provide a detergent composition which can be used in the washing process even at low temperatures and short wash times, which can offset whiteness loss and / or remove complex stains, for example, removing dark stains, deep cleaning, removing yellowing, cleaning collars and cuffs in particular and / or improving whiteness / offsetting whiteness loss, and which can be useful even at low temperatures and short wash times.The present invention particularly relates to laundry detergent compositions. [Means for solving the problem]

[0005] The present invention provides a detergent composition comprising 0.00005-5% by weight of an alginate lyase enzyme (active enzyme protein) and a cleaning adjunct, wherein the alginate lyase enzyme comprises an alginate lyase selected from: SEQ ID NO: 1 and at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%; or at least 99%, or 100% sequence identity to SEQ ID NO:2; an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:3 .... an alginate lyase having at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:4; an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:5;an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 6; an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 7; or a mixture thereof;

[0006] Preferably the cleaning adjunct comprises a surfactant, most preferably an anionic surfactant. Preferably the alginate lyase enzyme comprises an alginate lyase enzyme from polysaccharide lyase family 7.

[0007] The present invention also provides a method of treating a surface, the method comprising contacting the surface with an aqueous cleaning solution comprising an alginate lyase enzyme and a cleaning adjunct, wherein the alginate lyase enzyme is selected from: an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:1; an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:2; an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:3; an alginate lyase having at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:4; an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:5;an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 6; an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 7; or a mixture thereof;

[0008] Preferably, the aqueous cleaning solution comprises a surfactant, preferably an anionic surfactant, and preferably the surfactant is present in an amount of from 0.05 g / L to 5 g / L, preferably from 0.01 g / L to 3 g / L.

[0009] Preferably, the surface is contacted with the aqueous wash solution at a temperature of 60° C. or less, or more preferably at a temperature of 40° C. or less or 35° C. or less, and most preferably at a temperature of 30° C. or less or even 25° C. or less, and (iii) the surface is rinsed. The compositions and methods herein are particularly useful for treating laundry, for example, synthetic or natural fabrics, including cotton, wool, silk, polyester, nylon, elastane, or blended fabrics such as polycotton.

[0010] The present invention also relates to the use of the composition or method for improving the whiteness of fabrics or counteracting the loss of whiteness; improving stain removal from fabrics; removing dark stains; deep cleaning; removing or reducing yellowing; cleaning collars and / or cuffs; reducing or eliminating malodors from fabrics; preventing wrinkles on fabrics; improving drying of fabrics; preventing redeposition of stains. DETAILED DESCRIPTION OF THE INVENTION

[0011] definition Parent or Parent Alginate Lyase Enzyme: The term "parent" or "parent alginate lyase" refers to an alginate lyase that is modified to generate an enzyme variant. The parent can be a naturally occurring (wild-type) polypeptide or a variant thereof. For example, the parent can be any of SEQ ID NOs: 1, 2, 3, 4, 5, 6, or 7 listed herein.

[0012] Sequence identity: The relatedness between two amino acid sequences or two nucleotide sequences is expressed by the parameter "sequence identity." For the purposes of the present invention, the degree of sequence identity between two amino acid sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48:443-453), preferably version 3.0.0 or later, 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). Optional parameters used are a gap opening penalty of 10, a gap extension penalty of 0.5, and an EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. The output of Needle labeled "longest identity" (obtained using the -nobrief option) is used as the percentage identity, which is calculated as follows: (Identical residues × 100) / (length of alignment − total number of gaps in the alignment) Alternatively, optional parameters used can be a gap opening penalty of 10, a gap extension penalty of 0.5, and an EDNAFULL (the EMBOSS version of NCBI NUC4.4) substitution matrix. The Needle output labeled "Longest Identity" (obtained using the -nobrief option) is used as the percent identity, which is calculated as follows: (identical deoxyribonucleotides × 100) / (length of alignment − total number of gaps in alignment)

[0013] Variant: The term "variant" refers to a polypeptide having alginate lyase activity that contains modifications / mutations, i.e., substitutions, insertions, and / or deletions, at one or more (e.g., several) positions relative to a parent alginate lyase. A substitution refers to the replacement of an amino acid occupying a position with a different amino acid; a deletion refers to the removal of an amino acid occupying a position; and an insertion refers to the addition of 1 to 3 amino acids immediately adjacent to and following the amino acid occupying a position.

[0014] Wild-type enzyme: The term "wild-type" alginate lyase refers to an alginate lyase expressed by a naturally occurring microorganism, such as a bacterium, algae, yeast, or filamentous fungus, found in nature.

[0015] Alginate lyase enzyme The alginate lyase enzymes include alginate lyase enzymes selected from: an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:1; an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:2; an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:3. or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:4; or an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:5;an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 6; an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 7; or a mixture thereof;

[0016] The alginate lyase is preferably of microbial origin, preferably bacterial or algal (e.g. brown seaweeds (Phaeophyceae) such as Ascophyllum, Laminara, Macrocystis, etc.), most preferably bacterial. The alginate lyase enzyme may be derived from Aeromonas sp., Azotobacter sp., Bacillus sp., Flavobacterium sp., Klebsiella sp., Pseudomonas sp., Sphingomonas sp., Vibrio sp., Zobellia galactanivorans, etc. galactaivorans, and most preferably Flavobacterium sp.

[0017] Preferably, the alginate lyase enzyme comprises an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80% sequence identity to SEQ ID NO: 1, and / or an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80% sequence identity to SEQ ID NO: 5, or a mixture thereof. Preferably, the alginate lyase enzyme comprises an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80% sequence identity to SEQ ID NO: 6, and / or an alginate lyase having at least 60%, or at least 70%, or at least 75%, or at least 80% sequence identity to SEQ ID NO: 7, or a mixture thereof. Thus, preferred alginate lyase enzymes include alginate lyase enzymes corresponding to wild-type or, preferably, wild-type variants of any one of SEQ ID NOs: 1, 2, 3, 4, 5, 6, or 7 listed herein.

[0018] Where the alginate lyase enzyme is a variant of a parent amino acid sequence, the parent alginate lyase enzyme preferably has at least 50%, or at least 60%, or at least 70%, or at least 80%, such as at least 85%, at least 90%, for example, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99, or 100% sequence identity to one or more polypeptides of SEQ ID NOs: 1, 2, 3, 4, 5, 6, or 7, and has alginate lyase enzymatic activity. It may be preferred that the variant amino acid sequence differs from the parent alginate lyase by no more than 15, or no more than 10 amino acids, or no more than 5, or 4, or 3, or 2, or 1 amino acid from one or more polypeptides of SEQ ID NOs: 1, 2, 3, 4, 5, 6, or 7.

[0019] When the alginate lyase enzyme is a variant of a parent amino acid sequence, the parent can be obtained from any genus of microorganism. For purposes of the present invention, the term "obtained from," when used herein in reference to a given source, means that the parent encoded by the polynucleotide is produced by that source or by a cell into which a polynucleotide from that source has been inserted. In one embodiment, the parent is secreted extracellularly. Variants can be prepared using any mutagenesis technique known in the art, such as site-directed mutagenesis, synthetic gene construction, semi-synthetic gene construction, random mutagenesis, shuffling, etc.

[0020] Preferably, the alginate lyase enzyme has activity toward poly(β-D-mannuronic acid) (polyM activity) and activity toward poly(α-L-guluronic acid) (polyG activity). The alginate lyase enzyme may comprise a single alginate lyase enzyme to provide polyM activity and polyG activity, or may comprise two or more alginate lyase enzymes that, in combination, provide polyM activity and polyG activity. Preferably, the alginate lyase comprises an enzyme having both polyM activity and polyG activity. Preferably, polyM activity, as defined in the Test section herein, is at least 0.1 absorbance units, preferably at least 0.15 absorbance units, and most preferably at least 2 absorbance units. Preferably, polyG activity, as defined herein, is at least 0.3 absorbance units, preferably at least 0.4 absorbance units, or at least 0.5, or even at least 0.6 absorbance units. PolyM activity and polyG activity can be measured according to the tests described below.

[0021] The alginate lyase enzyme can be incorporated into the cleaning compositions and methods of the present invention in the form of a substantially pure enzyme, or, particularly if the enzyme is a variant of a wild-type enzyme, the variant is not recovered and a host cell expressing the enzyme is used as the source of the alginate lyase enzyme.

[0022] The alginate lyase enzyme may be in the form of a liquid or dry composition. For example, the composition may be in the form of particles or microparticles. The alginate lyase enzyme may be stabilized by encapsulation according to methods known in the art.

[0023] The alginate lyase enzyme is preferably present in the composition in an amount of 0.00005 to 5% by weight active enzyme protein, preferably 0.0001 to 2% by weight active protein, or 0.0005 or 0.001 to 1% by weight active protein, or 0.5% by weight or 0.1% by weight or 0.05% by weight active enzyme protein.

[0024] surfactants The inventors have found that while enzymes provide good soil degradation, the removal of the products of substrate degradation and soils containing them is improved by the presence of a surfactant. Therefore, the detergent composition preferably contains 1 to 80 wt. % of a surfactant. Preferably, the weight ratio of surfactant to active alginate lyase enzyme protein is at least 500:1, preferably at least 1000:1, or at least 1500:1, or at least 2000:1, and is preferably no greater than 500,000:1, preferably no greater than 400,000:1, or no greater than 200,000:1, or at most 150,000:1, or 100,000:1, or 50,000:1, or 10,000:1.

[0025] The surfactant may include anionic, non-ionic, including semi-polar, and / or cationic, and / or zwitterionic, and / or ampholytic, and / or amphoteric, and / or semi-polar non-ionic surfactants, and / or mixtures thereof.

[0026] Preferably, the surfactant comprises an anionic surfactant. Preferred anionic surfactants are sulfonate surfactants and sulfate surfactants, preferably alkylbenzene sulfonates and / or (optionally alkoxylated) alkyl sulfates. Particularly preferred anionic surfactants include linear alkylbenzene sulfonates (LAS). Preferred alkyl sulfates include alkyl ether sulfates, particularly C9-15 alcohol ether sulfates, especially those having an average degree of ethoxylation of 0.5-7, preferably 1-5, C8-C16 ester sulfates, and C10-C14 ester sulfates, such as monododecyl ester sulfate. In preferred compositions, the anionic surfactant comprises an alkylbenzene sulfonate and, optionally, an optionally ethoxylated alkyl sulfate, preferably having an ethoxylation degree of 0-7, more preferably 0.5-3. Isomers of LAS, branched alkylbenzene sulfonates (BABS), phenylalkane sulfonates, alpha-olefin sulfonates (AOS), olefin sulfonates, alkenesulfonates, alkane-2,3-diylbis(sulfates), hydroxyalkane sulfonates and disulfonates, alkyl sulfates (AS), such as sodium dodecyl sulfate (SDS), fatty alcohol sulfates (FAS), primary alcohol sulfates (PAS), alcohol ether sulfates (AES or AEOS or FES, alcohol ethoxy sulfates or aliphatic Also suitable anionic surfactants are alcohol ether sulfates), secondary alkane sulfonates (SAS), paraffin sulfonates (PS), ester sulfonates, sulfonated fatty acid glycerol esters, alpha-sulfofatty acid methyl esters (alpha-SFMe or SES) (including methyl ester sulfonates (MES)), alkyl- or alkenyl succinic acids, dodecenyl / tetradecenyl succinic acid (DTSA), fatty acid derivatives of amino acids, di- and monoesters of sulfosuccinic acid, or salts of fatty acids (soaps), and combinations thereof.

[0027] Anionic surfactants are preferably added to detergent compositions in the form of salts. Preferred cations are alkali metal ions such as sodium and potassium. However, salt forms of anionic surfactants may be formed in situ by neutralizing the acid form of the surfactant with an alkali, such as sodium hydroxide, or an amine, such as mono-, di-, or triethanolamine. The composition preferably contains 1 to 60%, or 1 to 50%, or 2, or 5 to 40% by weight of the composition of anionic surfactant. Preferably, the composition comprises a surfactant system including an anionic surfactant and one or more additional surfactants, which may be nonionic, including semi-polar, and / or cationic, and / or zwitterionic, and / or amphoteric, and / or amphoteric, and / or semi-polar nonionic surfactants, and / or mixtures thereof. Preferably, the cleaning composition of the present invention comprises 0.00005 to 5% by weight (active enzyme protein) of an alginate lyase enzyme as defined above and a surfactant comprising an anionic surfactant and a nonionic surfactant, preferably with a weight ratio of anionic surfactant to nonionic surfactant of 30:1 to 1:2, preferably 20:1 to 2:3 or 20:1 to 1:1.

[0028] Suitable nonionic surfactants include alcohol ethoxylates (AE), alcohol propoxylates, propoxylated fatty alcohols (PFAs), alkoxylated fatty acid alkyl esters, such as ethoxylated and / or propoxylated fatty acid alkyl esters, alkylphenol ethoxylates (APE), nonylphenol ethoxylates (NPE), alkyl polyglycosides (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), and products available under the trade names SPAN and TWEEN®, and combinations thereof. Particularly preferred are alcohol ethoxylates, preferably having a C9-18, or preferably a C12-15 alkyl chain, and preferably having an average degree of ethoxylation of 3 to 9, more preferably 3 to 7. Commercially available nonionic cleaning surfactants include Plurafac™, Lutensol™, and Pluronic™ from BASF, the Dehypon™ series from Cognis, and the Genapol™ series from Clariant.

[0029] The detergent composition preferably comprises from 0.5% to about 40% nonionic surfactant, preferably from 1 to 30% nonionic surfactant, by weight of the composition.

[0030] Cleaning Composition The detergent compositions of the present invention are particularly useful as laundry detergents. The compositions may be in the form of compositions for use in the main wash step, or as pre-treatment or rinse-additive cleaning compositions for consumer or institutional use.

[0031] The composition includes a cleaning adjuvant. Typically, the cleaning adjuvant is present in the composition in an amount of 1 to 98.9% by weight, more typically 5 to 90% by weight, of the cleaning adjuvant. Suitable cleaning adjuvants include additional surfactants, builders, bleaching components, colorants, chelating agents, dye transfer agents, deposition aids, dispersants, additional enzymes and enzyme stabilizers, catalytic materials, optical brighteners, photoactivators, fluorescent agents, fabric hueing agents (shading agents), fabric conditioners, preformed peracids, polymeric dispersants, mud stain removal / anti-redeposition agents, filler salts, hydrotropes, whitening agents, suds suppressors, structural elasticity agents, fabric softeners, preservatives, antioxidants, shrinkage inhibitors, bactericides, mildew inhibitors, anti-tarnish agents, corrosion inhibitors, alkalinity sources, solubilizers, carriers, processing aids, pigments, dyes, fragrances, and pH adjusters, encapsulating agents, polymers, and mixtures thereof.For example, these may include bleaching components such as bleach activators, bleach boosters such as imine bleach boosters, bleach catalysts, hydrogen peroxide, hydrogen peroxide sources such as percarbonates and / or perborates, particularly percarbonates coated with materials such as carbonates and / or sulfates, silicates, borosilicates, and any mixtures thereof, preformed peracids, including encapsulated forms of preformed peracids, transition metal catalysts; foam suppressors or foam suppressor systems, such as silicone-based foam suppressors and / or fatty acid-based foam suppressors; clays, silicones, and / or quaternary ammonium compounds. fabric softeners; flocculants such as polyethylene oxide; dye transfer inhibitors such as polyvinylpyrrolidone, poly 4-vinylpyridine N-oxide, and / or copolymers of vinylpyrrolidone and vinylimidazole; fabric integrity components such as oligomers produced by the condensation of imidazole with epichlorohydrin; soil dispersants and soil anti-redeposition aids such as alkoxylated polyamines and ethoxylated ethyleneimine polymers; anti-redeposition components such as polyesters; carboxylate polymers, e.g., maleic acid polymers, or copolymers of maleic acid and acrylic acid; fragrances, such as fragrance microcapsules, starch-encapsulated accords, fragrance sprays; soap rings; cosmetic particles; aesthetic dyes; fillers such as sodium sulfate and / or citrus fibers (although it may be preferred that the composition is substantially free of fillers); silicates such as sodium silicates, including 1.6R and 2.0R sodium silicates, or sodium metasilicate; copolyesters of dicarboxylic acids and diols; cellulosic polymers such as methylcellulose, carboxymethylcellulose, hydroxyethoxycellulose, or other alkyl or alkylalkoxycelluloses; solvents such as 1,2 propanediol, monoethanolamine; diethylene glycol, ethanol, and any mixtures thereof; hydrotropes, such as sodium cumene sulfonate, sodium xylene sulfonate, sodium toluene sulfonate, and any mixtures thereof; organic acids and their salts, such as citric acid / citrates; and any combinations thereof.The composition may be such that the cleaning adjunct comprises one or more selected from the group consisting of: (i) perfume microcapsules, (ii) fabric hueing agents, (iii) proteases, (iv) amphiphilic cleaning polymers, (v) lipases, or (vi) mixtures thereof.

[0032] The detergent composition preferably comprises one or more additional enzymes. Thus, preferred compositions comprise (a) an alginate lyase and (b) one or more additional enzymes selected from the group consisting of aminopeptidase, amylase, carbohydrase, carboxypeptidase, catalase, cellulase, chitinase, cutinase, cyclodextrin glycosyltransferase, deoxyribonuclease, esterase, alpha-galactosidase, beta-galactosidase, glucoamylase, alpha-glucosidase, beta-glucosidase, haloperoxidase, invertase, laccase, lipase, mannanase, mannosidase, oxidase, pectolytic enzyme, peptidoglutaminase, peroxidase, phytase, polyphenol oxidase, proteolytic enzyme, ribonuclease, transglutaminase, xylanase, xanthan lyase, xanthanase, endo-β-1,3-glucanase, and mixtures thereof. Preferably, the cleaning composition comprises an additional enzyme selected from amylases, nucleases such as DNases and RNases and mixtures thereof, hexosaminidases, mannanases, xanthan lyases, xanthanases, amylases, and mixtures thereof.

[0033] Preferably, the composition comprises an additional enzyme selected from xanthan lyase, xanthanase, mannanase, and mixtures thereof, with mannanase being particularly preferred.

[0034] The additional enzyme may be derived from, for example, Aspergillus species, such as Aspergillus aculetus, Aspergillus awamori, Aspergillus foetidus, Aspergillus fumigatus, Aspergillus japonicus, Aspergillus nidulans, Aspergillus niger, or Aspergillus oryzae; Fusarium species, such as Fusarium bactridioides, Fusarium cerealis, Fusarium cloquewellens, Fusarium culmorum, Fusarium graminearum, Fusarium graminum, Fusarium heterosporum, Fusarium negunzi, or Fusarium oxysporum. The fungus may be produced by a microorganism belonging to the genus Humicola, such as Humicola insolens or Humicola lanuginosa; or a microorganism belonging to the genus Trichoderma, such as Trichoderma harzianum, Trichoderma koningii, Trichoderma longibrachiatum, Trichoderma reesei, or Trichoderma viride.

[0035] Preferably, the composition comprises a protease or a mixture of one or more proteases, a lipase or a mixture of one or more lipases, a peroxidase or a mixture of one or more peroxidases, one or more starch-degrading enzymes, such as α-amylase, glucoamylase, maltogenic amylase, and / or cellulase, or a mixture thereof.

[0036] Generally, the properties of the selected enzyme must be compatible with the selected detergent (i.e., pH optimum, compatibility with other enzymatic and non-enzymatic ingredients, etc.), and the enzyme must be present in an effective amount. Preferably, the products of the present invention contain at least 0.01 mg, preferably from about 0.05 to about 10 mg, more preferably from about 0.1 to about 6 mg, and especially from about 0.2 to about 5 mg of additional active enzyme per gram of composition.

[0037] Protease: The compositions of the present invention preferably include a protease. A mixture of two or more proteases can contribute to improved cleaning over a wider temperature, cycle duration, and / or substrate range. Suitable proteases include metalloproteases and serine proteases, including, for example, neutral or alkaline microbial serine proteases such as subtilisin (EC 3.4.21.62). Suitable proteases include those of animal, vegetable, or microbial origin. In one aspect, such suitable proteases may be of microbial origin. Suitable proteases include chemically or genetically modified variants of the aforementioned suitable proteases. In one aspect, suitable proteases may be serine proteases, such as alkaline microbial proteases and / or trypsin-type proteases. Examples of suitable neutral or alkaline proteases include: Subtilisins (EC 3.4.21.62), in particular those described in WO 2004067737, WO 2015091989, WO 2015091990, WO 2015024739, WO 2015143360, U.S. Pat. Nos. 6,312,936(B1), 5,679,630, and 4,760,025, German Patent Publication No. 102006022216(A1), Bacillus species (Bacillus spp.) described in International Publication No. 102006022224(A1), International Publication No. 2015089447, International Publication No. 2015089441, International Publication No. 2016066756, International Publication No. 2016066757, International Publication No. 2016069557, International Publication No. 2016069563, International Publication No. 2016069569, and International Publication No. 2016174234 Those derived from the genus Bacillus, such as Bacillus sp., B. lentus, B. alkalophilus, B. subtilis, B. amyloliquefaciens, B. pumilus, B. gibsonii, and B. akibaii, specifically those with mutations S9R, A15T, V66A, A188P, V199I, Q239R, and N255D (savinase numbering system). Trypsin- or chymotrypsin-type proteases such as trypsin (e.g., of porcine or bovine origin), including the Fusarium proteases described in WO 89 / 06270, and the chymotrypsin proteases from Cellumonas described in WO 05 / 052161 and WO 05 / 052146. Metalloproteases, in particular those derived from Bacillus amyloliquefaciens as described in WO 07 / 044993(A2); those derived from Bacillus, Brevibacillus, Thermoactinomyces, Geobacillus, Paenibacillus, Lysinibacillus or Streptomyces species as described in WO 2014194032, WO 2014194054 and WO 2014194117, those derived from Kluyvera aluminosa as described in WO 2015193488, and those derived from Streptomyces and Lysobacter as described in WO 2016075078. A protease having at least 90% identity to the subtilase from Bacillus sp. TY145, NCIMB 40339, described in WO 92 / 17577 (Novozymes A / S), including variants of this Bacillus sp. TY145 subtilase described in WO 2015024739 and WO 2016066757.

[0038] Particularly preferred proteases for cleaning compositions of the present invention have at least 90%, preferably at least 95%, more preferably at least 98%, even more preferably at least 99%, especially 100% identity to the wild-type enzyme from Bacillus lentus and have the following positions: S9R, A15T, V68A, N76D, N87S, S99D, S99SD, S99A, S101G, S101M, S103A, V104N / I, G118V, G118R, S128L, P and / or M222S (using the BPN numbering system and amino acid abbreviations exemplified in WO 00 / 37627, incorporated herein by reference).

[0039] Most preferably, the protease is selected from the group of proteases comprising the following mutations (BPN numbering system) relative to either PB92 wild type (SEQ ID NO: 2 of WO 08 / 010925) or subtilisin 309 wild type (sequence according to the PB92 backbone except containing the natural mutation N87S): (i) G118V + S128L + P129Q + S130A (ii)S101M+G118V+S128L+P129Q+S130A (iii)N76D+N87R+G118R+S128L+P129Q+S130A+S188D+N248R (iv)N76D+N87R+G118R+S128L+P129Q+S130A+S188D+V244R (v)N76D+N87R+G118R+S128L+P129Q+S130A (vi) V68A+N87S+S101G+V104N (vii)S99AD (viii)S9R+A15T+V68A+N218D+Q245R

[0040] Suitable commercially available protease enzymes include those sold by Novozymes A / S (Denmark) under the trade names Alcalase®, Savinase®, Primase®, Durazym®, Polarzyme®, Kannase®, Liquanase®, Liquase Ultra®, Savinase Ultra®, Ovozyme®, Neutrase®, Everlase®, Coronase®, Blaze®, Blaze Ultra® and Esperase®; and those sold under the trade names Maxatase®, Maxacal®, Maxapem®, Properase®, Purafect®, Purafect Prime®, Purafect Ox®, FN3®, FN4®, Excellase®, Ultimase® and Purafect those sold by DuPont under the trade names Opticlean® and Optimase® by Solvay Enzymes; and those available from Henkel / Kemira, namely, BLAP (the sequence of which is shown in Figure 29 of U.S. Pat. No. 5,352,604 and which has the following mutations S99D+S101R+S103A+V104I+G159S, hereinafter referred to as BLAP), BLAP R (BLAP with S3T+V4I+V199M+V205I+L217D), BLAP X (BLAP with S3T+V4I+V205I) and BLAP F49 (BLAP with S3T+V4I+A194P+V199M+V205I+L217D); and KAP (Bacillus alkalophilus subtilisin with mutations A230V+S256G+S259N) from Kao.

[0041] Commercially available proteases selected from the group consisting of Properase®, Blaze®, Ultimase®, Everlase®, Savinase®, Excellase®, Blaze Ultra®, BLAP and BLAP variants are particularly preferred for use herein.

[0042] Preferred concentrations of protease in the products of the present invention include about 0.05 to about 10 mg, more preferably about 0.5 to about 7 mg, and especially about 1 to about 6 mg of active protease per gram of composition.

[0043] Lipase: The composition preferably comprises a lipase. The presence of oils and / or fats can further increase the recovery of stains containing mannans and other polysaccharides. Therefore, the presence of lipase in the enzyme package can further improve the removal of such stains. Suitable lipases include those of bacterial, fungal or synthetic origin, including chemically modified or genetically engineered variants of the protein. Examples of useful lipases include lipases from the genus Humicola (also known as Thermomyces), such as H. lanuginosa (T. lanuginosus) or H. insolens; lipases from the genus Pseudomonas, such as P. alcaligenes or P. pseudoalcaligenes, P. cepacia, P. stutzeri, P. fluorescens, Pseudomonas sp. SD705, and P. wisconsinensis; lipases from B. subtilis (Dartois et al. (1993), Biochemica et Biophysica Acta, 1131, 253-360), B. stearothermophilus, or B. pumilus.

[0044] The lipase may be a "first cycle lipase" as described in U.S. Patent No. 6,939,702 (B1) and U.S. Patent Application Publication No. 2009 / 0217464. In one aspect, the lipase is a first wash lipase, preferably a variant of the wild-type lipase from Thermomyces lanuginosus, containing the T231R and N233R mutations. The wild-type sequence is Swiss-Prot Accession No. Swiss-Prot O59952 (269 amino acids (amino acids 23-291) from Thermomyces lanuginosus (Humicola lanuginosa)). Preferred lipases include those sold under the tradenames Lipex®, Lipolex®, and Lipoclean®.

[0045] Other suitable lipases include, for example, Liprl 139 as described in WO 2013 / 171241, TfuLip2 as described in, for example, WO 2011 / 084412 and WO 2013 / 033318, Pseudomonas stutzeri lipase as described in, for example, WO 2018228880, Microbulbifer thermotolerans lipase as described in, for example, WO 2018228881, Sulfobacillus acidocaldarius lipase as described in, for example, EP 3299457, acidocaldarius lipases, such as LIP062 lipase as described in WO2018209026, PinLip lipase as described in WO2017036901, and Absidia sp. lipases as described in WO2017005798.

[0046] Suitable lipases are variants of SEQ ID NO: 5, including: (a) Substitution T231R and (b) Substitution N233R or N233C and (c) at least three additional substitutions selected from E1C, D27R, N33Q, G38A, F51V, G91Q, D96E, K98L, K98I, D111A, G163K, H198S, E210Q, Y220F, D254S, I255A, and P256T; wherein the positions correspond to those of SEQ ID NO: 5, and the lipase variant has at least 90% but less than 100% sequence identity to a polypeptide having the amino acid sequence of SEQ ID NO: 5, and the variant has lipase activity.

[0047] One preferred lipase is a variant of SEQ ID NO:5 containing the following substitutions: T231R, N233R, D27R, G38A, D96E, D111A, G163K, D254S, and P256T.

[0048] One preferred lipase is a variant of SEQ ID NO:5 containing the following substitutions: T231R, N233R, N33Q, G91Q, E210Q, I255A.

[0049] Suitable lipases are commercially available from Novozymes, for example as Lipex Evity 100L, Lipex Evity 200L (both liquid ingredients) and Lipex Evity 105T (granules), which have a different structure to the products Lipex 100L, Lipex 100T and Lipex Evity 100T, which are outside the scope of the present invention.

[0050] Cellulases: Suitable cellulases include those of bacterial or fungal origin, including chemically modified or genetically engineered variants of proteins. Suitable cellulases include cellulases from the genera Bacillus, Pseudomonas, Humicola, Fusarium, Thielavia, and Acremonium, such as fungal cellulases produced by Humicola insolens, Myceliophthora thermophila, and Fusarium oxysporum, as disclosed in U.S. Patent Nos. 4,435,307, 5,648,263, 5,691,178, 5,776,757, and 5,691,178.

[0051] In one aspect, preferred enzymes include endoglucanases from microorganisms exhibiting endo-beta-1,4-glucanase activity (EC 3.2.1.4), preferably selected from the group comprising: (a) a bacterial polypeptide endogenous to a member of the genus Bacillus having a sequence that is at least 90%, 94%, 97%, and even 99% identical to the amino acid sequence of SEQ ID NO:2 in U.S. Patent No. 7,141,403(B2), with preferred substitutions including one or more positions corresponding to positions 292, 274, 266, 265, 255, 246, 237, 224, and 221 of the mature polypeptide of SEQ ID NO:2, wherein the variant has cellulase activity; (b) a glycosyl hydrolase having enzymatic activity on both xyloglucan and amorphous cellulose substrates, the glycosyl hydrolase being selected from GH family 5, 7, 12, 16, 44, or 74; (c) glycosyl hydrolases having a sequence that is at least 90%, 94%, 97%, and even 99% identical to the amino acid sequence of SEQ ID NO: 3 of WO 09 / 148983; (d) A variant exhibiting at least 70% identity to SEQ ID NO: 5 in WO2017106676. Preferred substitutions include one or more positions corresponding to positions 4, 20, 23, 29, 32, 36, 44, 51, 77, 80, 87, 90, 97, 98, 99, 102, 112, 116, 135, 136, 142, 153, 154, 157, 161, 163, 192, 194, 204, 208, 210, 212, 216, 217, 221, 222, 225, 227, and 232; (e) and mixtures thereof.

[0052] Suitable endoglucanases are sold under the trade names Celluclean® and Whitezyme® (Novozymes A / S, Bagsvaerd, Denmark). Examples include Celluclean® 5000L, Celluclean® Classic 400L, Celluclean® Classic 700T, Celluclean® 4500T, Whitezyme® 1.5T, and Whitezyme® 2.0L.

[0053] Other commercially available cellulases include Celluzyme®, Carezyme®, Carezyme® Premium (Novozymes A / S), Clazinase®, Puradax HA®, Revitalenz® 1000, Revitalenz® 2000 (Genencor International Inc.), KAC-500(B)® (Kao Corporation), Biotouch® FCL, Biotouch® DCL, Biotouch® DCC, Biotouch® NCD, Biotouch® FCC, Biotouch® FLX1 (AB Enzymes).

[0054] Suitable glucanases include endo-β-1,3-glucanases, preferably from EC class 3.2.1.39, preferably obtained from a microorganism of the genus Paenibacillus, Zobellia galactanivorans, Thermotoga petrophila, or Trichoderma sp., preferably Paenibacillus or Zobellia galactanivorans, most preferably Paenibacillus.

[0055] Amylase: Preferably, the compositions of the present invention comprise an amylase. Suitable α-amylases include those of bacterial or fungal origin, including chemically or genetically modified variants. Preferred alkaline α-amylases are derived from Bacillus species, such as Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus stearothermophilus, Bacillus subtilis, or other Bacillus species, such as Bacillus species NCBI 12289, NCBI 12512, NCBI 12513, DSM 9375 (U.S. Pat. No. 7,153,818), DSM 12368, DSM Z No. 12649, KSM AP1378 (WO 97 / 00324), KSM K36, or KSM K38 (EP 1,022,334). Preferred amylases include: (a) Variants described in U.S. Pat. No. 5,856,164 and WO 99 / 23211, WO 96 / 23873, WO 00 / 60060, WO 06 / 002643, and WO 2017 / 192657, in particular variants having one or more substitutions at the following positions relative to the AA560 enzyme described as SEQ ID NO: 12 in WO 06 / 002643: 26, 30, 33, 82, 37, 106, 118, 128, 133, 149, 150, 160, 178, 182, 186, 193, 202, 214, 231, 246, 256, 257, 258, 269, 270, 272, 283, 295, 296, 298, 299, 303, 304, 305, 311, 314, 315, 318, 319, 339, 345, 361, 378, 383, 419, 421, 437, 441, 444, 445, 446, 447, 450, 461, 471, 482, 484, preferably D183 * and G184 * Variants that also contain deletions. (b) SEQ ID NO: 4 in WO 06 / 002643, the wild-type enzyme from Bacillus sp. SP722, in particular variants in which positions 183 and 184 are deleted, and variants as described in WO 00 / 60060, WO 2011 / 100410 and WO 2013 / 003659, in particular variants which have one or more substitutions at the following positions relative to SEQ ID NO: 4 in WO 06 / 002643, which are incorporated herein by reference: 51, 52, 54, 109, 304, 140, 189, 134, 195, 206, 243, 260, 262, 284, 347, 439, 469, 476, and 477. (c) Variants exhibiting at least 90% identity to the wild-type enzyme from Bacillus sp. 707 (SEQ ID NO: 7 in U.S. Pat. No. 6,093,562), particularly those containing one or more of the following mutations: M202, M208, S255, R172, and / or M261. Preferably, the amylase contains one or more of M202L, M202V, M202S, M202T, M202I, M202Q, M202W, S255N, and / or R172Q. Particularly preferred are those containing the M202L or M202T mutation. Additional relevant mutations / deletions based on the SP707 backbone are W48, A51, V103, V104, A113, R118, N125, V131, T132, E134, T136, E138, R142, S154, V165, R182, G182, H183, E190, D192, T193, I206, M208, D209, E212, V213, V214, N215, L217, R218, N219 Includes 9, V222, T225, T227, G229, I235, K242, Y243, S244, F245, T246, I250, S255, A256, H286, V291, T316, V317, V318, N417, T418, A419, H420, P421, I428, M429, F440, R443, N444, K445, Q448, S451, A465, N470, S472. (d) a variant described in WO 09 / 149130, preferably SEQ ID NO: 1 or SEQ ID NO: 2 in WO 09 / 149130, which shows at least 90% identity to the wild-type enzyme from Geobacillus Stearophermophilus, or a truncated version thereof. (e) a variant described in WO 10 / 115021, in particular SEQ ID NO: 2 in WO 10 / 115021, which exhibits at least 75%, or at least 85%, or at least 90%, or at least 95% identity with the alpha-amylase from Bacillus sp. TS-23. (f) Variants exhibiting at least 89% identity to SEQ ID NO: 1 in WO2016091688, in particular those containing a deletion at positions H183+G184 and further containing one or more mutations at positions 405, 421, 422, and / or 428. (g) Variants described in WO 2014099523, in particular those exhibiting at least 60% amino acid sequence identity with "PcuAmyl α-amylase" from Paenibacillus curdlanolyticus YK9 (SEQ ID NO: 3 in WO 2014099523). (h) Variants described in WO 2014099523, in particular "CspAmy2 amylase" from Cytophaga sp. (showing at least 60% amino acid sequence identity with SEQ ID NOs: 1 and 6 in WO 2014164777, in particular those containing one or more of the following deletions and / or mutations based on SEQ ID NO: 1 in WO 2014164777: R178 * , G179 * , T38N, N88H, N126Y, T129I, N134M, F153W, L171R, T180D, E187P, I203Y, G476K, G477E, Y303D. (i) A variant showing at least 85% identity with Bacillus subtilis (SEQ ID NO: 1 in WO 2009149271). (j) A variant showing at least 90% identity with the wild-type amylase derived from Bacillus sp. KSM-K38 under accession number AB051102. (k) variants described in WO 2016180748, in particular those exhibiting at least 80% identity with the mature amino acid sequence of AAI10 from Bacillus sp. as set forth in SEQ ID NO: 7 in WO 2016180748; those exhibiting at least 80% identity with the mature amino acid sequence of Alicyclobacillus sp. amylase as set forth in SEQ ID NO: 8 in WO 2016180748, and those exhibiting at least 80% identity with the mature amino acid sequence of SEQ ID NO: 13 in WO 2016180748, in particular those having the following mutations: * , N54S, V56T, K72R, G109A, F113Q, R116Q, W167F, Q172G, A174S, G184T, N195F, V206L, K391A, P473R, G476K. (l) Variants described in WO2018060216, in particular those exhibiting at least 70% identity to the mature amino acid sequence of SEQ ID NO: 4 in WO2018060216, fusion molecules of Bacillus amyloliquefaciens and Bacillus licheniformis. In particular, those containing one or more substitutions at positions H1, N54, V56, K72, G109, F113, R116, T134, W140, W159, W167, Q169, Q172, L173, A174, R181, G182, D183, G184, W189, E194, N195, V206, G255, N260, F262, A265, W284, F289, S304, G305, W347, K391, Q395, W439, W469, R444, F473, G476, and G477.

[0056] A preferred amylase is a genetically engineered enzyme in which one or more amino acids susceptible to bleaching oxidation are replaced with amino acids less susceptible to oxidation. In particular, it is preferred that methionine residues are replaced with any other amino acid. In particular, it is preferred that the methionine most susceptible to oxidation is replaced. Preferably, the methionine at position 202 in SEQ ID NO: 11 is replaced. Preferably, the methionine at this position is replaced with threonine or leucine, preferably leucine.

[0057] Suitable commercially available alpha-amylases include DURAMYL®, LIQUEZYME®, TERMAMYL®, TERMAMYL ULTRA®, NATALASE®, SUPRAMYL®, STAINZYME®, STAINZYME PLUS®, FUNGAMYL®, ATLANTIC®, ACHIEVE ALPHA®, AMPLIFY® PRIME, INTENSA®, and BAN® (Novozymes A / S, Bagsvaerd, Denmark), KEMZYM® AT 9000 (Biozym Biotech Trading GmbH, Wehlistrasse 27b A-1200 Wien, Austria), RAPIDASE®, PURASTAR®, ENZYSIZE®, OPTISIZE HT PLUS®, POWERASE®, and PREFERENZ. Examples include the S (registered trademark) series (including PREFERENZ S1000 (registered trademark) and PREFERENZ S2000 (registered trademark)), PURASTAR OXAM (registered trademark) (DuPont, Palo Alto, California), and KAM (registered trademark) (Kao, 14-10 Nihonbashi Kayabacho, 1-chome, Chuo-ku, Tokyo 103-8210, Japan).

[0058] Preferably, the composition contains at least 0.01 mg, preferably about 0.05 to about 10, more preferably about 0.1 to about 6, especially about 0.2 to about 5 mg of active amylase per gram of composition.

[0059] Peroxidases / Oxidases: Suitable peroxidases / oxidases include those of plant, bacterial, or fungal origin, including chemically modified or genetically engineered variants of the protein. Examples of useful peroxidases include peroxidases from the genus Coprinus, such as those from C. cinereus and variants thereof, as described in WO 93 / 24618, WO 95 / 10602, and WO 98 / 15257.

[0060] Commercially available peroxidases include GUARDZYME® (Novozymes A / S).

[0061] Pectate lyases: Suitable pectate lyases include those sold under the trade names Pectawash®, Pectaway®, X-Pect® (all Novozymes A / S, Bagsvaerd, Denmark) and Preferenz® F1000 (DuPont Industrial Biosciences).

[0062] Mannanase. The composition preferably contains one or more additional mannanase enzymes. As used herein, the term "mannanase" or "galactomannanase" refers to a mannanase enzyme, defined in accordance with what is known in the art as mannan endo-1,4-beta-mannosidase, also known as beta-mannanase and endo-1,4-mannanase, that catalyzes the hydrolysis of 1,4-beta-D-mannosidic linkages in mannans, galactomannans, glucomannans, and galactoglucomannans. Mannanases are classified as EC 3.2.1.78 according to enzyme nomenclature and belong to glycosyl hydrolase families 5, 26, and 113. Many suitable mannanases belong to glycosyl hydrolase family 5. Commercially available mannanases include all those sold under the trade name Mannaway® (Novozymes A / S), such as Mannaway® 200L and Mannaway Evity 4.0T. Other commercially available mannanases include Effectene® M1000, Mannastar® 375, Preferenz M100, and Purabrite® (all from DuPont Industrial Biosciences) and Biotouch M7 (AB enzyme). Other suitable mannanases belong to glycosyl hydrolase family 26, such as those described in WO 2018 / 191135, WO 2015040159, WO 2017 / 021515, WO 2017 / 021516, WO 2017 / 021517, and WO 2019 / 081515. Suitable mixtures of mannanases include the combination of glycosyl hydrolase family 5 mannanase and glycosyl hydrolase family 26 mannanase described in WO 2019 / 081515.

[0063] Xanthan gum-degrading enzymes: The compositions may contain one or more xanthan gum-degrading enzymes. Suitable enzymes for degrading xanthan gum-based stains include xanthan endoglucanases, optionally in combination with xanthan lyases. As used herein, the term "xanthan endoglucanase" refers to an enzyme that exhibits endo-β-1,4-glucanase activity, optionally in combination with a suitable xanthan lyase enzyme, capable of catalyzing the hydrolysis of the 1,4-linked β-D-glucose polymer backbone of xanthan gum. Suitable xanthan endoglucanases are described in WO 2013 / 167581, WO 2015 / 181299, WO 2015 / 181292, WO 2017 / 046232, WO 2017 / 046260, WO 2018 / 37062, WO 2018 / 37065, WO 2019 / 038059, and WO 2019 / 162000. As used herein, the term "xanthan lyase" refers to an enzyme that cleaves the β-D-mannosyl-β-D-1,4-glucuronosyl bond in xanthan gum. Such enzymes belong to the EC 4.2.2.12 enzyme class. Suitable xanthan lyases are described in WO 2015 / 001017, WO 2018 / 037061, WO 2018 / 37064, WO 2019 / 038060, WO 2019 / 162000, and WO 2019 / 038057.

[0064] Nuclease: Preferably, the composition comprises a nuclease, such as RNase or DNase, or a mixture thereof. A nuclease enzyme is an enzyme capable of cleaving phosphodiester bonds between nucleotide subunits of nucleic acids. The nuclease enzyme herein is preferably a deoxyribonuclease or ribonuclease enzyme or a functional fragment thereof. A functional fragment or portion refers to a portion of a nuclease enzyme that catalyzes the cleavage of phosphodiester bonds in the DNA backbone, and thus is a region of the nuclease protein that retains catalytic activity. It therefore includes truncated but functional versions of the enzyme and / or variants and / or derivatives and / or homologs in which its function is maintained.

[0065] Preferably, the nuclease enzyme is a deoxyribonuclease preferably selected from any of the following classes: EC 3.1.21.x (where x = 1, 2, 3, 4, 5, 6, 7, 8 or 9), EC 3.1.22.y (where y = 1, 2, 4 or 5), EC 3.1.30.z (where z = 1 or 2), EC 3.1.31.1, and mixtures thereof.

[0066] DNase: Suitable DNases include those set forth in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, and 9 of WO 2017162836 (Novozymes) and SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 110, DNase wild-type and variants defined by 4, 25, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, and 54, as well as variants of Bacillus cibi DNase, including those described in WO 2018011277 (Novozymes). Preferred DNases are as claimed in co-pending European Patent Application No. 18202967.

[0067] RNases: Suitable RNases include wild-type and variants of DNases as defined by SEQ ID NOs: 3, 6, 9, 12, 15, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 72, and 73 in WO2018178061 (Novozymes), and SEQ ID NOs: 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, and 104 in WO2020074499 (Novozymes), which are incorporated herein by reference.

[0068] Hexosaminidase: The composition may contain one or more hexosaminidases. The term "hexosaminidase" includes "dispersant" and the abbreviation "Dsp," meaning a polypeptide (EC 3.2.1) having hexosaminidase activity that catalyzes the hydrolysis of β-1,6-glycosidic bonds in N-acetyl-glucosamine polymers found in soils of microbial origin. The term "hexosaminidase" includes polypeptides having N-acetylglucosaminidase activity and β-N-acetylglucosaminidase activity. Hexosaminidase activity can be determined according to Assay II described in WO2018184873. Suitable hexosaminidases include those described in International Publication Nos. 2017186936, 2017186937, 2017186943, 2017207770, 2018184873, 2019086520, 2019086528, 2019086530, 2019086532, 2019086521, and 2019086522. Examples of suitable hexosaminidases include those disclosed in WO 2020207944, WO 2020002604, WO 2020002608, WO 2020007863, WO 2020007875, WO 2020008024, WO 2020070063, WO 2020070249, WO 2020088957, WO 2020088958, and WO 2020207944. Variants of the Teribacillus saccharophilus hexosaminidase defined by SEQ ID NO: 1 of WO 2020207944 may be preferred, particularly variants with improved thermostability disclosed therein.

[0069] Galactanase: Preferably, the composition comprises a galactanase, i.e., an extracellular polymer-degrading enzyme, including an endo-beta-1,6-galactanase enzyme. The term "endo-beta-1,6-galactanase" or "polypeptide having endo-beta-1,6-galactanase activity" refers to endo-beta-1,6-galactanase activity (EC 3.2.1.164) from glycoside hydrolase family 30 that catalyzes the hydrolytic cleavage of 1,6-3-D-galactooligosaccharides with a degree of polymerization (DP) greater than 3 and their acidic derivatives bearing a 4-O-methyl glucosyluronate or glucosyluronate group at the non-reducing end. For purposes of this disclosure, endo-beta-1,6-galactanase activity is determined in Assay I according to the procedure described in WO2015185689. Suitable examples from class EC3.2.1.164 are described in WO2015185689, e.g., mature polypeptide SEQ ID NO:2.

[0070] The additional enzymes may be included in the detergent composition by adding a separate enzyme additive containing the additional enzymes or a mixed enzyme additive containing two, some, or all of the additional enzymes. Such enzyme additives may be in the form of granules, liquids, or slurries, and preferably further include an enzyme stabilizer.

[0071] Preferably, the or each additional enzyme is present in the composition in an amount of at least 0.0001% to about 0.1% by weight of pure active enzyme protein, such as from about 0.0001% to about 0.01% by weight, from about 0.001% to about 0.01% by weight, or from about 0.001% to about 0.01% by weight, based on the weight of the composition.

[0072] Fabric Hueing Agents. The composition may include a fabric hueing agent (sometimes referred to as a tint, bluing, or whitening agent / dye). Typically, the hueing agent imparts a blue or purple hue to the fabric. Hueing agents can be used either alone or in combination to create a particular hue and / or tint different types of fabric. This can be achieved, for example, by mixing a red and a green-blue dye to produce a blue or purple hue. The hueing agent may be selected from any known chemical class of dyes, including, but not limited to, acridines, anthraquinones (including polycyclic quinones), azines, azos including premetallized azos (e.g., monoazos, diazos, trisazos, tetrakisazos, polyazos), benzodifurans and benzodifuranones, carotenoids, coumarins, cyanines, diazahemicyanines, diphenylmethanes, formazans, hemicyanines, indigoids, methanes, naphthalimides, naphthoquinones, nitro and nitroso, oxazines, phthalocyanines, pyrazoles, stilbenes, styryls, triarylmethanes, triphenylmethanes, xanthenes, and mixtures thereof. Azo dyes, particularly monoazo or bis-azo dyes, triarylmethane dyes, and anthraquinone dyes are preferred.

[0073] Suitable fabric hueing agents include dyes, dye-clay conjugates, and organic and inorganic pigments. Suitable dyes include small molecule dyes and polymeric dyes. Suitable small molecule dyes include small molecule dyes selected from the group consisting of dyes classified in the Color Index (CI) classification of direct dyes, basic dyes, reactive dyes, or hydrolysis reactive dyes, solvent dyes, or disperse dyes. Examples of suitable small molecule dyes include, for example, the Color Index (Society of Dyes and Colourists (Bradford, UK) numbers include direct violet dyes such as 9, 35, 48, 51, 66, and 99; direct blue dyes such as 1, 71, 80, and 279; acid red dyes such as 17, 73, 52, 88, and 150; acid violet dyes such as 15, 17, 24, 43, 49, 50, and 51; acid blue dyes such as 15, 17, 25, 29, 40, 45, 75, 80, 83, 90, and 113. The dyes include small molecule dyes selected from the group consisting of: dyes, acid black dyes such as 1; basic violet dyes such as 1, 3, 4, 10, and 35; basic blue dyes such as 3, 16, 22, 47, 66, 75, and 159; disperse or solvent dyes such as those described in EP 1794275 or EP 1794276; or dyes disclosed in U.S. Pat. No. 7,208,459 B2; and mixtures thereof.

[0074] Preferred are polymeric dyes, including polymers containing covalently attached (sometimes referred to as conjugated) chromogens (dye-polymer conjugates), such as polymers having chromogens copolymerized into the backbone of the polymer, and mixtures thereof, including those described in WO 2011 / 98355, WO 2011 / 47987, U.S. Patent Application Publication No. 2012 / 090102, WO 2010 / 145887, WO 2006 / 055787, and WO 2010 / 142503.

[0075] Preferred polymeric dyes include alkoxylated, preferably ethoxylated, azo, anthraquinone, or triarylmethane dyes. Particularly preferred are polymeric dyes selected from the group consisting of ethoxylated thiophene azo dyes, such as the fabric substantive colorant sold under the name Liquitint® (Milliken, Spartanburg, South Carolina, USA), and dye-polymer conjugates formed from at least one reactive dye and a polymer selected from the group consisting of polymers containing a moiety selected from the group consisting of hydroxyl moieties, primary amine moieties, secondary amine moieties, thiol moieties, and mixtures thereof. Suitable polymeric dyes include polymeric dyes selected from the group consisting of Liquitint® Violet CT, carboxymethyl cellulose (CMC) covalently bonded to a reactive blue, reactive violet, or reactive red dye, such as CMC conjugated with CI Reactive Blue 19 sold under the trade name AZO-CM-CELLULOSE, product code S-ACMC by Megazyme (Wicklow, Ireland), alkoxylated triphenyl-methane polymeric colorants, alkoxylated thiophene polymeric colorants, and mixtures thereof.

[0076] Preferred hueing dyes include the alkoxylated thiophene azo brighteners found in U.S. Patent Application Publication No. 2008 / 0177090, and optionally can be anionic, such as those selected from Examples 1-42 in Table 5 of WO 2011 / 011799. Other preferred dyes are disclosed in U.S. Patent No. 8,138,222.

[0077] Suitable pigments include pigments selected from the group consisting of Ultramarine Blue (CI Pigment Blue 29), Ultramarine Violet (CI Pigment Violet 15), and mixtures thereof. Pigments and / or dyes may also be added to add color for cosmetic reasons. Preferred are organic blue, purple, and / or green pigments.

[0078] Builders: The detergent composition may further contain builders such as builders based on carbonates, bicarbonates or silicates, which may be zeolites such as zeolite A, zeolite MAP (Maximum Aluminum type P). Laundry-safe zeolites are preferably of the formula Na 12 (AlO2) 12 (SiO2) 12 ·27H2O, with particle sizes typically ranging from 1 to 10 μm for zeolite A and 0.7 to 2 μm for zeolite MAP. Another builder is the strongly alkaline sodium metasilicate (Na2SiO3·nH2O or Na2SiO5·nH2O), preferably used in dishwashing. In preferred embodiments, the amount of detergent builder may be greater than 5%, greater than 10%, greater than 20%, greater than 30%, greater than 40%, or greater than 50%, or may be less than 80% or less than 65%. In dishwashing detergents, the builder concentration is typically 40 to 65%, particularly 50 to 65%, or even 75 to 90%.

[0079] Encapsulated Agent: The composition may comprise an encapsulated benefit agent comprising a core and a shell having an inner surface and an outer surface, the shell encapsulating the core. The core may comprise a material selected from the group consisting of fragrances; whitening agents; dyes, insect repellents; silicones; waxes; fragrances; vitamins; fabric softeners; skin care agents, and in one embodiment, paraffin; enzymes; antimicrobial agents; bleaching agents; sensates; and mixtures thereof. The shell may comprise a material selected from the group consisting of polyethylene; polyamide, polystyrene; polyisoprene; polycarbonate; polyester; polyacrylate; aminoplast (in one aspect, the aminoplast may comprise polyurea, polyurethane, and / or polyureaurethane, and in one aspect, the polyurea may comprise polyoxymethylene urea and / or melamine formaldehyde); polyolefin; polysaccharide (in one aspect, the polysaccharide may comprise alginate and / or chitosan); gelatin; shellac; epoxy resin; vinyl polymer; water-insoluble inorganic material; silicone; and mixtures thereof. A preferred encapsulant comprises a core containing a perfume. Such an encapsulant is a perfume microcapsule.

[0080] Enzyme Stabilizers: The compositions may include an enzyme stabilizer. Suitable enzyme stabilizers may be selected from the group consisting of: (a) inorganic salts selected from the group consisting of calcium salts, magnesium salts, and mixtures thereof; (b) carbohydrates selected from the group consisting of oligosaccharides, polysaccharides, and mixtures thereof, and sugars or sugar alcohols; (c) phenylboronic acid and derivatives thereof, such as aromatic borate esters, or phenylboronic acid derivatives, such as 4-formylphenylboronic acid, or peptide aldehydes, such as di-, tri-, or tetrapeptide aldehydes or aldehyde analogs (any of the form B1-B0-R, where R is H, CH3, CX3, CHX2, or CH2X (X = halogen), and B0 is a single amino acid residue, preferably having an optionally substituted aliphatic or aromatic side chain). (b) a protease inhibitor selected from the group consisting of: (a) a protease inhibitor selected from the group consisting of: (b) a protease inhibitor selected from the group consisting of: (c) a protease inhibitor selected from the group consisting of: (d) a protease inhibitor selected from the group consisting of: (e) a protease inhibitor selected from the group consisting of: (a) a protease inhibitor selected from the group consisting of: (b) a protease inhibitor selected from the group consisting of: (c) a protease inhibitor selected from the group consisting of: (d) a protease inhibitor selected from the group consisting of: (e) a protease inhibitor selected from the group consisting of: (a) a protease inhibitor selected from the group consisting of: (a) a protease inhibitor selected from the group consisting of: (b ...

[0081] Structural Agents: In one aspect, the composition may include a structuring agent selected from the group consisting of diglycerides and triglycerides, ethylene glycol distearate, microcrystalline cellulose, cellulosic materials, microfiber cellulose, biopolymers, xanthan gum, gellan gum, and mixtures thereof.

[0082] Polymer: The composition preferably comprises one or more polymers. Preferred examples are carboxymethylcellulose, poly(vinyl-pyrrolidone), poly(ethylene glycol), poly(vinyl alcohol), poly(vinylpyridine-N-oxide), poly(vinylimidazole), polycarboxylates such as polyacrylates, maleic acid / acrylic acid copolymers, and lauryl methacrylate / acrylic acid copolymers, and amphiphilic polymers, and mixtures thereof.

[0083] Amphiphilic cleaning polymers: Preferably, the amphiphilic cleaning polymers have the following general structure: bis((C2H5O)(C2H4O)n)(CH3)-N + -C x H 2x -N + Compounds having the formula -(CH3)-bis((C2H5O)(C2H4O)n) where n=20-30 and x=3-8, or sulfated or sulfonated variants thereof.

[0084] The amphiphilic alkoxylated grease cleaning polymer of the present invention refers to any alkoxylated polymer that has a balance of hydrophilic and hydrophobic properties to remove grease particles from fabrics and surfaces. Specific embodiments of the amphiphilic alkoxylated grease cleaning polymer of the present invention comprise a core structure and a plurality of alkoxylate groups attached to the core structure. These may comprise alkoxylated polyalkyleneimines, preferably having an inner polyethylene oxide block and an outer polypropylene oxide block.

[0085] The core structure may comprise a polyalkyleneimine structure comprising repeat units of formulae (I), (II), (III), and (IV) in condensed form.

[0086] [ka] where # in each case indicates a link between a nitrogen atom and a group A of two adjacent repeating units of formula (I), (II), (III) or (IV). 1indicates half of the binding between the free binding sites of * indicates in each case half of a bond with one of the alkoxylate groups, A 1 are independently selected from straight-chain or branched C2-C6-alkylene; the polyalkyleneimine structure consists of 1 repeating units of formula (I), x repeating units of formula (II), y repeating units of formula (III), and y+1 repeating units of formula (IV), where x and y in each instance have values ​​ranging from 0 to about 150; and the average weight average molecular weight Mw of the polyalkyleneimine core structure has a value ranging from about 60 to about 10,000 g / mol.

[0087] Alternatively, the core structure may comprise a polyalkanolamine structure that is a condensation product of at least one compound selected from N-(hydroxyalkyl)amines of formula (Ia) and / or (Ib).

[0088] [ka] wherein A is independently selected from C1-C6-alkylene; R 1 , R 1* , R 2 , R 2* , R 3 , R 3* , R 4 , R 4* , R 5 and R 5* are independently selected from hydrogen, alkyl, cycloalkyl, or aryl, the last three of which may be optionally substituted; R 6 is selected from hydrogen, alkyl, cycloalkyl, or aryl, the last three of which may be optionally substituted.

[0089] The multiple alkyleneoxy groups attached to the core structure are independently selected from alkyleneoxy units of formula (V).

[0090] [ka] During the ceremony,* represents in each case half of the bond to the nitrogen atom of a repeating unit of formula (I), (II), or (IV), and A 2 is in each occurrence independently selected from 1,2-propylene, 1,2-butylene, and 1,2-isobutylene; A 3 is 1,2-propylene; R in each occurrence is independently selected from hydrogen and C1-C4-alkyl; m has an average value ranging from 0 to about 2; n has an average value ranging from about 20 to about 50; and p has an average value ranging from about 10 to about 50.

[0091] Carboxylate polymer: The composition also preferably includes one or more carboxylate polymers, such as a maleate / acrylate random copolymer or a polyacrylate homopolymer. In one embodiment, the carboxylate polymer is a polyacrylate homopolymer having a molecular weight of 4,000 Da to 9,000 Da or 6,000 Da to 9,000 Da.

[0092] Soil Release Polymers: The composition also preferably includes one or more soil release polymers having a structure defined by one of the following structures (I), (II), or (III): (I)-[(OCHR 1 -CHR 2 ) a -O-OC-Ar-CO-] d (II)-[(OCHR 3 -CHR 4 ) b -O-OC-sAr-CO-] e (III)-[(OCHR 5 -CHR 6 ) c -OR 7 ] f During the ceremony, a, b, and c are 1 to 200; d, e, and f are 1 to 50; Ar is 1,4-substituted phenylene; sAr is 1,3-substituted phenylene substituted at the 5-position with SO3Me; Me is Li, K, Mg / 2, Ca / 2, Al / 3, ammonium, mono-, di-, tri-, or tetra-alkylammonium (the alkyl groups are C1-C 18 Alkyl or C2-C 10 hydroxyalkyl), or mixtures thereof; R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are independently H or C1 to C 18 n- or iso-alkyl; R 7 is a straight chain or branched C1-C 18 Alkyl, or straight or branched C2-C 30 Alkenyl or cycloalkyl groups having 5 to 9 carbon atoms, or C8 to C 30 Aryl group or C6-C 30 It is an arylalkyl group.

[0093] Suitable soil release polymers are polyester soil release polymers such as Repel-o-tex polymers, including Repel-o-tex SF, SF-2, and SRP6, supplied by Rhodia. Other suitable soil release polymers include Texcare polymers, including Texcare SRA100, SRA300, SRN100, SRN170, SRN240, SRN300, and SRN325, supplied by Clariant. Other suitable soil release polymers are Marloquest polymers, such as Marloquest SL, supplied by Sasol.

[0094] Cellulose polymer: The composition also preferably includes one or more cellulosic polymers, including those selected from alkyl celluloses, alkyl alkoxyalkyl celluloses, carboxyalkyl celluloses, and alkyl carboxyalkyl celluloses. In one embodiment, the cellulosic polymer is selected from the group consisting of carboxymethyl cellulose, methyl cellulose, methylhydroxyethyl cellulose, methylcarboxymethyl cellulose, and mixtures thereof. In one embodiment, the carboxymethyl cellulose has a degree of carboxymethyl substitution of 0.5 to 0.9 and a molecular weight of 100,000 Da to 300,000 Da.

[0095] Bleaching System: The compositions may contain a bleaching system comprising a source of HO, such as a perborate or percarbonate, which may be combined with a peracid-forming bleach activator, such as tetraacetylethylenediamine or nonanoyloxybenzenesulfonate. Alternatively, the bleaching system may comprise, for example, an amide, imide, or sulfone-type peroxyacid. Generally, when a bleaching agent is used, the compositions of the present invention may comprise from about 0.1% to about 30%, or even from about 0.1% to about 25%, by weight of the subject cleaning composition, of bleaching agent.

[0096] Chelating Agent: The composition comprises a chelating agent, preferably in an amount of from 0.005% to about 15%, or even from about 3.0% to about 10%, by weight of the composition. Suitable chelating agents include copper, iron, and / or manganese chelating agents, and mixtures thereof. Preferred chelating (complexing) agents include DTPA (diethylenetriaminepentaacetic acid), HEDP (hydroxyethanediphosphonic acid), DTPMP (diethylenetriaminepenta(methylenephosphonic acid)), 1,2-dihydroxybenzene-3,5-disulfonic acid disodium salt hydrate, ethylenediamine, diethylenetriamine, ethylenediaminedisuccinic acid (EDDS), N-hydroxyethylethylenediaminetriacetic acid (HEDTA), triethylenetetraaminehexaacetic acid (TTHA), N-hydroxyethyliminodiacetic acid (HEIDA), dihydroxyethylglycine (DHEG), ethylenediaminetetrapropionic acid (EDTP), methylglycinediacetic acid (MGDA), glutamic acid-N,N-diacetic acid (GLDA), iminodisuccinic acid (IDS), carboxymethylinulin, and salt derivatives and mixtures thereof. Preferred chelating agents are selected from the group consisting of methyl-glycine-diacetic acid (MGDA), its salts and derivatives, glutamic acid-N,N-diacetic acid (GLDA), its salts and derivatives, iminodisuccinic acid (IDS), its salts and derivatives, carboxymethyl inulin, its salts and derivatives, and mixtures thereof. MGDA and its salts, especially the trisodium salt of MGDA, are particularly preferred.

[0097] The compositions may contain other conventional detergent ingredients, such as fabric conditioners including viscosity, suds boosters, suds suppressors, corrosion inhibitors, soil suspending agents, dyes, disinfectants, optical brighteners, hydrotropes, anti-fog agents, organic solvents such as ethanol, or perfumes.

[0098] How to use The present invention also provides a method for treating a surface, the method comprising, in a contacting step, contacting the surface with an aqueous cleaning solution comprising the above-mentioned alginate lyase enzyme, preferably in an amount of 0.01 ppm to 10 ppm, preferably 0.1 ppm to 1 ppm, and a cleaning adjunct, optionally comprising a surfactant, preferably in an amount of 0.05 to 50 g / L, more preferably 0.2 g / L to 5 g / L or 0.5 g / L to 3 g / L, wherein the alginate lyase enzyme comprises an alginate lyase having at least 60% sequence identity to any of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, or SEQ ID NO:5.

[0099] An aqueous wash solution can be formed by adding the above-described composition to water, for example, in a washing machine or hand washing process. Alternatively, the aqueous wash solution can be formed by adding the alginate lyase enzyme and cleaning adjuvants as separate components to water to form the wash solution. The surface can then optionally be washed, and / or rinsed, and / or dried.

[0100] The alginate lyase enzyme and any additional enzymes may be present in the wash solution in an amount corresponding to 0.001 to 100 mg of active enzyme protein per liter of wash solution, preferably 0.005 to 5 mg of active enzyme protein per liter of wash solution, more preferably 0.01 to 1 mg of enzyme protein per liter of wash solution, specifically 0.1 to 1 mg of enzyme protein per liter of wash solution.

[0101] It may be preferable to use mechanical agitation during the contacting step or a subsequent step to facilitate cleaning and removal of the decomposed soil by-products from the surface. The wash solution preferably has a pH of about 7 or 8 to about 10.5. The composition may typically be used at a concentration of about 500 ppm to about 15,000 ppm in solution to form the wash solution. The wash solution preferably has a temperature of about 5°C to about 40°C, or preferably 10 to 35°C or 30°C. The water to fabric ratio is typically about 1:1 to about 30:1. Preferably, the surface comprises fabric and the method is a laundry treatment method.

[0102] test Enzyme activity towards β-D-mannuronic acid block (polyM activity) and α-L-guluronic acid block (polyG activity) Alginate lyase activity was measured using mannuronic acid block oligosaccharides DP20-DP35 (product code: ALG601) and guluronic acid oligosaccharides DP2-DP45 (product code: ALG610) manufactured by Elicityl (France) as substrates. Mannuronic acid block oligosaccharides DP20-DP35 were used to measure polyM activity, and guluronic acid oligosaccharides DP2-DP45 were used to measure polyG activity.

[0103] A 2.5% solution of each substrate was suspended in Tris buffer, pH 8.3, and incubated with 3 ppm of each alginate lyase of interest in a 96-well plate for 60 minutes at 25°C.

[0104] Activity for each substrate is given as the delta absorbance at 235 nm in a spectrophotometer relative to a sample without enzyme when the enzyme is contacted with each substrate. These values ​​are then used to evaluate the activity of each enzyme toward β-D-mannuronic acid blocks (polyM) and / or α-L-guluronic acid blocks (polyG). Enzymes with activity toward poly(β-D-mannuronic acid) (polyM activity) preferably provide a delta absorbance relative to the absence of enzyme of at least 0.1 absorbance units, more preferably at least 0.15 absorbance units, and more preferably at least 0.2 absorbance units. Enzymes with activity toward poly(α-L-guluronic acid) (polyG activity) preferably provide a delta absorbance relative to the absence of enzyme of at least 0.3 absorbance units, preferably at least 0.4, or even 0.5 or 0.6 absorbance units. [Example]

[0105] Example 1: Cleaning performance of liquid detergent compositions containing different alginate lyases The cleaning performance of alginate lyase for removing soiled collar and cuff stains from fabrics was determined for laundry detergents as follows: Stain removal tests were performed using a Talboys Professional Incubating Microplate Shaker supplied by Cole-Parmer Instrument Co Ltd, UK. Dark collar and cuff stains were cut into 2 cm x 2 cm sections and added to individual wells of a 6-well plate (VWR International Ltd, Leicester, UK).

[0106] To each container was added 6 mL of a solution containing 1.5 g / L of Ariel unit dose liquid (no enzyme). Stains were added and incubated at 35°C for 45 minutes at an agitation speed of 600 rpm with and without the alginate lyase with the specified SEQ ID NO: and unit dose liquid solution at a wash concentration of 2 ppm.

[0107] After the wash cycle, the wash water was drained and the fabrics were rinsed with cold tap water (19 grains per US gallon) at an agitation speed of 300 rpm, after which the washed stain swatches were laid flat on a rack to dry under ambient conditions.

[0108] This process was repeated three more times, resulting in a total of four washed stains per treatment, i.e., four external replicates containing one stain each.

[0109] Pre- and post-wash analysis of the stains was completed using image analysis (Illuminant D65 / 10) to calculate the difference in stain removal between the test and reference formulations.

[0110] The stain removal index (SRI) was calculated using the following formula: AB is the color difference between the stain-free area of ​​the fabric before washing and the stain before washing, and ΔE AD is the color difference between the stain-free area of ​​the fabric before washing and the stain after washing. SRI = 100 × (ΔE AB -ΔE AD ) / ΔEAB ,

[0111] The average test results are presented in the table below and show that the addition of alginate lyase of SEQ ID NO: 6 significantly improved stain removal by 41.8 SRI units for the soiled collar and cuffs. This improvement is statistically significant (p<0.001) versus the no enzyme control according to Student's T-test, i.e., greater than the 99.9% confidence level.

[0112] [Table 1] * contains an equal volume of buffer carried by SEQ ID NO:6.

[0113] Example 2: Cleaning performance of liquid detergent compositions containing defined alginate lyases The cleaning performance of alginate lyase for the removal of chocolate-containing stains from cotton was determined in the context of a liquid laundry detergent as follows: Stains with a diameter of approximately 3 cm were prepared on 5 cm x 5 cm swatches of knitted cotton supplied by Warwick Equest Ltd (Consett, UK) by pipetting 0.5 g of chocolate mousse or ice cream into the centre of each swatch. The stained swatches were left to hang dry for 24 hours before use and imaged prior to testing (see below).

[0114] Stain removal tests were performed using a Tergotometer Detergent Tester supplied by Copley Scientific Ltd (Nottingham, UK). Four pots were filled with 1000 mL of hard water (18 grains per US gallon) heated to 40°C, and 2.75 mL of Ariel liquid (purchased from Asda Stores Ltd in the UK in February 2019, product code: 6016650) was added to each pot. Four stains were added to each pot, along with 5 cm x 5 cm swatches of clean white knitted cotton ballast (Warwick Equest Ltd) and eight 5 cm x 5 cm swatches of WfK SBL 2004 stain sheets per pot, to reach a total fabric load of 60 g.

[0115] Five milligrams of active alginate lyase containing SEQ ID NO:6 and SEQ ID NO:7 was added to one of the two pots to give a wash concentration of 5 ppm active alginate lyase. The temperature was maintained at 35°C for the duration of the test. After 40 minutes, the wash water was drained and the fabrics were rinsed twice for 5 minutes with 1000 mL of cold tap water (18 grains per US gallon), after which the washed stain swatches were laid flat on a rack and allowed to dry under ambient conditions.

[0116] This process was repeated four more times, alternating treatments between the two pots, resulting in a total of 16 washed stains per treatment, i.e., four external replicates containing four stains each.

[0117] The average test results are presented in the table below and show that the addition of the combination of SEQ ID NO: 6 and SEQ ID NO: 7 significantly improved stain removal by 8.78 SRI units for chocolate ice cream stains and 8.54 SRI units for mousse mix stains. These improvements are statistically significant (p<0.01) versus the no enzyme control according to Student's T-test, i.e., at a confidence level of greater than 99%.

[0118] [Table 2]

[0119] Example 3: Cleaning performance of liquid detergents containing different alginate lyases The cleaning performance of different sequences of alginate lyase for the removal of alginate-containing stains from cotton was determined in liquid laundry detergent as follows: Circular stains with a diameter of approximately 3 cm were prepared on 5 cm x 5 cm swatches of knitted cotton supplied by Warwick Equest Ltd (Consett, UK) by pipetting 1 mL of a solution of 3% low viscosity sodium alginate (Carbosynth Ltd (Compton, UK) batch number Y8317361901) and 0.6% carbon black (Alfa Aesar (Lancashire, UK), batch number N21E016) into the centre of each swatch. The stained swatches were left to hang dry for 24 hours before use and imaged prior to testing (see below).

[0120] Stain removal tests were performed using a Talboys Professional Incubating Microplate Shaker supplied by Cole-Parmer Instrument Co., Ltd., UK. A circular carbon black / sodium alginate stain was added to each well of a 6-well plate (VWR International Ltd., Leicester, UK). A stock solution of alginate lyase was prepared by adding 50 ppm of active alginate lyase enzyme to tap water (6 gallons per well). In each well, 8 mL of a solution containing 5 ppm of active alginate lyase and 10 g / L Ariel liquid (purchased from Asda Stores Ltd., UK, in February 2019, product code: 6016650) was added. The stain and alginate lyase solution were incubated with the Ariel liquid solution at 40°C for 1 hour at a stirring speed of 600 rpm. A blank was obtained by adding only Ariel liquid and tap water (6 gallons per well) to six wells to obtain an Ariel concentration equivalent to 10 g / L. Six replicates were collected for each alginate lyase. At the end of the incubation period, the wash water was drained and the washed stains were rinsed once with 8 mL of cold tap water (6 gallons per gallon) at 20°C for 10 minutes, after which the washed stain swatches were placed flat on a rack and allowed to dry under ambient conditions.

[0121] This process was repeated once more, resulting in a total of 12 washed stains per treatment, i.e., two external replicates containing six stains each.

[0122] Pre- and post-analysis of the stains was completed using image analysis (Illuminant D65 / 10) to calculate the difference in stain removal between the test and reference formulations.

[0123] The stain removal index (SRI) was calculated using the following formula: AB is the color difference between the stain-free area of ​​the fabric before washing and the stain before washing, and ΔE AD is the color difference between the stain-free area of ​​the fabric before washing and the stain after washing. SRI = 100 × (ΔE AB -ΔE AD ) / ΔE AB ,

[0124] The average test results are presented in the table below. They show that the present invention significantly improves stain removal by 30.0 SRI units for sodium alginate-containing stains. This improvement is statistically significant (p<0.001) versus the no-alginate lyase control according to Student's T-test, i.e., greater than a 99.9% confidence level. Addition of alginate lyases having less than 25% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and SEQ ID NO:5 removes 2.8 SRI units of stain. This change is not statistically significant (p>0.1) versus the no-alginate lyase control according to Student's T-test, i.e., less than a 90% confidence level.

[0125] [Table 3]

[0126] Detergent Example Examples 1-6. Granular laundry detergent compositions designed for hand washing or top loading washing machines.

[0127] [Table 4]

[0128] Examples 7-13. Granular laundry detergent compositions designed for front-loading automatic washing machines.

[0129] [Table 5] * DNase is given as mg of active enzyme per 100 g of detergent.

[0130] Examples 14 to 21. Heavy-duty liquid laundry detergent compositions

[0131] [Table 6-1]

[0132] [Table 6-2]

[0133] Examples 22-28. Unit Dose Laundry Detergent Compositions Such unit dose formulations may contain one or more compartments.

[0134] [Table 7]

[0135] Example 29. Multi-compartment unit dose composition Multi-compartment unit dose laundry detergent formulations of the present invention are provided below. In these examples, the unit dose has three compartments, but similar compositions can also be made with two, four, or five compartments. The film used to encapsulate the compartments is polyvinyl alcohol.

[0136] [Table 8]

[0137] [Table 9]

[0138] Examples 30 to 33. Fabric softener compositions of the present invention

[0139] [Table 10]

[0140] Ingredients and Notes for Composition Examples 1-33 Linear alkylbenzene sulfonates with an average aliphatic carbon chain length of C11 to C18 C12-18 Dimethylhydroxyethylammonium Chloride AE3S is a C12-15 alkyl ethoxy (3) sulfate. AE7 is a C12-15 alcohol ethoxylate with an average degree of ethoxylation of 7. AE9 is a C12-16 alcohol ethoxylate with an average degree of ethoxylation of 9. HSAS is a mid-chain branched primary alkyl sulfate having a carbon chain length of about 16-17, as disclosed in US Pat. Nos. 6,020,303 and 6,060,443. Polyacrylate MW4500 is supplied by BASF. The carboxymethylcellulose is Finnfix® V supplied by CP Kelco (Arnhem, Netherlands). CHEC is a cationically modified hydroxyethyl cellulose polymer. An example of a phosphonate chelating agent is diethylenetetraaminepentaacetic acid (DTPA) hydroxyethane diphosphonate (HEDP). Savinase®, Natalase®, Stainzyme®, Lipex®, Celluclean™, Mannaway®, and Whitezyme® are all products of Novozymes (Bagsvaerd, Denmark). Purafect® and Purafect Prime® are products of Genencor International (Palo Alto, California, USA). Optical Brightener 1 is Tinopal® AMS, Optical Brightener 2 is Tinopal® CBS-X, Direct Violet 9 is Pergasol® Violet BN-Z, and NOBS is sodium nonanoyloxybenzenesulfonate. TAED is tetraacetylethylenediamine. S-ACMC is carboxymethylcellulose conjugated with CI Reactive Blue 19, trade name AZO-CM-CELLULOSE. The soil release agent is Repel-o-tex® PF. The acrylic acid / maleic acid copolymer has a molecular weight of 70,000 and an acrylate:maleate ratio of 70:30. EDDS is the sodium salt of ethylenediamine-N,N'-disuccinic acid, (S,S) isomer, a suds suppressor aggregate supplied by Dow Corning (Midland, Michigan, USA). HSAS is a medium-chain branched alkyl sulfate. Liquitint® Violet CT is a polymeric color pigment supplied by Milliken (Spartanburg, South Carolina, USA). The polyethoxylated azothiophene dye is Violet DD™ polymeric hue dye supplied by Milliken (Spartanburg, South Carolina, USA). 1 Random graft copolymers are polyvinyl acetate-grafted polyethylene oxide copolymers having a polyethylene oxide backbone and multiple polyvinyl acetate side chains. The molecular weight of the polyethylene oxide backbone is about 6000, the weight ratio of polyethylene oxide to polyvinyl acetate is about 40-60, and there is no more than one grafting point per 50 ethylene oxide units. 2 Polyethyleneimine (MW=600) with 20 ethoxylate groups per —NH. 3 The amphiphilic alkoxylated polymer is polyethyleneimine (MW600) prepared from a polymer derivatized to contain 24 ethoxylate groups per —NH and 16 propoxylate groups per —NH. 4Amylase is given as mg of active enzyme per 100g of detergent. 5 DNase in all these examples is given as mg of active enzyme per 100 g of detergent. DNase may contain trace amounts of superoxide dismutase impurity. 6 Defined alginate lyase (given in all examples as mg of active enzyme per 100 g of detergent). a Proxel GXL, a 20% aqueous dipropylene glycol solution of 1,2-benzisothiazolin-3-one supplied by Lonza. b N,N-bis(hydroxyethyl)-N,N-dimethylammonium chloride fatty acid ester. The iodine value of the parent fatty acid of this material is 18-22. The material obtained from Evonik contains impurities in the form of free fatty acids, the monoester form of N,N-bis(hydroxyethyl)-N,N-dimethylammonium chloride fatty acid ester, and fatty acid esters of N,N-bis(hydroxyethyl)-N-methylamine. c MP10®, supplied by Dow Corning, 8% active d Expressed as 100% encapsulated perfume oil as described in U.S. Pat. No. 8,765,659 e Rheovis® CDE, a cationic polymeric thickener supplied by BASF f N,N-dimethyloctanamide and N,N-dimethyldecanamide in an approximately 55:45 weight ratio, Steposol® M-8-10 from Stepan Company

[0141] Dimensions and values ​​disclosed herein should not be understood as being strictly limited to the exact numerical values ​​recited. Instead, unless otherwise indicated, each such dimension is intended to mean both the recited value and a functionally equivalent range surrounding that value. For example, a dimension disclosed as "40 mm" is intended to mean "about 40 mm."

Claims

1. 1. A detergent composition comprising an alginate lyase enzyme and a cleaning adjunct, wherein the alginate lyase enzyme comprises an alginate lyase selected from an alginate lyase having at least 90% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, and mixtures thereof.

2. 2. The detergent composition of claim 1, wherein the alginate lyase is selected from (i) an alginate lyase having at least 90% sequence identity to SEQ ID NO:1, and (ii) an alginate lyase having at least 90% sequence identity to SEQ ID NO:5, and (iii) mixtures thereof.

3. 2. The detergent composition of claim 1, wherein the alginate lyase is selected from (i) an alginate lyase having at least 90% sequence identity to SEQ ID NO:6, and (ii) an alginate lyase having at least 90% sequence identity to SEQ ID NO:7, and (iii) mixtures thereof.

4. 4. The detergent composition according to any one of claims 1 to 3, wherein the alginate lyase enzyme comprises two or more alginate lyase enzymes.

5. 5. A detergent composition according to any one of claims 1 to 4, wherein the alginate lyase enzyme is obtainable from Flavobacterium sp., Sphingomonas sp., Zobellia galactaivorans.

6. 6. A detergent composition according to any preceding claim, wherein the alginate lyase enzyme is from polysaccharide lyase family 7.

7. 10. The detergent composition of claim 1, wherein the alginate lyase enzyme provides activity against poly(β-D-mannuronic acid) (polyM activity) and activity against poly(α-L-guluronic acid) (polyG activity).

8. The detergent composition according to any one of claims 1 to 7, further comprising a surfactant.

9. The detergent composition of claim 8, wherein the weight ratio of surfactant to active alginate lyase enzyme protein is at least 500:

1.

10. 10. The detergent composition of claim 8 or 9, wherein the surfactant comprises a nonionic surfactant.

11. 11. The detergent composition of claim 10, wherein the surfactant further comprises an anionic surfactant.

12. A detergent composition according to any preceding claim, comprising an additional enzyme.

13. 1. A method of treating a surface, comprising contacting the surface with an aqueous cleaning solution comprising (i) an alginate lyase enzyme, and (ii) a cleaning adjunct, wherein the alginate lyase enzyme is selected from an alginate lyase having at least 90% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, and mixtures thereof.

14. 13. Use of a composition according to any one of claims 1 to 12 for improving the whiteness of fabrics, and / or for improving stain removal from fabrics, and / or for removing malodours from fabrics, and / or for removing stains on collars and / or cuffs, and / or for anti-wrinkle effect on fabrics, and / or for improving drying of fabrics.

15. 14. Use of the method according to claim 13 for improving the whiteness of fabrics, and / or for improving stain removal from fabrics, and / or for removing malodours from fabrics, and / or for removing stains on collars and / or cuffs, and / or for anti-wrinkle effect of fabrics, and / or for improving drying of fabrics.

Images