Compositions comprising a hexosaminidase and a protease

Abstract

The present invention relates to compositions such as detergent or cleaning compositions comprising a polypeptide having hexosaminidase activity and a polypeptide having protease activity, as well as methods of using the compositions for e.g. laundry.

Description

[0001] COMPOSITIONS COMPRISING A HEXOSAMINIDASE AND A PROTEASE

[0002] Reference to a Sequence Listing

[0003] This application contains a Sequence Listing in computer readable form, which is incorporated herein by reference.

[0004] Field of the Invention

[0005] The present invention relates to compositions such as detergent or cleaning compositions comprising a polypeptide having hexosaminidase activity and a polypeptide having protease activity, as well as methods of using the compositions for e.g. laundry.

[0006] Background of the Invention

[0007] Detergent or cleaning compositions comprising enzymes such as proteases, amylases, lipases, cellulases, mannanases and pectate lyases have been used for decades in e.g. laundry, dishwashing and industrial cleaning. More recently, newer enzyme classes have been disclosed for use in e.g. laundry to provide additional cleaning benefits that are not necessarily provided by the conventional cleaning enzymes. Such benefits include deep cleaning of soil that may not be removed by conventional enzymes, for example removal of organic materials that may include components of biofilm, such as polysaccharides, PNAG (poly-N-acetylglucosamine), proteins, DNA or other components present in biofilm as well as e.g. sebum and skin debris.

[0008] Despite the benefits of existing enzymatic cleaning solutions, there is still a need for improved solutions that may be provide by new combinations of enzymes.

[0009] Summary of the Invention

[0010] The present invention provides cleaning compositions comprising a polypeptide having hexosaminidase activity, a polypeptide having protease activity and a cleaning adjunct ingredient, in particular wherein the polypeptide having protease activity comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 1 and comprises a substitution in position 209 of SEQ ID NO: 1 selected from the group consisting of A209K, A209R, A209Q, A209N, A209S and A209T.

[0011] The invention further provides methods for laundering textiles using the cleaning compositions disclosed herein as well as use of the cleaning compositions in a cleaning process, such as laundry or hard surface cleaning such as dishwashing

[0012] Overview of sequences

[0013] SEQ I D NO: 1 is the sequence of the Savinase® protease polypeptide from Bacillus lentus. SEQ ID NO: 2 is a mature hexosaminidase polypeptide obtained from Terribacillus saccharophilus.

[0014] SEQ ID NO: 3 is a mature hexosaminidase polypeptide obtained from Terribacillus saccharophilus.

[0015] SEQ ID NO: 4 is a mature hexosaminidase polypeptide obtained from Terribacillus goriensis.

[0016] SEQ ID NO: 5 is a mature hexosaminidase polypeptide obtained from Terribacillus saccharophilus.

[0017] SEQ ID NO: 6 is a mature hexosaminidase polypeptide obtained from Terribacillus saccharophilus.

[0018] SEQ ID NO: 7 is a variant of the hexosaminidase polypeptide of SEQ ID NO: 2.

[0019] Definitions

[0020] In accordance with this detailed description, the following definitions apply. Note that the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0021] Unless defined otherwise or clearly indicated by context, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0022] Hexosaminidase: The term “polypeptide having hexosaminidase activity” or “hexosaminidase” includes the term "dispersin'1and the abbreviation "Dsp", and refers to a polypeptide having hexosaminidase activity, EC 3.2.1., that catalyzes the hydrolysis of (3-1 ,6- glycosidic linkages of N-acetyl-glucosamine polymers found in, e.g., biofilm. The term hexosaminidase includes polypeptides having N-acetylglucosaminidase activity and p-N- acetylglucosaminidase activity. The term "polypeptide having hexosaminidase activity" may be used interchangeably with the term "hexosaminidase". Hexosaminidase activity may be determined according to the procedure described in Assay 1 or 2.

[0023] Protease: The term “polypeptide having protease activity” or “protease” refers to an enzyme that hydrolyses peptide bonds and includes enzymes belonging to the EC 3.4 enzyme group. The most widely used proteases in the detergent industry for e.g. laundry and dishwashing are the serine proteases or serine peptidases, which are a subgroup of proteases characterised by having a serine in the active site that forms a covalent adduct with the substrate. Subtilases (and serine proteases) are also characterised by having two active site amino acid residues apart from the serine residue, namely a histidine and an aspartic acid residue, i.e. an Asp / Ser / His catalytic triad. Subtilase refers to a sub-group of serine protease according to Siezen et al., Protein Engng. 4 (1991) 719-737 and Siezen et al. Protein Science 6 (1997) 501-523. The subtilases may be divided into six sub-divisions, i.e. the Subtilisin family, the Thermitase family, the Proteinase K family, the Lantibiotic peptidase family, the Kexm family and the Pyrolysm family. Proteases have been used in detergents for decades, the proteases used in detergents being mainly endopeptidases (EC 3.4.21).

[0024] Adjunct ingredients: The term "adjunct ingredient" or “adjunct material” refers to any ingredient used in a cleaning or detergent composition of the invention in combination with the hexosaminidase and protease enzymes. More detailed information on adjunct ingredients is provided further below.

[0025] Biofilm: A biofilm is any group of microorganisms in which cells stick to each other on a surface, such as a textile or a dishware or other hard surface. These adherent cells are frequently embedded within a self-produced matrix of extracellular polymeric substance (EPS). Biofilm EPS is a polymeric conglomeration generally composed of extracellular DNA, proteins and polysaccharides. Biofilms may form on living or non-living surfaces. The microbial cells growing in a biofilm are physiologically distinct from planktonic cells of the same organism, which, by contrast, are single cells that may float or swim in a liquid medium. Bacteria living in a biofilm usually have significantly different properties from free-floating bacteria of the same species, as the dense and protected environment of the film allows them to cooperate and interact in various ways. One benefit of this environment is increased resistance to detergents and antibiotics, as the dense extracellular matrix and the outer layer of cells protect the interior of the community. On laundry, biofilm producing bacteria can be found among the following species: Acinetobacter sp., Aeromicrobium sp., Brevundimonas sp., Micro bacterium sp., Micrococcus luteus, Pseudomonas sp., Staphylococcus epidermidis, and Stenotrophomonas sp.

[0026] Deep cleaning: The term “deep cleaning” refers to reduction or removal of organic materials including components of biofilm, such as EPS or parts hereof, polysaccharides, PNAG (poly-N-acetylglucosamine), proteins, DNA, soil or other components present in biofilm. Deep cleaning encompasses not only removal of visible soiling on e.g. textiles, but also e.g. organic material or malodor within a textile that may not be removed using conventional detergent compositions and enzymes. Such organic material may for example include dead cell material, skin debris, sebum, sweat, grease and other stains derived from e.g. humans (body soils) or microbes.

[0027] Detergent composition: The term "detergent composition" (or “cleaning composition”) includes unless otherwise indicated any form of detergent or cleaning composition. These include granular or powder-form all-purpose or heavy-duty washing agents, especially cleaning detergents; liquid, gel or paste-form all-purpose washing agents, especially the so-called heavy- duty liquid (HDL) types; single unit dose (SUD) compositions such as pods, capsules, tabs, etc. with one or more chambers; liquid fine-fabric detergents; hand dishwashing agents or light duty dishwashing agents, especially those of the high-foaming type; machine dishwashing agents, including the various tablet, granular, liquid and rinse-aid types for household and institutional use; liquid cleaning and disinfecting agents, including antibacterial hand-wash types, cleaning bars, soap bars, mouthwashes, denture cleaners, car or carpet shampoos, bathroom cleaners; hair shampoos and hair-rinses; shower gels, foam baths; metal cleaners; as well as cleaning auxiliaries such as bleach additives and "stain-stick" or pre-treat types. The terms "detergent composition" and "detergent formulation" are used in reference to mixtures which are intended for use in a wash medium for the cleaning of soiled objects. In some embodiments, the term is used in reference to laundering fabrics and / or garments (e.g., "laundry detergents"). In alternative embodiments, the term refers to other detergents, such as those used to clean dishes, cutlery, etc. (e.g., "dishwashing detergents"). It is not intended that the present invention be limited to any particular detergent formulation or composition. The term “detergent composition” is not intended to be limited to compositions that contain surfactants. It is intended that in addition to the polypeptides of the invention, the term encompasses detergents that may contain, e.g., surfactants, builders, chelators or chelating agents, bleach system or bleach components, polymers, fabric conditioners, foam boosters, suds suppressors, dyes, perfume, tannish inhibitors, optical brighteners, bactericides, fungicides, soil suspending agents, anti-corrosion agents, enzyme inhibitors or stabilizers, enzyme activators, transferases, hydrolytic enzymes, oxido reductases, bluing agents and fluorescent dyes, antioxidants, and solubilizers.

[0028] Effective amount of enzyme: The term "effective amount of enzyme" refers to the quantity of enzyme necessary to achieve the enzymatic activity required in the specific application, e.g., in a defined detergent composition. Such effective amounts are readily ascertained by one of ordinary skill in the art and are based on many factors, such as the particular enzyme used, the cleaning application, the specific composition of the detergent composition, and whether a liquid or dry (e.g., granular, bar) composition is required, and the like. Exemplary concentrations of enzyme in a detergent composition and in a wash liquor are provided elsewhere herein.

[0029] Expression: The term “expression” includes any step involved in the production of a enzyme including, but not limited to, transcription, post-transcriptional modification, translation, post-translational modification and secretion.

[0030] Extension: The term “extension” means an addition of one or more amino acids to the amino and / or carboxyl terminus of a hexosaminidase or a protease, wherein the “extended” enzyme has hexosaminidase activity or protease activity.

[0031] Fabric: The term "fabric" encompasses any textile material. Thus, it is intended that the term encompass garments, as well as fabrics, yarns, fibers, non-woven materials, natural materials, synthetic materials, and any other textile material.

[0032] Fragment: The term “fragment” means an enzyme having one or more amino acids absent from the amino and / or carboxyl terminus of the enzyme, wherein the fragment has enzyme activity, i.e. hexosaminidase activity or protease activity. Hard surface cleaning: The term hard surface cleaning comprises, in addition to dishwashing, other domestic or industrial hard surfaces, such as but not limited to domestic hard surfaces including floors, walls, tabletops, kitchen surfaces including kitchen appliance surfaces, bathroom surfaces, etc.

[0033] Host cell: A "host cell" is an organism into which an expression vector, phage, virus, or other DNA construct, including a polynucleotide encoding an enzyme, has been introduced. Exemplary host strains are microorganism cells (e.g., bacteria, filamentous fungi, and yeast) capable of expressing the polypeptide of interest.

[0034] Improved wash performance: The term “improved wash performance” may be defined as improved cleaning, for example improved deep cleaning of e.g. textiles. Wash performance may be expressed as a remission value of the stained swatches. After washing and rinsing the swatches are spread out flat and allowed to air dry at room temperature overnight. All washed swatches are evaluated the day after washing. Light reflectance evaluations of the swatches is performed using a suitable instrument such as a Macbeth Color Eye 7000 reflectance spectrophotometer with a very small aperture. The measurements are made without UV in the incident light and remission value typically at 460 nm is extracted.

[0035] Laundering: The term “laundering” relates to both household laundering and industrial laundering and means the process of treating textiles with a solution containing e.g. a cleaning or detergent composition of the present invention. The laundering process can for example be carried out using a household or an industrial washing machine or can be carried out by hand.

[0036] Low / medium / high detergent concentration: The term “low detergent concentration” system includes detergents where less than about 800 ppm of detergent components is present in the wash water. Asian, e.g., Japanese detergents are typically considered low detergent concentration systems. The term “medium detergent concentration” system includes detergents wherein between about 800 ppm and about 2000 ppm of detergent components is present in the wash water. North American detergents are generally considered to be medium detergent concentration systems. The term “high detergent concentration” system includes detergents wherein greater than about 2000 ppm of detergent components is present in the wash water. European detergents are generally considered to be high detergent concentration systems.

[0037] Mature polypeptide: The term “mature polypeptide” means a polypeptide in its mature form following N-terminal processing and / or C-terminal processing (e.g., removal of signal peptide).

[0038] It is known in the art that a host cell may produce a mixture of two of more different mature polypeptides (i.e., with a different C terminal and / or N terminal amino acid) expressed by the same polynucleotide. It is also known that different host cells process polypeptides differently, and thus, one host cell expressing a polynucleotide may produce a different mature polypeptide (e.g., having a different C terminal and / or N terminal amino acid) as compared to another host cell expressing the same polynucleotide.

[0039] Parent: The term “parent”, “parent enzyme” or “parent polypeptide” means an enzyme, in particular a hexosaminidase or a protease, in which an alteration is made to produce a variant having the same enzymatic function, e.g. a hexosaminidase variant will have hexosaminidase activity. The alterations in the parent to result in the variant will typically include one or more substitutions and may also include one or more insertions and / or deletions, and / or N- or C- terminal extensions or truncations. The parent may be a naturally occurring (wild-type) polypeptide or a variant or fragment thereof.

[0040] Polypeptide: The enzymes disclosed herein, including hexosaminidases and proteases, may be referred to interchangeably as a “polypeptide” or an “enzyme”.

[0041] Recombinant: The term "recombinant" is used in its conventional meaning to refer to the manipulation, e.g., cutting and rejoining, of nucleic acid sequences to form constellations different from those found in nature. The term recombinant refers to a cell, nucleic acid, polypeptide or vector that has been modified from its native state. Thus, for example, recombinant cells express genes that are not found within the native (non-recombinant) form of the cell, or express native genes at different levels or under different conditions than found in nature. The term “recombinant” is synonymous with “genetically modified” and “transgenic”.

[0042] Sequence identity: The relatedness between two amino acid sequences or between two nucleotide sequences is described by the parameter “sequence identity”.

[0043] For purposes of the present invention, the sequence identity between two amino acid sequences is determined as the output of “longest identity” using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16: 276-277), preferably version 6.6.0 or later. The parameters used are a gap open penalty of 10, a gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. In order for the Needle program to report the longest identity, the -nobrief option must be specified in the command line. The output of Needle labeled “longest identity” is calculated as follows:

[0044] (Identical Residues x 100) / (Length of Alignment - Total Number of Gaps in Alignment)

[0045] Textile: The term "textile" refers to woven fabrics, as well as staple fibers and filaments suitable for conversion to or use as yarns, woven, knit, and non-woven fabrics. The term encompasses yarns made from natural, as well as synthetic (e.g., manufactured) fibers. The term "textile materials" is a general term for fibers, yarn intermediates, yarn, fabrics, and products made from fabrics (e.g., garments and other articles).

[0046] Variant: The term “variant” means a polypeptide of a parent enzyme having hexosaminidase activity or protease activity comprising a substitution, an insertion (including extension), and / or a deletion (e.g., truncation), at one or more positions relative to the parent. A substitution means replacement of the amino acid occupying a position with a different amino acid; a deletion means removal of the amino acid occupying a position; and an insertion means adding 1-5 amino acids (e.g., 1-3 amino acids, in particular one amino acid) adjacent to and immediately following the amino acid occupying a position.

[0047] Wash liquor: The term “wash liquor” (or “wash water”) refers to an aqueous solution comprising a hexosaminidase and protease of the invention. A wash liquor is a solution, e.g. found in a washing machine or dishwasher, containing water and a detergent composition comprising the hexosaminidase and protease. The detergent composition, prior to being mixed with water to form a wash liquor, may be in any suitable form as described elsewhere herein, for example a liquid or powder.

[0048] Water hardness: The term “water hardness” or “degree of hardness” or “dH” or “°dH” as used herein refers to German degrees of hardness. One degree is defined as 10 milligrams of calcium oxide per liter of water.

[0049] Wild-type: The term "wild-type" in reference to an amino acid sequence or nucleic acid sequence means that the amino acid sequence or nucleic acid sequence is a native or naturally occurring sequence. As used herein, the term "naturally occurring" refers to anything (e.g., proteins, amino acids, or nucleic acid sequences) that is found in nature. Conversely, the term "non-naturally occurring" refers to anything that is not found in nature (e.g., recombinant nucleic acids and protein sequences produced in the laboratory or by modification of the wild-type sequence).

[0050] Conventions for Designation of Variants

[0051] In describing enzyme variants in the context of the present invention, the nomenclature described below is adapted for ease of reference. The accepted IIIPAC single letter amino acid abbreviation is employed.

[0052] Substitutions. For an amino acid substitution, the following nomenclature is used: Original amino acid, position, substituted amino acid. Accordingly, the substitution of threonine at position 226 with alanine is designated as “T226A”. Multiple mutations may be separated by addition marks (“+”), e.g., “G205R + S411 F”, representing substitutions at positions 205 and 411 of glycine (G) with arginine (R) and serine (S) with phenylalanine (F), respectively. Multiple mutations may alternatively be indicated by a space, a comma, or a plus sign, e.g. “G205R S411 F”, “G205R, S411 F” or “G205R+S411 F”.

[0053] Deletions. For an amino acid deletion, the following nomenclature is used: Original amino acid, position, *. Accordingly, the deletion of glycine at position 195 is designated as “G195*”. Multiple deletions are separated by addition marks (“+”), e.g., “G195* + S411*”. Insertions. For an amino acid insertion, the following nomenclature is used: Original amino acid, position, original amino acid, inserted amino acid. Accordingly, the insertion of lysine after glycine at position 195 is designated “G195GK”. An insertion of multiple amino acids is designated [Original amino acid, position, original amino acid, inserted amino acid #1 , inserted amino acid #2; etc.]. For example, the insertion of lysine and alanine after glycine at position 195 is indicated as “G195GKA”.

[0054] Multiple alterations. Variants comprising multiple alterations are separated by addition marks (“+”) as explained above, e.g., “R170Y+G195E” representing a substitution of arginine and glycine at positions 170 and 195 with tyrosine and glutamic acid, respectively. Alternatively, the multiple alterations may be separated by a space or comma as mentioned above.

[0055] Different alterations. Where different alterations can be introduced at a position, the different alterations are separated by a comma, e.g., “R170Y.E” represents a substitution of arginine at position 170 with tyrosine or glutamic acid. Thus, “Y167G.A + R170G.A” designates the following variants: “Y167G+R170G”, “Y167G+R170A”, “Y167A+R170G” and “Y167A+R170A”.

[0056] Detailed Description of the Invention

[0057] One aspect of the invention relates to a cleaning composition comprising a polypeptide having hexosaminidase activity, a polypeptide having protease activity and a cleaning adjunct ingredient, wherein the polypeptide having protease activity comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 1 and comprises a substitution in position 209 of SEQ ID NO: 1 selected from the group consisting of A209K, A209R, A209Q, A209N, A209S and A209T.

[0058] The polypeptide having hexosaminidase activity preferably has beta-1 , 6-N- acetylglucosaminidase activity, and preferably comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 6.

[0059] Other aspects of the invention relate to methods for laundering a textile using the cleaning compositions, as well as use of the compositions in a cleaning process.

[0060] Polypeptides having hexosaminidase activity

[0061] The cleaning compositions of the invention comprise a polypeptide having hexosaminidase activity, preferably having beta-1 , 6-N-acetylglucosaminidase activity, and a polypeptide having protease activity.

[0062] In some preferred embodiments, the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 70% sequence identity to 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, for example at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94% or at least 95% sequence identity to 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.

[0063] In one preferred embodiment, the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 2, for example at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94% or at least 95% sequence identity to SEQ ID NO: 2.

[0064] In another preferred embodiment, the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 7.

[0065] In another preferred embodiment, the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 2, such as least 75%, at least 80%, at least 85% or at least 90% sequence identity to SEQ ID NO: 2, and comprises a substitution at one or more positions selected from positions 3, 15, 49, 59, 163, 186, 225, 227, 232, 235, 252, 260, 272, 279, 281 , 308, 309 and 312 of SEQ ID NO: 2. Preferably, the polypeptide having hexosaminidase activity comprises comprises a substitution at two or more positions, such as at three, four, five or more positions, selected from positions 3, 15, 49, 59, 163, 186, 225, 227, 232, 235, 252, 260, 272, 279, 281 , 308, 309 and 312 of SEQ ID NO: 2. More preferably, the polypeptide having hexosaminidase activity comprises comprises a substitution at six or more positions, such as at seven, eight, nine, ten or more, positions selected from positions 3, 15, 49, 59, 163, 186, 225, 227, 232, 235, 252, 260, 272, 279, 281 , 308, 309 and 312 of SEQ ID NO: 2. Still more preferably, the polypeptide having hexosaminidase activity comprises comprises a substitution at 11 or more positions, such as at 12, 13, 14, 15, 16, 17 or all 18 positions selected from positions 3, 15, 49, 59, 163, 186, 225, 227, 232, 235, 252, 260, 272, 279, 281 , 308, 309 and 312 of SEQ ID NO: 2.

[0066] In a further preferred embodiment, the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 2, such as least 75%, at least 80%, at least 85% or at least 90% sequence identity to SEQ ID NO: 2, and comprises at least one substitution selected from the group consisting of Q3F, Q3I, H15Y, A49W, N59E, S163P, S186R, S225G, N227T, E232D, G235W, N252P, N260Q, H272V, H272P, S279D, Y281 P, K308Q, K308E, K309E, K312Q and K312E, wherein numbering is based on SEQ ID NO: 2. Preferably, the polypeptide having hexosaminidase activity comprises two, three, four, five or more substitutions selected from the group consisting of Q3I, H15Y, A49W, N59E, S163P, S186R, S225G, N227T, E232D, G235W, N252P, N260Q, H272V, S279D, Y281 P, K308Q, K309E and K312Q, wherein numbering is based on SEQ ID NO: 2. More preferably, the polypeptide having hexosaminidase activity comprises six, seven, eight, nine, ten or more substitutions selected from the group consisting of Q3l, H15Y, A49W, N59E, S163P, S186R, S225G, N227T, E232D, G235W, N252P, N260Q, H272V, S279D, Y281 P, K308Q, K309E and K312Q, wherein numbering is based on SEQ ID NO: 2. Still more preferably, the polypeptide having hexosaminidase activity comprises eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen or all eighteen substitutions selected from the group consisting of Q3I, H15Y, A49W, N59E, S163P, S186R, S225G, N227T, E232D, G235W, N252P, N260Q, H272V, S279D, Y281 P, K308Q, K309E and K312Q, wherein numbering is based on SEQ ID NO: 2.

[0067] In another preferred embodiment, the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 7. In this embodiment, the polypeptide preferably has one or more of I in position 3, Y in position 15, W in position 49, E in position 59, P in position 163, R in position 186, G in position 225, T in position 227, D in position 232, Win position 235, P in position 252, Q in position 260, V in position 272, D in position 279, P in position 281 , Q in position 308, E in position 309 or Q in position 312. More preferably, the polypeptide has 2, 3, 4, 5, 6, 7, 8, 9 or 10 or more of the indicated amino acid residues in the indicated positions, for example 11 , 12, 13, 14, 15, 16, 17 or even all 18 of the indicated amino acid residues in the indicated positions.

[0068] In another embodiment, the polypeptide having hexosaminidase activity may be a variant of SEQ ID NO: 2, wherein the variant comprises, compared to SEQ ID NO: 2: a substitution at one or more positions, preferably two, three or four positions, selected from positions 163, 227, 252 and 309; and a substitution at one or more positions corresponding to positions 111 , 120, 106, 124, 127, 150, 170, 171 , 178, 199, 208, 254, 255 and 278. Preferred variants of this embodiment comprise two, three or four substitutions selected from the group consisting of S163P, N227T, N252P and K309E; and further comprise at least one substitution selected from the group consisting of D111 R, T120V, V106M, V106N, Y124I, Y124K, Y124L, R127P, E150N, L170G, D171 F, D171 H, Q178K, S199W, S208W, F254I, T255D and I278V. Other preferred variants of this embodiment comprise, compared to SEQ ID NO: 2, at least one substitution selected from the group consisting of H15Y, S186R, S225G, E232D, G235W, N260Q, H272V, S279D, Y281 P, K308Q and K312Q, for example 2, 3, 4, 5, 6, 7, 8, 9, 10 or all of said substitutions; and optionally further comprise one or more substitutions selected from the group consisting of Q3I, A49W and N59E, preferably 2 or all 3 of said substitutions; or one or more substitutions selected from the group consisting of Q3F, V140I, Q215K and N267T, preferably 2, 3 or all 4 of said substitutions. Hexosaminidase variants of this embodiment may further comprise at least one substitution selected from the group consisting of D111 R, T120V, V106M / N, E150N, D171 F and I278V, for example at least two of said substitutions; preferably wherein the variant comprises at least one substitution selected from the group consisting of D111 R, T120V and E150N. In further embodiments, the polypeptide having hexosaminidase activity may be a variant of a parent hexosaminidase selected from SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, wherein the variant comprises, compared to the parent hexosaminidase, a substitution at one or more positions, for example two, three, four, five, six, seven, eight, nine, ten or more positions, selected from positions 3, 49, 59, 186, 225, 227, 232, 235, 252, 260, 272, 279, 281 , 309 and 312, wherein position numbers are those of the parent hexosaminidase. Preferred substitutions in these positions include one or more of Q3F, Q3I, A49W, X59E, S186R, S225G, X227T, E232D, X235W, X252P, T260Q, Y272V, Y272P, S279D, Y281 P, G309E and K312E, for example two, three, four, five, six, seven, eight, nine, ten or more of these substitutions, where “X” indicates the amino acid residue present in the parent hexosaminidase.

[0069] Polypeptides having protease activity

[0070] The polypeptide having protease activity in the compositions of the invention comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 1 , such as at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95% or at least 96%, and comprises a substitution in position 209 of SEQ ID NO: 1 selected from the group consisting of A209K, A209R, A209Q, A209N, A209S and A209T.

[0071] Preferred subsitutions in position 209 of SEQ ID NO: 1 are selected from the group consisting of A209K, A209Q, A209N, A209S and A209T; more preferred substitutions are selected from the group consisting of A209K, A209Q, A209N and A209T; still more preferred substitutions are A209K and A209N; and most preferably the substitution is A209K.

[0072] In one preferred embodiment, the polypeptide having protease activity comprises at least one substitution selected from the group consisting of S3T, V4I, S97D, S99E, S101A, V102I, S154D, G157S, V199I, Q200L, Y203W and L256E, wherein numbering is based on SEQ ID NO: 1. Preferably, the polypeptide comprises two or more, such as three, four, five, six or more, substitutions selected from the group consisting of S3T, V4I, S97D, S99E, S101A, V102I, S154D, G157S, V199I, Q200L, Y203W, A209K and L256E. More preferably, the polypeptide of this embodiment comprises seven or more, such as eight, nine, ten, eleven or all twelve, substitutions selected from the group consisting of S3T, V4I, S97D, S99E, S101A, V102I, S154D, G157S, V199I, Q200L, Y203W and L256E.

[0073] An example of a preferred polypeptide having protease activity of this embodiment is thus one comprising the substitutions S3T, V4I, S97D, S99E, S101A, V102I, S154D, G157S, V199I, Q200L, Y203W, A209X and L256E, wherein X in A209X is K, R, Q, N, S or T, preferably wherein X is K or N, more preferably wherein X is K, and wherein numbering is based on SEQ ID NO: 1 .

[0074] In another preferred embodiment, the polypeptide having protease activity comprises at least one substitution selected from the group consisting of S9E, N42R, N74D, V199I, Q200L, Y203W, S253D, N255W and L256E, wherein numbering is based on SEQ ID NO: 1. Preferably, the polypeptide comprises two or more, such as three, four, five or more, substitutions selected from the group consisting of S9E, N42R, N74D, V199I, Q200L, Y203W, S253D, N255W and L256E, wherein numbering is based on SEQ ID NO: 1. More preferably, the polypeptide of this embodiment comprises six or more, such as seven, eight or all nine, substitutions selected from the group consisting of S9E, N42R, N74D, V199I, Q200L, Y203W, S253D, N255W and L256E, wherein numbering is based on SEQ ID NO: 1.

[0075] An example of a preferred polypeptide having protease activity of this embodiment is thus one comprising the substitutions S9E, N42R, N74D, V199I, Q200L, Y203W, A209X, S253D, N255W and L256E, wherein X in A209X is K, R, Q, N, S or T, preferably wherein X is K or N, more preferably wherein X is K, and wherein numbering is based on SEQ ID NO: 1.

[0076] In any of the embodiments outlined above or elsewhere herein, the polypeptide having protease activity may further comprise the substitution G95D. The polypeptide may, for example, comprise at least one substitution selected from the group consisting of S9E, N42R, N74D, V199I, Q200L, Y203W, S253D, N255W and L256E, such as two, three, four, five, six, seven, eight or all nine of said substitutions, together with the substitution G95D and a substitution in position 209 such as A209K. An example of a preferred polypeptide having protease activity of this embodiment is thus one comprising the substitutions S9E, N42R, N74D, G95D, V199I, Q200L, Y203W, A209X, S253D, N255W and L256E, wherein X in A209X is K, R, Q, N, S or T, preferably wherein X is K or N, more preferably wherein X is K, and wherein numbering is based on SEQ ID NO: 1.

[0077] Methods of Production

[0078] Expression and purification of the hexosaminidase and protease polypeptides of the invention may be performed by standard methods known in the art for production of recombinant enzymes and other recombinant proteins. For example, a hexosaminidase may suitably be produced e.g. as described in WO 2023 / 194204. Methods for production fo recombinant proteases are well-known; see, for example, WO 2021 / 239818.

[0079] Compositions

[0080] The present invention provides cleaning compositions comprising a polypeptide having hexosaminidase activity and a polypeptide having protease activity as disclosed elsewhere herein and at least one cleaning adjunct ingredient.

[0081] The cleaning compositions contain one or more cleaning adjunct ingredients selected from the group consisting of surfactants, builders, flocculating aid, chelating agents, dye transfer inhibitors, enzymes, enzyme stabilizers, enzyme inhibitors, catalytic materials, bleach activators, hydrogen peroxide, sources of hydrogen peroxide, preformed peracids, polymeric dispersing agents, clay soil removal / anti-redeposition agents, brighteners, suds suppressors, dyes, perfumes, structure elasticizing agents, fabric softeners, carriers, hydrotropes, builders and cobuilders, fabric hueing agents, anti-foaming agents, dispersants, processing aids, and / or pigments.

[0082] In the cleaning compositions of the invention, the cleaning adjunct ingredient will typically comprise at least one surfactant, builder or bleach component. The compositions will normally comprise at least one surfactant, preferably at least one anionic surfactant, along with other cleaning adjunct ingredients such as a builder, other enzymes, etc.

[0083] The cleaning adjunct ingredient may include one or more enzymes other than the hexosaminidase and protease disclosed herein. The one or more enzymes may e.g. be selected from the group consisting of amylases, lipases, cutinases, cellulases, endoglucanases, xyloglucanases, pectinases, pectin lyases, xanthanases, peroxidases, haloperoxygenases, catalases and mannanases. It may also be desirable to include a second protease having a different specificity than the protease disclosed herein in compositions of the invention, and / or a second hexosaminidase, typically a second protease. Specific enzymes suitable for the detergent compositions of the invention are described below.

[0084] The cleaning composition may be formulated in any suitable form, such as a bar, a homogenous tablet, a tablet having two or more layers, a pouch having one or more compartments, a regular or compact powder, a granule, a paste, a gel, or a regular, compact or concentrated liquid. The cleaning composition can thus e.g. be a liquid detergent or a powder or granular detergent, optionally in “concentrated” or "compact" form. It may also be in the form of a single unit dose composition.

[0085] The amount of hexosaminidase in the cleaning composition may vary depending on factors such as the degree of concentration or compactness of the composition and the desired enzyme concentration in the wash liquor. The hexosaminidase will normally be included in the cleaning composition in an amount of up to about 10,000 ppm, typically up to about 5000 ppm or up to about 2000 ppm. The hexosaminidase can e.g. be included in the cleaning composition at a level of from 1 ppm to 10,000 ppm, such as from 10 ppm to 5000 ppm, from 20 ppm to 2000 ppm, from 50 ppm to 1000 ppm, from 80 ppm to 600 ppm, or from 100 ppm to 500 ppm. The unit “ppm” in this context is intended to refer to mg / l for an enzyme added to a liquid composition (e.g. liquid, gel, etc.), or mg / kg for an enzyme added to a solid composition (e.g. powder, granulate, tablet, etc.).

[0086] The protease may be used in amount similar to the amounts of hexosaminidase disclosed above. The protease may thus be included in the cleaning composition in an amount of up to about 10,000 ppm, typically up to about 5000 ppm or up to about 2000 ppm, for example at a level of from 1 ppm to 10,000 ppm, such as from 10 ppm to 5000 ppm, from 20 ppm to 2000 ppm, from 50 ppm to 1000 ppm, from 80 ppm to 600 ppm, or from 100 ppm to 500 ppm. In some aspects, the detergent composition is a liquid or powder laundry detergent, suitable for e.g. washing at high temperature and / or pH, such as at or above 40°C and / or at or above pH 8. In some aspects, the detergent composition is a liquid or powder laundry detergent, suitable for e.g. washing at low temperature and / or pH, such as at or below 20°C and / or pH 6. The detergent may also be formulated as a unit dose detergent and / or compact detergent optionally with minimum or no water. The detergent may also be a dish wash detergent. The laundry and dish wash detergents may be phosphate-free.

[0087] Surfactants

[0088] A surfactant may be selected among nonionic, anionic and / or amphoteric surfactants as described above, preferably anionic or nonionic surfactants but also amphoteric surfactants may be used. In general, bleach-stable surfactants are preferred. Preferred anionic surfactants are sulphate surfactants and in particular alkyl ether sulphates, especially C-9-15 alcohol ethersulfates, C12-15 primary alcohol ethoxylate, C8-C16 ester sulphates and C10-C14 ester sulphates, such as mono dodecyl ester sulphates Non-limiting examples of anionic surfactants include sulfates and sulfonates, in particular, linear alkylbenzenesulfonates (LAS), isomers of LAS, branched alkylbenzenesulfonates (BABS), phenylalkanesulfonates, alpha-olefinsulfonates (AOS), olefin sulfonates, alkene sulfonates, alkane-2,3-diylbis(sulfates), hydroxyalkanesulfonates and disulfonates, alkyl sulfates (AS) such as sodium dodecyl sulfate (SDS), fatty alcohol sulfates (FAS), primary alcohol sulfates (PAS), alcohol ethersulfates (AES or AEOS or FES, also known as alcohol ethoxysulfates or fatty alcohol ether sulfates), secondary alkanesulfonates (SAS), paraffin sulfonates (PS), ester sulfonates, sulfonated fatty acid glycerol esters, alpha-sulfo fatty acid methyl esters (alpha-SFMe or SES) including methyl ester sulfonate (MES), alkyl- or alkenylsuccinic acid, dodecenyl / tetradecenyl succinic acid (DTSA), fatty acid derivatives of amino acids, diesters and monoesters of sulfo-succinic acid or salt of fatty acids (soap), and combinations thereof.

[0089] The anionic surfactants are preferably added to the detergent in the form of salts. Suitable cations in these salts are alkali metal ions, such as sodium, potassium and lithium and ammonium salts, for example (2-hydroxyethyl) ammonium, bis(2-hydroxyethyl) ammonium and tris(2- hydroxyethyl) ammonium salts. Non-limiting examples of nonionic surfactants include alcohol ethoxylates (AE or AEO), alcohol propoxylates, propoxylated fatty alcohols (PFA), alkoxylated fatty acid alkyl esters, such as ethoxylated and / or propoxylated fatty acid alkyl esters, alkylphenol ethoxylates (APE), nonylphenol ethoxylates (NPE), alkylpolyglycosides (APG), alkoxylated amines, fatty acid monoethanolamides (FAM), fatty acid diethanolamides (FADA), ethoxylated fatty acid monoethanolamides (EFAM), propoxylated fatty acid monoethanolamides (PFAM), polyhydroxyalkyl fatty acid amides, or N-acyl N-alkyl derivatives of glucosamine (glucamides, GA, or fatty acid glucamides, FAGA), as well as products available under the trade names SPAN and TWEEN, and combinations thereof. Commercially available nonionic surfactants include Plurafac™, Lutensol™ and Pluronic™ from BASF, Dehypon™ series from Cognis and Genapol™ series from Clariant.

[0090] Builders

[0091] A builder is preferably selected among phosphates, sodium citrate builders, sodium carbonate, sodium silicate, sodium aluminosilicate (zeolite). Suitable builders are alkali metal or ammonium phosphates, polyphosphates, phosphonates, polyphosphates, carbonates, bicarbonates, borates, citrates, and polycarboxylates. Citrate builders, e.g., citric acid and soluble salts thereof (particularly sodium salt), are polycarboxylate builders. Citrates can be used in combination with zeolite, silicates like the BRITESIL types, and / or layered silicate builders. The builder is preferably added in an amount of about 0-65% by weight, such as about 5% to about 50% by weight. In a laundry detergent, the level of builder is typically about 40-65% by weight, particularly about 50-65% by weight, particularly from 20% to 50% by weight. The builder and / or co-builder may particularly be a chelating agent that forms water-soluble complexes with Ca and Mg. Any builder and / or co-builder known in the art for use in cleaning detergents may be utilized. Non-limiting examples of builders include zeolites, diphosphates (pyrophosphates), triphosphates such as sodium triphosphate (STP or STPP), carbonates such as sodium carbonate, soluble silicates such as sodium metasilicate, layered silicates (e.g., SKS-6 from Hoechst), and (carboxymethyl)inulin (CMI), and combinations thereof. Further non-limiting examples of builders include citrate, chelators such as aminocarboxylates, aminopolycarboxylates and phosphonates, and alkyl- or alkenylsuccinic acid. Additional specific examples include 2,2’,2”-nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), iminodisuccinic acid (IDS), ethylenediamine-N,N’-disuccinic acid (EDDS), methylglycine-N,N- diacetic acid (MGDA), glutamic acid-N,N -di acetic acid (GLDA), 1-hydroxyethane-1 ,1-diphosphonic acid, N-(2-hydroxyethyl)iminodiacetic acid (EDG), aspartic acid-N-monoacetic acid (ASMA), aspartic acid-N,N-diacetic acid (ASDA), aspartic acid-N-monopropionic acid (ASMP), iminodisuccinic acid (IDA), N-(sulfomethyl)aspartic acid (SMAS), N-(2-sulfoethyl)-aspartic acid (SEAS), N- (sulfomethylglutamic acid (SMGL), N-(2-sulfoethyl)-glutamic acid (SEGL), N-methyliminodiacetic acid (MIDA), serine-N,N-diacetic acid (SEDA), isoserine-N,N-diacetic acid (ISDA), phenylalanine- N,N-diacetic acid (PHDA), anthranilic acid-N,N-diacetic acid (ANDA), sulfanilic acid-N,N -di acetic acid (SLDA) , taurine-N,N-diacetic acid (TLIDA) and N'-(2-hydroxyethyl)ethylenediamine-N,N,N’- triacetic acid (HEDTA), diethanolglycine (DEG), and combinations and salts thereof. Phosphonates suitable for use herein include 1-hydroxyethane-1 ,1-diphosphonic acid (HEDP), ethylenediaminetetrakis (methylenephosphonicacid) (EDTMPA), diethylenetriaminepentakis (methylenephosphonic acid) (DTMPA or DTPMPA or DTPMP), nitrilotris (methylenephosphonic acid) (ATMP or NTMP), 2-phosphonobutane-1 ,2,4-tricarboxylic acid (PBTC), hexamethylenediaminetetrakis (methylenephosphonic acid) (HDTMP). The composition may also contain 0-50% by weight, such as about 5% to about 30%, of a detergent co-builder. The detergent composition may include a co-builder alone, or in combination with a builder, for example a zeolite builder. Non-limiting examples of co-builders include homopolymers of polyacrylates or copolymers thereof, such as poly (acrylic acid) (PAA) or copoly (acrylic acid / maleic acid) (PAA / PMA) or polyaspartic acid. Further exemplary builders and / or co-builders are described in, e.g., WO 09 / 102854, US 5977053. In some aspects, the builder is a non-phosphorus based builder such as citric acid and / or methylglycine-N, N-diacetic acid (MGDA) and / or glutamic-N, N-diacetic acid (GLDA) and / or salts thereof. The liquid composition may also be phosphate free in that instance the preferred builders includes citrate and / or methylglycine-N, N-diacetic acid (MGDA) and / or glutamic- N, N-diacetic acid (GLDA) and I or salts thereof.

[0092] Bleach components

[0093] The cleaning composition may contain 0-30% by weight, such as about 1% to about 20%, of a bleaching system. Any bleaching system comprising components known in the art for use in cleaning detergents may be utilized. Suitable bleaching system components include sources of hydrogen peroxide; sources of peracids; and bleach catalysts or boosters.

[0094] Sources of hydrogen peroxide: Suitable sources of hydrogen peroxide are inorganic persalts, including alkali metal salts such as sodium percarbonate and sodium perborates (usually mono- or tetrahydrate), and hydrogen peroxide-urea (1 / 1).

[0095] Sources of peracids: Peracids may be (a) incorporated directly as preformed peracids or (b) formed in situ in the wash liquor from hydrogen peroxide and a bleach activator (perhydrolysis) or (c) formed in situ in the wash liquor from hydrogen peroxide and a perhydrolase and a suitable substrate for the latter, e.g., an ester. a) Suitable preformed peracids include, but are not limited to, peroxycarboxylic acids such as peroxybenzoic acid and its ring-substituted derivatives, peroxy-a-naphthoic acid, peroxyphthalic acid, peroxylauric acid, peroxystearic acid, e-phthalimidoperoxycaproic acid [phthalimidoperoxyhexanoic acid (PAP)], and o-carboxybenzamidoperoxycaproic acid; aliphatic and aromatic diperoxydicarboxylic acids such as diperoxydodecanedioic acid, diperoxyazelaic acid, diperoxysebacic acid, diperoxybrassylic acid, 2-decyldiperoxybutanedioic acid, and diperoxyphthalic, -isophthalic and -terephthalic acids; perimidic acids; peroxymonosulfuric acid; peroxydisulfuric acid; peroxyphosphoric acid; peroxysilicic acid; and mixtures of said compounds. It is understood that the peracids mentioned may in some cases be best added as suitable salts, such as alkali metal salts (e.g., Oxone®) or alkaline earth-metal salts. b) Suitable bleach activators include those belonging to the class of esters, amides, imides, nitriles or anhydrides and, where applicable, salts thereof. Suitable examples are tetraacetylethylenediamine (TAED), sodium 4-[(3,5,5-trimethylhexanoyl)oxy]benzene-1-sulfonate (ISONOBS), sodium 4-(dodecanoyloxy)benzene-1-sulfonate (LOBS), sodium 4- (decanoyloxy)benzene-l -sulfonate, 4-(decanoyloxy)benzoic acid (DOBA), sodium 4- (nonanoyloxy)benzene-l -sulfonate (NOBS), and / or those disclosed in WO98 / 17767. A particular family of bleach activators of interest was disclosed in EP624154 and particularly preferred in that family is acetyl triethyl citrate (ATC). ATC or a short chain triglyceride like triacetin has the advantage that they are environmentally friendly. Furthermore, acetyl triethyl citrate and triacetin have good hydrolytical stability in the product upon storage and are efficient bleach activators. Finally, ATC is multifunctional, as the citrate released in the perhydrolysis reaction may function as a builder.

[0096] Bleach catalysts and boosters: The bleaching system may also include a bleach catalyst or booster. Some non-limiting examples of bleach catalysts that may be used in the compositions of the present invention include manganese oxalate, manganese acetate, manganese-collagen, cobalt-amine catalysts and manganese triazacyclononane (MnTACN) catalysts; particularly preferred are complexes of manganese with 1 ,4,7-trimethyl-1 ,4,7-triazacyclononane (Me3-TACN) or 1 ,2,4,7-tetramethyl-1 ,4,7-triazacyclononane (Me4-TACN), in particular Me3-TACN, such as the dinuclear manganese complex [(Me3-TACN)Mn(O)3Mn(Me3-TACN)](PF6)2, and [2, 2', 2"- nitrilotris(ethane-1 ,2-diylazanylylidene-KN-methanylylidene)triphenolato-K3O]manganese(lll). The bleach catalysts may also be other metal compounds, such as iron or cobalt complexes. Other suitable bleach catalysts are acylhydrazone catalysts such as those disclosed in US 2014 / 0323381.

[0097] In some aspects, where a source of a peracid is included, an organic bleach catalyst or bleach booster may be used having one of the following formulae:

[0098] (iii) and mixtures thereof; wherein each R1 is independently a branched alkyl group containing from 9 to 24 carbons or linear alkyl group containing from 11 to 24 carbons, preferably each R1 is independently a branched alkyl group containing from 9 to 18 carbons or linear alkyl group containing from 11 to 18 carbons, more preferably each R1 is independently selected from the group consisting of 2-propyl heptyl, 2-butyloctyl, 2-pentylnonyl, 2-hexyldecyl, dodecyl, tetradecyl, hexadecyl, octadecyl, isononyl, isodecyl, isotridecyl and isopentadecyl.

[0099] Other exemplary bleaching systems are described, e.g. in WO 2007 / 087258, WO 2007 / 087244, WO 2007 / 087259, EP 1867708 (Vitamin K) and WO 2007 / 087242. Suitable photobleaches may for example be sulfonated zinc or aluminium phthalocyanines.

[0100] The choice of detergent components may include, for textile care, the consideration of the type of textile to be cleaned, the type and / or degree of soiling, the temperature at which cleaning is to take place, and the formulation of the detergent product. Although components mentioned below are categorized by general header according to a functionality, this is not to be construed as a limitation, as a component may comprise additional functionalities as will be appreciated by the skilled artisan, including the exemplary non-limiting components shown in below.

[0101] The detergent composition may contain 0-10% by weight, for example 0-5% by weight, such as about 0.5 to about 5%, or about 3% to about 5%, of a hydrotrope. Any hydrotrope known in the art for use in detergents may be utilized. Non-limiting examples of hydrotropes include sodium benzenesulfonate, sodium p-toluene sulfonate (STS), sodium xylene sulfonate (SXS), sodium cumene sulfonate (SCS), sodium cymene sulfonate, amine oxides, alcohols and polyglycolethers, sodium hydroxynaphthoate, sodium hydroxynaphthalene sulfonate, sodium ethylhexyl sulfate, and combinations thereof.

[0102] The detergent composition may contain 0-10% by weight, such as 0.5-5%, 2-5%, 0.5-2% or 0.2-1 % of a polymer. Any polymer known in the art for use in detergents may be utilized. The polymer may function as a co-builder as mentioned above, or may provide antiredeposition, fibre protection, soil release, dye transfer inhibition, grease cleaning and / or anti-foaming properties. Some polymers may have more than one of the above-mentioned properties and / or more than one of the below-mentioned motifs. Exemplary polymers include (carboxymethyl)cellulose (CMC), poly(vinyl alcohol) (PVA), poly(vinylpyrrolidone) (PVP), poly(ethyleneglycol) or poly(ethylene oxide) (PEG), ethoxylated poly(ethyleneimine), carboxymethyl inulin (CMI), and polycarboxylates such as PAA, PAA / PMA, poly-aspartic acid, and lauryl methacrylate / acrylic acid copolymers , hydrophobically modified CMC (HM-CMC) and silicones, copolymers of terephthalic acid and oligomeric glycols, copolymers of polyethylene terephthalate) and poly(oxyethene terephthalate) (PET-POET), PVP, poly(vinylimidazole) (PVI), poly(vinylpyridine- / V-oxide) (PVPO or PVPNO) and polyvinylpyrrolidone-vinylimidazole (PVPVI). Further exemplary polymers include sulfonated polycarboxylates, polyethylene oxide and polypropylene oxide (PEO-PPO) and diquaternium ethoxy sulfate. Other exemplary polymers are disclosed in, e.g., WO 2006 / 130575. Salts of the above- mentioned polymers are also contemplated.

[0103] Fabric hueing agents

[0104] The detergent composition of the present invention may also include fabric hueing agents such as dyes or pigments, which when formulated in detergent compositions can deposit onto a fabric when said fabric is contacted with a wash liquor comprising said detergent compositions and thus altering the tint of said fabric through absorption / reflection of visible light. Fluorescent whitening agents emit at least some visible light. In contrast, fabric huemg agents alter the tint of a surface as they absorb at least a portion of the visible light spectrum. Suitable fabric hueing agents include dyes and dye-clay conjugates, and may also include 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 falling into the Colour Index (C.l.) classifications of Direct Blue, Direct Red, Direct Violet, Acid Blue, Acid Red, Acid Violet, Basic Blue, Basic Violet and Basic Red, or mixtures thereof, for example as described in WO 2005 / 03274, WO 2005 / 03275, WO 2005 / 03276 and EP 1876226 (hereby incorporated by reference). The detergent composition preferably comprises from about 0.00003 wt% to about 0.2 wt%, from about 0.00008 wt% to about 0.05 wt%, or even from about 0.0001 wt% to about 0.04 wt% fabric hueing agent. The composition may comprise from 0.0001 wt% to 0.2 wt% fabric hueing agent, this may be especially preferred when the composition is in the form of a unit dose pouch. Suitable hueing agents are also disclosed in, e.g. WO 2007 / 087257 and WO 2007 / 087243.

[0105] Enzymes

[0106] The detergent composition may comprise one or more additional enzymes such as a lipase, cutinase, amylase, carbohydrase, cellulase, DNase, pectinase, mannanase, arabinase, galactanase, xylanase, licheninase, xyloglucanase, oxidase, e.g., a laccase, and / or peroxidase. The composition may also include one or more additional proteases.

[0107] In general, the properties of the selected enzyme(s) should be compatible with the selected detergent, ( / .e., pH-optimum, compatibility with other enzymatic and non-enzymatic ingredients, etc.), and the enzyme(s) should be present in effective amounts.

[0108] Cellulases

[0109] Suitable cellulases include those of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Suitable cellulases include cellulases from the genera Bacillus, Pseudomonas, Humicola, Fusarium, Thielavia, Acremonium, e.g., the fungal cellulases produced from Humicola insolens, Myceliophthora thermophila and Fusarium oxysporum disclosed in US 4,435,307, US 5,648,263, US 5,691 ,178, US 5,776,757 and WO 89 / 09259.

[0110] Especially suitable cellulases are the alkaline or neutral cellulases having colour care benefits. Examples of such cellulases are cellulases described in EP 0 495 257, EP 0 531 372, WO 96 / 11262, WO 96 / 29397, WO 98 / 08940. Other examples are cellulase variants such as those described in WO 94 / 07998, EP 0 531 315, US 5,457,046, US 5,686,593, US 5,763,254, WO 95 / 24471 , WO 98 / 12307 and WO 99 / 001544.

[0111] Other cellulases are endo-beta-1 , 4-glucanase enzyme having a sequence of at least 97% identity to the amino acid sequence of position 1 to position 773 of SEQ ID NO: 2 of WO 2002 / 099091 or a family 44 xyloglucanase, which a xyloglucanase enzyme having a sequence of at least 60% identity to positions 40-559 of SEQ ID NO: 2 of WO 2001 / 062903.

[0112] Commercially available cellulases include Celluzyme™, and Carezyme™ (Novozymes A / S) Carezyme Premium™ (Novozymes A / S), Celluclean™ (Novozymes A / S), Celluclean Classic™ (Novozymes A / S), Cellusoft™ (Novozymes A / S), Whitezyme™ (Novozymes A / S), Clazinase™, and Puradax HA™ (Genencor International Inc.), and KAC-500(B)™ (Kao Corporation).

[0113] Proteases

[0114] In addition to the proteases disclosed elsewhere herein, detergent compositions may include one or more additional proteases, e.g. a protease having a different specificity. Suitable proteases may be of any origin, but are preferably of bacterial or fungal origin, optionally in the form of protein engineered or chemically modified mutants. The protease may be an alkaline protease, such as a serine protease or a metalloprotease. A serine protease may for example be of the S1 family, such as trypsin, or the S8 family such as a subtilisin. A metalloprotease may for example be a thermolysin, e.g. from the M4 family, or another metalloprotease such as those from the M5, M7 or M35 families.

[0115] The term "subtilases" refers to a sub-group of serine proteases according to Siezen et al., Protein Eng. 4 (1991) 719-737 and Siezen et al., Protein Sci. 6 (1997) 501-523. Serine proteases are a subgroup of proteases characterized by having a serine in the active site, which forms a covalent adduct with the substrate. The subtilases may be divided into six subdivisions, the Subtilisin family, the Thermitase family, the Proteinase K family, the Lantibiotic peptidase family, the Kexin family and the Pyrolysin family.

[0116] Although proteases suitable for detergent use may be obtained from a variety of organisms, including fungi such as Aspergillus, detergent proteases have generally been obtained from bacteria and in particular from Bacillus. Examples of Bacillus species from which subtilases have been derived include Bacillus lentus, Bacillus alkalophilus, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus pumilus and Bacillus gibsonii. Particular subtilisins include subtilisin lentus, subtilisin Novo, subtilisin Carlsberg, subtilisin BPN’, subtilisin 309, subtilisin 147 and subtilisin 168 and e.g. protease PD138 (described in WO 93 / 18140). Other useful proteases are e.g. those described in WO 01 / 16285 and WO 02 / 16547.

[0117] Examples of trypsin-like proteases include the Fusarium protease described in WO 94 / 25583 and WO 2005 / 040372, and the chymotrypsin proteases derived from Cellumonas described in WO 2005 / 052161 and WO 2005 / 052146.

[0118] Examples of metalloproteases include the neutral metalloproteases described in WO 2007 / 044993 such as those derived from Bacillus amyloliquefaciens, as well as e.g. the metalloproteases described in WO 2015 / 158723 and WO 2016 / 075078. Examples of useful proteases are the protease variants described in WO 89 / 06279 WO 92 / 19729, WO 96 / 34946, WO 98 / 20115, WO 98 / 20116, WO 99 / 11768, WO 01 / 44452, WO 03 / 006602, WO 2004 / 003186, WO 2004 / 041979, WO 2007 / 006305, WO 2011 / 036263, WO 2014 / 207227, WO 2016 / 087617 and WO 2016 / 174234.

[0119] Suitable commercially available protease enzymes include those sold under the trade names Alcalase®, Duralase™, Durazym™, Relase®, Relase® Ultra, Savinase®, Savinase® Ultra, Primase™, Polarzyme®, Kannase®, Liquanase®, Liquanase® Ultra, Ovozyme®, Coronase®, Coronase® Ultra, Blaze®, Blaze Evity® 100T, Blaze Evity® 125T, Blaze Evity® 150T, Blaze Evity® 200T, Neutrase®, Everlase®, Esperase®, Progress® Uno, Progress® In, Progress® Key and Progress® Excel (Novozymes A / S), those sold under the tradename Maxatase™, Maxacai™, Maxapem®, Purafect® Ox, Purafect® OxP, Puramax®, FN2™, FN3™, FN4ex™, Excellase®, Excellenz™ P1000, Excellenz™ P1250, Eraser™, Preferenz® P100, Preferenz® P300, Preferenz® P400, Purafect Prime, Preferenz P110™, Effectenz P1000™, Purafect®, Effectenz P1050™, Purafect® Ox, Effectenz ™ P2000, Purafast™, Properase®, Opticlean™ and Optimase® (Danisco / DuPont / IFF), BLAP (sequence shown in Figure 29 of US 5352604) and variants hereof (Henkel AG), and KAP (Bacillus alkalophilus subtilisin) from Kao.

[0120] Lipases and Cutinases

[0121] Suitable lipases and cutinases include those of bacterial or fungal origin. Chemically modified or protein engineered mutant enzymes are included. Examples include lipase from Thermomyces, e.g. from T. lanuginosus (previously named Humicola lanuginosa) as described in EP258068 and EP305216, cutinase from Humicola, e.g. H. insolens (WO96 / 13580), lipase from strains of Pseudomonas (some of these now renamed to Burkholderia), e.g. P. alcaligenes or P. pseudoalcaligenes (EP218272), P. cepacia (EP331376), P. sp. strain SD705 (W095 / 06720 & W096 / 27002), P. wisconsinensis (WO96 / 12012), GDSL-type Streptomyces lipases (W010 / 065455), cutinase from Magnaporthe grisea (WO10 / 107560), cutinase from Pseudomonas mendocina (US5,389,536), lipase from Thermobifida fusca (W011 / 084412), Geobacillus stearothermophilus lipase (W011 / 084417), lipase from Bacillus subtilis (W011 / 084599), and lipase from Streptomyces griseus (WO11 / 150157) and S. pristinaespiralis (W012 / 137147).

[0122] Other examples are lipase variants such as those described in EP407225, WO92 / 05249, WO94 / 01541 , WO94 / 25578, WO95 / 14783, WO95 / 30744, WO95 / 35381 , WO95 / 22615, W096 / 00292, W097 / 04079, W097 / 07202, WO00 / 34450, WO00 / 60063, W001 / 92502, W007 / 87508 and WO09 / 109500.

[0123] Preferred commercial lipase products include Lipolase™, Lipex™; Lipolex™ and Lipoclean™ (Novozymes A / S), Lumafast (originally from Genencor) and Lipomax (originally from Gist-Brocades). Still other examples are lipases sometimes referred to as acyltransferases or perhydrolases, e.g. acyltransferases with homology to Candida antarctica lipase A (WO10 / 111143), acyltransferase from Mycobacterium smegmatis (WO05 / 56782), perhydrolases from the CE 7 family (WO09 / 67279), and variants of the M. smegmatis perhydrolase in particular the S54V variant used in the commercial product Gentle Power Bleach from Huntsman Textile Effects Pte Ltd (W010 / 100028).

[0124] Amylases

[0125] Suitable amylases which can be used together with the hexosaminidase and protease of the invention may be an alpha-amylase or a glucoamylase and may be of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Amylases include, for example, alpha-amylases obtained from Bacillus, e.g., a special strain of Bacillus licheniformis, described in more detail in GB 1 ,296,839.

[0126] Suitable amylases include amylases having SEQ ID NO: 2 in WO 95 / 10603 or variants having 90% sequence identity to SEQ ID NO: 1 thereof. Preferred variants are described in WO 94 / 02597, WO 94 / 18314, WO 97 / 43424 and SEQ ID NO: 4 of WO 99 / 019467, such as variants with substitutions in one or more of the following positions: 15, 23, 105, 106, 124, 128, 133, 154, 156, 178, 179, 181 , 188, 190, 197, 201 , 202, 207, 208, 209, 211 , 243, 264, 304, 305, 391 , 408, and 444.

[0127] Different suitable amylases include amylases having SEQ ID NO: 6 in WO 02 / 010355 or variants thereof having 90% sequence identity to SEQ ID NO: 6. Preferred variants of SEQ ID NO: 6 are those having a deletion in positions 181 and 182 and a substitution in position 193.

[0128] Other amylases which are suitable are hybrid alpha-amylase comprising residues 1-33 of the alpha-amylase obtained from B. amyloliquefaciens shown in SEQ ID NO: 6 of WO 2006 / 066594 and residues 36-483 of the B. licheniformis alpha-amylase shown in SEQ ID NO: 4 of WO 2006 / 066594 or variants having 90% sequence identity thereof. Preferred variants of this hybrid alpha-amylase are those having a substitution, a deletion or an insertion in one of more of the following positions: G48, T49, G107, H156, A181 , N190, M197, 1201 , A209 and Q264.

[0129] Other examples are amylase variants such as those described in WO 2011 / 098531 , WO 2013 / 001078 and WO 2013 / 001087.

[0130] Commercially available amylases include Duramyl™, Termamyl™, Fungamyl™, Stainzyme™, Stainzyme Plus™, Natalase™, Liquozyme X and BAN™ (from Novozymes A / S), and Rapidase™, Purastar™ / Effectenz™, Powerase and Preferenz S100 (from Genencor International Inc. / DuPont).

[0131] Peroxidases / Oxidases A peroxidase may be comprised by the enzyme classification EC 1.11.1.7, as set out by the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUBMB), or any fragment obtained therefrom, exhibiting peroxidase activity. Suitable peroxidases include those of plant, bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Examples of useful peroxidases include peroxidases from Coprinopsis, e.g., from C. cinerea (EP 179,486), and variants thereof as those described in WO 93 / 24618, WO 95 / 10602, and WO 98 / 15257. A peroxidase may also include a haloperoxidase enzyme, such as chloroperoxidase, bromoperoxidase and compounds exhibiting chloroperoxidase or bromoperoxidase activity. Haloperoxidases are classified according to their specificity for halide ions. Chloroperoxidases (E.C. 1.11.1.10) catalyze formation of hypochlorite from chloride ions. The haloperoxidase may be a chloroperoxidase. Preferably, the haloperoxidase is a vanadium haloperoxidase, i.e., a vanadate-containing haloperoxidase. In a preferred method the vanadate- containing haloperoxidase is combined with a source of chloride ion. Haloperoxidases have been isolated from many different fungi, in particular from the fungus group dematiaceous hyphomycetes, such as Caldariomyces, e.g., C. fumago, Alternaria, Curvularia, e.g., C. verruculosa and C. inaequalis, Drechslera, Ulocladium and Botrytis. Haloperoxidases have also been isolated from bacteria such as Pseudomonas, e.g., P. pyrrocinia and Streptomyces, e.g., S. aureofaciens. In a preferred aspect, the haloperoxidase is derivable from Curvularia sp., in particular Curvularia verruculosa or Curvularia inaequalis, such as C. inaequalis CBS 102.42 as described in WO 95 / 27046; or C. verruculosa CBS 147.63 or C. verruculosa CBS 444.70 as described in WO 97 / 04102; or from Drechslera hartlebii as described in WO 01 / 79459, Dendryphiella salina as described in WO 01 / 79458, Phaeotrichoconis crotalarie as described in WO 01 / 79461 , or Geniculosporium sp. as described in WO 01 / 79460.

[0132] An oxidase includes any laccase enzyme comprised by the enzyme classification EC 1 .10.3.2, or any fragment obtained therefrom exhibiting laccase activity, or a compound exhibiting a similar activity, such as a catechol oxidase (EC 1.10.3.1), an o-aminophenol oxidase (EC 1.10.3.4), or a bilirubin oxidase (EC 1.3.3.5).

[0133] Preferred laccase enzymes are enzymes of microbial origin. The enzymes may be obtained from plants, bacteria or fungi (including filamentous fungi and yeasts).

[0134] Suitable examples from fungi include a laccase derivable from a strain of Bacillus, Neurospora, e.g., N. crassa, Podospora, Botrytis, Collybia, Pomes, Lentinus, Pleurotus, Trametes, e.g., T. villosa and T. versicolor, Rhizoctonia, e.g., R. solani, Coprinopsis, e.g., C. cinerea, C. comatus, C. friesii, and C. plicatilis, Psathyrella, e.g., P. condelleana, Panaeolus, e.g., P. papilionaceus, Myceliophthora, e.g., M. thermophila, Schytalidium, e.g., S. thermophilum, Polyporus, e.g., P. pinsitus, Phlebia, e.g., P. radiata (WO 92 / 01046), or Coriolus, e.g., C. hirsutus (JP 2238885).

[0135] Suitable examples from bacteria include a laccase derivable from a strain of Bacillus. A laccase obtained from Coprinopsis or Myceliophthora is preferred; a laccase obtained from Coprinopsis cinerea, as disclosed in WO 97 / 08325; or from Myceliophthora thermophila, as disclosed in WO 95 / 33836.

[0136] DNases

[0137] The term “DNase” means a polypeptide having DNase (deoxyribonuclease) activity that catalyzes the hydrolytic cleavage of phosphodiester linkages in a DNA backbone, thus degrading DNA. DNase polypeptides have been found to be useful for deep cleaning of microbial biofilm that may be present on surfaces such as textiles or dishware or other hard surfaces, and which consists of a matrix of extracellular polymeric substance (EPS) composed of extracellular DNA, proteins, and polysaccharides.

[0138] The DNase polypeptide is typically a microbial enzyme, preferably of fungal or bacterial origin, or a genetically engineered variant of a microbial DNase.

[0139] Suitable bacterial DNases may, for example, be obtained from species of Bacillus and related genera (cf. Patel and Gupta, int. J. Syst. Evol. Microbiol. 2020; 70:406-438, who proposed six new Bacillaceae genera from species formerly classified as belonging to the genus Bacillus), e.g. from Bacillus, Cytobacillus, Metabacillus, Alkalihalobacillus, Rossellomorea or Mesobacillus. Examples of species from which DNases may be obtained include Bacillus licheniformis, Bacillus subtilis, Bacillus horikoshii, Cytobacillus horneckiae, Metabacillus indicus, Alkalihalobacillus algicola, Rossellomorea vietnamensis, Alkalihalobacillus hwajinpoensis, Metabacillus indicus, Mesobacillus campisalis, Bacillus idriensis, Bacillus algicola, Bacillus marisflavi and Bacillus luciferensis. Preferred bacterial DNases include those obtained from Metabacillus indicus (previously known as Bacillus cibi) and variants thereof.

[0140] DNases may also be obtained from a fungal species. Examples of preferred fungal DNases are those obtained from Aspergillus, for example from Aspergillus oryzae, from Trichoderma, for example from Trichoderma harzianum, from Vibressa, for example from Vibressea flavovirens, from Morchella, for example from Morchella costata, and from Rhizoctonia, for example from Rhizoctonia solani, as well as variants thereof. Preferred fungal DNases include those obtained from Aspergillus oryzae and variants thereof.

[0141] Suitable DNases, DNase variants, and use thereof in detergent compositions are disclosed, for example, in WO 2014 / 087011 , WO 2015 / 155350, WO 2015 / 155351 , WO 2017 / 060475, WO 2017 / 060493, WO 2017 / 060505, WO 2017 / 064269, WO 2018 / 011277, WO 2018 / 177203, WO 2018 / 177936, WO 2018 / 177938, WO 2019 / 081724, WO 2019 / 081721 , WO 2021 / 130167, WO 2022 / 194668 and WO 2022 / 194673. Dispersants

[0142] The detergent compositions of the present invention can also contain dispersants. Powdered detergents may comprise dispersants. Suitable water-soluble organic materials include the homo- or co-polymeric acids or their salts, in which the polycarboxylic acid comprises at least two carboxyl radicals separated from each other by not more than two carbon atoms. Suitable dispersants are for example described in Powdered Detergents, Surfactant science series volume 71 , Marcel Dekker, Inc.

[0143] Dye Transfer Inhibiting Agents

[0144] The cleaning compositions of the present invention may also include one or more dye transfer inhibiting agents. Suitable polymeric dye transfer inhibiting agents include, but are not limited to, polyvinylpyrrolidone polymers, polyamine / V-oxide polymers, copolymers of / V- vinylpyrrolidone and / V-vinylimidazole, polyvinyloxazolidones and polyvinylimidazoles or mixtures thereof. When present in a subject composition, the dye transfer inhibiting agents may be present at levels from about 0.0001 % to about 10%, from about 0.01 % to about 5% or even from about 0.1 % to about 3% by weight of the composition.

[0145] Fluorescent whitening agents

[0146] The detergent composition may preferably also contain additional components that may tint articles being cleaned, such as fluorescent whitening agent or optical brighteners. Where present the brightener is preferably at a level of about 0.01% to about 0.5%. Any fluorescent whitening agent suitable for use in a laundry detergent composition may be used in the composition of the present invention. The most commonly used fluorescent whitening agents are those belonging to the classes of diaminostilbene-sulfonic acid derivatives, diarylpyrazoline derivatives and bisphenyl-distyryl derivatives. Examples of the diaminostilbene-sulfonic acid derivative type of fluorescent whitening agents include the sodium salts of: 4,4'-bis-(2- diethanolamino-4-anilino-s-triazin-6-ylamino) stilbene-2,2'-disulfonate, 4,4'-bis-(2,4-dianilino-s- triazin-6-ylamino) stilbene-2.2'-disulfonate, 4,4'-bis-(2-anilino-4-( / V-methyl- / \ / -2-hydroxy- ethylamino)-s-triazin-6-ylamino) stilbene-2,2'-disulfonate, 4,4'-bis-(4-phenyl-1 ,2,3-triazol-2- yl)stilbene-2,2'-disulfonate and sodium 5-(2 / 7-naphtho[1 ,2-d][1 ,2,3]triazol-2-yl)-2-[(E)-2- phenylvinyl]benzenesulfonate. Preferred fluorescent whitening agents are Tinopal DMS and Tinopal CBS available from Ciba-Geigy AG, Basel, Switzerland. Tinopal DMS is the disodium salt of 4,4'-bis-(2-morpholino-4-anilino-s-triazin-6-ylamino) stilbene-2,2'-disulfonate. Tinopal CBS is the disodium salt of 2,2'-bis-(phenyl-styryl)-disulfonate. Also preferred are fluorescent whitening agents is the commercially available Parawhite KX, supplied by Paramount Minerals and Chemicals, Mumbai, India. Tinopal CBS-X is a 4.4'-bis-(sulfostyryl)-biphenyl disodium salt also known as Disodium Distyrylbiphenyl Disulfonate. Other fluorescers suitable for use in the invention include the 1 -3-diaryl pyrazolines and the 7-alkylaminocoumarins.

[0147] Suitable fluorescent brightener levels include lower levels of from about 0.01 , from 0.05, from about 0.1 or even from about 0.2 wt % to upper levels of 0.5 or even 0.75 wt%.

[0148] Soil release polymers

[0149] The detergent compositions may also include one or more soil release polymers which aid the removal of soils from fabrics such as cotton and polyester based fabrics, the removal of hydrophobic soils from polyester-based fabrics. The soil release polymers may for example be nonionic or anionic terephthalte based polymers, polyvinyl caprolactam and related copolymers, vinyl graft copolymers, polyester polyamides see for example Chapter 7 in Powdered Detergents, Surfactant science series volume 71 , Marcel Dekker, Inc. Another type of soil release polymers is amphiphilic alkoxylated grease cleaning polymers comprising a core structure and a plurality of alkoxylate groups attached to that core structure. The core structure may comprise a polyalkylenimine structure or a polyalkanolamine structure as described in detail in WO 2009 / 087523 (hereby incorporated by reference). Furthermore, random graft co-polymers are suitable soil release polymers. Suitable graft co-polymers are described in more detail in WO 2007 / 138054, WO 2006 / 108856 and WO 2006 / 113314 (hereby incorporated by reference). Other soil release polymers are substituted polysaccharide structures especially substituted cellulosic structures such as modified cellulose deriviatives such as those described in EP 1867808 or WO 2003 / 040279 (both are hereby incorporated by reference). Suitable cellulosic polymers include cellulose, cellulose ethers, cellulose esters, cellulose amides and mixtures thereof. Suitable cellulosic polymers include anionically modified cellulose, nonionically modified cellulose, cationically modified cellulose, zwitterionically modified cellulose, and mixtures thereof. Suitable cellulosic polymers include methyl cellulose, carboxy methyl cellulose, ethyl cellulose, hydroxyl ethyl cellulose, hydroxyl propyl methyl cellulose, ester carboxy methyl cellulose, and mixtures thereof.

[0150] Anti-redeposition agents

[0151] The detergent compositions of the present invention may also include one or more antiredeposition agents such as carboxymethylcellulose (CMC), polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyoxyethylene and / or polyethyleneglycol (PEG), homopolymers of acrylic acid, copolymers of acrylic acid and maleic acid, and ethoxylated polyethyleneimines. The cellulose-based polymers described under soil release polymers above may also function as antiredeposition agents. Rheology Modifiers

[0152] The detergent compositions of the present invention may also include one or more rheology modifiers, structurants or thickeners, as distinct from viscosity reducing agents. The rheology modifiers are selected from the group consisting of non-polymeric crystalline, hydroxyfunctional materials, polymeric rheology modifiers which impart shear thinning characteristics to the aqueous liquid matrix of a liquid detergent composition. The rheology and viscosity of the detergent can be modified and adjusted by methods known in the art, for example as shown in EP 2169040.

[0153] Other suitable adjunct materials include, but are not limited to, anti-shrink agents, antiwrinkling agents, bactericides, binders, carriers, dyes, enzyme stabilizers, fabric softeners, fillers, foam regulators, hydrotropes, perfumes, pigments, sod suppressors, solvents, and structurants for liquid detergents and / or structure elasticizing agents.

[0154] Other materials

[0155] Any detergent components known in the art for use in the cleaning composition of the invention may also be utilized. Other optional detergent components include anti-corrosion agents, anti-shrink agents, anti-soil redeposition agents, anti-wrinkling agents, bactericides, binders, corrosion inhibitors, disintegrants / disintegration agents, dyes, enzyme stabilizers (including boric acid, borates, CMC, and / or polyols such as propylene glycol), fabric conditioners including clays, fillers / processing aids, fluorescent whitening agents / optical brighteners, foam boosters, foam (suds) regulators, perfumes, soil-suspending agents, softeners, suds suppressors, tarnish inhibitors, and wicking agents, either alone or in combination. Any ingredient known in the art for use in detergents may be utilized. The choice of such ingredients is well within the skill of the artisan.

[0156] Formulation of detergent products

[0157] The detergent composition may be in any convenient form, e.g., a bar, a homogenous tablet, a tablet having two or more layers, a regular or compact powder, a granule, a paste, a gel, or a regular, compact or concentrated liquid. Other detergent formulation forms include single unit dose forms such as layered forms and pouches.

[0158] Pouches can be configured as single or multicompartments. They can be of any form, shape and material which is suitable for hold the composition, e.g. without allowing release of the composition from the pouch prior to water contact. The pouch is made from water soluble film which encloses an inner volume, which can be divided into compartments. Preferred films are polymeric materials, preferably polymers which are formed into a film or sheet. Preferred polymers, copolymers or derivates thereof are selected from polyacrylates and water-soluble acrylate copolymers, methyl cellulose, carboxy methyl cellulose, sodium dextrin, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, maltodextrin, polymethacrylates, most preferably polyvinyl alcohol copolymers and hydroxypropyl methyl cellulose (HPMC). Preferably the level of polymer in a film such as is at least about 60%. Preferred average molecular weight will typically be about 20,000 to about 150,000. Films can also be of blend compositions comprising hydrolytically degradable and water-soluble polymer blends such as polylactide and polyvinyl alcohol plus plasticisers such as glycerol, ethylene glycerol, propylene glycol, sorbitol and mixtures thereof. The pouches can comprise a solid laundry cleaning composition or part components and / or a liquid cleaning composition or part components separated by the water-soluble film. Compartments for liquid components can be different in composition than compartments containing solids; see e.g. US 2009 / 0011970 A1.

[0159] Detergent ingredients can be separated physically from each other by compartments in water dissolvable pouches or in different layers of tablets, thereby avoiding negative storage interaction between components. Different dissolution profiles of each of the compartments can also give rise to delayed dissolution of selected components in the wash solution.

[0160] A liquid or gel detergent which is not unit dosed may be aqueous, typically containing at least 20% by weight and up to 95% water, such as up to about 70% water, up to about 65% water, up to about 55% water, up to about 45% water, or up to about 35% water. Concentrated liquid detergents may have lower water contents, for example not more than about 30% or not more than about 20%, e.g. in the range of about 1% to about 20%, such as from about 2% to about 15%. Other types of liquids, including without limitation, alkanols, amines, diols, ethers and polyols may be included in an aqueous liquid or gel. An aqueous liquid or gel detergent may contain from 0-30% organic solvent. A liquid or gel detergent may alternatively be non-aqueous.

[0161] Liquid detergent compositions may be formulated to have a moderate pH of e.g. from about 6 to about 10, such as about pH 7, about pH 8 or about pH 9, or they may be formulated to have a higher pH of e.g. from about 10 to about 12, such as about pH 10, about pH 11 or about pH 12.

[0162] Unless indicated otherwise, the term “liquid” as used herein should be understood to encompass any kind of liquid detergent composition, for example concentrated liquids, gels, or the liquid or gel part of e.g. a pouch with one or more compartments.

[0163] Detergent enzymes may be included in a detergent composition by adding separate additives containing one or more enzymes, or by adding a combined additive comprising these enzymes. A detergent additive, i.e., a separate additive or a combined additive, can be formulated, for example, as a granulate, liquid, slurry, etc. Preferred detergent additive formulations are granulates, non-dusting granulates, liquids, stabilized liquids and slurries.

[0164] Non-dusting granulates may be produced, e.g. as disclosed in US 4,106,991 and 4,661,452 and may optionally be coated by methods known in the art. Examples of waxy coating materials are poly (ethylene oxide) products (polyethyleneglycol, PEG) with mean molar weights of 1000 to 20000; ethoxylated nonylphenols having from 16 to 50 ethylene oxide units; ethoxylated fatty alcohols in which the alcohol contains from 12 to 20 carbon atoms and in which there are 15 to 80 ethylene oxide units; fatty alcohols; fatty acids; and mono- and di- and triglycerides of fatty acids. Examples of film-forming coating materials suitable for application by fluid bed techniques are given in GB 1483591. Liquid enzyme preparations may, for instance, be stabilized by adding a polyol such as propylene glycol, a sugar or sugar alcohol, lactic acid or boric acid according to established methods. Protected enzymes may be prepared according to the method disclosed in EP 238216.

[0165] Granular detergent formulations

[0166] Enzymes in the form of granules, comprising an enzyme-containing core and optionally one or more coatings, are commonly used in granular (powder) detergents. Various methods for preparing the core are well-known in the art and include, for example, a) spray drying of a liquid enzyme-containing solution, b) production of layered products with an enzyme coated as a layer around a pre-formed inert core particle, e.g. using a fluid bed apparatus, c) absorbing an enzyme onto and / or into the surface of a pre-formed core, d) extrusion of an enzyme-containing paste, e) suspending an enzyme-containing powder in molten wax and atomization to result in prilled products, f) mixer granulation by adding an enzyme-containing liquid to a dry powder composition of granulation components, g) size reduction of enzyme-containing cores by milling or crushing of larger particles, pellets, etc., and h) fluid bed granulation. The enzyme-containing cores may be dried, e.g. using a fluid bed drier or other known methods for drying granules in the feed or enzyme industry, to result in a water content of typically 0.1 -10% w / w water.

[0167] The enzyme-containing cores are optionally provided with a coating to improve storage stability and / or to reduce dust formation. One type of coating that is often used for enzyme granulates for detergents is a salt coating, typically an inorganic salt coating, which may e.g. be applied as a solution of the salt using a fluid bed. Other coating materials that may be used are, for example, polyethylene glycol (PEG), methyl hydroxy-propyl cellulose (MHPC) and polyvinyl alcohol (PVA). The granules may contain more than one coating, for example a salt coating followed by an additional coating of a material such as PEG, MHPC or PVA.

[0168] For further information on enzyme granules and production thereof, see WO 2013 / 007594 as well as e.g. WO 2009 / 092699, EP 1705241 , EP 1382668, WO 2007 / 001262, US 6,472,364, WO 2004 / 074419 and WO 2009 / 102854.

[0169] Formulation of enzyme in co-qranule

[0170] The hexosaminidase and protease may optionally be formulated as a co-granule that combines the two enzymes, optionally together with one or more additional enzymes. However, since proteases can tend to reduce the stability of other enzymes, it may be preferable to have two or more different types of granules in the compositions. This can, for example, be granules that contain the protease as the sole enzyme and other granules that comprise the hexosaminidase and one or more other (non-protease) enzymes, where the latter may be in the form of a co-granule comprising the hexosaminidase and one or more other (non-protease) enzymes.

[0171] An advantage of co-granules is that each enzyme will then be present in more granules, securing a more uniform distribution of enzymes in the detergent. This also reduces the physical segregation of different enzymes due to different particle sizes. Methods for producing multi-enzyme co-granulate for the detergent industry is disclosed in the IP.com disclosure IPCOM000200739D.

[0172] Another example of formulation of enzymes using co-granulates are disclosed in WO 2013 / 188331 , which relates to a detergent composition comprising (a) a multi-enzyme co-granule; (b) less than 10 wt zeolite (anhydrous basis); and (c) less than 10 wt phosphate salt (anhydrous basis), wherein said enzyme co-granule comprises from 10 to 98 wt% moisture sink components and the composition additionally comprises from 20 to 80 wt% detergent moisture sink components. WO 2013 / 188331 also relates to a method of treating and / or cleaning a surface, preferably a fabric surface comprising the steps of (i) contacting said surface with the detergent composition as claimed and described herein aqueous wash liquor, (ii) rinsing and / or drying the surface.

[0173] Liquid Formulations

[0174] The present invention also relates to liquid compositions comprising a hexosaminidase and protease of the invention. The composition may comprise an enzyme stabilizer (examples of which include polyols such as propylene glycol or glycerol, sugar or sugar alcohol, lactic acid, reversible protease inhibitor, boric acid, or a boric acid derivative, e.g., an aromatic borate ester, or a phenyl boronic acid derivative such as 4-formylphenyl boronic acid).

[0175] In some embodiments, fillers or carrier materials are included to increase the volume of such compositions. Suitable filler or carrier materials include, but are not limited to, various salts of sulfate, carbonate and silicate as well as talc, clay and the like. Suitable filler or carrier materials for liquid compositions include, but are not limited to, water or low molecular weight primary and secondary alcohols including polyols and diols. Examples of such alcohols include, but are not limited to, methanol, ethanol, propanol and isopropanol. In some embodiments, the compositions contain from about 5% to about 90% of such materials.

[0176] In an aspect, the liquid formulation comprises 20-80% w / w of polyol. In one embodiment, the liquid formulation comprises 0.001-2% w / w preservative.

[0177] In another embodiment, the invention relates to liquid formulations comprising:

[0178] (A) 0.001-25% w / w, such as 0.001-5%, of a hexosaminidase disclosed herein, and optionally 0.001-25% w / w, such as 0.001-5%, of a protease disclosed herein;

[0179] (B) 20-80% w / w of polyol;

[0180] (C) optionally 0.001-2% w / w preservative; and

[0181] (D) water. In another embodiment, the invention relates to liquid formulations comprising:

[0182] (A) 0.001-25% w / w, such as 0.001-5%, of a hexosaminidase disclosed herein, and optionally 0.001-25% w / w, such as 0.001-5%, of a protease disclosed herein;

[0183] (B) 0.001-2% w / w preservative;

[0184] (C) optionally 20-80% w / w of polyol; and

[0185] (D) water.

[0186] In another embodiment, the liquid formulation comprises one or more formulating agents, such as a formulating agent selected from the group consisting of polyol, sodium chloride, sodium benzoate, potassium sorbate, sodium sulfate, potassium sulfate, magnesium sulfate, sodium thiosulfate, calcium carbonate, sodium citrate, dextrin, glucose, sucrose, sorbitol, lactose, starch, PVA, acetate and phosphate, preferably selected from the group consisting of sodium sulfate, dextrin, cellulose, sodium thiosulfate, kaolin and calcium carbonate. In one embodiment, the polyols is selected from the group consisting of glycerol, sorbitol, propylene glycol (MPG), ethylene glycol, diethylene glycol, triethylene glycol, 1 ,2-propylene glycol or 1 ,3-propylene glycol, dipropylene glycol, polyethylene glycol (PEG) having an average molecular weight below about 600 and polypropylene glycol (PPG) having an average molecular weight below about 600, more preferably selected from the group consisting of glycerol, sorbitol and propylene glycol (MPG) or any combination thereof.

[0187] In another embodiment, the liquid formulation comprises 20-80% polyol ( / .e., total amount of polyol), e.g., 25-75% polyol, 30-70% polyol, 35-65% polyol, or 40-60% polyol. In one embodiment, the liquid formulation comprises 20-80% polyol, e.g., 25-75% polyol, 30-70% polyol, 35-65% polyol, or 40-60% polyol, wherein the polyol is selected from the group consisting of glycerol, sorbitol, propylene glycol (MPG), ethylene glycol, diethylene glycol, triethylene glycol, 1 ,2-propylene glycol or 1 ,3-propylene glycol, dipropylene glycol, polyethylene glycol (PEG) having an average molecular weight below about 600 and polypropylene glycol (PPG) having an average molecular weight below about 600. In one embodiment, the liquid formulation comprises 20-80% polyol ( / .e., total amount of polyol), e.g., 25-75% polyol, 30-70% polyol, 35-65% polyol, or 40-60% polyol, wherein the polyol is selected from the group consisting of glycerol, sorbitol and propylene glycol (MPG).

[0188] In another embodiment, the preservative is selected from the group consisting of sodium sorbate, potassium sorbate, sodium benzoate and potassium benzoate or any combination thereof. In one embodiment, the liquid formulation comprises 0.02-1.5% w / w preservative, e.g., 0.05-1% w / w preservative or 0.1-0.5% w / w preservative. In one embodiment, the liquid formulation comprises 0.001-2% w / w preservative ( / .e., total amount of preservative), e.g., 0.02- 1.5% w / w preservative, 0.05-1% w / w preservative, or 0.1-0.5% w / w preservative, wherein the preservative is selected from the group consisting of sodium sorbate, potassium sorbate, sodium benzoate and potassium benzoate or any combination thereof. In another embodiment, the liquid formulation further comprises one or more additional enzymes, e.g. as described above.

[0189] Use of compositions of the invention

[0190] The compositions of the invention comprising a hexosaminidase and a protease are suitable for use in a cleaning process, for example for laundry or hard surface cleaning, in particular for laundry.

[0191] Thus, one aspect of the invention relates a method for laundering an item, wherein the item is a textile, the method comprising: a) exposing the item to a wash liquor comprising a polypeptide having hexosaminidase activity and a polypeptide having protease activity defined elsewhere herein; b) completing at least one wash cycle; and optionally c) rinsing the item.

[0192] In a related embodiment, the invention relates to a method for laundering an item, wherein the item is a textile, the method comprising: a) exposing the item to a wash liquor comprising a cleaning composition as defined elsewhere herein comprising a polypeptide having hexosaminidase activity and a polypeptide having protease activity; b) completing at least one wash cycle; and optionally c) rinsing the item.

[0193] The cleaning compositions of the invention may further be advantageous for preventing, reducing and / or removing a biofilm from a hard surface such as the interior surfaces of a washing machine or dishwasher. It is known that microbial biofilm can accumulate on interior surfaces of e.g. washing machines or dishwashers that are regularly subject to water or moisture as well as heat, even where these surfaces are also subject to the action of detergents. This can lead to development of odor, which can remain after use of the machine and which in turn can be a source of malodor. This for example result in odor being transferred from biofilm on the interior surfaces of a washing machine to textiles in a subsequent wash load.

[0194] It is thus contemplated that use of the cleaning compositions of the invention comprising a hexosaminidase and a protease can help prevent, reduce and / or remove biofilm from the interior surfaces of a washing machine or dishwasher. The surfaces in question include any type of surface that is regularly subjected to heat and moisture, for example surfaces made of steel or other metal, plastic, rubber, glass, etc. Examples of such surfaces include soap intake units, walls, windows, baskets, racks, nozzles, pumps, sump, filters, pipelines, tubes, joints, seals, gaskets, fittings, impellers, drums, drains, and coin traps inlets and outlets. The polypeptide having hexosaminidase activity for use in these methods herein is preferably a hexosaminidase polypeptide as disclosed elsewhere herein. Similarly, the polypeptide having protease activity is also preferably a protease polypeptide as defined elsewhere herein.

[0195] The pH of the liquid wash liquor solution is typically in the range about 5.5 to about 10, more typically in the range of about 7 to about 9, such as in the range of about 7 to about 8.5 or about 7 to about 8.

[0196] The wash liquor may have a temperature in the range of 5°C to 95°C, or in the range of 10°C to 80°C, in the range of 10°C to 70°C, in the range of 10°C to 60°C, in the range of 10°C to 50°C, in the range of 15°C to 40°C or in the range of 20°C to 30°C.

[0197] The concentration of the hexosaminidase in the wash liquor is typically in the range of from 0.0001 mg / l to 10 mg / l enzyme protein, from 0.0002 mg / l to 10 mg / l, from 0.001 mg / l to 10 mg / l, from 0.002 mg / l to 10 mg / l, from 0.01 mg / l to 10 mg / l, from 0.02 mg / l to 10 mg / l, from 0.1 mg / l to 10 mg / l, from 0.2 mg / l to 10 mg / l, or from 0.2 mg / l to 5 mg / l. The concentration of a protease in the wash liquor will typically be in the same ranges as for the hexosaminidase.

[0198] The invention is further defined by the following numbered paragraphs:

[0199] 1. A cleaning composition comprising a polypeptide having hexosaminidase activity, a polypeptide having protease activity and a cleaning adjunct ingredient, wherein the polypeptide having protease activity comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 1 and comprises a substitution in position 209 of SEQ ID NO: 1 selected from the group consisting of A209K, A209R, A209Q, A209N, A209S and A209T.

[0200] 2. The cleaning composition of paragraph 1 , wherein the polypeptide having protease activity comprises a substitution in position 209 of SEQ ID NO: 1 selected from the group consisting of A209K, A209Q, A209N, A209S and A209T, preferably selected from the group consisting of A209K, A209Q, A209N and A209T, more preferably selected from the group consisting of A209K and A209N, most preferably A209K.

[0201] 3. The cleaning composition of paragraph 1 or 2 wherein the polypeptide having hexosaminidase activity has beta-1 , 6-N-acetylglucosaminidase activity.

[0202] 4. The cleaning composition of any of the preceding paragraphs, wherein the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 70% sequence identity to 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.

[0203] 5. The cleaning composition of paragraph 4, wherein the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94% or at least 95% sequence identity to 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.

[0204] 6. The cleaning composition of paragraph 5, wherein the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 2 and comprises a substitution at one or more positions selected from positions 3, 15, 49, 59, 163, 186, 225, 227, 232, 235, 252, 260, 272, 279, 281 , 308, 309 and 312 of SEQ ID NO: 2.

[0205] 7. The cleaning composition of paragraph 6, wherein the polypeptide having hexosaminidase activity comprises comprises a substitution at two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen or all positions selected from positions 3, 15, 49, 59, 163, 186, 225, 227, 232, 235, 252, 260, 272, 279, 281 , 308, 309 and 312 of SEQ ID NO: 2.

[0206] 8. The cleaning composition of paragraph 6 or 7, wherein the polypeptide having hexosaminidase activity comprises at least one substitution selected from the group consisting of Q3F, Q3I, H15Y, A49W, N59E, S163P, S186R, S225G, N227T, E232D, G235W, N252P, N260Q, H272V, H272P, S279D, Y281 P, K308Q, K308E, K309E, K312Q and K312E, wherein numbering is based on SEQ ID NO: 2.

[0207] 9. The cleaning composition of paragraph 8, wherein the polypeptide having hexosaminidase activity comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen or all eighteen substitutions selected from the group consisting of Q3I, H15Y, A49W, N59E, S163P, S186R, S225G, N227T, E232D, G235W, N252P, N260Q, H272V, S279D, Y281 P, K308Q, K309E and K312Q, wherein numbering is based on SEQ ID NO: 2.

[0208] 10. The cleaning composition of paragraph 9, wherein the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 7. The cleaning composition of paragraph 4, wherein the polypeptide having hexosaminidase activity comprises: a substitution at one or more positions, preferably two, three or four positions, corresponding to positions 163, 227, 252 and 309 of SEQ ID NO: 2; and a substitution at one or more positions corresponding to positions 111 , 120, 106, 124, 127, 150, 170, 171 , 178, 199, 208, 254, 255 and 278 of SEQ ID NO: 2; wherein the polypeptide at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% sequence identity to SEQ ID NO: 2. The cleaning composition of paragraph 11 , wherein the polypeptide having hexosaminidase activity comprises two, three or four substitutions selected from the group consisting of S163P, N227T, N252P and K309E, wherein numbering is based on SEQ ID NO: 2; and further comprises at least one substitution selected from the group consisting of D111 R, T120V, V106M, V106N, Y124I, Y124K, Y124L, R127P, E150N, L170G, D171 F, D171 H, Q178K, S199W, S208W, F254I, T255D and I278V, wherein numbering is based on SEQ ID NO: 2. The cleaning composition of paragraph 11 or 12, wherein the polypeptide having hexosaminidase activity further comprises at least one substitution selected from the group consisting of H15Y, S186R, S225G, E232D, G235W, N260Q, H272V, S279D, Y281 P, K308Q and K312Q, wherein numbering is based on SEQ ID NO: 2, for example 2, 3, 4, 5, 6, 7, 8, 9, 10 or all of said substitutions; and wherein the polypeptide optionally further comprises one or more substitutions selected from the group consisting of Q3I, A49W and N59E, preferably 2 or all 3 of said substitutions; or one or more substitutions selected from the group consisting of Q3F, V140I, Q215K and N267T, preferably 2, 3 or all 4 of said substitutions; wherein numbering is based on SEQ ID NO: 2. The cleaning composition of any of paragraphs 11-13, wherein the polypeptide having hexosaminidase activity comprises at least one substitution selected from the group consisting of D111 R, T120V, V106M / N, E150N, D171 F and I278V, for example at least two of said substitutions; preferably wherein the polypeptide comprises at least one substitution selected from the group consisting of D111 R, T120V and E150N; wherein numbering is based on SEQ ID NO: 2. 15. The cleaning composition of any of paragraphs 1-5, wherein the polypeptide having hexosaminidase activity is a variant of a parent hexosaminidase selected from SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, wherein the variant comprises, compared to the parent hexosaminidase, a substitution at one or more positions, for example two, three, four, five, six, seven, eight, nine, ten or more positions, selected from positions 3, 49, 59, 186, 225, 227, 232, 235, 252, 260, 272, 279, 281 , 309 and 312, wherein position numbers are those of the parent hexosaminidase.

[0209] 16. The cleaning composition of paragraph 15, wherein the hexosaminidase variant comprises one or more substitutions selected from the group consisting of Q3F, Q3I, A49W, X59E, S186R, S225G, X227T, E232D, X235W, X252P, T260Q, Y272V, Y272P, S279D, Y281 P, G309E and K312E, for example two, three, four, five, six, seven, eight, nine, ten or more of said substitutions, where “X” indicates the amino acid residue present in the parent hexosaminidase.

[0210] 17. The cleaning composition of any of the preceding paragraphs, wherein the polypeptide having protease activity has at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95% or at least 96% sequence identity to SEQ ID NO: 1.

[0211] 18. The cleaning composition of any of paragraphs 1-17, wherein the polypeptide having protease activity comprises at least one substitution compared to SEQ ID NO: 1 selected from the group consisting of S3T, V4I, S97D, S99E, S101A, V102I, S154D, G157S, V199I, Q200L, Y203W and L256E, wherein numbering is based on SEQ ID NO: 1.

[0212] 19. The cleaning composition of paragraph 18, wherein the polypeptide having protease activity comprises two, three, four, five, six, seven, eight, nine, ten, eleven or all twelve substitutions selected from the group consisting of S3T, V4I, S97D, S99E, S101A, V102I, S154D, G157S, V199I, Q200L, Y203W and L256E.

[0213] 20. The cleaning composition of paragraph 19, wherein the polypeptide having protease activity comprises the substitutions S3T, V4I, S97D, S99E, S101A, V102I, S154D, G157S, V199I, Q200L, Y203W, A209X and L256E, wherein X in A209X is K, R, Q, N, S or T, preferably wherein X is K or N, more preferably wherein X is K, and wherein numbering is based on SEQ ID NO: 1. 21. The cleaning composition of any of paragraphs 1-17, wherein the polypeptide having protease activity comprises at least one substitution selected from the group consisting of S9E, N42R, N74D, V199I, Q200L, Y203W, S253D, N255Wand L256E, wherein numbering is based on SEQ ID NO: 1.

[0214] 22. The cleaning composition of paragraph 21 , wherein the polypeptide having protease activity comprises two, three, four, five, six, seven, eight or all nine substitutions selected from the group consisting of S9E, N42R, N74D, V199I, Q200L, Y203W, S253D, N255Wand L256E, wherein numbering is based on SEQ ID NO: 1.

[0215] 23. The cleaning composition of paragraph 22, wherein the polypeptide having protease activity comprises the substitutions S9E, N42R, N74D, V199I, Q200L, Y203W, A209X, S253D, N255W and L256E, wherein X in A209X is K, R, Q, N, S or T, preferably wherein X is K or N, more preferably wherein X is K, and wherein numbering is based on SEQ ID NO: 1.

[0216] 24. The cleaning composition of any of paragraphs 17-23, wherein the polypeptide having protease activity further comprises the substitution G95D.

[0217] 25. The cleaning composition of paragraph 24, wherein the polypeptide having protease activity comprises the substitutions S9E, N42R, N74D, G95D, V199I, Q200L, Y203W, A209X, S253D, N255W and L256E, wherein X in A209X is K, R, Q, N, S or T, preferably wherein X is K or N, more preferably wherein X is K, and wherein numbering is based on SEQ ID NO: 1.

[0218] 26. The cleaning composition of any of the preceding paragraphs, wherein the cleaning adjunct ingredient comprises at least one surfactant, builder or bleach component.

[0219] 27. The cleaning composition of paragraph 26, comprising at least one surfactant, preferably comprising at least one anionic surfactant

[0220] 28. The cleaning composition of paragraph 26 or 27, comprising at least one builder.

[0221] 29. The cleaning composition of any of paragraphs 26-28, comprising at least one bleach component.

[0222] 30. The cleaning composition of any of the preceding paragraphs, further comprising at least one additional enzyme selected from the group consisting of lipases, cutinases, amylases, carbohydrases, cellulases, DNases, pectinases, mannanases, arabmases, galactanases, xylanases, licheninases, xyloglucanases, oxidases, e.g., a laccase or peroxidase, and additional proteases.

[0223] 31. A method for laundering a textile item, comprising: a) exposing the item to a wash liquor comprising a polypeptide having hexosaminidase activity and a polypeptide having protease activity, wherein the polypeptide having hexosaminidase activity and the polypeptide having protease activity are as defined in any of the preceding paragraphs; b) completing at least one wash cycle; and optionally c) rinsing the item.

[0224] 32. A method for laundering a textile item, comprising: a) exposing the item to a wash liquor comprising a cleaning composition as defined in any of paragraphs 1-30; b) completing at least one wash cycle; and optionally c) rinsing the item.

[0225] 33. Use of a cleaning composition as defined in any of paragraphs 1-30 in a cleaning process, such as laundry or hard surface cleaning such as dishwashing.

[0226] 34. Use of a cleaning composition as defined in any of paragraphs 1-30 for preventing, reducing and / or removing biofilm from the interior surfaces of a washing machine or dishwasher.

[0227] The present invention is further described by the following examples that should not be construed as limiting the scope of the invention.

[0228] Examples

[0229] Materials and methods

[0230] Hexosaminidase activity assays

[0231] Hexosaminidase activity may e.g. be determined using one of the following assays.

[0232] Assay 1

[0233] The hexosaminidase activity of hexosaminidase polypeptides may be determined using 4-nitrophenyl N-acetyl-p-D-glucosaminide (Sigma-Aldrich) as substrate. The enzymatic reaction is performed in triplicate in a 96 well flat bottom polystyrene microtiter plate (Thermo Scientific) with the following conditions: 50 mM 2-(N-morpholino) ethanesulfonic acid pH 6 buffer, 1.5 mg / ml 4-nitrophenyl N-acetyl-p-D-glucosaminide and 20 pg / ml purified enzyme sample in a total reaction volume of 100 pl. Blank samples without enzyme are run in parallel. The reactions are carried out at 37°C in a Thermomixer comfort (Eppendorf). After 10 minutes of incubation, 5 pl 1 M NaOH is added to each reaction mixture to stop the enzymatic reaction. The absorbance is read at 405 nm using a POLARstar Omega plate reader (BMG LABTECH) to estimate the formation of 4- nitrophenolate ion released due to enzymatic hydrolysis of the 4-nitrophenyl N-acetyl-p-D- glucosaminide substrate. An increased absorbance over the blank (difference in absorbance at 405 nm between the sample and the blank) indicates hexosaminidase activity.

[0234] Assay 2

[0235] The hexosaminidase activity of hexosaminidase polypeptides may be determined using 4-methylumbeliferyl N-acetyl-p-D-glucosaminide (Sigma-Aldrich) as substrate. The enzymatic reaction is performed in triplicate in a 96 well flat bottom polystyrene microtiter plate (Thermo Scientific) with the following conditions: 20 mM 3-morpholinopropane-l-sulfonic acid pH 7 buffer, 5 mM 4-methylumbeliferyl N-acetyl-p-D-glucosaminide and 20 nM purified enzyme sample in a total reaction volume of 200 pl. Blank samples without enzyme are run in parallel. The reactions are carried out at ambient temperature 20-25°C. The reaction kinetics are followed immediately after mixing of enzyme and substrate using a SpectraMax M2e plate reader. Excitation wavelength is set to 368 nm and fluorescence emission reading is done at 448 nm. The reaction is followed for 30 min at 60 second intervals. The increase in fluorescence signal is used to estimate the formation of 4-methylumbeliferyl ion released due to enzymatic hydrolysis of the 4- methylumbeliferyl N-acetyl-p-D-glucosaminide substrate.

[0236] The results are expressed as an average initial rate of reaction measured as relative fluorescence units per minute (RFU / min) using excitation at 368 nm and fluorescence emission at 448 nm for each reaction performed in triplicate. An increased value over the blank (A RFU / min) indicates hexosaminidase activity.

[0237] Protease activity assay

[0238] Protease activity may be determined using assays known in the art, for example the Suc- AAPF-pNA activity assay disclosed in WO 2016 / 001450.

[0239] Example 1

[0240] Synergistic effect of hexosaminidase and protease on deep-cleaning in liquid model detergent on EPS swatches A PNAG-producing Pseudomonas fluorescens isolate was restreaked on Tryptone Soya Agar (TSA) (pH 7.3) (CM0131 ; Oxoid Ltd, Basingstoke, UK) and incubated at 23°C. A pre-culture was then established in Tryptone Soy Broth (TSB), grown for 16h at 30°C, 200 rpm, and used to inoculate biofilm cultures in TSBG (TSB with 1% glucose) in sterile 6-well microplates (Thermo scientific, #150239). The biofilm cultures were incubated statically for 3 days at 30°C prior to harvesting. The biofilm cultures were subsequently retrieved and pelleted by centrifugation (10min, 8000g, 21 °C), resuspended in 3M NaCI and incubated for 15min at 30°C. The cells were then re-pelleted, resuspended in 3M NaCI and incubated for 15min at 30°C. The EPS-containing supernatants obtained after a final centrifugation step (10min, 8000g, 21 °C) were pooled and stored at -20°C until further use (termed crude EPS).

[0241] For testing wash performance, 50 pl aliquots of the crude EPS were spotted on sterile textile swatches (WFK20A; Wfk-Testgewebe GmbH, Germany) and incubated for 15 min at ambient temperature. The swatches (sterile or with EPS) were placed in 50 mL test tubes and 10 mL of wash liquor containing 15°dH water with 0.7 g / L WFK 09V pigment soil (Wfk-Testgewebe GmbH, #00500) and 3.15 g / L liquid model A2 detergent (12% sodium salt of linear alkylbenzene sulphonates (Na-LAS), 12% alcohol ethoxylate (AEG) Biosoft N25-7 (Nl), 4% alkyl ethoxysulfate / sodium laureth sulfate (AEOS / SLES), 2% MPG (monopropylene glycol), 3.1 % ethanol, 2% triethanolamine (TEA), 3% palm kernel oil soap, 2% sodium hydroxide, 3.9% sodium citrate, 1.5% diethylenetriaminepenta(methylenephosphonic acid) (DTMPA) and 0.5% phenoxyethanol (all percentages are w / w)). Enzymes were added as to each tube as 0.02 pg / ml hexosaminidase, 0.02 pg / ml protease or a mix of 0.01 pg / ml or 0.02 pg / ml of each. Washes without enzyme were included as controls.

[0242] The protease used in this example is a variant of SEQ ID NO: 1 having the substitutions S3T, V4I, S97D, S99E, S101A, V102I, S154D, G157S, V199I, Q200L, Y203W, A209K and L256E. Hexosaminidase 1 is the hexosaminidase of SEQ ID NO: 2, and hexosaminidase 2 is the hexosaminidase of SEQ ID NO: 7.

[0243] The test tubes were placed in a Stuart rotator and incubated for 1 hour at 30°C at 20 rpm. The wash liquor was then removed, and the swatches were rinsed twice with 15°dH water and dried on filter paper over night. The tristimulus light intensity (Y) values were measured using a Datacolor 800V spectrophotometer and are displayed in Table 1 below. Wash performance, WP, (AY— Y (swatch washed with enzyme) Y (swatch washed without enzyme)) and the W3Sh performance Synergies, WP syn (AY(cocktail) " AY (sum of individual enzyme treatments)) are alSO indicated. Table 1. Synergistic effect of hexosaminidase and protease on EPS swatches in model A2 detergent.

[0244] As seen in Table 1 , an enzyme cocktail comprising hexosaminidase and protease provides superior deep-cleaning properties in model A2 detergent as compared to the individual enzymes. The wash performances of the low dose cocktails (containing 0.01 pg / ml hexosaminidase + 0.01 pg / ml protease) are better than either of the individual enzymes when dosed individually at a comparable concentration, and are also better than the sum of the wash performances of the protease and the hexosaminidase (WPsyn> 0), even though the total enzyme concentration of 0.02 pg / ml is the same. The wash performance of the high dose enzyme cocktail (0.02 pg / ml hexosaminidase + 0.02 pg / ml protease) is even better, exceeding the sum of the wash performances of the individual hexosaminidase or protease enzymes (WPsyn> 0) by a greater amount. This is consistent with there being a synergistic effect between the two enzymes on the deep-cleaning properties in model A2. This is believed to be due to the different EPS components targeted by these enzymes being localized in complex macromolecular structures which shield each other from enzymatic hydrolysis, whereby the two different enzymes can increase the accessibility to other matrix components, either by exposing shielded target substances or by allowing an increased penetration of the other enzyme into the matrix.

Claims

Claims1. A cleaning composition comprising a polypeptide having hexosaminidase activity, a polypeptide having protease activity and a cleaning adjunct ingredient, wherein the polypeptide having protease activity comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 1 and comprises a substitution in position 209 of SEQ ID NO: 1 selected from the group consisting of A209K, A209R, A209Q, A209N, A209S and A209T.

2. The cleaning composition of claim 1 , wherein the polypeptide having protease activity comprises a substitution in position 209 of SEQ ID NO: 1 selected from the group consisting of A209K, A209Q, A209N, A209S and A209T, preferably selected from the group consisting of A209K, A209Q, A209N and A209T, more preferably selected from the group consisting of A209K and A209N, most preferably A209K.

3. The cleaning composition of claim 1 or 2, wherein the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 70% sequence identity to 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; for example at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94% or at least 95% sequence identity to 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.

4. The cleaning composition of claim 3, wherein the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 2 and comprises a substitution at one or more positions selected from positions 3, 15, 49, 59, 163, 186, 225, 227, 232, 235, 252, 260, 272, 279, 281 , 308, 309 and 312 of SEQ ID NO: 2; for example a substitution at two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen or all of said positions.

5. The cleaning composition of claim 4, wherein the polypeptide having hexosaminidase activity comprises at least one substitution selected from the group consisting of Q3F, Q3I, H15Y, A49W, N59E, S163P, S186R, S225G, N227T, E232D, G235W, N252P, N260Q, H272V, H272P, S279D, Y281 P, K308Q, K308E, K309E, K312Q and K312E, wherein numbering is based on SEQ ID NO: 2; preferably two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen or all substitutions selected from the group consisting of Q3I, H15Y, A49W, N59E, S163P, S186R, S225G,N227T, E232D, G235W, N252P, N260Q, H272V, S279D, Y281 P, K308Q, K309E and K312Q.

6. The cleaning composition of claim 5, wherein the polypeptide having hexosaminidase activity comprises an amino acid sequence having at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 7.

7. The cleaning composition of claim 3, wherein the polypeptide having hexosaminidase activity comprises: a substitution at one or more positions, preferably two, three or four positions, corresponding to positions 163, 227, 252 and 309 of SEQ ID NO: 2; and a substitution at one or more positions corresponding to positions 111 , 120, 106, 124, 127, 150, 170, 171 , 178, 199, 208, 254, 255 and 278 of SEQ ID NO: 2; wherein the polypeptide at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% sequence identity to SEQ ID NO: 2.

8. The cleaning composition of claim 7, wherein the polypeptide having hexosaminidase activity comprises two, three or four substitutions selected from the group consisting of S163P, N227T, N252P and K309E, wherein numbering is based on SEQ ID NO: 2; and further comprises at least one substitution selected from the group consisting of D111 R, T120V, V106M, V106N, Y124I, Y124K, Y124L, R127P, E150N, L170G, D171 F, D171 H, Q178K, S199W, S208W, F254I, T255D and I278V, wherein numbering is based on SEQ ID NO: 2.

9. The cleaning composition of any of the preceding claims, wherein the polypeptide having protease activity has at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95% or at least 96% sequence identity to SEQ ID NO: 1.

10. The cleaning composition of any of claims 1-9, wherein the polypeptide having protease activity comprises at least one substitution compared to SEQ ID NO: 1 selected from the group consisting of S3T, V4I, S97D, S99E, S101A, V102I, S154D, G157S, V199I, Q200L, Y203W and L256E, wherein numbering is based on SEQ ID NO: 1 ; for example two, three, four, five, six, seven, eight, nine, ten, eleven or all of said substitutions.

11. The cleaning composition of claim 10, wherein the polypeptide having protease activity comprises the substitutions S3T, V4I , S97D, S99E, S101A, V102I, S154D, G157S, V199I, Q200L, Y203W, A209X and L256E, wherein X in A209X is K, R, Q, N, S or T, preferably wherein X is K or N, more preferably wherein X is K, and wherein numbering is based on SEQ ID NO: 1.

12. The cleaning composition of any of claims 1-9, wherein the polypeptide having protease activity comprises at least one substitution selected from the group consisting of S9E, N42R, N74D, V199I, Q200L, Y203W, S253D, N255W and L256E, wherein numbering is based on SEQ I D NO: 1 ; for example two, three, four, five, six, seven, eight or all of said substitutions.

13. The cleaning composition of any of claims 9-12, wherein the polypeptide having protease activity further comprises the substitution G95D; for example wherein the polypeptide having protease activity comprises the substitutions S9E, N42R, N74D, G95D, V199I, Q200L, Y203W, A209X, S253D, N255W and L256E, wherein X in A209X is K, R, Q, N, S or T, preferably wherein X is K or N, more preferably wherein X is K, and wherein numbering is based on SEQ ID NO: 1.

14. A method for laundering a textile item, comprising: a) exposing the item to a wash liquor comprising a cleaning composition as defined in any of claims 1-13; b) completing at least one wash cycle; and optionally c) rinsing the item.

15. Use of a cleaning composition as defined in any of claims 1-13 in a cleaning process, such as laundry or hard surface cleaning such as dishwashing.

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