Compositions comprising a protease and a xyloglucanase

Abstract

The present invention relates to compositions such as detergent or cleaning compositions comprising a protease and a cellulase, in particular a protease and a xyloglucanase, as well as cleaning methods using the compositions and use of the compositions for cleaning an item such as a textile.

Description

[0001] COMPOSITIONS COMPRISING A PROTEASE AND A XYLOGLUCANASE REFERENCE TO A SEQUENCE LISTING

[0002] This application contains a Sequence Listing in computer readable form, which is incorporated herein by reference.

[0003] FIELD OF THE INVENTION

[0004] The present invention relates to compositions, in particular detergent or cleaning compositions, comprising a protease and a cellulase, in particular a protease and a xyloglucanase, as well as cleaning methods using the compositions and use of the compositions for cleaning an item such as a textile.

[0005] BACKGROUND OF THE INVENTION

[0006] Various enzymes have been used in detergent compositions for many decades. These include enzymes such as proteases, lipases, amylases, cellulases, mannanases as well as other enzymes and mixtures thereof. Commercially, the most important enzymes are proteases.

[0007] Many commercially available proteases used for, e.g., laundry and dishwashing detergents are engineered variants of naturally occurring wild-type proteases. Protease variants have been described in the art with alterations relative to a parent protease, resulting in improvements such as better wash performance, thermal stability, storage stability and catalytic activity.

[0008] In addition to proteases, many detergents contain other enzymes ( / .e., companion enzymes) that offer complementary cleaning benefits, and various enzymes have been disclosed for potential use in detergent compositions. An example of a class of enzymes finding beneficial use in combination with proteases are cellulolytic enzymes ( / .e., cellulases). For example, xyloglucanases belonging to glycosyl hydrolase family 44 and use thereof in detergent compositions are disclosed in WO 01 / 62903. Variants of a glycosyl hydrolase family 44 xyloglucanase are disclosed in WO 2009 / 147210 and in WO 2022 / 043321.

[0009] Xyloglucan is a hemicellulose which is a major structural polysaccharide in the primary (growing) cell wall of plants. Structurally, xyloglucans consist of a cellulose-like beta-1, 4-linked glucose backbone, which is frequently substituted with various side chains, notably with 1,6 linked xylose sidechains. Xyloglucan is believed to function in the primary wall of plants by cross-linking cellulose microfibrils, forming a cellulose-xyloglucan network. Xyloglucanses are capable of catalyzing the solubilization of xyloglucan to xyloglucan oligosaccharides.

[0010] It has now surprisingly been found that a beneficial cleaning effect may be obtained, e.g., in laundry by combining a protease and a cellulase, in particular a beneficial cleaning effect may be obtained, e.g., in laundry by combining a protease and a glycosyl hydrolase family 44 xyloglucanase.SUMMARY OF THE INVENTION

[0011] The present invention relates to a composition, in particular a detergent or cleaning composition, comprising a protease and a cellulase, such as, e.g., a protease and a glycoside hydrolase family 44 xyloglucanase, as well as use of the composition for cleaning an object such as a textile, and a method of cleaning an object using the composition.

[0012] SEQUENCE OVERVIEW SEQ ID NO: 1 is subtilisin 309 (Savinase®) from Bacillus lentus.

[0013] SEQ ID NO: 2 is subtilisin BPN’ from Bacillus amyloliquefaciens.

[0014] SEQ ID NO: 3 is subtilisin Carlsberg (Alcalase®) from Bacillus licheniformis.

[0015] SEQ ID NO: 4 is Bacillus lentus alkaline protease (BLAP) from Bacillus lentus DSM 5483. SEQ ID NO: 5 is a subtilisin protease from Bacillus gibsonii DSM 14391.

[0016] SEQ ID NO: 6 is a subtilisin protease from Bacillus gibsonii DSM 8722.

[0017] SEQ ID NO: 7 is a mature xyloglucanase polypeptide obtained from Paenibacillus polymyxa.

[0018] SEQ ID NO: 8 is a variant of SEQ ID NO: 7.

[0019] SEQ ID NO: 9 is a variant of SEQ ID NO: 7.

[0020] SEQ ID NO: 10 is the cellulase disclosed in Example 2.

[0021] DEFINITIONS

[0022] 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.

[0023] 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.

[0024] Protease: The term “protease” means an enzyme having peptidase activity (EC 3.4; also known as proteolytic activity or protease activity) that catalyzes the hydrolysis of peptide bonds. The EC 3.4 group includes several sub-groups, including EC 3.4.21 (serine endopeptidase), which contains further sub-groups, including EC. 3.4.21.62 (subtilisin). The terms “protease” and the expression “polypeptide having protease activity” are used interchangeably herein. Protease activity may be determined according to methods known in the art, for example as described in Protease Activity Assay I and Protease Activity Assay II in WO 2024 / 121070.

[0025] Cellulolytic enzyme or Cellulase: The terms “cellulolytic enzymes” and “cellulase” are used interchangeably and means one or more (e.g., several) enzymes that hydrolyze a cellulosic material. Cellulases may be selected from the group consisting of cellulases belonging to GH5, GH44, GH45, EC 3.2.1.4, EC 3.2.1.21, EC 3.2.1.91 and EC 3.2.1.172. Such enzymes includeendoglucanase(s) (e.g., EC 3.2.1.4), cellobiohydrolase(s), beta-glucosidase(s), or combinations thereof.

[0026] Suitable cellulases include mono-component and mixtures of enzymes of bacterial or fungal origin. Chemically modified or protein engineered mutants are also contemplated. The cellulase may for example be a mono-component or a mixture of mono-component endo-1,4-beta-glucanase also referred to as endoglucanase.

[0027] Suitable cellulases include those from the genera Bacillus, Pseudomonas, Humicola, Myceliophthora, Fusarium, Thielavia, Trichoderma, and Acremonium. Exemplary cellulases include a fungal cellulase from Humicola insolens (US 4,435,307) or from Trichoderma, e.g., T. reesei or T. viride. Other suitable cellulases are from Thielavia, e.g., Thielavia terrestris as described in WO 96 / 29397 or the fungal cellulases produced from Myceliophthora thermophila and Fusarium oxysporum disclosed in US 5,648,263, US 5,691 ,178, US 5,776,757, WO 89 / 09259 and WO 91 / 17244. Also relevant are cellulases from Bacillus as described in WO 02 / 099091 and JP 2000210081. Suitable cellulases are alkaline or neutral cellulases having care benefits. Examples of cellulases are 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. The cellulases applied in the present invention may be expressed with or without signal peptide or with part of the signal peptide depending on the actual expression conditions.

[0028] Examples of commercially available cellulases include Carezyme®, Carezyme® Premium, Celluzyme®, Celluclean®, Celluclast®, Endolase®, Renozyme®, Whitezyme®, Celluclean® Classic, Cellusoft® (Novozymes A / S); Puradax®, Puradax HA, Puradax EG, Revitalenz 1000, Revitalenz 200, Revitalenz 2000 (Dupont Industrial Biosciences); KAC-500(B)™ (Kao Corporation); Biotouch DCL, Biotouch FLX1 (AB enzymes).

[0029] The two basic approaches for measuring cellulolytic enzyme activity may include: (1) measuring the total cellulolytic enzyme activity, and (2) measuring the individual cellulolytic enzyme activities (endoglucanases, cellobiohydrolases, and beta-glucosidases) as reviewed in Zhang et al., 2006, Biotechnology Advances 24: 452-481. Total cellulolytic enzyme activity can be measured using insoluble substrates, including Whatman N°1 filter paper, microcrystalline cellulose, bacterial cellulose, algal cellulose, cotton, pretreated lignocellulose, etc. The most common total cellulolytic activity assay is the filter paper assay using Whatman N°1 filter paper as the substrate. The assay was established by the International Union of Pure and Applied Chemistry (IUPAC) (Ghose, 1987, Pure Appl. Chem. 59: 257-68).

[0030] Family GH45 cellulase: The term “family GH45 cellulase” as used herein, refers to Glycosyl hydrolases are enzymes that catalyze the hydrolysis of the glycosyl bond. There are over 100 classes of Glycosyl hydrolases which have been classified, see Henrissat etal. (1991), A classification of glycosyl hydrolases based on amino acid sequence similarities, J. Biochem.

[0031] 280: 309-316 and the CAZY website at www.cazy.org. The glycoside hydrolases of family 45(GH45) have so far been identified as endoglucanase (EC 3.2.1.4). Within the definition falls enzymes which are commonly known as “cellulases”. Such enzymes comprise also enzymes that may be known as endoglucanases.

[0032] Cellulosic material: The term “cellulosic material” means any material containing cellulose. The predominant polysaccharide in the primary cell wall of biomass is cellulose, the second most abundant is hemicellulose, and the third is pectin. The secondary cell wall, produced after the cell has stopped growing, also contains polysaccharides and is strengthened by polymeric lignin covalently cross-linked to hemicellulose. Cellulose is a homopolymer of anhydrocellobiose and thus a linear beta-(1-4)-D-glucan, while hemicelluloses include a variety of compounds, such as xylans, xyloglucans, arabinoxylans, and mannans in complex branched structures with a spectrum of substituents. Although generally polymorphous, cellulose is found in plant tissue primarily as an insoluble crystalline matrix of parallel glucan chains. Hemicelluloses usually hydrogen bond to cellulose, as well as to other hemicelluloses, which help stabilize the cell wall matrix.

[0033] Xyloglucanase: The term “xyloglucanase” means a polypeptide having xyloglucanase activity, i.e., the ability to catalyze the solubilization of xyloglucan to xyloglucan oligosaccharides. Some xyloglucanases only exhibit xyloglucanase activity, whereas others exhibit both xyloglucanase and cellulase activity. The xyloglucanases of interest in the context of the present invention belong to the glycoside hydrolase family 44 (abbreviated Glyco_hydro_44 or GH44), which was formerly known as cellulase family J. The GH44 family includes enzymes with endoglucanase (EC 3.2.1.4) and xyloglucanase (EC 3.2.1.151) activities. The terms “xyloglucanase” and the expression “polypeptide having xyloglucanase activity” are used interchangeably herein. Xyloglucanase activity may be determined by methods known in the art, for example using the xyloglucanase assay described in Example 2 of WO 2022 / 043321 or the xyloglucanase assay described on page 60 of WO 01 / 62903.

[0034] Together, the protease and the xyloglucanase polypeptides / enzymes disclosed herein may be referred to as “the enzymes of the invention”. Similarly, when reference is made to “the enzymes”, if nothing else is apparent from the context, this should be understood as referring to the enzymes in a composition disclosed herein, including at least the protease and the xyloglucanase.

[0035] AlphaFold structure calculation: AlphaFold is a computational method for calculating the three-dimensional structure of a polypeptide from its amino acid sequence (Jumper et al., 2021, Nature 596: 583-589). Predicted structures for millions of polypeptides deposited in the UniProt database have been deposited in the AlphaFold Protein Structure Database, using the AlphaFold Monomer v2.0 algorithm (Varadi etal., 2021, Nucleic Acids Res. 50(D1):D439-D444). In the AlphaFold Protein Structure Database, the three-dimensional structure of a polypeptide can be obtained by searching for the UniProt accession number of the polypeptide.In addition to the many three-dimensional structures that are already publicly available, code is available for reproducing and predicting structures of new polypeptides at source code repositories such as Github.com under deepmind / alphafold / , using notebooks / AlphaFold.ipynb, which uses AlphaFold v2.3.1 or newer. Additionally, it can be found in Github.com under sokrypton / ColabFold using v1.5.2 or newer, using AlphaFold2.ipynb. For technical details, please see Jumper et al. (vide supra).

[0036] AlphaFold produces a per-residue estimate of its confidence on a scale from 0 to 100. This confidence measure is called pLDDT and corresponds to the model’s predicted score on the IDDT-Ca metric. It is stored in the B-factor fields of the mmCIF and PDB files available for download (although unlike a B-factor, higher pLDDT is better). Regions with pLDDT score of more than 90 are expected to be modelled to high accuracy. These should be suitable for any application that benefits from high accuracy (e.g., characterization of binding sites). Regions with a pLDDT score between 70 and 90 are expected to be modelled well, corresponding to a generally good backbone prediction.

[0037] Cleaning / detergent composition: The terms “cleaning composition”, “detergent composition” or “detergent or cleaning composition”, which may be used interchangeably, refer to a composition suitable for cleaning, e.g., of laundry / textiles and / or hard surfaces, and which comprise the protease and cellulase, in particular the protease and xyloglucanase, enzymes disclosed herein as well as at least one detergent adjunct ingredient, typically at least one surfactant or builder. Detergent adjunct ingredients are disclosed in more detail below and include components typically used in detergent compositions, for example surfactants, builders, bleach components, other enzymes, etc. Typical detergent compositions include laundry detergent compositions and dishwashing compositions, which include automatic dishwashing (ADW) compositions and hand dishwashing (HDW) compositions.

[0038] Detergent compositions 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.

[0039] 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 oneof 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.

[0040] Expression: The term “expression” includes any step involved in the production of a variant including, but not limited to, transcription, post-transcriptional modification, translation, post-translational modification and secretion.

[0041] Extension: The term “extension” means an addition of one or more amino acids to the amino and / or carboxyl terminus of a polypeptide, wherein the “extended” polypeptide has the relevant enzymatic activity, e.g., protease, cellulase, orxyloglucanase activity.

[0042] Fragment: The term “fragment” means a polypeptide having one or more amino acids absent from the amino and / or carboxyl terminus of the polypeptide, wherein the fragment has relevant enzymatic activity, e.g., protease, cellulase, orxyloglucanase activity.

[0043] Hard surface cleaning: The term “hard surface cleaning” comprises, in addition to automatic and hand 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.

[0044] Improved wash performance: The term “improved wash performance” may be defined as improved cleaning, for example improved cleaning of, e.g., textiles. Wash performance may, for example, be expressed as a remission value of stained swatches. After washing and rinsing the swatches are spread out flat and allowed to air dry at room temperature, typically overnight. Washed swatches are typically evaluated the day after washing by measuring light reflectance of the swatches using a suitable spectrophotometer. The measurements are made without UV in the incident light and the remission value typically at 460 nm is extracted.

[0045] Improved property: The term “improved property” means a characteristic associated with a variant that is improved compared to a reference polypeptide, e.g., a reference protease, a reference cellulase or a reference xyloglucanase. The improved property may also be a characteristic associated with a composition comprising one or more variants, such as a protease variant and a cellulase variant, compared to a corresponding composition comprising a reference protease and / or reference cellulase. The improved property may also be a characteristic associated with a composition comprising one or more variants, such as a protease variant and a xyloclucanase variant, compared to a corresponding composition comprising a reference protease and / or reference xyloglucanase. Such improved properties may include, but are not limited to, storage stability, mildness (of a protease towards other enzymes) and wash performance.

[0046] Isolated: The term “isolated” means a polypeptide, nucleic acid, cell, or other specified material or component that is separated from at least one other material or component, including but not limited to, other proteins, nucleic acids, cells, etc. An isolated polypeptide, nucleic acid,cell or other material is thus in a form that does not occur in nature. An isolated polypeptide includes, but is not limited to, a culture broth containing the secreted variant expressed in a host cell.

[0047] 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.

[0048] 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).

[0049] Mildness: The term “mildness” refers to the influence of a protease variant on the storage stability of a companion enzyme, i.e., another enzyme that is present in a detergent composition together with a protease variant. The term “improved mildness” means that a variant is less aggressive in terms of proteolytic activity towards a companion enzyme, thereby providing improved residual activity of a companion enzyme after storage with a variant in a detergent composition. Mildness may, e.g., be measured using a lipase as described in European patent application No. 24203579.8.

[0050] Parent or parent enzyme: The term “parent”, e.g., “parent protease”, “parent cellulase” or “parent xyloglucanase” means an enzyme, e.g., a protease, a cellulase, or a xyloglucanase, to which an alteration is made to produce an enzyme variant having the same enzymatic function, e.g., a protease variant will have protease activity. The parent may be a naturally occurring (wildtype) polypeptide or a variant or fragment thereof. The parent may, e.g., be a reference enzyme, for example SEQ ID NO: 1 herein in the case of a protease or SEQ ID NO: 8 herein in the case of a xyloglucanase, or an enzyme having the identical amino acid sequence of a specific variant but not having the alterations (e.g., substitutions) at one or more specified positions in the variant.

[0051] Purified: The term “purified” means a nucleic acid, polypeptide or cell that is substantially free from other components as determined by analytical techniques well known in the art (e.g., a purified polypeptide or nucleic acid may form a discrete band in an electrophoretic gel, chromatographic eluate, and / or a media subjected to density gradient centrifugation). A purified nucleic acid or polypeptide is at least about 50% pure, usually at least about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5%, about 99.6%, about 99.7%, about 99.8% or more pure (e.g., percent by weight or on a molar basis). In a related sense, a composition is enriched for a molecule when there is a substantial increase in the concentration of the molecule after application of a purification or enrichment technique. The term "enriched" refers to a compound, polypeptide, cell, nucleic acid, amino acid, or other specified material or component that is present in a composition at a relative or absolute concentration that is higher than a starting composition.In one aspect, the term purified as used herein refers to the polypeptide or cell being essentially free from components (especially insoluble components) from the production organism. In other aspects, the term "purified" refers to the polypeptide being essentially free of insoluble components (especially insoluble components) from the native organism from which it is obtained. In one aspect, the polypeptide is separated from some of the soluble components of the organism and culture medium from which it is recovered. The polypeptide may be purified ( / .e., separated) by one or more of the unit operations filtration, precipitation, or chromatography.

[0052] Accordingly, the polypeptide may be purified such that only minor amounts of other proteins, in particular, other polypeptides, are present. The term "purified" as used herein may refer to removal of other components, particularly other proteins and most particularly other enzymes present in the cell of origin of the polypeptide. The polypeptide may be "substantially pure", i.e., free from other components from the organism in which it is produced, e.g., a host organism for recombinantly produced polypeptide. In one aspect, the polypeptide is at least 40% pure by weight of the total polypeptide material present in the preparation. In one aspect, the polypeptide is at least 50%, 60%, 70%, 80% or 90% pure by weight of the total polypeptide material present in the preparation. As used herein a "substantially pure polypeptide" may denote a polypeptide preparation that contains at most 10%, preferably at most 8%, more preferably at most 6%, more preferably at most 5%, more preferably at most 4%, more preferably at most 3%, even more preferably at most 2%, most preferably at most 1%, and even most preferably at most 0.5% by weight of other polypeptide material with which the polypeptide is natively or recombinantly associated.

[0053] It is, therefore, preferred that the substantially pure polypeptide is at least 92% pure, preferably at least 94% pure, more preferably at least 95% pure, more preferably at least 96% pure, more preferably at least 97% pure, more preferably at least 98% pure, even more preferably at least 99% pure, most preferably at least 99.5% pure by weight of the total polypeptide material present in the preparation. The polypeptides used in the present invention are preferably in a substantially pure form i.e., the preparation is essentially free of other polypeptide material with which it is natively or recombinantly associated). This can be accomplished, for example by preparing the polypeptides by well-known recombinant methods or by classical purification methods.

[0054] 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, variant 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”.Recover: The terms recover or recovery means the removal of a polypeptide from at least one fermentation broth component selected from a cell, a nucleic acid, or other specified material, e.g., recovery of the polypeptide from the whole fermentation broth, or from the cell-free fermentation broth, by polypeptide crystal harvest, by filtration, e.g., depth filtration (by use of filter aids or packed filter medias, cloth filtration in chamber filters, rotary-drum filtration, drum filtration, rotary vacuum-drum filters, candle filters, horizontal leaf filters or similar, using sheed or pad filtration in framed or modular setups) or membrane filtration (using sheet filtration, module filtration, candle filtration, microfiltration, ultrafiltration in either cross flow, dynamic cross flow or dead end operation), or by centrifugation (using decanter centrifuges, disc stack centrifuges, hyrdo cyclones or similar), or by precipitating the polypeptide and using relevant solid-liquid separation methods to harvest the polypeptide from the broth media by use of classification separation by particle sizes. Recovery encompasses isolation and / or purification of the polypeptide.

[0055] Sequence difference: The term "sequence difference" means the percentage amino acid differences between two polypeptides, e.g., between a variant and the parent, and is calculated as follows:

[0056] (Number of different residues x 100) / (Number of residues in parent protease) wherein the term “different residues” means the total number of amino acid residues that have been substituted, deleted, and / or inserted in the variant compared to the parent.

[0057] For example, for a variant of SEQ ID NO: 1 having the insertion S97SE and the substitutions A188P, V199I, Q200L, Y203W, and L256E ( / .e., six different residues), the sequence difference is calculated as follows:

[0058] (6 x 100) / 269 = 2.2%

[0059] Sequence identity: The relatedness between two amino acid sequences or between two nucleotide sequences is described by the parameter “sequence identity”.

[0060] 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:

[0061] (Identical Residues x 100) / (Length of Alignment -Total Number of Gaps in Alignment) Structural Similarity: For purposes of the present invention, the relatedness between the three-dimensional structure of two polypeptides is described by the parameter “structural similarity”.A three-dimensional structure of any polypeptide may be obtained experimentally via, e.g. , X-ray crystallography or using in silico methods such as AlphaFold (vide supra). The structural similarity between three-dimensional structures may then be determined by the TM-score, which is calculated using the following general formula (Zhang & Skolnick, Proteins 57:702-710, 2004):

[0062] TM-score = Max

[0063]

[0064] where LN is the length of the native structure, LT is the length of the aligned residues to the template structure, d, is the distance between pair / of aligned residues and do is a scale to normalize the match difference. ‘Max’ denotes the maximum value after optimal spatial superposition.

[0065] For the purposes of the present invention, LN is the length of the reference polypeptide:

[0066] 1 'V 1

[0067] TM-score = - > - =•

[0068] fd -\2

[0069] l=1l + hr

[0070]

[0071] \do /

[0072] A structural alignment of the three-dimensional structures of two polypeptides is necessary before the TM-score can be calculated. This is achieved via algorithms that optimize the structural overlap, and several methods are available, such as CEalign (Shindyalov and Bourne, Protein Eng., 11, 739-747, 1998), DALI (Holm and Sander, Trends Biochem. Sci., 20, 478-480, 1995), orTM-align (Nucleic Acids Res. 33:2302-2309, 2005).

[0073] For the purposes of the present invention, TM-align is applied. For convenience, TM-score is integrated in the TM-align software, which is available from the author’s website (zhanggroup.org / TM-score / ). The version of TM-align is preferably updated 2019-08-22 or later, and the TM-score between a reference and a query protein is determined by running this command:

[0074] TMalign <query.pdb> <ref erence . pdb> -L <length of reference> Where <query.pdb> is the name of the PDB file containing coordinates of the query polypeptide, and <reference.pdb> is the name of the PDB file containing coordinates of the reference polypeptide. The TM-score is calculated and reported in the output, along with several other parameters from the alignment.

[0075] The maximal TM-score is 1, e.g., 1.0, corresponding to identical three-dimensional structures.

[0076] 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).Variant: The term variant means a polypeptide having enzyme activity, e.g., protease, cellulase, endoglucanase, or xyloglucanase activity, comprising a substitution, an insertion (including extension), and / or a deletion (e.g., truncation), at one or more positions relative to a parent enzyme or reference sequence. 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, 1 amino acid) adjacent to and immediately following the amino acid occupying a position.

[0077] Wash liquor: The term “wash liquor” refers to an aqueous solution comprising a composition of the invention, including a cellulase and a protease, e.g., a xyloglucanase and a protease, as described herein. A wash liquor is a solution, e.g., found in a washing machine or dishwasher, containing water and a detergent composition comprising the cellulase and protease, such as the xyloglucanase 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.

[0078] Wash performance: The term "wash performance" describes the ability of one or more enzymes to remove relevant stains present on an object to be cleaned during a cleaning process, for example laundry or hard surface cleaning such as dishwashing, e.g., automatic dishwashing (ADW) or hand dishwashing (HDW). Relevant stains in the context of the present invention will typically contain a protein and / or a xyloglucan. Non-limiting examples of relevant stains are, e.g., the commercially available stains used in the examples herein. Wash performance may be quantified according to the TOM assay described in the Examples herein. An improvement in wash performance of an enzyme, e.g., a protease variant, may be determined relative to the wash performance of a reference or parent enzyme, e.g., a reference protease such as a parent protease. In an embodiment, the improvement in wash performance of the protease variant according to the invention is determined relative to the wash performance of an otherwise identical protease without the insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1. In an embodiment, the improvement in wash performance of the protease variant according to the invention is determined relative to the wash performance of an otherwise identical protease without the insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and without substitutions at at least two, e.g., at least three, at least four, or five, positions corresponding to any of positions 188, 199, 200, 203 and 256 of SEQ ID NO: 1.

[0079] 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.

[0080] 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) 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 a laboratory or modification of the wild-type sequence).

[0081] Conventions for Designation of Variants

[0082] For purposes of the present invention, the polypeptide disclosed in SEQ ID NO: 1 is used to determine the corresponding amino acid positions in another protease. The amino acid sequence of another protease is aligned with the polypeptide disclosed in SEQ ID NO: 1, and based on the alignment, the amino acid position number corresponding to any amino acid residue in the polypeptide disclosed in SEQ ID NO: 1 is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16: 276-277), preferably version 5.0.0 or later. The parameters used are gap open penalty of 10, gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix.

[0083] Examples of alignments of different protease sequences may be seen in Figures 1-7 of WO 2024 / 121070. For example, Figure 1 of WO 2024 / 121070 is an alignment of SEQ ID NO: 1 and SEQ ID NO: 2 therein, corresponding to SEQ ID NO: 1 and SEQ ID NO: 2 of the present application. Similarly, Figure 2 of WO 2024 / 121070 is an alignment of SEQ ID NO: 1 and SEQ ID NO: 3 therein, corresponding to SEQ ID NO: 1 and SEQ ID NO: 3 of the present application. Alignments of other protease sequences can be prepared in a similar manner.

[0084] Similarly, the corresponding amino acid positions in a xyloglucanase may be determined based on SEQ ID NO: 7, 8 or 9, as these three sequences have the same number of amino acids and have only minor variations relative to each other. For other xyloglucanases, alignments of with SEQ ID NO: 7, 8 or 9 may be prepared in the same manner as alignments between different protease sequences.

[0085] In describing the variants of the present invention, the nomenclature described below is adapted for ease of reference. The accepted IIIPAC single letter or three letter amino acid abbreviation is employed.

[0086] Substitutions: For an amino acid substitution, the following nomenclature is used: Original amino acid, position, substituted amino acid. Accordingly, the substitution of glycine at position 195 with lysine is designated as “G195K”. Multiple mutations may be separated by addition marks (“+”) or by commas, e.g., “G195K, S411F” or “G195K+S411F”, or simply by a space between the individual mutations, e.g., “G195K S411F”, representing substitutions at positions 195 and 411 of glycine (G) with lysine (K) and serine (S) with phenylalanine (F), respectively. Because the amino acid residue at a given position varies from parent to parent, the original amino acid may be indicated with X, e.g., “X195K”.Deletions: For an amino acid deletion, the following nomenclature is used: Original amino acid, position, *. Accordingly, the deletion of the amino acid glycine at position 195 is designated as “G195*”. Multiple deletions may be separated by addition marks (“+”) or by commas, e.g., “G195*+S411*” or“G195*, S411*”, or by a space between the deletions. Because the amino acid residue at a given position varies from parent to parent, the original amino acid may be indicated with X, e.g., “X195*”.

[0087] Insertions: Foran amino acid insertion, the following nomenclature is used: Original amino acid, position, original amino acid, inserted amino acid. Accordingly, the insertion of lysine after the amino acid glycine at position 195 is designated “G195GK”. An insertion of multiple amino acids is designated using the following nomenclature: 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 the amino acid glycine at position 195 is indicated as “G195GKA”. Multiple insertions are separated by addition marks (“+”) or by commas, e.g., “G195GK+L217LY” or ““G195GK,L217LY”, or by a space between the insertions. Because the amino acid residue at a given position varies from parent to parent, the original amino acid may be indicated with X, e.g., “X195XK”.

[0088] With respect to insertions, the inserted amino acid residue(s) are numbered by the addition of lower-case letters to the position number of the amino acid residue preceding the inserted amino acid residue(s). In the example G195GKA, the numbering would thus be:

[0089] Parent: Variant:

[0090] 195 195 195a 195b

[0091] G G - K - A

[0092]

[0093] Alternatively, an insertion of an amino acid residue such as lysine after the amino acid at position 195 may be indicated by “195aK”, and the insertion of two or more additional amino acid residues such as lysine and alanine after the amino acid at position 195 may be indicated by “195aK,195bA”.

[0094] DETAILED DESCRIPTION OF THE INVENTION

[0095] The present invention provides compositions, in particular detergent or cleaning compositions, comprising a protease and a cellulase, such as in particular, a protease and a glycosyl hydrolase family 44 xyloglucanase.

[0096] In one aspect, the invention relates to a composition, in particular a detergent composition, comprising a protease and a xyloglucanase, wherein

[0097] a) the protease is a variant of a parent protease, wherein the variant has protease activity and comprises an insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises substitutions at at least two, e.g., at least three, at least four, or five, positions corresponding to any of positions 188, 199, 200, 203, and 256 of SEQ ID NO: 1;wherein position numbering is based on the numbering of SEQ ID NO: 1; wherein the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of the parent protease, wherein the three-dimensional structure is calculated using AlphaFold; and

[0098] b) the xyloglucanase is a variant of a parent xyloglucanase, wherein the variant has xyloglucanase activity and comprises substitutions at positions 111, 123, 129 and 159 of SEQ ID NO: 8, wherein the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of the parent xyloglucanase, wherein the three-dimensional structure is calculated using AlphaFold.

[0099] In one embodiment, the parent protease is SEQ ID NO: 1 and the variant has a TM-score of at least 0.80, e.g., at least 0.81 , at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 1, wherein the three-dimensional structure is calculated using AlphaFold. Preferably, the parent is SEQ ID NO: 1 , and the variant has a TM-score of at least 0.980, e.g., at least 0.985, at least 0.990, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 1, wherein the three-dimensional structure is calculated using AlphaFold. Most preferably, the parent is SEQ ID NO: 1, and the variant has a TM-score of at least 0.990, e.g., at least 0.991, at least 0.992, at least 0.993, at least 0.994, at least 0.995, at least 0.996, at least 0.997, at least 0.998, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 1, wherein the three-dimensional structure is calculated using AlphaFold.

[0100] In another embodiment, the parent protease is SEQ ID NO: 2 and the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 2, wherein the three-dimensional structure is calculated using AlphaFold. Preferably, the parent is SEQ ID NO: 2, and the variant has a TM-score of at least 0.980, e.g., at least 0.985, at least 0.990, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 2, wherein the three-dimensional structure is calculated using AlphaFold. Most preferably, the parent is SEQ ID NO: 2, and the variant has a TM-score of at least 0.990,e.g., at least 0.991, at least 0.992, at least 0.993, at least 0.994, at least 0.995, at least 0.996, at least 0.997, at least 0.998, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 2, wherein the three-dimensional structure is calculated using AlphaFold.

[0101] In another embodiment, the parent protease is SEQ ID NO: 3 and the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 3, wherein the three-dimensional structure is calculated using AlphaFold. Preferably, the parent is SEQ ID NO: 3, and the variant has a TM-score of at least 0.980, e.g., at least 0.985, at least 0.990, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 3, wherein the three-dimensional structure is calculated using AlphaFold. Most preferably, the parent is SEQ ID NO: 3, and the variant has a TM-score of at least 0.990, e.g., at least 0.991, at least 0.992, at least 0.993, at least 0.994, at least 0.995, at least 0.996, at least 0.997, at least 0.998, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 3, wherein the three-dimensional structure is calculated using AlphaFold.

[0102] In another embodiment, the parent protease is SEQ ID NO: 4 and the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 4, wherein the three-dimensional structure is calculated using AlphaFold. Preferably, the parent is SEQ ID NO: 4, and the variant has a TM-score of at least 0.980, e.g., at least 0.985, at least 0.990, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 4, wherein the three-dimensional structure is calculated using AlphaFold. Most preferably, the parent is SEQ ID NO: 4, and the variant has a TM-score of at least 0.990, e.g., at least 0.991, at least 0.992, at least 0.993, at least 0.994, at least 0.995, at least 0.996, at least 0.997, at least 0.998, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 4, wherein the three-dimensional structure is calculated using AlphaFold.

[0103] In another embodiment, the parent protease is SEQ ID NO: 5 and the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 5, wherein the three-dimensional structure is calculated using AlphaFold. Preferably, the parent is SEQ ID NO: 5, and the variant has a TM-score of at least 0.980, e.g., at least 0.985, at least 0.990, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 5, wherein the three-dimensional structure is calculated using AlphaFold.Most preferably, the parent is SEQ ID NO: 5, and the variant has a TM-score of at least 0.990, e.g., at least 0.991, at least 0.992, at least 0.993, at least 0.994, at least 0.995, at least 0.996, at least 0.997, at least 0.998, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 5, wherein the three-dimensional structure is calculated using AlphaFold.

[0104] In another embodiment, the parent protease is SEQ ID NO: 6 and the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 6, wherein the three-dimensional structure is calculated using AlphaFold. Preferably, the parent is SEQ ID NO: 6, and the variant has a TM-score of at least 0.980, e.g., at least 0.985, at least 0.990, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 6, wherein the three-dimensional structure is calculated using AlphaFold. Most preferably, the parent is SEQ ID NO: 6, and the variant has a TM-score of at least 0.990, e.g., at least 0.991, at least 0.992, at least 0.993, at least 0.994, at least 0.995, at least 0.996, at least 0.997, at least 0.998, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 6, wherein the three-dimensional structure is calculated using AlphaFold.

[0105] For some parent proteases, a three-dimensional structure is publicly available. A three-dimensional structure of SEQ ID NO: 1 (Savinase®) is available under UniProt accession number P29600 or, alternatively, PDB accession number 1SVN. A three-dimensional structure of SEQ ID NO: 2 (BPN’) is available under UniProt accession number P00782. A three-dimensional structure of SEQ ID NO: 3 (Alcalase®) is available under UniProt accession number P00780.

[0106] In one embodiment, the parent xyloglucanase is SEQ ID NO: 7 and the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 7, wherein the three-dimensional structure is calculated using AlphaFold. Preferably, the parent is SEQ ID NO: 7, and the variant has a TM-score of at least 0.980, e.g., at least 0.985, at least 0.990, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 7, wherein the three-dimensional structure is calculated using AlphaFold. Most preferably, the parent is SEQ ID NO: 7, and the variant has a TM-score of at least 0.990, e.g., at least 0.991, at least 0.992, at least 0.993, at least 0.994, at least 0.995, at least 0.996, at least 0.997, at least 0.998, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 7, wherein the three-dimensional structure is calculated using AlphaFold.

[0107] In one embodiment, the parent xyloglucanase is SEQ ID NO: 8 and the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, atleast 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 8, wherein the three-dimensional structure is calculated using AlphaFold. Preferably, the parent is SEQ ID NO: 8, and the variant has a TM-score of at least 0.980, e.g., at least 0.985, at least 0.990, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 8, wherein the three-dimensional structure is calculated using AlphaFold. Most preferably, the parent is SEQ ID NO: 8, and the variant has a TM-score of at least 0.990, e.g., at least 0.991, at least 0.992, at least 0.993, at least 0.994, at least 0.995, at least 0.996, at least 0.997, at least 0.998, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 8, wherein the three-dimensional structure is calculated using AlphaFold.

[0108] In one embodiment, the parent xyloglucanase is SEQ ID NO: 9 and the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 9, wherein the three-dimensional structure is calculated using AlphaFold. Preferably, the parent is SEQ ID NO: 9, and the variant has a TM-score of at least 0.980, e.g., at least 0.985, at least 0.990, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 9, wherein the three-dimensional structure is calculated using AlphaFold. Most preferably, the parent is SEQ ID NO: 9, and the variant has a TM-score of at least 0.990, e.g., at least 0.991, at least 0.992, at least 0.993, at least 0.994, at least 0.995, at least 0.996, at least 0.997, at least 0.998, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 9, wherein the three-dimensional structure is calculated using AlphaFold.

[0109] In another aspect, the invention relates to a composition, in particular a detergent composition, comprising a protease and a xyloglucanase, wherein

[0110] a) the protease is a variant of a parent protease, wherein the variant has protease activity and comprises an insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises substitutions at at least two, e.g., at least three, at least four, or five, positions corresponding to any of positions 188, 199, 200, 203, and 256 of SEQ ID NO: 1; wherein position numbering is based on the numbering of SEQ ID NO: 1; and wherein the variant has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98%, but less than 100% sequence identity to the parent protease;

[0111] b) the xyloglucanase is a variant of a parent xyloglucanase, wherein the variant has xyloglucanase activity and comprises substitutions at positions 111, 123, 129 and 159 of SEQ ID NO: 8, and wherein the variant has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity to the parent xyloglucanase.In one embodiment, the parent protease is SEQ ID NO: 1 and the variant has at least 60%, e.g., at least 65%, at least 70%, 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%, such as at least 96%, at least 97%, or at least 98%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1. In a preferred embodiment, the parent protease is SEQ ID NO: 1.

[0112] In one embodiment, the parent protease is SEQ ID NO: 2 and the variant has at least 60%, e.g., at least 65%, at least 70%, 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%, such as at least 96%, at least 97%, or at least 98%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 2. In a preferred embodiment, the parent protease is SEQ ID NO: 2.

[0113] In one embodiment, the parent protease is SEQ ID NO: 3 and the variant has at least 60%, e.g., at least 65%, at least 70%, 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%, such as at least 96%, at least 97%, or at least 98%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 3. In a preferred embodiment, the parent protease is SEQ ID NO: 3.

[0114] In one embodiment, the parent protease is SEQ ID NO: 4 and the variant has at least 60%, e.g., at least 65%, at least 70%, 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%, such as at least 96%, at least 97%, or at least 98%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 4. In a preferred embodiment, the parent protease is SEQ ID NO: 4.

[0115] In one embodiment, the parent protease is SEQ ID NO: 5 and the variant has at least 60%, e.g., at least 65%, at least 70%, 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%, such as at least 96%, at least 97%, or at least 98%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 5. In a preferred embodiment, the parent protease is SEQ ID NO: 5.

[0116] In one embodiment, the parent protease is SEQ ID NO: 6 and the variant has at least 60%, e.g., at least 65%, at least 70%, 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%, such as at least 96%, at least 97%, or at least 98%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 6. In a preferred embodiment, the parent protease is SEQ ID NO: 6.

[0117] In one embodiment, the parent protease is an otherwise identical protease without the insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1.

[0118] In one embodiment, the parent protease is an otherwise identical protease without the insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and without substitutions at at least two, e.g., at least three, at least four, or five, positions corresponding to any of positions 188, 199, 200, 203 and 256 of SEQ I D NO: 1.

[0119] In one embodiment, the parent xyloglucanase is SEQ ID NO: 7 and the variant has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, or at least 98%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 7. In a preferred embodiment, the parent xyloglucanase is SEQ ID NO: 7.

[0120] In one embodiment, the parent xyloglucanase is SEQ ID NO: 8 and the variant has at least 60%, e.g., at least 65%, at least 70%, 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%, such as at least 96%, at least 97%, or at least 98%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 8. In a preferred embodiment, the parent xyloglucanase is SEQ ID NO: 8.

[0121] In one embodiment, the parent xyloglucanase is SEQ ID NO: 9 and the variant has at least 60%, e.g., at least 65%, at least 70%, 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%, such as at least 96%, at least 97%, or at least 98%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 9. In a preferred embodiment, the parent xyloglucanase is SEQ ID NO: 9.

[0122] Protease variants

[0123] In one embodiment, the parent protease may be selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6.

[0124] In one embodiment, the protease variant comprises an insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises at least three substitutions at positions corresponding to any of positions 188, 199, 200, 203, and 256 of SEQ I D NO: 1.

[0125] In one embodiment, the protease variant comprises an insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises at least four substitutions at positions corresponding to any of positions 188, 199, 200, 203, and 256 of SEQ ID NO: 1.

[0126] In one embodiment, the protease variant comprises an insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises five substitutions at positions corresponding to each of positions 188, 199, 200, 203, and 256 of SEQ ID NO: 1.

[0127] In one embodiment, the protease variant comprises an insertion of Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises at least three substitutions at positions corresponding to any of positions 188, 199, 200, 203, and 256 of SEQ I D NO: 1.

[0128] In one embodiment, the protease variant comprises an insertion of Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises at least four substitutions at positions corresponding to any of positions 188, 199, 200, 203, and 256 of SEQ ID NO: 1.

[0129] In one embodiment, the protease variant comprises an insertion of Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises five substitutions at positions corresponding to each of positions 188, 199, 200, 203, and 256 of SEQ ID NO: 1.

[0130] In one embodiment, the protease variant comprises an insertion corresponding to S97SE of SEQ ID NO: 1 and further comprises at least two substitutions selected from the groupconsisting of substitutions corresponding to A188P, V199I, Q200L, Y203W, and L256E of SEQ ID NO: 1.

[0131] In one embodiment, the protease variant comprises an insertion corresponding S97SE of SEQ ID NO: 1 and further comprises at least three substitutions selected from the group consisting of substitutions corresponding to A188P, V199I, Q200L, Y203W, and L256E of SEQ ID NO: 1.

[0132] In one embodiment, the protease variant comprises an insertion corresponding to S97SE of SEQ ID NO: 1 and further comprises at least four substitutions selected from the group consisting of substitutions corresponding to A188P, V199I, Q200L, Y203W, and L256E of SEQ ID NO: 1.

[0133] In one embodiment, the protease variant comprises an insertion corresponding to S97SE of SEQ ID NO: 1 and further comprises five substitutions selected from the group consisting of substitutions corresponding to A188P, V199I, Q200L, Y203W, and L256E of SEQ ID NO: 1.

[0134] In one embodiment, any of the protease variants herein comprises an insertion Glu at a position corresponding to position 97 of SEQ ID NO: 1.

[0135] In another embodiment, any of the protease variants herein comprises at least two, e.g., at least three, at least four, or five, substitutions selected from the group consisting of substitutions corresponding to A188P, V199I, Q200L, Y203Wand L256E of SEQ ID NO: 1.

[0136] In another embodiment, any of the protease variants herein further comprises at least one, e.g., at least two, at least three, at least four, at least five, at least six, at least seven, or eight, substitutions at a position corresponding to any of positions 3, 4, 9, 95, 154, 206, 209 or 210 of SEQ ID NO: 1. In a preferred embodiment, any of the protease variants herein comprises at least one, e.g. , at least two, at least three, at least four, at least five, at least six, at least seven, or eight, substitutions selected from the group consisting of substitutions corresponding to S3T, V4I, S9R, G95D, S154D, S206G, A209K and S210V of SEQ ID NO: 1.

[0137] In another embodiment, any of the protease variants herein comprises at least one, e.g., two, three or four, substitutions selected from the group consisting of substitutions corresponding to S154D, S206G, A209K and S210V of SEQ ID NO: 1. In a preferred embodiment, any of the protease variants herein comprises the insertion S97SE and the substitutions S154D, A188P, V199I, Q200L, Y203W, S206G, A209K, S210V and L256E.

[0138] In one embodiment, the protease variant comprises, compared to the polypeptide of SEQ ID NO: 1, the insertion S97SE and the substitutions S154D, A188P, V199I, Q200L, Y203W, S206G, A209K, S210V and L256E, and has a TM-score of at least 0.90, e.g., at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of SEQ ID NO: 1 , wherein the three-dimensional structure is calculated using AlphaFold.

[0139] In another embodiment, the protease variant comprises, compared to the polypeptide of SEQ ID NO: 1, the insertion S97SE and the substitutions S154D, A188P, V199I, Q200L, Y203W,S206G, A209K, S210V and L256E, and has at least 60%, e.g., at least 65%, at least 70%, 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%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1.

[0140] In one embodiment, the protease variant has at least 90%, e.g., at least 95%, at least 96%, at least 97%, or at least 98%, but less than 100%, sequence identity to SEQ ID NO: 1 and comprises an insertion corresponding to S97SD or S97SE of SEQ ID NO: 1, and at least two, e.g., at least three, at least four, or five, substitutions selected from the group consisting of substitutions corresponding to A188P, V199I, Q200L, Y203W, and L256E of SEQ ID NO: 1. In a preferred embodiment, the variant comprisesan insertion corresponding to S97SEof SEQ ID NO: 1.

[0141] In one preferred embodiment, the protease variant is the polypeptide of SEQ ID NO: 1 comprising the insertion S97SE and the substitutions S154D, A188P, V199I, Q200L, Y203W, S206G, A209K, S210V and L256E.

[0142] In another preferred embodiment, the protease variant is the polypeptide of SEQ ID NO: 1 comprising the insertion S97SE and the substitutions A188P, V199l, Q200L, Y203W, S206G, A209K, S210V and L256E.

[0143] In another preferred embodiment, the protease variant is the polypeptide of SEQ ID NO: 1 comprising the insertion S97SE and the substitutions A188P, V199I, Q200L, Y203W, S206G, S210V and L256E.

[0144] It should be understood that any of the protease variants having the insertion and substitutions described herein preferably has a TM-score compared to the parent protease as described above, i.e., a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of the parent protease, wherein the three-dimensional structure is calculated using AlphaFold; and / or has a sequence identity to the parent protease as described above, i.e., at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98%, but less than 100% sequence identity to the parent protease.

[0145] Xyloglucanase variants

[0146] In one embodiment, the parent xyloglucanase may be selected from the group consisting of SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9. In some preferred embodiments, the parent xyloglucanase is SEQ ID NO: 8.

[0147] In a preferred embodiment, the parent xyloglucanase is SEQ ID NO: 8, and the variant comprises substitutions at positions 111, 123, 129 and 159 of SEQ ID NO: 8.In one embodiment, the xyloglucanase variant comprises one or more substitutions selected from the group consisting of P111Q, S123P, A129T and V159M, wherein position numbers correspond to positions of SEQ ID NO: 8. Preferably, the variant comprises two or more substitutions selected from the group consisting of P111Q, S123P, A129T and V159M, more preferably three or more substitutions selected from the group consisting of P111Q, S123P, A129T and V159M, most preferably all four substitutions P111 Q, S123P, A129T and V159M.

[0148] In one embodiment, the xycoglucanase variant further comprises at least one substitution at a position selected from the group consisting of 41, 82, 83, 87, 147, 203, 217, 240, 252, 256, 294, 347, 383 and 402, wherein position numbers correspond to positions of SEQ ID NO: 8. Preferably, the variant comprises a substitution at two or more of said positions, for example at three or more of said positions.

[0149] In preferred embodiments, the xyloglucanse variant comprises at least one substitution selected from the group consisting of A41L, Q82E, A83E, K87E, Q147K, V203T, K217R, K217T, K240F, K252E, S256E, S256Q, I294E, I294Q, K347E, N383E and S402Q. Preferably, the variant comprises two or more of said substitutions, for example three or more of said substitutions.

[0150] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 41, preferably A41L.

[0151] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 82, preferably Q82E.

[0152] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 83, preferably A83E.

[0153] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 87, preferably K87E.

[0154] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 147, preferably Q147K.

[0155] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two,three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 203, preferably V203T.

[0156] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 217, preferably K217R or K217T.

[0157] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 240, preferably K240F.

[0158] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 252, preferably K252E.

[0159] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 256, preferably S256E or S256Q.

[0160] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 294, preferably I294E or I294Q.

[0161] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 347, preferably K347E.

[0162] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 383, preferably N383E.

[0163] In one embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising substitutions at positions 111, 123, 129 and 159, preferably one, two, three or all of the substitutions P111Q, S123P, A129T and V159M, and further comprising a substitution at position 402, preferably S402Q.

[0164] In one preferred embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising the set of substitutions A41L, P111Q, S123P, A129T, Q147K, V159M and V203T.In one preferred embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising the set of substitutions A41L, P111Q, S123P, A129T, Q147K, V159M and K217R.

[0165] In one preferred embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising the set of substitutions P111Q, S123P, A129T, Q147K, V159M, I294Q and S402Q.

[0166] In one preferred embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising the set of substitutions A83E, P111Q, S123P, A129T, V159M, S256E and I294E.

[0167] In one preferred embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising the set of substitutions Q82E, P111Q, S123P, A129T, V159M, S256E and I294E.

[0168] In one preferred embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising the set of substitutions Q82E, P111Q, S123P, A129T, Q147K, V159M and I294E.

[0169] In one preferred embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising the set of substitutions K87E, P111Q, S123P, A129T, V159M, K217T and I294E.

[0170] In one preferred embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising the set of substitutions A83E, P111Q, S123P, A129T, V159M, K240F and K252E.

[0171] In one preferred embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising the set of substitutions K87E, P111Q, S123P, A129T, V159M, S256Q and I294E.

[0172] In one preferred embodiment, the polypeptide having xyloglucanase activity is a variant of SEQ ID NO: 7, 8 or 9 comprising the set of substitutions K87E, P111Q, S123P, A129T, V159M, K347E and N383E.

[0173] It should be understood that any of the xyloglucanase variants having the substitutions described herein preferably has a TM-score compared to the parent xyloglucanase as described above, i.e., a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of the parent xyloglucanase, wherein the three-dimensional structure is calculated using AlphaFold; and / or has a sequence identity to the parent xyloglucanase as described above, i.e., at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity to the parent xyloglucanase.The protease and / or xyloglucanase enzymes of the present invention may comprise one or more conservative amino acid substitutions that do not significantly alter their physico-chemical properties, such as thermal thermal stability, substrate specificity or pH optimum. Examples of conservative substitutions are within the groups of basic amino acids (arginine, lysine and histidine), acidic amino acids (glutamic acid and aspartic acid), polar amino acids (glutamine and asparagine), hydrophobic amino acids (leucine, isoleucine and valine), aromatic amino acids (phenylalanine, tryptophan and tyrosine), and small amino acids (glycine, alanine, serine, threonine and methionine). Amino acid substitutions that do not generally alter specific activity are well-known in the art and are described, for example, by H. Neurath and R.L. Hill, 1979, In, The Proteins, Academic Press, New York. Common substitutions are Ala / Ser, Val / lle, Asp / Glu, Thr / Ser, Ala / Gly, Ala / Thr, Ser / Asn, Ala / Val, Ser / Gly, Tyr / Phe, Ala / Pro, Lys / Arg, Asp / Asn, Leu / lle, Leu / Val, Ala / Glu, and Asp / Gly.

[0174] Methods of Production

[0175] Expression and purification of the protease and xyloglucanase 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 xyloglucanase may suitably be produced, e.g., as described in WO 2022 / 043321. Methods for production of recombinant proteases are well-known; see, for example, WO 2021 / 239818.

[0176] Compositions

[0177] The present invention provides cleaning / detergent compositions comprising a polypeptide having protease activity and a polypeptide having cellulase activity as disclosed elsewhere herein and at least one cleaning adjunct ingredient.

[0178] In particular, the present invention provides cleaning / detergent compositions comprising a polypeptide having protease activity and a polypeptide having xyloglucanase activity as disclosed elsewhere herein and at least one cleaning adjunct ingredient.

[0179] In an embodiment, the composition according to the invention has improved wash performance. In an embodiment, the composition according to the invention has improved wash performance at cold wash conditions. Preferably, the composition according to the invention has improved wash performance at a wash temperature of <20°C.

[0180] In an embodiment, the composition according to the invention has improved stain removal, preferably wherein the stain is a proteinaceous stain and / or a plant-derived, hemicellulosic food stain. In one embodiment, the composition according to the invention has improved broad stain removal activity, in particular improved stain removal activity against protein-, fruit-, vegetable-, and cereal-based stains. In a prefered embodiment, the composition according to the invention has improved stain removal, wherein the stain is a stain comprising at least a cereal-based material.In an embodiment, the composition according to the invention has improved wash performance as determined in Example 1 herein.

[0181] In an embodiment, the composition according to the invention has improved wash performance as determined in Example 2 herein.

[0182] 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.

[0183] 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.

[0184] The cleaning adjunct ingredient may include one or more enzymes other than the protease, cellulase or xyloglucanase 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, hexosaminidases, DNases, 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 xyloglucanase, typically a second protease. Specific enzymes suitable for the detergent compositions of the invention are described below.

[0185] 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 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 such as a tablet or pouch.

[0186] The amount of xyloglucanase 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 xyloglucanase 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 xyloglucanase 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.).

[0187] The protease may be used in amount similar to the amounts of xyloglucanase 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.

[0188] 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 dishwashing detergent. The laundry and dishwashing detergents may be phosphate-free.

[0189] Surfactants

[0190] The detergent composition may comprise one or more surfactants, which may be anionic and / or cationic and / or non-ionic and / or semi-polar and / or zwitterionic, or a mixture thereof. In a particular embodiment, the detergent composition includes a mixture of one or more nonionic surfactants and one or more anionic surfactants. The surfactant(s) is typically present at a level of from about 0.1% to 60% by weight, such as about 1% to about 40%, or about 3% to about 20%, or about 3% to about 10%. Surfactants may be chosen based on the desired cleaning application, and may include any conventional surfactants known in the art for for use in detergents. Surfactants lower the surface tension in the detergent, which allows the stain being cleaned to be lifted and dispersed and then washed away.

[0191] When included therein, the detergent will usually contain from about 1% to about 40% by weight, such as from about 5% to about 30%, including from about 5% to about 15%, or from about 20% to about 25% of an anionic surfactant. Non-limiting examples of anionic surfactants include sulfates and sulfonates, in particular, linear alkylbenzenesulfonates (LAS), isomers of LAS, branched alkylbenzenesulfonates (BABS), phenylalkanesulfonates, alpha-olefinsulfonates (AOS), olefin sulfonates, alkene sulfonates, alkane-2,3-diylbis(sulfates), hydroxyalkanesulfonates and disulfonates, alkyl sulfates (AS) such as sodium dodecyl sulfate (SDS), fatty alcohol sulfates (FAS), primary alcohol sulfates (PAS), alcohol ethersulfates (AES or AEOS or FES, also known as alcohol ethoxysulfates or fatty alcohol ether sulfates), secondary alkanesulfonates (SAS), paraffin sulfonates (PS), ester sulfonates, sulfonated fatty acid glycerol esters, alpha-sulfo fatty acid methyl esters (alpha-SFMe or SES) including methyl ester sulfonate (MES), alkyl- or alkenylsuccinic acid, dodecenyl / tetradecenyl succinic acid (DTSA), fatty acid derivatives of amino acids, diesters and monoesters of sulfo-succinic acid or soap, and combinations thereof.When included therein, the detergent will usually contain from about 0% to about 10% by weight of a cationic surfactant. Non-limiting examples of cationic surfactants include alklydimethylethanolamine quat (ADMEAQ), cetyltrimethylammonium bromide (CTAB), dimethyldistearylammonium chloride (DSDMAC), and alkylbenzyldimethylammonium, alkyl quaternary ammonium compounds, alkoxylated quaternary ammonium (AQA) compounds, and combinations thereof.

[0192] When included therein, the detergent will usually contain from about 0.2% to about 40% by weight of a non-ionic surfactant, for example from about 0.5% to about 30%, in particular from about 1% to about 20%, from about 3% to about 10%, such as from about 3% to about 5%, or from about 8% to about 12%. Non-limiting examples of non-ionic 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), polyhydroxy alkyl fatty acid amides, or N-acyl N-alkyl derivatives of glucosamine (glucamides, GA, or fatty acid glucamide, FAGA), as well as products available under the trade names SPAN and TWEEN, and combinations thereof.

[0193] When included therein, the detergent will usually contain from about 0% to about 10% by weight of a semipolar surfactant. Non-limiting examples of semipolar surfactants include amine oxides (AO) such as alkyldimethylamineoxide, N-(coco alkyl)-N,N-dimethylamine oxide and N-(tallow-alkyl)-N,N-bis(2-hydroxyethyl)amine oxide, fatty acid alkanolamides and ethoxylated fatty acid alkanolamides, and combinations thereof.

[0194] When included therein, the detergent will usually contain from about 0% to about 10% by weight of a zwitterionic surfactant. Non-limiting examples of zwitterionic surfactants include betaine, alkyldimethylbetaine, sulfobetaine, and combinations thereof.

[0195] Builders and co-builders

[0196] 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 orSTPP), carbonates such as sodium carbonate, soluble silicates such as sodium metasilicate, layered silicates (e.g., SKS-6from 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)iminodiaceticacid (EDG), aspartic acid-N-monoaceticacid (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.

[0197] Bleach components

[0198] 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 incleaning detergents may be utilized. Suitable bleaching system components include sources of hydrogen peroxide, sources of peracids, and bleach catalysts or boosters.

[0199] 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).

[0200] 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.

[0201] 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.

[0202] 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.

[0203] 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). Thebleach 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.

[0204] 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:

[0205]

[0206] (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.

[0207] 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.

[0208] 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.

[0209] Hydrotropes

[0210] 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.

[0211] Polymers

[0212] 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.

[0213] Fabric hueinq agents

[0214] 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 hueing 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.

[0215] Additional enzymes

[0216] The detergent composition may comprise one or more additional enzymes, such as an enzyme selected from the group consisting of lipases, cutinases, amylases, carbohydrases,cellulases, DNases, hexosaminidases, pectinases, mannanases, arabmases, galactanases, xylanases, licheninases, oxidases, e.g., a laccase or peroxidase. The composition may also include one or more additional proteases and / or one or more additional xyloglucanases.

[0217] 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.

[0218] Cellulases

[0219] Cellulases are enzymes that hydrolyze a cellulosic material. Cellulases may be selected from the group consisting of cellulases belonging to GH5, GH44, GH45, EC 3.2.1.4, EC 3.2.1.21, EC 3.2.1.91 and EC 3.2.1.172. Such enzymes include endoglucanases (e.g., EC 3.2.1.4), cellobiohydrolases, beta-glucosidases, or combinations thereof.

[0220] 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.

[0221] Especially suitable cellulases are the alkaline or neutral cellulases having colour care benefits. Examples of such cellulases are cellulases described in EP 0495257, EP 0531 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.

[0222] 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.

[0223] 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).

[0224] Proteases

[0225] 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 beof 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.

[0226] 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.

[0227] 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.

[0228] 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.

[0229] 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.

[0230] 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.

[0231] 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, Progress® Excel and Progress® Power (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 inFigure 29 of US 5352604) and variants hereof (Henkel AG), and KAP (Bacillus alkalophilus subtilisin) from Kao.

[0232] Lipases and Cutinases

[0233] 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).

[0234] 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.

[0235] Preferred commercial lipase products include Lipolase™, Lipex™; Lipolex™ and Lipoclean™ (Novozymes A / S), Lumafast (originally from Genencor) and Lipomax (originally from Gist-Brocades).

[0236] 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).

[0237] Amylases

[0238] Suitable amylases which can be used together with the protease and xyloglucanase of the invention may be an alpha-amylase ora 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.

[0239] 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 WO94 / 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.

[0240] 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.

[0241] 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 or more of the following positions: G48, T49, G107, H156, A181, N190, M197, 1201, A209 and Q264.

[0242] Other examples are amylase variants such as those described in WO 2011 / 098531, WO 2013 / 001078 and WO 2013 / 001087.

[0243] 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).

[0244] Peroxidases / Oxidases

[0245] Suitable peroxidases / oxidases include those of plant, bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Examples of useful peroxidases include peroxidases from Coprinus, e.g., from C. cinereus, and variants thereof as those described in WO 93 / 24618, WO 95 / 10602, and WO 98 / 15257.

[0246] Commercially available peroxidases include Guardzyme™ (Novozymes A / S).

[0247] DNases

[0248] 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.

[0249] The DNase polypeptide is typically a microbial enzyme, preferably of fungal or bacterial origin, or a genetically engineered variant of a microbial DNase.

[0250] 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 proposedsix 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.

[0251] 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.

[0252] 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, WO 2022 / 194673, WO 2022 / 189521 and WO 2023 / 165950.

[0253] Hexosaminidases

[0254] The detergent compositions of the invention may comprise one or more hexosaminidases. The term hexosaminidase includes “dispersin’’ and the abbreviation “Dsp”, which means 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 e.g., in biofilm. The term hexosaminidase includes polypeptides having N-acetylglucosaminidase activity and p-N-acetylglucosaminidase activity.

[0255] A polypeptide having hexosaminidase activity may be obtained from microorganisms of any genus, in particular from bacteria or fungi. Preferably the hexosaminidase, e.g., a dispersin, is obtained from Terribacillus, Curtobacterium, Aggregatibacter, Haemophilus or Actinobacillus, preferably Terribacillus. The hexosaminidase may also be a variant of a polypeptide obtained from any of these or other organisms.

[0256] Suitable hexosaminidases include those disclosed in WO2017186936, WO2017186937, WO2017186943, WO2017207770, WO2018184873, WO2019086520, WO2019086528 WO2019086530, WO2019086532, WO2019086521, WO2019086526, W02020002604 W02020002608, W02020007863, W02020007875, W02020008024, W02020070063 W02020070249, W02020088957, W02020088958, W02020207944 and WO2023194204.Dispersants

[0257] 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.

[0258] Dye Transfer Inhibiting Agents

[0259] 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.

[0260] Fluorescent whitening agents

[0261] 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.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%.

[0262] Soil release polymers

[0263] 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.

[0264] Anti-redeposition agents

[0265] 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.

[0266] Rheology Modifiers

[0267] 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, hydroxy-functional 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.

[0268] 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.

[0269] Other materials

[0270] 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.

[0271] Formulation of detergent products

[0272] 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.

[0273] Pouches can be configured as single or multiple compartments. 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 asglycerol, 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.

[0274] 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.

[0275] A liquid or gel detergent which is not unit dosed will typically 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 nonaqueous.

[0276] 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.

[0277] 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.

[0278] 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.

[0279] 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 suchas 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.

[0280] Granular detergent formulations

[0281] 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.

[0282] 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.

[0283] A layered granule may comprise (a) a core, which may be enzymatic or non-enzymatic; (b) a coating surrounding the core, wherein the coating comprises at least one enzyme; and (c) optionally a coating, such as a salt coating, consisting of one or more layer(s) surrounding the enzyme containing coating. In this embodiment, the core (a) may optionally comprise the protease, while the coating (b) comprises the xyloglucanase and possibly other enzymes; or the core (a) may optionally comprise the xyloglucanase and possibly other enzymes, while the coating (b) comprises the protease. Or, in this embodiment, the core (a) may optionally comprise the protease, while the coating (b) comprises the cellulase and possibly other enzymes; or the core (a) may optionally comprise the xyloglucanase and possibly other enzymes, while the coating (b) comprises the protease.

[0284] 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.Formulation of enzyme in co-qranule

[0285] The protease and cellulase or xyloglucanase 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 xyloglucanase and one or more other (non-protease) enzymes, where the latter may be in the form of a co-granule comprising the xyloglucanase and one or more other (non-protease) enzymes.

[0286] 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.

[0287] 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.

[0288] Liquid Formulations

[0289] The present invention also relates to liquid compositions comprising a protease and cellulase of the invention.

[0290] In particular, the present invention also relates to liquid compositions comprising a protease and xyloglucanase 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).

[0291] 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.

[0292] 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.In another embodiment, the invention relates to liquid formulations comprising:

[0293] (A) 0.001-25% w / w, such as 0.001-5%, of a xyloglucanase disclosed herein, and 0.001-25% w / w, such as 0.001-5%, of a protease disclosed herein;

[0294] (B) 20-80% w / w of polyol;

[0295] (C) optionally 0.001-2% w / w preservative; and

[0296] (D) water.

[0297] In another embodiment, the invention relates to liquid formulations comprising:

[0298] (A) 0.001-25% w / w, such as 0.001-5%, of a xyloglucanase disclosed herein, and 0.001-25% w / w, such as 0.001-5%, of a protease disclosed herein;

[0299] (B) 0.001-2% w / w preservative;

[0300] (C) optionally 20-80% w / w of polyol; and

[0301] (D) water.

[0302] 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.

[0303] 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).

[0304] 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.

[0305] In another embodiment, the liquid formulation further comprises one or more additional enzymes, e.g., as described above.

[0306] Laundry Soap Bars

[0307] The protease and cellulase enzymes of the invention may be added to laundry soap bars and used for hand washing laundry, fabrics and / or textiles.

[0308] The protease and xyloglucanase enzymes of the invention may be added to laundry soap bars and used for hand washing laundry, fabrics and / or textiles. The term laundry soap bar includes laundry bars, soap bars, combo bars, syndet bars and detergent bars. The types of bar usually differ in the type of surfactant they contain, and the term laundry soap bar includes those containing soaps from fatty acids and / or synthetic soaps. Laundry soap bars have a physical form which is solid at room temperature.

[0309] The laundry soap bar may contain one or more additional enzymes, protease inhibitors such as peptide aldehydes (or hydrosulfite adductor hemiacetal adduct), boric acid, borate, borax and / or phenylboronic acid derivatives such as 4-formylphenylboronic acid, one or more soaps or synthetic surfactants, polyols such as glycerin, pH controlling compounds such as fatty acids, citric acid, acetic acid and / or formic acid, and / or a salt of a monovalent cation and an organic anion wherein the monovalent cation may be for example Na+, K+, or NH4+and the organic anion may be for example formate, acetate, citrate, or lactate such that the salt of a monovalent cation and an organic anion may be, for example, sodium formate.

[0310] The laundry soap bar may also contain complexing agents such as EDTA and HEDP, perfumes and / or different type of fillers, surfactants, e.g., anionic synthetic surfactants, builders, polymeric soil release agents, detergent chelators, stabilizing agents, fillers, dyes, colorants, dye transfer inhibitors, alkoxylated polycarbonates, suds suppressers, structurants, binders, leaching agents, bleaching activators, clay soil removal agents, anti-redeposition agents, polymeric dispersing agents, brighteners, fabric softeners, perfumes and / or other compounds known in the art.

[0311] The laundry soap bar may be processed in conventional laundry soap bar making equipment such as, but not limited to, mixers, plodders, e.g., a two-stage vacuum plodder, extruders, cutters, logo-stampers, cooling tunnels and wrappers. A premix containing a soap, the enzyme of the invention, optionally one or more additional enzymes, a protease inhibitor, and a salt of a monovalent cation and an organic anion may be prepared and the mixture is then plodded. The enzyme and optional additional enzymes may be added at the same time as the proteaseinhibitor for example in liquid form. Besides the mixing step and the plodding step, the process may further comprise the steps of milling, extruding, cutting, stamping, cooling and / or wrapping.

[0312] Use of compositions of the invention

[0313] The compositions of the invention comprising a cellulase and a protease are suitable for use in a cleaning process, for example for laundry or hard surface cleaning, in particular for laundry.

[0314] The compositions of the invention comprising a xyloglucanase and a protease are suitable for use in a cleaning process, for example for laundry or hard surface cleaning, in particular for laundry.

[0315] One aspect of the invention thus relates to use of a detergent or cleaning composition as described herein for cleaning an object, for example wherein the object is a textile or a hard surface such as dishware, preferably wherein the object is a textile.

[0316] A related aspect relates to use of a polypeptide having protease activity as described herein and a polypeptide having cellulase activity, e.g., endoglucanase activity, as described herein for cleaning an object, for example wherein the object is a textile or a hard surface such as dishware, preferably wherein the object is a textile.

[0317] Another related aspect relates to use of a polypeptide having protease activity as described herein and a polypeptide having xyloglucanase activity as described herein for cleaning an object, for example wherein the object is a textile or a hard surface such as dishware, preferably wherein the object is a textile.

[0318] For hard surface cleaning, the compositions may be used to clean any suitable surface, for example floors, tables, walls, roofs etc. as well as surfaces of hard objects such as cars (car wash) and dishes (automatic or hand dishwashing), as well as, e.g., kitchen surfaces including kitchen appliance surfaces or bathroom surfaces.

[0319] For laundry, the compositions will typically be used as normal laundry detergent compositions, especially for machine washing, but may also be used, e.g., as a fabric pretreatment.

[0320] The invention further relates to a method for laundering a textile item, the method comprising:

[0321] a) exposing the item to a wash liquor comprising a detergent composition as disclosed herein comprising a polypeptide having protease activity and a polypeptide having cellulase activity, such as, e.g., endoglucanase activity;

[0322] b) completing at least one wash cycle; and optionally

[0323] c) rinsing the item.

[0324] The invention also further relates to a method for laundering a textile item, the method comprising:a) exposing the item to a wash liquor comprising a detergent composition as disclosed herein comprising a polypeptide having protease activity and a polypeptide having xyloglucanase activity;

[0325] b) completing at least one wash cycle; and optionally

[0326] c) rinsing the item.

[0327] Another aspect of the invention relates to a method for laundering a textile item, the method comprising:

[0328] a) exposing the item to a wash liquor comprising a polypeptide having protease activity as defined herein and a polypeptide having cellulase activity as defined herein; b) completing at least one wash cycle; and optionally

[0329] c) rinsing the item.

[0330] Another aspect of the invention relates to a method for laundering a textile item, the method comprising:

[0331] a) exposing the item to a wash liquor comprising a polypeptide having protease activity as defined hterein and a polypeptide having xyloglucanase activity as defined herein;

[0332] b) completing at least one wash cycle; and optionally

[0333] c) rinsing the item.

[0334] 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.

[0335] 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.

[0336] The concentration of the xyloglucanase in the wash liquor is typically in the range of from 0.00001 mg / l to 10 mg / l enzyme protein, 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 xyloglucanase, e.g., 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.

[0337] The invention described and claimed herein is not to be limited in scope by the specific aspects herein disclosed, since these aspects are intended as illustrations of several aspects of the invention. Any equivalent aspects are intended to be within the scope of this invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are alsointended to fall within the scope of the appended claims. In the case of conflict, the present disclosure including definitions will control.

[0338] The invention is further defined by the following numbered paragraphs:

[0339] 1. A detergent composition comprising a protease, a xyloglucanase and at least one detergent adjunct ingredient, wherein

[0340] a) the protease is variant of a parent protease, wherein the variant has protease activity and comprises an insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises substitutions at at least two, e.g., at least three, at least four, or five, positions corresponding to any of positions 188, 199, 200, 203, and 256 of SEQ ID NO: 1 ; wherein position numbering is based on the numbering of SEQ ID NO: 1; wherein the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91 , at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of the parent protease, wherein the three-dimensional structure is calculated using AlphaFold; and

[0341] b) the xyloglucanase is variant of a parent xyloglucanase, wherein the variant has xyloglucanase activity and comprises substitutions at positions 111, 123, 129 and 159 of SEQ ID NO: 8, wherein the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of the parent xyloglucanase, wherein the three-dimensional structure is calculated using AlphaFold.

[0342] 2. A detergent composition comprising a protease, a xyloglucanase and at least one detergent adjunct ingredient, wherein

[0343] a) the protease is variant of a parent protease, wherein the variant has protease activity and comprises an insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises substitutions at at least two, e.g., at least three, at least four, or five, positions corresponding to any of positions 188, 199, 200, 203, and 256 of SEQ ID NO: 1 ; wherein position numbering is based on the numbering of SEQ ID NO: 1 ; and wherein the variant has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98%, but less than 100% sequence identity to the parent protease; andb) the xyloglucanase is variant of a parent xyloglucanase, wherein the variant has xyloglucanase activity and comprises substitutions at positions 111, 123, 129 and 159 of SEQ ID NO: 8, and wherein the variant has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity to the parent xyloglucanase.

[0344] 3. The composition of paragraph 1 or 2, wherein the parent protease is selected from the group consisting of SEQ ID NO: 1 , SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; preferably wherein the parent protease is SEQ ID NO: 1.

[0345] 4. The composition of any of the preceding paragraphs, wherein the protease variant comprises an insertion Glu at a position corresponding to position 97 of SEQ ID NO: 1.

[0346] 5. The composition of any of the preceding paragraphs, wherein the protease variant comprises at least two, e.g., at least three, at least four, or five, substitutions selected from the group consisting of substitutions corresponding to A188P, V199I, Q200L, Y203W, and L256E of SEQ ID NO: 1.

[0347] 6. The composition of any of the preceding paragraphs, wherein the protease variant further comprises at least one, e.g., at least two, at least three, at least four, at least five, at least six, at least seven, or eight, substitutions at a position corresponding to any of positions 3, 4, 9, 95, 154, 206, 209 or 210 of SEQ ID NO: 1.

[0348] 7. The composition of paragraph 6, wherein the protease variant comprises at least one, e.g., at least two, at least three, at least four, at least five, at least six, at least seven, or eight, substitutions selected from the group consisting of substitutions corresponding to S3T, V4I, S9R, G95D, S154D, S206G, A209K and S210V of SEQ ID NO: 1.

[0349] 8. The composition of any of the preceding paragraphs, wherein the protease variant comprises at least one, e.g., two, three or four, substitutions selected from the group consisting of substitutions corresponding to S154D, S206G, A209K and S210V of SEQ ID NO: 1.

[0350] 9. The composition of any of the preceding paragraphs, wherein the protease variant comprises the insertion S97SE and the substitutions S154D, A188P, V199I, Q200L, Y203W, S206G, A209K, S210V and L256E.10. The composition of any of the preceding paragraphs, wherein the protease variant is the polypeptide of SEQ ID NO: 1 comprising the insertion S97SE and the substitutions S154D, A188P, V199I, Q200L, Y203W, S206G, A209K, S210V and L256E.

[0351] 11. The composition of any of the preceding paragraphs, wherein the parent xyloglucanase is selected from the group consisting of SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9; preferably wherein the parent xyloglucanase is SEQ ID NO: 8.

[0352] 12. The composition of any of the preceding paragraphs, wherein the xyloglucanase variant comprises comprises one or more, preferably two, three or four, substitutions selected from the group consisting of P111Q, S123P, A129T and V159M, wherein position numbers correspond to positions of SEQ ID NO: 8.

[0353] 13. The composition of any of the preceding paragraphs, wherein the xycoglucanase variant further comprises at least one substitution at a position selected from the group consisting of 41 , 82, 83, 87, 147, 203, 217, 240, 252, 256, 294, 347, 383 and 402, wherein position numbers correspond to positions of SEQ ID NO: 8, preferably wherein the variant comprises a substitution at two or more of said positions, for example at three or more of said positions.

[0354] 14. The composition of paragraph 13, wherein the xyloglucanase variant comprises at least one substitution selected from the group consisting of A41L, Q82E, A83E, K87E, Q147K, V203T, K217R, K217T, K240F, K252E, S256E, S256Q, I294E, I294Q, K347E, N383E and S402Q, preferably wherein the variant comprises two or more of said substitutions, for example three or more of said substitutions.

[0355] 15. The composition of any of the preceding paragraphs, wherein the xyloglucanase variant comprises a set of substitutions selected from the group consisting of:

[0356] A41 L+P111Q+S123P+A129T+Q147K+V159M+V203T,

[0357] A41 L+P111Q+S123P+A129T+Q147K+V159M+K217R,

[0358] P111Q+S123P+A129T+Q147K+V159M+I294Q+S402Q,

[0359] A83E+P111Q+S123P+A129T+V159M+S256E+I294E,

[0360] Q82E+P111Q+S123P+A129T+V159M+S256E+I294E,

[0361] Q82E+P111Q+S123P+A129T+Q147K+V159M+I294E,

[0362] K87E+P111Q+S123P+A129T+V159M+K217T+I294E,

[0363] A83 E+ P 111 Q+S 123 P+ A 129T+ V159M + K240 F+ K252 E,

[0364] K87E+P111Q+S123P+A129T+V159M+S256Q+I294E, and

[0365] K87E+P111Q+S123P+A129T+V159M+K347E+N383E.16. The composition of paragraph 15, wherein the xyloglucanase variant is the polypeptide of SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9 comprising one of said sets of substitutions.

[0366] 17. The composition of any of the preceding paragraphs, wherein the xyloglucanase variant is the polypeptide of SEQ ID NO: 8 comprising the substitutions A83E, P111Q, S123P, A129T, V159M, S256E and I294E.

[0367] 18. The composition of any of the preceding paragraphs, wherein the detergent adjunct ingredient comprises at least one surfactant, builder or bleach component.

[0368] 19. The composition of any of the preceding paragraphs, comprising at least one surfactant, preferably comprising at least one anionic surfactant

[0369] 20. The composition of any of the preceding paragraphs, comprising at least one builder.

[0370] 21. The composition of any of the preceding paragraphs, comprising at least one bleach component.

[0371] 22. The composition of any of the preceding paragraphs, further comprising at least one additional enzyme selected from the group consisting of proteases, lipases, cutinases, amylases, carbohydrases, cellulases, DNases, hexosaminidases, pectinases, mannanases, arabinases, galactanases, xylanases, licheninases, xyloglucanases and oxidases.

[0372] 23. The composition of any of the preceding paragraphs, wherein the composition is in the form of a bar, a homogenous tablet, a tablet having two or more layers, a unit dose product such as a pouch having one or more compartments, a regular or compact powder, a granule, a paste, a gel, or a regular or concentrated liquid.

[0373] 24. The composition of any of the preceding paragraphs, wherein the composition is a laundry detergent composition or an automatic dishwashing composition.

[0374] 25. The composition of any of the preceding paragraphs, wherein the composition provides an improved wash performance, e.g., relative to a corresponding composition without the polypeptide having protease activity and / or the polypeptide having xyloglucanase activity, in particular in laundry.25a. The composition of any of paragraphs 1-24, wherein the composition provides an improved wash performance, e.g., relative to a corresponding composition without the protease according to any of paragraphs 1-25, in particular in laundry.

[0375] 25b. The composition of any of the preceding paragraphs, wherein the composition provides an improved wash performance at cold wash conditions, e.g., at a wash temperature of < 20°C.

[0376] 26. Use of a detergent composition comprising a polypeptide having protease activity, a polypeptide having xyloglucanase activity and at least one detergent adjunct ingredient, wherein the composition is as defined in any of paragraphs 1-25b, for cleaning an object, wherein the object is a textile or a hard surface such as dishware, preferably wherein the object is a textile.

[0377] 27. A method for laundering a textile item, the method comprising:

[0378] a) exposing the item to a wash liquor a comprising detergent composition comprising a polypeptide having protease activity, a polypeptide having xyloglucanase activity and at least one detergent adjunct ingredient, wherein the composition is as defined in any of paragraphs 1-25b;

[0379] b) completing at least one wash cycle; and optionally

[0380] c) rinsing the item.

[0381] 28. A detergent composition comprising a protease, a cellulase and at least one detergent adjunct ingredient, wherein

[0382] i) the protease is variant of a parent protease, wherein the variant has protease activity and comprises an insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises substitutions at at least two, e.g., at least three, at least four, or five, positions corresponding to any of positions 188, 199, 200, 203, and 256 of SEQ ID NO: 1 ; wherein position numbering is based on the numbering of SEQ ID NO: 1; wherein the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91 , at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of the parent protease, wherein the three-dimensional structure is calculated using AlphaFold; or

[0383] ii) the protease is a variant of a parent protease, wherein the variant has protease activity and comprises an insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises substitutions at at least two, e.g., at least three, at least four, or five, positions corresponding to any of positions 188, 199, 200, 203, and 256 of SEQ ID NO: 1 ; wherein position numbering is based on the numbering of SEQ ID NO: 1 ; and wherein the variant has at least 60%, e.g., at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98%, but less than 100% sequence identity to the parent protease.

[0384] 29. The composition of paragraph 28, wherein the parent protease is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; preferably wherein the parent protease is SEQ ID NO: 1.

[0385] 30. The composition of any of paragraphs 28-29, wherein the protease variant comprises an insertion Glu at a position corresponding to position 97 of SEQ ID NO: 1.

[0386] 31. The composition of any of paragraphs 28-30, wherein the protease variant comprises at least two, e.g., at least three, at least four, or five, substitutions selected from the group consisting of substitutions corresponding to A188P, V199I, Q200L, Y203W, and L256E of SEQ ID NO: 1.

[0387] 32. The composition of any of paragraphs 28-31, wherein the protease variant further comprises at least one, e.g., at least two, at least three, at least four, at least five, at least six, at least seven, or eight, substitutions at a position corresponding to any of positions 3, 4, 9, 95, 154, 206, 209 or 210 of SEQ ID NO: 1.

[0388] 33. The composition of paragraph 32, wherein the protease variant comprises at least one, e.g., at least two, at least three, at least four, at least five, at least six, at least seven, or eight, substitutions selected from the group consisting of substitutions corresponding to S3T, V4I, S9R, G95D, S154D, S206G, A209K and S210V of SEQ ID NO: 1.

[0389] 34. The composition of any of paragraphs 28-33, wherein the protease variant comprises at least one, e.g., two, three or four, substitutions selected from the group consisting of substitutions corresponding to S154D, S206G, A209K and S210V of SEQ ID NO: 1.

[0390] 35. The composition of any of paragraphs 28-34, wherein the protease variant comprises the insertion S97SE and the substitutions S154D, A188P, V199I, Q200L, Y203W, S206G, A209K, S210V and L256E.

[0391] 36. The composition of any of paragraphs 28-35, wherein the protease variant is the polypeptide of SEQ ID NO: 1 comprising the insertion S97SE and the substitutions S154D, A188P, V199I, Q200L, Y203W, S206G, A209K, S210V and L256E.

[0392] 37. The composition of any of paragraphs 28-36, wherein the cellulase has endoglucanase activity, preferably wherein the cellulase is derived or obtained from Humicola insolens.38. The composition of any of paragraphs 28-37, wherein the cellulase is according to SEQ ID NO:10.

[0393] 39. The composition of any of paragraphs 28-38, wherein the detergent adjunct ingredient comprises at least one surfactant, builder or bleach component.

[0394] 40. The composition of any of paragraphs 28-39, comprising at least one surfactant, preferably comprising at least one anionic surfactant.

[0395] 41. The composition of any of paragraphs 28-40, comprising at least one builder and / or at least one bleach component.

[0396] 42. The composition of any of paragraphs 28-41, further comprising at least one additional enzyme selected from the group consisting of proteases, lipases, cutinases, amylases, carbohydrases, cellulases, DNases, hexosaminidases, pectinases, mannanases, arabinases, galactanases, xylanases, licheninases, xyloglucanases and oxidases.

[0397] 43. The composition of any of paragraphs 28-42, wherein the composition is in the form of a bar, a homogenous tablet, a tablet having two or more layers, a unit dose product such as a pouch having one or more compartments, a regular or compact powder, a granule, a paste, a gel, or a regular or concentrated liquid.

[0398] 44. The composition of any of paragraphs 28-43, wherein the composition is a laundry detergent composition or an automatic dishwashing composition.

[0399] 45. The composition of any of paragraphs 28-44, wherein the composition provides an improved wash performance, e.g., relative to a corresponding composition without the protease according to any of paragraphs 28-44, in particular in laundry.

[0400] 46. The composition of any of preceding paragraphs 28-44, wherein the composition provides an improved wash performance at cold wash conditions, e.g., at a wash temperature of < 20°C.

[0401] 47. Use of a detergent composition comprising a polypeptide having protease activity, a polypeptide having cellulase activity and at least one detergent adjunct ingredient, wherein the composition is as defined in any of paragraphs 28-46, for cleaning an object, wherein the object is a textile or a hard surface such as dishware, preferably wherein the object is a textile.48. A method for laundering a textile item, the method comprising:

[0402] a) exposing the item to a wash liquor a comprising detergent composition comprising a polypeptide having protease activity, a polypeptide having cellulase activity and at least one detergent adjunct ingredient, wherein the composition is as defined in any of paragraphs 28-46;

[0403] b) completing at least one wash cycle; and optionally

[0404] c) rinsing the item.

[0405] The present invention is further described by the following examples that should not be construed as limiting the scope of the invention.

[0406] EXAMPLES

[0407] Materials and Methods

[0408] Production and purification of enzyme variants

[0409] Enzymes and enzyme variants for use in the present invention are prepared by standard procedures, in brief, typically: Introducing random and / or site-directed mutations into a gene encoding an enzyme of interest, transforming relevant host cells, for example cells from a species of Bacillus such as Bacillus subtilis, with a mutated gene of interest, fermenting the transformed host cells, and obtaining the enzyme or variant from the fermentation broth using purification procedures known in the art.

[0410] Protease variants for use in the present invention may, for example, be produced and purified using the procedure described in European patent application No. 24203579.8 or in WO 2024 / 121070.

[0411] Xyloglucanase variants for use in the present invention may, for example, be produced and purified using the procedure described in WO 2022 / 043321.

[0412] Example 1 : Terg-o-tometer (TOM) Wash Assay

[0413] Materials and methods

[0414] The Terg-o-tometer (TOM) is a medium scale wash assay that can be applied to simultaneously test up to 16 different conditions at the same time. Briefly, it consists of 16 x 2 L metal beakers, each fitted with an agitator, which rotate in a back-and-forth manner at a controlled speed to simulate the agitation occurring in commercial top-loader washing machines. The beakers are partly submerged in thermostatic water baths where the temperature can be controlled. Each beaker is filled with 1 L detergent solution, and test swatches, ballast and enzymes are added to the requisite levels. After a timed wash period, the swatches are promptlyremoved from the beakers and rinsed thoroughly. The swatches are then spread out flat on a rack covered with filter paper, covered, and allowed to dry overnight at room temperature. All washes are evaluated the day after the wash. Light reflectance evaluations of the swatches are done using a DataColor® Model 800V reflectance spectrophotometer. The measurements are made without UV in the incident light and remission (REM) at 460 nm is extracted. Measurements are made on unwashed and washed swatches. The test swatch to be measured is placed on top of another swatch of the same type and color.

[0415] Table 1: TOM wash experimental conditions

[0416] European liquid model detergent (Ell liquid) or plant-based Detergent

[0417] green detergent (Green)

[0418] 1.7 g / L (Ell liquid detergent)

[0419] Detergent dosage

[0420] 2.5 g / L (Green detergent)

[0421] Test solution volume 1 L

[0422] Wash time 20 minutes

[0423] Agitation 120 rpm

[0424] Temperature 30°C

[0425] Water hardness 14°dH (4:1:7.5 Ca2+:Mg2+:HCO3’)

[0426] Protease concentration in 19.9 nM (EU liquid detergent)

[0427] test solution 29.2 nM (Green detergent)

[0428] Xyloglucanase concentration 0.88 nM (EU liquid detergent)

[0429] in test solution 1.29 nM (Green detergent)

[0430] 2 swatches (5 cm x 5 cm) per TOM beaker of PC-05 (Blood, milk, ink) and KC-S-54 (Oatmeal, chocolate, aged); and 1 swatch (5 cm x 9 cm) per TOM beaker of KC-H097 Test material

[0431] (Oatmeal, chocolate); all from Center for Testmaterials B.V., Stoomloggerweg 11, 3133 KT Vlaardingen, The

[0432] Netherlands

[0433]

[0434] The European model detergent was produced by mixing water, propylene glycol, triethanolamine and sodium hydroxide. Then topped palm kernel fatty acid, Na-LAS, SLES and AEO were added in portions and stirred fora few hours. Trisodium citrate dihydrate, DTPMP Na7 and 2-phenoxyethanol were added and pH was adjusted to the target after overnight stirring at room temperature. Commercial raw materials were used, and the dosage of the raw materialswas adjusted with the purity of the individual ingredients to achieve the listed active ingredient content.

[0435] Table 2. European liquid model detergent composition

[0436] Weight % of active Ingredient

[0437] ingredient

[0438] Sodium linear alkylbenzene sulfonate (Na-LAS) 12.0% Sodium lauryl ether sulfate (SLES) 4.1% Alcohol ethoxylate with 7 mol EO (AEO) 12.0% Topped Palm Kernel Fatty Acid 3.0% Sodium hydroxide 0.5% Ethanol 6.0% Propylene glycol (MPG) 2.0% Triethanolamine (TEA) 2.0% Trisodium citrate dihydrate 3.9% Diethylenetriaminepentakis(methylene)pentakis(phosphonic

[0439] 1.6%

[0440] acid), heptasodium salt (DTPMP Na7)

[0441] 2-Phenoxyethanol 0.5% Water (demineralized) Add to 100%

[0442] pH target 8.4

[0443]

[0444] The plant-based green liquid laundry detergent formulation was produced by mixing water, AES (SLES, 2EO), alkyl polyglucoside (APG), sodium citrate, soap, glycerol, sodium chloride and phenoxyethanol, all added in portions, and pH was adjusted to the target after overnight stirring at room temperature. Commercial raw materials were used, and the dosage of raw materials was adjusted with the purity of the individual ingredients to achieve the listed active ingredient content.

[0445] Table 3. Plant-based green detergent composition

[0446] Weight % of active

[0447] Ingredient

[0448] ingredient

[0449] SLES, 2EO 8

[0450] APG, C8-C14 glucoside 4

[0451] Sodium citrate 4

[0452] Phenoxyethanol 0.5

[0453] Soap, coco fatty acid 2

[0454] Glycerol 4

[0455]

[0456] Sodium chloride 0.5

[0457] Water (demineralized) Add to 100%

[0458] pH target 8.5

[0459]

[0460] The enzymes used were the following:

[0461] Xyloglucanase: SEQ ID NO: 8 with the substitutions A83E, P111Q, S123P, A129T, V159M, S256E and I294E

[0462] Protease 1 : SEQ ID NO: 1

[0463] Protease 2: SEQ ID NO: 1 with the substitutions Y161A, R164S, A188P

[0464] Protease 3: SEQ ID NO: 1 with the insertion S97SE and the substitutions S154D, A188P, V199I, Q200L, Y203W, S206G, A209K, S210V and L256E

[0465] The Xyloglucanase and Protease 3 correspond to enzymes disclosed and claimed herein.

[0466] Wash performance in Terg-o-tometer (TOM) assay

[0467] Wash experiments were performed in order to assess the wash performance of the protease variants in a laundry model detergent with different combinations of proteases and xyloglucanase in two different detergent formulations. The results are provided in Tables 3 and 4 below as remission values, where a higher remission value (more reflected light) indicates a better cleaning effect. Tables 5 and 6 show the same data as Tables 3 and 4, but expressed as delta remission values (AREM), calculated as the remission value for a treatment with a protease and / or a xyloglucanase minus the remission value without enzymes.

[0468] Table 4: Remission values on PC-05, KC-H097 and KC-S-54 stain swatches from TOM washes in European liquid model detergent with different combinations of protease and xyloglucanase Protease Xyloglucanase PC-05 KC-H097 KC-S-54 No protease No xyloglucanase 17.3 38.5 46.3 Protease 1 No xyloglucanase 18.7 41.2 51.0 Protease 2 No xyloglucanase 21.1 41.4 52.8 Protease 3 No xyloglucanase 22.2 41.6 54.7 No Protease Xyloglucanase 17.7 47.0 50.4 Protease 1 Xyloglucanase 18.0 48.9 53.9 Protease 2 Xyloglucanase 23.1 50.3 58.1 Protease 3 Xyloglucanase 24.5 54.2 63.9

[0469]

[0470] Table 5: Remission values on PC-05, KC-H097 and KC-S-54 stain swatches from TOM washes in plant-based green detergent with different combinations of protease and xyloqlucanase Protease Xyloglucanase PC-05 KC-H097 KC-S-54 No protease No xyloglucanase 16.9 37.0 43.9 Protease 1 No xyloglucanase 18.4 39.5 49.2 Protease 2 No xyloglucanase 20.2 39.3 51.2 Protease 3 No xyloglucanase 21.2 40.7 53.9 No Protease Xyloglucanase 16.7 48.2 47.7 Protease 1 Xyloglucanase 17.7 49.8 54.1 Protease 2 Xyloglucanase 20.8 50.1 57.4 Protease 3 Xyloglucanase 23.5 54.1 60.8

[0471]

[0472] Table 6: Delta remission values on PC-05, KC-H097 and KC-S-54 stain swatches from TOM washes in European liquid model detergent with different combinations of protease and xyloglucanase

[0473] Protease Xyloglucanase PC-05 KC-H097 KC-S-54 No protease No xyloglucanase 0.0 0.0 0.0 Protease 1 No xyloglucanase 1.4 2.7 4.7 Protease 2 No xyloglucanase 3.8 2.9 6.5 Protease 3 No xyloglucanase 4.9 3.1 8.4 No Protease Xyloglucanase 0.4 8.5 4.1 Protease 1 Xyloglucanase 0.7 10.4 7.6 Protease 2 Xyloglucanase 5.8 11.8 11.8 Protease 3 Xyloglucanase 7.2 15.7 17.6

[0474]

[0475] Table 7: Delta remission values on PC-05, KC-H097 and KC-S-54 stain swatches from TOM washes in plant-based green detergent with different combinations of protease and xyloglucanase Protease Xyloglucanase PC-05 KC-H097 KC-S-54 No protease No xyloglucanase 0.0 0.0 0.0 Protease 1 No xyloglucanase 1.5 2.5 5.3

[0476]

[0477] Protease Xyloglucanase PC-05 KC-H097 KC-S-54 Protease 2 No xyloglucanase 3.3 2.3 7.3 Protease 3 No xyloglucanase 4.3 3.7 10.0 No Protease Xyloglucanase

[0478] Protease 1 Xyloglucanase 0.8 12.8 10.2 Protease 2 Xyloglucanase 3.9 13.1 13.5 Protease 3 Xyloglucanase 6.6 17.1 16.9

[0479]

[0480] It may be seen from the above data that the three proteases and the xyloglucanase act differently on the three stains. The PC-05 stain (blood, milk, ink) is a protease-sensitive stain, and all three proteases are active on this stain, especially proteases 2 and 3, whereas little or no effect is seen by the xyloglucanase. The KC-H097 stain (oatmeal with chocolate), on the other hand, shows a good effect of the xyloglucanase and a smaller effect of the proteases. The proteases as well as the xyloglucanase all show an effect individually on the KC-S-54 stain (oatmeal with chocolate, aged), but with a smaller effect by the xyloglucanase.

[0481] Significantly, the data in Tables 4, 5, 6 and 7 shows that for each of the three stains and in both detergents, the combination of a protease and a xyloglucanase according to the invention, i.e., Protease 3 and the xyloglucanase, clearly provides the best result. Moreover, the combination of Protease 3 and xyloglucanase provides a synergistic effect in each case. This may be seen by comparing the delta remission values in Tables 6 and 7 for the combination of Protease 3 and xyloglucanase with the sum of the delta remission values for Protease 3 without xyloglucanase and for xyloglucanase without protease.

[0482] Example 2: Terg-o-tometer (TOM) Wash Assay testing cold wash conditions

[0483] Cold wash experiments were performed in order to assess the wash performance of the protease variants in a laundry model detergent with different combinations of proteases and xyloglucanase or cellulase in two different detergent formulations.

[0484] The enzymes used were the following:

[0485] Xyloglucanase: SEQ ID NO: 8 with the substitutions A83E, P111Q, S123P, A129T, V159M, S256E and I294E

[0486] Protease 1 : SEQ ID NO: 1.

[0487] Protease 2: SEQ ID NO: 1 with the substitutions Y161A, R164S, A188P.

[0488] Protease 3: SEQ ID NO: 1 with the insertion S97SE and the substitutions S154D, A188P, V199I, Q200L, Y203W, S206G, A209K, S210V and L256E.

[0489] Cellulase: SEQ ID NO: 10.The Xyloglucanase, Cellulase and Protease 3 correspond to enzymes disclosed and claimed herein.

[0490] To evaluate wash performance, the Terg-o-tometer (TOM) wash assay as described in Example 1 was used (see Table 1), except wash temperature was 20°C and water hardness 5.5 °dH. Wash performance was tested in the European model detergent according to Table 2 and the plant-based green liquid laundry detergent formulation according to Table 3. Cellulase was dosed at same concentration as the Xyloglucanase (see Table 1). In addition to the stains mentioned in Table 1, two swatches (5 cm x 5 cm) per TOM beaker of PC-03 (Chocolate milk with Carbon Black) (from Center for Testmaterials B.V., Stoomloggerweg 11, 3133 KT Vlaardingen, The Netherlands) was included in the tests.

[0491] The results are provided in Table 8 and Table 9 below as remission values, where a higher remission value (more reflected light) indicates a better cleaning effect.

[0492] Table 8: Remission values on PC-05, KC-H097, KC-S-54 and PC-03 stain swatches from TOM washes in European liquid model detergent with different combinations of protease and xyloglucanase or cellulase

[0493] Xyloglucanase

[0494] Protease PC-05 KC-H097 KC-S-54 PC-03

[0495] / Cellulase

[0496] Protease 1 No 19.0 38.5 44.4 50.5 Protease 2 No 18.9 39.4 43.9 49.8 Protease 3 No 22.4 42.4 45.5 52.5 No Xyloglucanase 18.0 47.8 44.4 50.3 Protease 1 Xyloglucanase 19.2 48.2 44.8 50.1 Protease 2 Xyloglucanase 20.2 47.4 45.8 49.8 Protease 3 Xyloglucanase 21.9 50.8 48.5 53.2 Protease 1 Cellulase 19.8 42.1 45.0 49.5 Protease 2 Cellulase 20.5 44.8 44.6 49.7 Protease 3 Cellulase 21.8 45.9 47.4 53.6

[0497]

[0498] Table 9: Remission values on PC-05, KC-H097, KC-S-54 and PC-03 stain swatches from TOM washes in plant-based green detergent with different combinations of protease and xyloqlucanase or cellulase

[0499] Xyloglucanase

[0500] Protease PC-05 KC-H097 KC-S-54 PC-03

[0501] / Cellulase

[0502] Protease 1 No 19.1 38.6 42.1 48.3 Protease 2 No 19.4 37.3 42.6 48.2 Protease 3 No 21.8 39.4 45.6 53.1 No Xyloglucanase 18.2 48.7 41.5 47.9 Protease 1 Xyloglucanase 19.0 48.2 43.8 48.4 Protease 2 Xyloglucanase 19.9 46.8 45.9 49.6 Protease 3 Xyloglucanase 21.4 51.4 46.7 52.4 Protease 1 Cellulase 19.3 42.6 44.6 49.0 Protease 2 Cellulase 19.8 43.5 46.0 50.1 Protease 3 Cellulase 22.4 45.7 49.6 53.2

[0503]

[0504] It may be seen from the above data that the three proteases and the xyloglucanase and cellulase act differently on the four stains.

[0505] The data shown in Tables 8 and 9 demonstrate that Protease 3 consistently outperforms benchmark Protease 1 and Protease 2 when combined with either of the tested cellulase or xyloglucanase across both detergent systems. In both the European model detergent and the plant-based green detergent, the combinations comprising Protease 3 with a cellulase or a xyloglucanase provide the highest remission values across the tested stains, including protease-sensitive (PC-05), polysaccharide-rich (KC-H097, KC-S-54) and mixed stains (PC-03).

[0506] Interestingly, the superior performance is observed under cold wash conditions (wash temperature of 20°C), demonstrating that Protease 3 provides enhanced cleaning performance at low temperature. The improvement is observed independently of the companion enzyme used, showing that Protease 3 interacts more effectively with the cellulase and xyloglucanase compared to the benchmark proteases. While Protease 1 and Protease 2 show incremental improvements when combined with a cellulase / xyloglucanase, the corresponding combinations with Protease 3 consistently result in higher absolute wash performance.

Claims

CLAIMS1. A detergent composition comprising a protease, a xyloglucanase and at least one detergent adjunct ingredient, whereina) the protease is a variant of a parent protease, wherein the variant has protease activity and comprises an insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises substitutions at at least two, e.g., at least three, at least four, or five, positions corresponding to any of positions 188, 199, 200, 203, and 256 of SEQ ID NO: 1 ; wherein position numbering is based on the numbering of SEQ ID NO: 1; wherein the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91 , at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of the parent protease, wherein the three-dimensional structure is calculated using AlphaFold; andb) the xyloglucanase is a variant of a parent xyloglucanase, wherein the variant has xyloglucanase activity and comprises substitutions at positions 111, 123, 129 and 159 of SEQ ID NO: 8, wherein the variant has a TM-score of at least 0.80, e.g., at least 0.81, at least 0.82, at least 0.83, at least 0.84, at least 0.85, at least 0.86, at least 0.87, at least 0.88, at least 0.89, at least 0.90, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 0.995, at least 0.999, but less than 1.0, compared to the three-dimensional structure of the parent xyloglucanase, wherein the three-dimensional structure is calculated using AlphaFold.

2. A detergent composition comprising a protease, a xyloglucanase and at least one detergent adjunct ingredient, whereina) the protease is a variant of a parent protease, wherein the variant has protease activity and comprises an insertion of Asp or Glu at a position corresponding to position 97 of SEQ ID NO: 1 and further comprises substitutions at at least two, e.g., at least three, at least four, or five, positions corresponding to any of positions 188, 199, 200, 203, and 256 of SEQ ID NO: 1 ; wherein position numbering is based on the numbering of SEQ ID NO: 1; and wherein the variant has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98%, but less than 100% sequence identity to the parent protease; and b) the xyloglucanase is a variant of a parent xyloglucanase, wherein the variant has xyloglucanase activity and comprises substitutions at positions 111, 123, 129 and 159 of SEQ ID NO: 8, and wherein the variant has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least97%, at least 98%, or at least 99%, but less than 100% sequence identity to the parent xyloglucanase.

3. The composition of claim 1 or 2, wherein the parent protease is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; preferably wherein the parent protease is SEQ ID NO: 1.

4. The composition of any of the preceding claims, wherein the protease variant comprises an insertion Glu at a position corresponding to position 97 of SEQ ID NO: 1.

5. The composition of any of the preceding claims, wherein the protease variant comprises at least two, e.g., at least three, at least four, or five, substitutions selected from the group consisting of substitutions corresponding to A188P, V199I, Q200L, Y203W, and L256E of SEQ ID NO: 1.

6. The composition of any of the preceding claims, wherein the protease variant further comprises at least one, e.g., at least two, at least three, at least four, at least five, at least six, at least seven, or eight, substitutions at a position corresponding to any of positions 3, 4, 9, 95, 154, 206, 209 or 210 of SEQ ID NO: 1; preferably wherein the protease variant comprises at least one, e.g., at least two, at least three, at least four, at least five, at least six, at least seven, or eight, substitutions selected from the group consisting of substitutions corresponding to S3T, V4I, S9R, G95D, S154D, S206G, A209K and S210V of SEQ ID NO: 1.

7. The composition of any of the preceding claims, wherein the protease variant comprises at least one, e.g., two, three or four, substitutions selected from the group consisting of substitutions corresponding to S154D, S206G, A209K and S210V of SEQ ID NO: 1.

8. The composition of any of the preceding claims, wherein the protease variant comprises the insertion S97SE and the substitutions S154D, A188P, V199I, Q200L, Y203W, S206G, A209K, S210V and L256E; e.g., wherein the protease variant is the polypeptide of SEQ ID NO: 1 comprising the insertion S97SE and the substitutions S154D, A188P, V199I, Q200L, Y203W, S206G, A209K, S210V and L256E.

9. The composition of any of the preceding claims, wherein the parent xyloglucanase is selected from the group consisting of SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9; preferably wherein the parent xyloglucanase is SEQ ID NO: 8.

10. The composition of any of the preceding claims, wherein the xyloglucanase variant comprises one or more, preferably two, three or four, substitutions selected from the group consisting of P111Q, S123P, A129T and V159M, wherein position numbers correspond to positions of SEQ ID NO: 8.

11. The composition of any of the preceding claims, wherein the xycoglucanase variant further comprises at least one substitution at a position selected from the group consisting of 41, 82, 83, 87, 147, 203, 217, 240, 252, 256, 294, 347, 383 and 402, wherein position numbers correspond to positions of SEQ ID NO: 8, preferably wherein the variant comprises a substitution at two or more of said positions; preferably wherein the xyloglucanse variant comprises at least one substitution selected from the group consisting of A41L, Q82E, A83E, K87E, Q147K, V203T, K217R, K217T, K240F, K252E, S256E, S256Q, I294E, I294Q, K347E, N383E and S402Q, preferably wherein the variant comprises two or more of said substitutions.

12. The composition of any of the preceding claims, wherein the xyloglucanase variant comprises a set of substitutions selected from the group consisting of:A41 L+P111 Q+S123P+A129T+Q147K+V159M+V203T,A41 L+P111Q+S123P+A129T+Q147K+V159M+K217R,P111 Q+S123P+A129T+Q147K+V159M+I294Q+S402Q,A83E+P111 Q+S123P+A129T+V159M+S256E+I294E,Q82E+P111Q+S123P+A129T+V159M+S256E+I294E,Q82E+P111Q+S123P+A129T+Q147K+V159M+I294E,K87E+P111Q+S123P+A129T+V159M+K217T+I294E,A83E+P111Q+S123P+A129T+V159M+K240F+K252E,K87E+P111 Q+S123P+A129T+V159M+S256Q+I294E, andK87E+P111Q+S123P+A129T+V159M+K347E+N383E.

13. The composition of any of the preceding claims, comprising at least one detergent adjunct ingredient selected from surfactants, builders and bleach components, and optionally comprising at least one additional enzyme.

14. Use of a detergent composition comprising a polypeptide having protease activity, a polypeptide having xyloglucanase activity and at least one detergent adjunct ingredient, wherein the composition is as defined in any of claims 1-13, for cleaning an object, wherein the object is a textile or a hard surface such as dishware, preferably wherein the object is a textile.

15. A method for laundering a textile item, the method comprising:a) exposing the item to a wash liquor comprising a detergent composition comprising a polypeptide having protease activity, a polypeptide having xyloglucanase activity and at least one detergent adjunct ingredient, wherein the composition is as defined in any of claims 1-13;b) completing at least one wash cycle; and optionallyc) rinsing the item.

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