Composition

Abstract

The present invention relates to a detergent composition comprising from 0.0005 to 6% by weight of a lipase from Mortiella mucilaginosa and from 1 to 60% by weight of a detersive surfactant. The invention also relates to a method of treating textiles having fat stains wherein the textiles are treated with a detergent composition according to any of claims 1-10 to provide enhanced lipolytic cleaning against fat stains; relates to a method of treating a textile having fat stains wherein the textile is pretreated with a detergent composition according to claim 1 or claim 2 to provide enhanced lipolytic cleaning against fat stains, the textile being subsequently washed with a laundry main washing composition; the invention also relates to the use of a lipase from Mosla mucilaginosa for improving the cleaning of fat stains on textiles.

Description

TECHNICAL FIELD

[0001] The present invention relates to compositions, in particular detergent compositions, comprising a specific lipase. BACKGROUND

[0002] Lipases incorporated in cleaning compositions, in particular detergent compositions, are useful ingredients. They perform particularly well in cleaning fatty stains. However, lipases are expensive ingredients and there is always a need to improve their performance.

[0003] It is an object of the present invention to improve the performance of lipases against fatty stains, in particular margarine.

[0004] It is also an object of the present invention to improve the stability of lipases in combination with other common detergent ingredients, in particular proteases and soil release polymers. SUMMARY

[0005] We have found that a lipase from Moritella viscosa solves these problems.

[0006] In one aspect, the present invention provides a detergent composition comprising:

[0007] (a) 0.0005 to 6 wt.%, preferably 0.005 to 4 wt.%, more preferably 0.001 to 2 wt.% of a lipase from Moritella viscosa; and

[0008] (b) 1 to 60 wt.%, preferably 1 to 50 wt.%, more preferably 1 to 35 wt.% of a soil release surfactant.

[0009] Preferably, the present invention provides a detergent composition comprising:

[0010] (a) 0.0005 to 6 wt.%, preferably 0.005 to 4 wt.%, more preferably 0.001 to 2 wt.% of a lipase from Moritella viscosa; and

[0011] (b) 1 to 60 wt.%, preferably 2 to 50 wt.%, more preferably 4 to 35 wt.% of a soil release surfactant.

[0012] Preferably, the lipase from Moritella viscosa has at least 70% sequence identity to SEQ. ID. 1. Preferably, the lipase from Moritella viscosa has at least 75%, preferably 80%, more preferably 85%, even more preferably 90%, even more preferably 95%, even more preferably 98%, even more preferably 99%, most preferably 100% sequence identity to SEQ. ID. 1.

[0013] Preferably, the anionic surfactant is present in an amount of 1-50 wt%, preferably 2-40 wt%, more preferably 3-30 wt%, and is preferably selected from linear alkylbenzene sulfonates, secondary alkane sulfonates, sodium laureth sulfate, sodium lauryl sulfate, sodium oleyl sulfate and sodium oleyl ether sulfate, methyl ester sulfonates, secondary alkyl sulfates (SALS), cardanol ether sulfate and rhamnolipids.

[0014] Preferably, the nonionic surfactant is present in an amount of 1-30 wt%, preferably 2-20 wt%, more preferably 3-15 wt%, and is preferably selected from alcohol ethoxylates, alcohol propoxylates, methyl ester ethoxylates and alkyl polyglycosides.

[0015] Preferred detergent compositions are laundry detergent compositions. Preferably, the laundry detergent composition is in the form of a liquid, solid, powder, pastille, bead or paste. More preferably the composition is a liquid or powder, most preferably a liquid detergent.

[0016] The laundry detergent preferably comprises an alkoxylated polyamine, preferably in an amount of 0.1 to 8 wt%, more preferably 0.2 to 6 wt%, most preferably 0.5 to 5 wt%.

[0017] The laundry detergent preferably comprises a soil release polymer, preferably selected from copolyesters of dicarboxylic acids and polyglycols, more preferably copolyesters formed by the condensation of terephthalate and 1,2-propylene glycol, preferably present in an amount of 0.1-8 wt%, more preferably 0.2-6 wt%, most preferably 0.5-5 wt%.

[0018] Preferred detergent compositions, especially laundry detergent compositions additionally comprise one or more additional enzymes selected from the group consisting of proteases, cellulases, alpha-amylases, peroxidases / oxidases, pectate lyases and / or mannanases.Preferably, the one or more enzymes comprise a protease.

[0019] In another aspect, the present invention provides a method of treating a textile having fatty stains, wherein the textile is treated with a detergent composition according to the first aspect of the invention to provide enhanced lipolytic cleaning of fatty stains, preferably margarine stains, the textile being preferably subsequently rinsed and dried.

[0020] In another aspect, the present invention provides a method of treating textiles having fatty stains, wherein the textiles are pre-treated with a composition according to the first aspect of the invention to provide enhanced lipolytic cleaning of fatty stains, preferably margarine stains, and the textiles are then washed with a laundry main wash composition and then preferably rinsed and dried.

[0021] In another aspect, the present application provides a lipase from Mycobacterium adhesivum, preferably a lipase from Mycobacterium adhesivum having at least 70% sequence identity to SEQ. ID. 1, for use in improving cleaning of fatty stains, preferably margarine stains, on a textile.

[0022] More preferably, the present application provides a detergent composition comprising a lipase from Mycobacterium adhesivum, preferably a lipase from Mycobacterium adhesivum having at least 70% sequence identity to SEQ. ID. 1, for use in improving cleaning of fatty stains, preferably margarine stains, on a textile. DETAILED DESCRIPTION

[0023] As used herein, the indefinite articles "a" or "an," and their corresponding definite article "the," mean at least one, or one or more, unless indicated otherwise.

[0024] All % amounts of ingredients in the compositions (formulations) listed herein are % by weight based on the total formulation, unless otherwise specified.

[0025] It will be appreciated that any reference to preferred ingredients of the detergent composition is contemplated in combination with any other preferred ingredients of the detergent compositions disclosed herein.

[0026] The detergent composition can be applied to any suitable substrate. A particularly preferred substrate is a textile. A particularly preferred detergent composition is a laundry detergent composition.

[0027] The laundry detergent composition can take any suitable form. Preferably, the laundry detergent composition is in the form of a liquid, solid, powder, tablet, bead or paste, preferably the composition is a liquid or a powder, more preferably a liquid detergent.

[0028] Lipase from Mycobacterium adhesivum MorvLip

[0029] Lipases (E.C. 3.1.1.3) are hydrolytic enzymes known to cleave ester bonds in lipids. The lipases of the present application are from the species Mycobacterium adhesivum. These lipases from Mycobacterium adhesivum are referred to herein as MorvLip.

[0030] A preferred lipase from Mycobacterium adhesivum has at least 70% sequence identity to SEQ. ID. 1. Preferably, the lipase from Mycobacterium adhesivum has at least 75%, preferably 80%, more preferably 85%, even more preferably 90%, even more preferably 95%, even more preferably 98%, even more preferably 99%, most preferably 100% sequence identity to SEQ. ID. 1.

[0031] The composition comprises 0.0005 to 6 wt%, preferably 0.005 to 4 wt%, more preferably 0.001 to 2 wt% of the lipase from Mycobacterium adiacens. Other preferred amounts include 0.001 to 1 wt% of the lipase.

[0032] Percent sequence identity

[0033] Percent (%) sequence identity is defined as the percent of amino acid residues in a candidate sequence that are identical with the residues in the given sequence (indicated by SEQ ID No.) after aligning the sequences and introducing gaps, if necessary, to achieve the maximum sequence identity, and not considering any conservative substitutions as part of the sequence identity. Sequence identity is calculated over the entire length of the respective sequences.

[0034] In the case of aligned sequences of different lengths, the sequence identity of the shorter comparison sequence can be determined over the entire length of the longer given sequence, or in the case of a comparison sequence that is longer than the given sequence, the sequence identity of the comparison sequence can be determined over the entire length of the shorter given sequence.

[0035] For example, in the case of a given sequence comprising 100 amino acids and a candidate sequence comprising 10 amino acids, the candidate sequence can have a maximum identity of only 10% over the entire length of the given sequence. This is further illustrated in the following examples:

[0036] (A)

[0037] Given seq: XXXXXXXXX XXXXXXXX (15 amino acids)

[0038] Comparison seq: XXXXX YYYY YYYYYY (12 amino acids)

[0039] % sequence identity = number of identical matching amino acid residues after alignment divided by the total number of amino acid residues in the longer given sequence, i.e. (5 divided by 15) x 100 = 33.3%

[0040] In the case of a comparison sequence that is longer than the given sequence, the sequence identity can be determined over the entire length of the given sequence. For example:

[0041] (B)

[0042] Given seq: XXXXXXXXX (10 amino acids)

[0043] Comparison seq: XXXXX YYYY YYY ZZZ ZZZ ZZZZ (20 amino acids)

[0044] % sequence identity = (number of identical amino acids after alignment divided by the total number of amino acid residues in the given sequence) x 100 = (5 divided by 10) x 100 = 50%.

[0045] Alignments for the purpose of determining percent amino acid sequence identity can be achieved in various ways known to persons skilled in the art, for example, using publicly available computer software such as ClustalW 1.82.T-coffee or Megalign (DNASTAR) software. When using such software, the default parameters are preferably used, for example for gap penalty and extension penalty. The default parameters for ClustalW 1.82 are: Protein Gap Open Penalty = 10.0, Protein Gap Extension Penalty = 0.2, Protein Matrix = Gonnet, Protein / DNA ENDGAP = -1, Protein / DNA GAPDIST = 4.

[0046] Identity of nucleic acid sequences can be determined in a similar manner, including aligning the sequences and introducing gaps, if necessary, to achieve maximum sequence identity, and calculating the sequence identity over the entire length of the respective sequences. In case the sequences to be aligned have different lengths, the sequence identity can be determined as described above and illustrated in examples (A) and (B).

[0047] The most preferred lipase is given by Sequence ID No. 1 (SEQ.ID.1). The letters refer to the amino acids of the protein sequence.

[0048] SEQ. ID. 1

[0049] MHKKIISPTDSLKTQHLELLESQVPHYRQAYSDRTAWLMACMSELAYVKFNPLYPNKFTEEAISSTLQKLSSTTLDPRVGKILSTLSSMSYDHVEEKKLLESELSELNGAILLDTFDCNGTQAMLVELASFNILAFRGTEATSLKDIKADIKAVTTRCETGGQVHSGFKDAFDEVCDSIVGALKAINNNKPLMITGHSLGGALATIAAKRLTFKNGIAGCYTFGSPRVGDDKWISTVKTPIYRVVNAADVVTMLPPNSIAIESASAVFSLVPYAGEVIKNTLLSKFNGYIHGGNMRYLTNCEAGNYANVKLLYSVSFLYRIKALLMGKLPVKKLAADHSISIYRQKLALVALQRKSLGDQTIKTGTKQ

[0050] Surfactant

[0051] The composition is a detergent composition. The detergent composition comprises a surfactant (which can include a mixture of two or more surfactants). The composition comprises 1-60 wt%, more preferably 1-50 wt%, most preferably 1-35 wt% of detersive surfactant.

[0052] More preferably, the composition comprises 1-60 wt%, more preferably 2-50 wt%, most preferably 4-35 wt% of detersive surfactant. Even more preferred levels of surfactant are 6-35 wt%, more preferably 8-35 wt%.

[0053] The detergent composition (preferably a laundry detergent composition) comprises anionic and / or nonionic surfactant, preferably comprises both anionic and nonionic surfactant.

[0054] Anionic surfactants are described in Anionic Surfactants Organic Chemistry (Surfactant Science Series Vol. 56) edited by H.W. Stache (Marcel Dekker 1996).

[0055] Preferably, the composition comprises 1-50 wt%, preferably 2-40 wt%, more preferably 3-30 wt% of anionic surfactant, based on the total weight of the composition. Non-soap anionic surfactants useful in the present application are generally salts of organic sulphates and sulphonates having alkyl groups containing from about 8 to about 22 carbon atoms, the term "alkyl" being used to include the alkyl portion of higher acyl groups. Examples of such materials include alkyl sulphates, alkyl ether sulphates, alkyl sulphonates, alpha-olefinsulphonates and mixtures thereof. The alkyl groups preferably contain from 10 to 18 carbon atoms and can be unsaturated. The alkyl ether sulphates can contain from 1 to 10 ethylene or propylene oxide units per molecule, preferably from 1 to 3 ethylene oxide units. The counterions of the anionic surfactants are usually alkali metals such as sodium or potassium; or ammonium counterions such as monoethanolamine (MEA), diethanolamine (DEA) or triethanolamine (TEA). Mixtures of these counterions can also be used.

[0056] The composition according to the present application can comprise an alkyl benzene sulfonate, in particular a linear alkyl benzene sulfonate (LAS) having an alkyl chain length of 10 to 18 carbon atoms. Commercial LAS are mixtures of closely related isomers and homologous alkyl chains, each containing an aromatic ring sulfonated in the "para" position and attached to a linear alkyl chain at any position other than the terminal carbon. The linear alkyl chain typically has a chain length of 11 to 15 carbon atoms, with the major species having a chain length of about C12. Each alkyl chain homolog is composed of a mixture of all possible sulfophenyl isomers except the 1-phenyl isomer. LAS are typically formulated into the composition in the acid form (i.e. HLAS) and then at least partially neutralized in situ.

[0057] Some alkyl sulfate surfactants (PAS) can be used, such as non-ethoxylated primary and secondary alkyl sulfate salts with alkyl chain lengths of 10 to 18.

[0058] Also typically used in laundry liquid compositions are alkyl ether sulfates having a linear or branched alkyl group containing 10 to 18, more preferably 12 to 14 carbon atoms and containing on average 1 to 3 EO units per molecule. A preferred example is sodium lauryl ether sulfate (SLES) in which the major C12 lauryl alkyl group is ethoxylated with an average of 3 EO units per molecule.

[0059] The alkyl ether sulfates can be provided as a single raw material component or through a mixture of components.

[0060] Preferred anionic surfactants also include C16 / 18 alkyl ether sulfates.

[0061] Preferred anionic surfactants also include rhamnolipids.

[0062] The anionic surfactant is preferably selected from linear alkyl benzene sulfonate, secondary alkane sulfonate, sodium laureth sulfate, sodium lauryl sulfate, sodium oleyl sulfate and sodium oleyl ether sulfate, methyl ester sulfonate, secondary alkyl sulfate (SALS), cardanol ether sulfate, and rhamnolipids.

[0063] Mixtures of any of the above materials can also be used.

[0064] Preferably, the detergent composition comprises a non-ionic surfactant, preferably 1-30 wt%, preferably 2-20 wt%, more preferably 3-15 wt% of a non-ionic surfactant.

[0065] Suitable non-ionic detergent compounds which can be used include, in particular, compounds having aliphatic hydrophobic groups and reactive hydrogen atoms, such as aliphatic alcohols, acids or amides, especially the reaction products of ethylene oxide or a mixture of ethylene oxide and propylene oxide. The non-ionic surfactant is preferably selected from alcohol ethoxylates, alcohol propoxylates, methyl ester ethoxylates and alkyl polyglycosides.

[0066] Preferred nonionic detergent compounds are aliphatic C8-C 18 The condensation products of straight-chain or branched primary or secondary alcohols with ethylene oxide.

[0067] Most preferably, the nonionic detergent compound is an alkyl ethoxylated nonionic surfactant, which is a C8-C10 alkyl ethoxylated surfactant having an average ethoxylation of 7EO to 9EO units. 18 Primary alcohol.

[0068] Preferably the surfactant used is saturated.

[0069] The compositions of the present invention may contain one or more amphoteric surfactants (e.g. zwitterionic surfactants), preferably wherein the amphoteric surfactant, if present, is present at a level of from 0.1 to 15 wt%, preferably from 0.5 to 10 wt%, more preferably from 1 to 5 wt%, and is preferably selected from alkyl betaines and alkyl sultaines (sultaines), more preferably carbobetaines and laurylamine oxide.

[0070] Further specific amphoteric surfactants include alkylamine oxides, alkylamidopropyl betaines, alkyl glycinates, alkylcarboxyglycinates, alkyl amphoacetates, alkyl amphopropionates, alkyl amphoglycinates, alkylamidopropyl hydroxysulfobetaines, acyl taurates and acyl glutamates.

[0071] The amphoteric surfactant preferably comprises an alkyl group containing from about 8 to about 22 carbon atoms, preferably selected from C12, C14, C16, C18 and C18:1, the term "alkyl" being used to include the alkyl portion of higher acyl groups.

[0072] Mixtures of any of the above materials may also be used.

[0073] Detergent polymer

[0074] When the detergent composition is in the form of a laundry composition, it is preferred to include a soil release polymer.

[0075] The soil release polymer is preferably present at a level of from 0.1 to 10% by weight.

[0076] The soil release polymer is preferably present in an amount of from 0.1 to 8 wt%, more preferably from 0.2 to 6 wt%, most preferably from 0.5 to 5 wt%.

[0077] Preferably, the soil release polymer is a polyester-based soil release polymer. More preferably, the polyester soil release polymer is selected from copolyesters of a dicarboxylic acid and a polyglycol. More preferably, the soil release polymer is a soil release polymer based on polyethylene terephthalate and / or polypropylene terephthalate, most preferably a soil release polymer based on polypropylene terephthalate, most preferably a copolyester formed by condensation of terephthalate and 1,2-propanediol.

[0078] Suitable polyester-based soil release polymers are described in WO2014 / 029479 and WO2016 / 005338.

[0079] Alkoxylated polyamine

[0080] When the detergent composition is in the form of a laundry composition, it is preferred to include an alkoxylated polyamine.

[0081] The preferred level of the alkoxylated polyamine is in the range of 0.1 to 8 wt%, preferably 0.2 to 6 wt%, more preferably 0.5 to 5 wt%. Another preferred level is 1 to 4 wt%.

[0082] The alkoxylated polyamine can be linear or branched. It can be branched to the extent that it is a dendrimer. The alkoxylation can typically be ethoxylation or propoxylation, or a mixture of the two. When the nitrogen atoms are alkoxylated, the preferred average degree of alkoxylation is 10 to 30, preferably 15 to 25.

[0083] A preferred material is an alkoxylated polyethyleneimine, most preferably an ethoxylated polyethyleneimine, having an average degree of ethoxylation of 10 to 30, preferably 15 to 25, in which the nitrogen atoms are ethoxylated.

[0084] Additional enzymes

[0085] Additional enzymes other than the specified lipase can be present in the detergent composition. It is preferred that additional enzymes are present in the preferred laundry detergent compositions.

[0086] If present, each additional enzyme is present in the laundry composition of the application at a level of from 0.0001 wt% to 0.1 wt%.

[0087] The level of enzyme present in the composition is preferably related to the level of enzyme as pure protein.

[0088] Preferred additional enzymes include those in the group consisting of proteases, cellulases, alpha-amylases, peroxidases / oxidases, pectate lyases and / or mannanases. The preferred additional enzymes include mixtures of two or more of these enzymes.

[0089] Preferably, the additional enzyme is selected from the group consisting of: a protease, a cellulase and / or an alpha-amylase. Most preferably, the additional enzyme comprises a protease.

[0090] Proteases hydrolyse bonds within peptides and proteins, which in the context of laundry results in enhanced removal of protein or peptide containing stains. Examples of suitable protease families include aspartic proteases; cysteine proteases; glutamic acid proteases; aspartic peptide lyases; serine proteases and threonine proteases. These protease families are described in the MEROPS peptidase database (http: / / merops.sanger.ac.uk / ). Serine proteases are preferred. Subtilase-type serine proteases are more preferred. The term "subtilase" refers to a sub-group of serine proteases according to Siezen et al., Protein Engng. 4 (1991) 719-737 and Siezen et al. Protein Science 6 (1997) 501-523. Serine proteases are a sub-group of proteases characterized by a serine in the active site that forms a covalent adduct with the substrate. Subtilases can be divided into six sub-classes, namely the subtilisin family, the thermitase family, the proteinase K family, the lanthinopeptidase family, the Kexin family and the Pyrolysin family.

[0091] Examples of subtilases are those derived from Bacillus, such as Bacillus lentus, Bacillus alkalophilus, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus pumilus and Bacillus gilii as described in US 7262042 and WO 09 / 021867, and Bacillus lentus protease lentus, Bacillus lentus protease Novo, Bacillus lentus protease Carlsberg, Bacillus licheniformis, Bacillus lentus protease BPN', Bacillus lentus protease 309, Bacillus lentus protease 147 and Bacillus lentus protease 168 described in WO 89 / 06279, and protease PD138 described in (WO 93 / 18140). Other useful proteases can be those described in WO 92 / 175177, WO 01 / 016285, WO 02 / 026024 and WO 02 / 016547. Examples of trypsin-like proteases are trypsin (e.g. of porcine or bovine origin) and the fusarium proteases described in WO 89 / 06270, WO 94 / 25583 and WO 05 / 040372, and the chymotrypsin derived from cellulomonas described in WO 05 / 052161 and WO 05 / 052146.

[0092] Most preferably, the protease is a subtilisin (EC 3.4.21.62).

[0093] Examples of subtilases are those derived from Bacillus, such as Bacillus lentus, Bacillus alkalophilus, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus pumilus, and Bacillus gibsonii, as described in US 7262042 and WO 09 / 021867, and Bacillus lentus protease lentinus, Bacillus lentus protease Novo, Bacillus lentus protease Carlsberg, Bacillus licheniformis, Bacillus lentus protease BPN', Bacillus lentus protease 309, Bacillus lentus protease 147, and Bacillus lentus protease 168 described in WO 89 / 06279, and protease PD138 described in (WO 93 / 18140). Preferably, the subtilisin is derived from Bacillus, preferably Bacillus lentus, Bacillus alkalophilus, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus pumilus, and Bacillus gibsonii, preferably as described in US 6,312,936 B1, US 5,679,630, US 4,760,025, US 7,262,042 and WO 09 / 021867. Most preferably, the subtilisin is derived from Bacillus gibsonii or Bacillus lentus.

[0094] Suitable commercially available proteases include those sold under the trade names: DuralaseTm, DurazymTm, Ultra, Ultra, Ultra, Ultra, and are sold as or (Novozymes A / S).

[0095] The composition can use a cutinase classified as EC 3.1.1.74. The cutinase used according to the present application can be of any origin. Preferably, the cutinase is of microbial origin, in particular bacterial, fungal or yeast origin.

[0096] Suitable amylases (alpha and / or beta) include those of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Amylases include, for example, alpha-amylases TM Termamyl TM Termamyl Ultra TM NatalaseTM 、Stainzyme TM 、Amplify TM 、Fungamyl TM and BAN TM (Novozymes A / S), Rapidase TM and Purastar TM (From Genencor International Inc.).

[0097] Suitable cellulases include those of bacterial or fungal origin. Chemically modified or protein engineered mutants are also included. Suitable cellulases include cellulases from Bacillus, Pseudomonas, Humicola, Fusarium, Thielavia, Acremonium, for example, the fungal cellulases produced by Humicola insolens, Thielavia terrestris, Myceliophthora thermophila and Fusarium oxysporum disclosed in US 4,435,307, US 5,648,263, US 5,691,178, US 5,776,757, WO 89 / 09259, WO 96 / 029397 and WO 98 / 012307. Commercially available cellulases include Celluzyme TM 、Carezyme TM 、Celluclean TM 、Endolase TM 、Renozyme TM (Novozymes A / S), Clazinase TM and Puradax HA TM (Genencor International Inc.) and KAC-500(B) TM (Kao Corporation). Celluclean TM is preferred.

[0098] 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, for example from C. cinereus, and variants thereof, such as those described in WO 93 / 24618, WO 95 / 10602 and WO 98 / 15257. Commercially available peroxidases include Guardzyme TM and Novozym TM 51004 (Novozymes A / S).

[0099] Further enzymes suitable for use are discussed in WO 2009 / 087524, WO 2009 / 090576, WO 2009 / 107091, WO 2009 / 1 1258 and WO 2009 / 148983.

[0100] Enzyme stabilizer

[0101] Any enzyme present in the composition can be stabilized using conventional means, for example, by the use of a polyol such as propylene glycol or glycerol, a sugar or sugar alcohol, lactic acid, boric acid or a boric acid derivative, e.g., an aromatic borate ester, or a phenyl boronic acid derivative such as 4-formylphenyl boric acid, and the composition can be formulated as described in, for example, WO 92 / 19709 and WO 92 / 19708.

[0102] Chelating agent

[0103] Chelating agents can be present or absent from the detergent composition.

[0104] If present, the chelating agent is present at a level of from 0.01 to 5 wt. %.

[0105] Preferred chelating agents are phosphonic acid (or salts thereof) chelating agents, preferably selected from: 1 -hydroxyethylidene- 1, 1 -diphosphonic acid (HEDP); diethylenetriaminepenta(methylene phosphonic acid) (DTPMP); hexamethylenediaminetetra(methylene phosphonic acid) (HDTMP); amino tri(methylene phosphonic acid) (ATMP); ethylenediaminetetra(methylene phosphonic acid) (EDTMP); tetramethylenediaminetetra(methylene phosphonic acid) (TDTMP); and phosphonobutane tricarboxylic acid (PBTC).

[0106] Fragrance / flavor

[0107] As used herein, the terms fragrance and perfume are used interchangeably.

[0108] The composition preferably comprises a fragrance. Many examples of suitable fragrances are provided in the CTFA (Cosmetic, Toiletry and Fragrance Association) 1992 International Buyers Guide, published by CFTA Publications, and OPD 1993 Chemicals Buyers Directory 80th Annual Edition, published by Schnell Publishing Co.

[0109] Preferably, the laundry composition comprises a fragrance, preferably wherein the fragrance is comprised at 0.001 to 2.0 wt.%, more preferably 0.01 to 1.5 wt.%, most preferably 0.1 to 1.0 wt.%. Preferably, the fragrance comprises greater than 50 wt.% biodegradable material, more preferably greater than 60 wt.% biodegradable material, more preferably greater than 70 wt.% biodegradable material, more preferably greater than 80 wt.% biodegradable material, more preferably greater than 90% biodegradable material, and most preferably the fragrance consists of 100 wt.% biodegradable material.

[0110] Preferably, the fragrance comprises at least one note (compound) selected from the group consisting of: a- isomethyl ionone, benzyl salicylate; citronellol; coumarin; hexyl cinnamal; linalool; ethyl 2-methyl valerate; octanal; benzyl acetate; 3,7-dimethyl-1,6-octadien-3-ol 3-acetate; 2-(1,1-dimethyl ethyl)-cyclohexanol 1 -acetate; delta-damascone; beta-ionone; tricyclo decenyl acetate; dodecanal; hexyl cinnamal; cyclopentadecanolide; phenyl acetic acid, 2-phenylethyl ester; amyl salicylate; beta-caryophyllene; ethyl undecylenate; geranyl o-aminobenzoate; alpha-irone; beta-phenyl ethyl benzoate; alpha-santalol; cedrol; cedryl acetate; cedryl formate; cyclohexyl salicylate; gamma-dodecalactone; and beta-phenyl ethyl phenyl acetate.

[0111] Polymer

[0112] The composition can comprise one or more additional polymers. Examples are carboxymethylcellulose, poly(ethylene glycol), poly(vinyl alcohol), polycarboxylates such as polyacrylates, maleic / acrylic acid copolymers and lauryl methacrylate / acrylic acid copolymers.

[0113] Optional ingredients

[0114] The compositions of the present application can contain optional ingredients to enhance performance and / or consumer acceptability. Examples of these ingredients include antifoams, fluorescers, shading dyes, preservatives, antimicrobial agents (e.g., bactericides), suds boosters, polyelectrolytes, anti-shrinkage agents, anti-wrinkle agents, antioxidants, sunscreens, anti-corrosion agents, drape imparting agents, anti-static agents, ironing aids, dyes / colorants, shading dyes, pearlizing agents and / or opacifiers, and microcapsules. Each of these ingredients is present in an amount effective to perform its intended purpose. Typically, these optional ingredients are included individually in amounts up to 5% (by weight based on the total weight of the composition).

[0115] If included, the antifoam agent is preferably a fatty acid soap. In the context of the present application, suitable fatty acids include aliphatic carboxylic acids of the formula RCOOH, wherein R is a linear or branched alkyl or alkenyl chain containing 6 to 24, more preferably 10 to 22, most preferably 12 to 18 carbon atoms and 0 or 1 double bond. Preferred examples of such materials include saturated C12-18fatty acids, such as lauric acid, myristic acid, palmitic acid or stearic acid; and fatty acid mixtures wherein 50 to 100% (by weight based on the total weight of the mixture) consist of saturated C12-18fatty acids. Such mixtures can typically be derived from natural fats and / or optionally hydrogenated natural oils (such as coconut oil, palm kernel oil or tallow).

[0116] Examples

[0117] The present application is illustrated by the following non-limiting examples.

[0118] MorvLip referred to is an enzyme from Moricella viscosa having the sequence of SEQ. ID. 1.

[0119] PinLip referred to is an enzyme from Psychromonas ingrahamii as described in WO2017 / 036901.

[0120] Production of MorvLip

[0121] Materials and methods

[0122] The gene encoding the protein of interest MorvLip with a C-terminal polyhistidine tag (His6 tag) was obtained in a pET28a vector (with kanamycin resistance) from TWIST Biosciences, San Francisco. MorvLip-pET28a was transformed into E. coli BL21 (DE3) cells. Positive clones from the transformation were used to overexpress MorvLip in 1 L LB media in 2.5 L flasks at 12°C for 39 hours after IPTG (1 mM). The cell slurry was collected and stored at -20°C before use. The cells were lysed and purified using Ni-IMAC chromatography followed by dialysis into storage buffer. Protein concentration was determined using Qubit TM 4 Fluorometer (Thermo Fisher Scientific) and Qubit TM Protein concentration was determined using Qubit

[0123] Buffer

[0124] Purification buffer (shown in Table 1) was filtered through a 0.2 μΜ nylon membrane filter (Whatman, Maidstone, UK) under vacuum. The buffer was cooled to 4 °C before adjusting the pH.

[0125] HEPPS is 4-(2-hydroxyethyl)-1-piperazinepropanesulfonic acid, 4-(2- hydroxyethyl)piperazine-1-propanesulfonic acid, N-(2-hydroxyethyl)piperazine-N'-(3- propanesulfonic acid)

[0126] SLES is sodium lauryl ether sulfate

[0127] TCEP is tris(2-carboxyethyl)phosphine

[0128] EDTA is ethylenediaminetetraacetic acid

[0129] Table 1. Buffers used for purification of MorvLip.

[0130]

[0131] Transformation

[0132] An aliquot of E. coli BL21(DE3) (50 μΐ^) was transferred from a -80 °C freezer to ice and allowed to thaw for 5 minutes. The plasmid encoding the gene of interest was mixed with the cells (1 μΐ^ for plasmids with a concentration of 100 ng / μΐ^). One or two plasmids were added for single or double transformation, respectively, and then added to the cells under gentle mixing and placed on ice for 30 minutes. The cells were heat shocked at 42 °C in a water bath for 45 seconds and then returned to ice for 5 minutes. Pre-warmed (37 °C) SOC medium (200 μΐ^) was added to the cells and then incubated at 37 °C and 220 rpm for 1 hour. The transformed cells were then plated on LB agar plates supplemented with kanamycin (50 mg / L). The plates were incubated at 37 °C overnight, or at 20 °C for 3 nights. Non-transformed cells were used as a negative control, while control plasmids pUC18 or pUC19 were used as a positive control.

[0133] Overexpression

[0134] Pre-warmed (37 °C) LB medium (1 L), starter culture (10 mL), and kanamycin (50 mg / L) were added to a sterile, fully baffled 2.5 L Tunair TM flask and incubated at 37 °C at 250 rpm until OD 600To achieve: 0.6-0.8. The flask was then placed on ice for 20-30 minutes before the addition of β-D-1 -thiogalactopyranoside (IPTG) to a final concentration of 1 mM. The flask was then incubated at 12°C for 39h. The cells were harvested by centrifugation at 4500 rpm for 20 minutes at 4°C. The cell pellet was washed by resuspension in phosphate buffered saline. The resuspension was decanted into a 50 mL sterile Falcon tube and the cell pellet was pelleted by centrifugation at 4500 rpm for 20 minutes at 4°C. The supernatant was discarded and the cell pellet was stored at -20°C.

[0135] Sonication

[0136] Lysis buffer was prepared by adding (1 U / 50 mL) and lysozyme (0.1 mg / mL) to IMAC buffer A. The cell paste (2-15 g) was transferred from -20°C storage to ice. The appropriate ice-cold lysis buffer (20-50 mL) was added and the cells were thawed with occasional inversion of the Falcon tube to begin resuspension of the cells. Once the cells were thawed, the cells were mixed vigorously with a metal spatula until a uniform suspension was obtained. The cell suspension was decanted into a narrow 75 mL glass beaker and firmly placed on a bed of wet ice. The cells were then sonicated using a Fisherbrand Model 550 Sonic Dismembrator (Fisher Scientific, Loughborough) with a 6.3 mm probe. The program was set to 80% intensity with 5 seconds on and 10 seconds off for a total sonication time of 10 minutes to prevent heating of the sample. The sonicated cells were centrifuged at 14500 rpm for 30 minutes at 4°C after which the supernatant containing the soluble proteins was separated from the pelleted insoluble fraction for downstream purification. The supernatant fraction was then clarified using a 28 mm syringe filter with Sartorius Stedim, minisart 0.2 μιη disposable filtration units (Stonehouse, UK). TM 120The cell suspension was sonicated using a Fisherbrand Model 550 Sonic Dismembrator (Fisher Scientific, Loughborough). The program was set to 80% intensity with 5 seconds on and 10 seconds off for a total sonication time of 10 minutes to prevent heating of the sample. The sonicated cells were centrifuged at 14500 rpm for 30 minutes at 4°C after which the supernatant containing the soluble proteins was separated from the pelleted insoluble fraction for downstream purification. The supernatant fraction was then clarified using a 28 mm syringe filter with Sartorius Stedim, minisart 0.2 μιη disposable filtration units (Stonehouse, UK).

[0137] Purification

[0138] The protein sample was purified using an AKTA Purifier (GE Healthcare, Cincinnati, OH, USA) maintained at 7°C.

[0139] Nickel-NTA affinity chromatography

[0140] ​A 5 mL nickel-NTA (Ni-NTA) column (Cytiva) stored in 20% (v / v) ethanol was equilibrated with 10 column volumes (CV) of ddH2O and then equilibrated with 10 CV of IMAC buffer A. The clear supernatant was then loaded and the flow-through was collected, where the protein of interest was expected to bind to the Ni-NTA resin until the elution buffer was applied. The column was then washed with 20 CV of IMAC buffer A, or until the absorbance at 280 nm (A 280 ) to a baseline close to that of the lysis buffer. The protein of interest was then eluted with 5 CV of IMAC buffer B. Fractions were analyzed by SDS-PAGE and Western blotting. Fractions containing the gene of interest were pooled for downstream processing and experiments.

[0141] Dialysis

[0142] The soluble protein sample was dialyzed using a dialysis tubing membrane (Thermo Fisher Scientific) with a molecular weight cutoff (MWCO) of 8 kDa. The membrane was rinsed with ddH2O, a knot was tied to seal the tubing at one end, and the protein sample was added. Another knot was tied to completely seal the tubing, and plastic clamps were attached to both ends. The tubing was carefully placed in 4 L of storage buffer at 4°C and left for 18 hours with gentle agitation.

[0143] Protein concentration determination

[0144] Using the manufacturer's recommended conditions, use Qubit TM 4 Fluorometer (Thermo Fisher Scientific) and Qubit TM Protein concentration was determined using a Qubit protein assay kit. TM Protein assays are performed by reading the fluorescence of a detergent-based dye that responds quantitatively to the presence of protein in solution.

[0145] Before performing the assay, all reagents were equilibrated to room temperature. Each time the assay was performed, calibration was performed using the three protein standards provided in the kit. TM Dilute Qubit 1:200 in protein buffer TM Protein reagents are used to prepare Qubit TM Working solution. Prepare calibration standards by adding 190 μL of working solution to 10 μL of protein standards 1, 2, and 3. Prepare protein samples by adding 190 μL of working solution to 10 μL of protein sample. Vortex all tubes for 2-3 seconds and then allow them to incubate for 15 minutes. Then calibrate the Qubit with the three protein standards before reading each protein sample. TM 4 Fluorometer.

[0146] Protein purity determination by SDS-PAGE analysis

[0147] SDS-PAGE was performed using a Bio-Rad mini-Protean cell. After dilution in Laemmli loading buffer 257 , protein samples were boiled at 100°C for 10 minutes to produce denatured samples, which were ready for loading after centrifugation at 12000 RPM for 5 minutes. After cooling to room temperature, samples (2-20 μL) were loaded onto 4-12% ExpressPlus TM PAGE gels (GenScript, Piscatawat, NJ, USA). 2 μL of Spectra broad range multicolour protein ladder (Bio-Rad) was loaded onto the gels to enable determination of protein molecular weight.

[0148] Protein separation was achieved using MOPS running buffer (50 mM Tris-Base, 50 mM MOPS, 1 mM EDTA and 0.1% SDS (w / v), pH 7.7) and electrode conditions of 140 V, 400 mA, RT for 60 minutes. Protein bands were stained on SDS-PAGE gels using rapid Coomassie staining (Generon, Slough, UK) according to the manufacturer’s protocol. SDS-PAGE gels were transferred to ddH2O at room temperature for 1 hour, replenished with ddH2O and the gels left for a further hour to remove all unbound dye.

[0149] Protein characterization by Western blot analysis

[0150] SDS-PAGE gels were prepared with the sample of interest, but the gels were not stained after protein separation.

[0151] Protein bands on SDS-PAGE gels were transferred to nitrocellulose membranes (Sartorius Stedim) in a Pierce G2 Fast Blotter (Thermo Fisher Scientific) according to the manufacturer’s instructions (25 V, 1.3 A, 7 minutes). The iBind Western system (Thermo Fisher Scientific) was used for antibody application and washing. The iBind Western system utilises lateral flow capillary diffusion to perform all western blot steps in a single procedure.

[0152] A 1X iBind master mix was prepared by adding 5X iBind buffer (6 mL), additive (300 μί), and ddH2O (23.7 mL) into a sterile 50 mL Falcon tube. The nitrocellulose membrane was soaked in the iBind master mix (6 mL) for 5 minutes. Meanwhile, the iBind card was pre-equilibrated with 6 mL of iBind master mix without liquid touching the kit. The antibody solution was prepared to a final volume of 2 mL in iBind master mix according to the manufacturer's recommended dilution. After adding an additional 1 mL of iBind master mix to the center of the card, the soaked nitrocellulose membrane was placed so that the protein bands were in contact with the card and the lowest molecular weight band was at the bottom. The iBind system was then closed and the following solutions were added to the slots:

[0153] 1.2 mL 1X primary antibody solution

[0154] 2.2 mL iBind master mix

[0155] 3.2 mL 1X secondary antibody solution

[0156] 4.6 mL iBind master mix

[0157] Monoclonal anti-polyhistidine mouse IgG (Sigma) and 800CW goat anti-mouse IgG (LI-COR) were used as primary and secondary antibodies, respectively.

[0158] The iBind system was incubated at room temperature for 4-18 hours. The nitrocellulose membrane was then rinsed in ddH2O to remove any residual unbound antibody. The membrane was then imaged using an Odyssey CLx imaging system (LI-COR, Lincoln, NE, USA) and analyzed using Image Studio TM Lite Light (LI-COR).

[0159] The identity of the MorvLip full-length sequence was confirmed by SDS-PAGE iBind Western blot analysis and the purity of MorvLip was determined by SDS-PAGE analysis of the MorvLip Ni-IMAC purified fraction. This analysis showed satisfactory production and purification of MorvLip enzyme.

[0160] Example 1 - wash application test: MorvLip and 200L comparison

[0161] Materials and methods

[0162] Morvlip was compared to commercial enzymes 200L (Novozymes) were compared for cleaning efficacy. Due to the large amount of protein required for this study (~100 mg), comparison with PinLip was not feasible due to its low overexpression and purification yield.

[0163] Stained fabric sets "Lipase Set 2" were used to determine stain removal on a wide range of stains. Pre-soaked fabrics with particularly stubborn stains can increase the cleaning efficacy of laundry formulations and are a habit of consumers. Therefore, the efficacy of MorvLip was investigated using a pre-soaking method. All stained fabrics were pre-soaked overnight at 2.5 g / L in either a custom formulation F4 containing 200L or 50 mM Tris-HCl (pH 8.5). 200L and 50 mM Tris-HCl (pH 8.5). Pre-soaked stained fabrics were then washed in a Tergo-O-Tometer with 0.1 g / L Persil UK Non-Bio at 30 °C for 45 minutes, followed by a rinse in water. The 0.1 g / L of detergent is at the lower threshold of the laundry cycle and was chosen to highlight the positive effect of using lipase in the formulation to reduce the amount of detergent required for effective stain removal. The washed fabric stains were then measured to determine the Stain Removal Index (SRI) for each pre-soaking treatment method.

[0164] Pre-soak method

[0165] A custom formulation was used to pre-soak the stained fabrics before Tergo-O-Tometer washing with MorvLip or 200L. A working pre-soak solution was prepared by adding custom formulation 2X (200 mL), water (150 mL) and 50 mL MorvLip 8X, A working pre-soak solution was prepared by adding custom formulation 2X (200 mL), water (150 mL) and 50 mL MorvLip 8X,

[0166] Table 2. Solutions required for pre-soak wash methods.

[0167]

[0168] Custom formulation 2X is a pH 8.5 buffered di-rhamnolipid (derived from Evonik) cocamidopropyl betaine surfactant composition (2: 1 weight ratio) dosed at 5 g / L.

[0169] Terg-O-Tometer wash performance

[0170] Actual wash conditions found in a washing machine were then simulated using a Terg-O-Tometer (Test fabric Inc., West Pittson, PA, US). The Terg-O-Tometer is a laboratory scale washing apparatus used to evaluate the cleaning efficacy of laundry washing liquids and powders. The instrument is a beater-type washer with 6 temperature-controlled beakers, allowing experiments to be easily performed in replicates. Each wash was performed using a combination of soiled fabric and a mixture of clean woven cotton and knitted polyester squares (10 x 10 cm) called ballast. 1 L of wash liquor was used to wash each piece of fabric. After washing, the soiled fabric was removed from the wash liquor and the ballast fabric was discarded. The remaining soiled fabric was then rinsed in 1 L of water and allowed to dry overnight at room temperature on a horizontal rack in the dark. The dried soiled fabric was then measured in an X-Rite instrument (Pantone, Carlstadt, NJ, US) with a UV excitation light source to determine the delta E and SRI relative to the un-washed white fabric.

[0171] SRI calculation

[0172] All fabric stain removal was evaluated using the Stain Removal Index (SRI) method. The soiled fabrics were digitally scanned to measure their color difference from the control white fabric using the following equation:

[0173] Equation 1.

[0174] Where delta L, delta a and delta b are the differences in darkness, redness and yellowness between the white fabric and the soiled fabric, respectively.

[0175] Equation 1 tells us that lower values of delta L, delta a and delta b result in lower values of delta E, with a value of 0 corresponding to a perfectly white fabric.

[0176] The cleaning efficacy was then expressed as:

[0177] Equation 2. SRI = 100 - delta E

[0178] Where higher values of SRI correspond to cleaner fabrics, while 100 corresponds to a perfectly white fabric.

[0179] The raw values of the results from Equations 1 and 2 were normalized to give 100 for the undyed fabric. The results for the margarine fat soiled fabric are given in Table 3 below. There were 4 replicates.

[0180] Table 3

[0181]

[0182] It can be seen that MorvLip performed much better than commercial lipase (Lipex® Evity) in terms of stain removal on fatty margarine stains.

[0183] Example 2 - Stability in commercial additives for cold-adapted lipases

[0184] Materials and methods

[0185] This experiment shows that MorvLip is more stable relative to PinLip as measured by residual efficacy when the lipase is present in formulations containing certain useful general laundry composition additives.

[0186] MorvLip is a quite different enzyme sequence to PinLip. The MorvLip of SEQ. ID. 1 in this specification has only 35% sequence identity to the PinLip enzyme of WO2017 / 036901.

[0187] Long-term stability of MorvLip and PinLip was tested in the presence of soil release polymer (SRP) and protease, which are commonly used in commercial laundry formulations. Enzymes were incubated at 1 mg / mL in the presence of 2% SRP or 0.95 mg / mL protease in SEC buffer (50 mM HEPPS pH 8.0, 1 M sodium chloride, 50 mM maltose, 5 mM EDTA, 0.005% (w / v) SLES, 0.3 mM TCEP) for 4 weeks. PNp-laurate assay was performed to determine hydrolytic activity. Initial activity was recorded in the absence of any additives, then activity was measured again after 4 weeks. Residual activity after 4 weeks was calculated as a percentage of initial activity. All activity assays were performed in 50 mM Tris-HCl pH 8.6 at 25 °C.

[0188] PNp-laurate hydrolysis assay

[0189] ​A pNp laurate stock solution was prepared at 8 mM in methanol (51.4 mg / 20 mL) and stored at -20 °C for future use. The stock solution was diluted to 0.25 mM in ddH2O, protected from light exposure by aluminium foil and kept on ice. All solutions were used within 24 hours of dilution. The diluted pNp ester was added to the vessel and allowed to equilibrate to the required temperature. The TECAN Infinite M200 PRO plate reader was turned on at least 15 minutes before the experiment. Diluted protein samples were prepared by diluting the protein sample (1 mg / mL) 10-fold in Tis-HCl buffer (100 mM, pH 8.6). The samples were prepared by adding 100 mM Tris-HCl pH 8.0 (100 pL) and the diluted protein solution (20 pL) to a 96-well microtitre plate in triplicate. Instead of the protein solution, ddH2O (20 pL) was added in triplicate as a control, which was used to measure the background pNp ester hydrolysis. Where different additives were used, each additive was added in the absence of protein to measure the individual background activity of each. The plate was allowed to equilibrate to the temperature of the plate reader for 5 minutes. Using a 12-channel P100 pipette, 0.25 mM pNp laurate (80 pL) was added to all wells and the plate was transferred to the plate reader. Activity was followed by detecting the absorbance at 347 nm (A 347 ) every 60 seconds for 10 minutes. The enzymatic steady-state hydrolysis rate of the pNp ester was calculated in PRISM 8.0.

[0190] For the kinetic measurements at a wide range of pH conditions, the method described by Lucia et al. was employed for non-pH dependence. For the set-up of the samples, the same method as described in the previous section was followed, except that the buffer can be interchanged with any buffer system. Activity was measured by detecting the absorbance at 347 nm (A 347 ) every 60 seconds for 10 minutes. The enzymatic hydrolysis rate of the pNp-laurate was calculated in excel and PRISM 8.0.

[0191] The results of the stability experiments are given in Table 4. With 3 replicates.

[0192] Table 4 - Relative activity (%) as percentage of the "no additive" condition after 4 weeks of incubation

[0193]

[0194] UL 50 corresponds to a soil release polymer. The soil release polymer is commercially available from Clariant. CE16L corresponds to a protease and relates to Carnival Evity 16L, which is commercially available from Novozymes.

[0195] When incorporated into compositions containing common laundry detergent additives such as proteases and soil release polymers, MorvLip lipase retains much higher activity compared to PinLip.

[0196] Examples Residual stability after 3-4 weeks as a percentage of initial activity

[0197] Test MorvLip, PinLip, 100L and Stability of 200 L in buffer. The residual activity after 4 weeks is given as a percentage of the initial activity in Table 5.

[0198] Table 5 - Residual activity (%) after 4 weeks of incubation as a percentage of the activity on day 0

[0199]

[0200] MorvLip and two The enzymes generally retained their initial activity levels, however the MorvLip enzyme retained much higher activity levels compared to the PinLip lipase.

Claims

1. A detergent composition comprising: (a) 0.0005-6 wt%, preferably 0.005-4 wt%, more preferably 0.001-2 wt% of a lipase from Morellia viscosus; and (b) 1-60 wt%, preferably 1-50 wt%, more preferably 1-35 wt% of a detersive surfactant.

2. A detergent composition as claimed in claim 1, wherein the lipase from Moriella viscosus has a sequence identity of at least 70%, preferably 75%, more preferably 80%, more preferably 85%, even more preferably 90%, even more preferably 95%, even more preferably 98%, even more preferably 99%, most preferably 100% to SEQ.ID.

1.

3. A detergent composition according to claim 1 or claim 2, wherein the detergent composition comprises an anionic surfactant and / or a nonionic surfactant, preferably comprises both anionic surfactant and nonionic surfactant.

4. A detergent composition according to claim 3, wherein the anionic surfactant is present in an amount of 1-50 wt%, preferably 2-40 wt%, more preferably 3-30 wt%, and is preferably selected from linear alkylbenzene sulfonates, secondary alkane sulfonates, sodium laureth sulfate, sodium lauryl sulfate, sodium oleyl sulfate and sodium oleyl ether sulfate, methyl ester sulfonates, secondary alkyl sulfates (SALS), cardanol ether sulfate and rhamnolipids.

5. A detergent composition according to claim 3 or claim 4, wherein the nonionic surfactant is present in an amount of 1-30 wt%, preferably 2-20 wt%, more preferably 3-15 wt%, and is preferably selected from alcohol ethoxylates, alcohol propoxylates, methyl ester ethoxylates and alkyl polyglycosides.

6. A detergent composition according to any one of the preceding claims, wherein the detergent composition is a laundry detergent composition.

7. A laundry detergent composition according to claim 6, wherein the laundry detergent composition is in the form of a liquid, solid, powder, pastille, beads or paste, preferably the composition is a liquid or powder, more preferably a liquid detergent.

8. A laundry detergent composition according to claim 6 or claim 7, wherein the laundry detergent composition comprises an alkoxylated polyamine, preferably at a level of from 0.1 to 8 wt%, more preferably from 0.2 to 6 wt%, most preferably from 0.5 to 5 wt%.

9. A laundry detergent composition according to any one of claims 6 to 8, wherein the laundry detergent composition comprises a soil release polymer, preferably selected from copolyesters of dicarboxylic acids and polyglycols, more preferably copolyesters formed from the condensation of terephthalate and 1,2-propylene glycol, preferably present at a level of 0.1 to 8 wt%, more preferably 0.2 to 6 wt%, most preferably 0.5 to 5 wt%.

10. A detergent composition according to any preceding claim, further comprising one or more additional enzymes selected from the group consisting of proteases, cellulases, alpha-amylases, peroxidases / oxidases, pectate lyases and / or mannanases, preferably the one or more enzymes include a protease.

11. A method of treating a textile having fatty stains, wherein the textile is treated with a composition according to any one of claims 1 to 10 to provide enhanced lipolytic cleaning of fatty stains, preferably margarine stains, the textile being preferably subsequently rinsed and dried.

12. A method of treating textiles having fatty stains, wherein the textiles are pre-treated with a composition according to claim 1 or claim 2 to provide enhanced lipolytic cleaning of fatty stains, preferably margarine stains, and the textiles are then washed with a laundry main wash composition, and then preferably rinsed and dried.

13. Use of a lipase from Morella viscosus, preferably a lipase from Morella viscosus having at least 70% sequence identity to SEQ. ID. 1, for improving the cleaning of fatty stains, preferably margarine stains, on textiles.

14. Use of a detergent composition comprising a lipase from Morella viscosus, preferably a lipase from Morella viscosus having at least 70% sequence identity to SEQ. ID. 1, for improved cleaning of fatty stains, preferably margarine stains, on textiles.

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