Liquid anti-biofilm composition

Abstract

To provide a detergent combination having anti-biofilm activity (that means biofilm formation on a device and / or an article to be cleaned is inhibited or reduced).SOLUTION: A liquid composition contains (a) at least one chlorinated hydroxy diphenyl ether represented by formula (I) (where R1, R2 and R3 are selected from H and Cl, and at least one of R1, R2 and R3 is Cl), (b) at least one compound demonstrating DNAse activity, and (c) at least one compound selected from an anionic surfactant and a nonionic surfactant.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a liquid composition comprising at least one compound selected from the group of chlorinated hydroxydiphenyl ethers, at least one compound exhibiting DNAse activity, and at least one compound selected from anionic surfactants and nonionic surfactants. The present invention also relates to the use of the liquid composition to (a) reduce and / or inhibit biofilm formation on surfaces of equipment that regularly comes into contact with water, e.g., cleaning equipment such as washing machines, and / or (b) reduce and / or inhibit microbial growth on materials that regularly come into contact with water, e.g., items to be washed, such as hard surfaces and / or textiles. [Background technology]

[0002] Microbial growth distinguishes between planktonic and biofilm modes of microbial growth. Microbial cells growing in biofilms are physiologically distinct from planktonic cells of the same organism, which, in contrast, are single cells that may drift or swim in a liquid medium. A "biofilm" typically comprises a consortium of microorganisms in which cells adhere to each other and, frequently, to surfaces. In one aspect, biofilm formation is said to begin with the attachment of planktonic microorganisms to a surface. Once colonization begins, biofilms grow by a combination of cell division and recruitment. Cells within biofilms are typically embedded in an extracellular matrix, often referred to as extracellular polymeric substances (EPS). EPS typically contains exopolysaccharides, proteins, lipids, and DNA. These bacteria growing in biofilms are a source of hygiene problems, e.g., microbial contamination, potentially resulting in disease or at least a foul odor.

[0003] Biofilms can form on solid surfaces as well as on soft surfaces, such as textiles, and are generally resistant to conventional disinfection methods. Increased resistance to detergent application, UV irradiation, and antibiotics has also been observed. In particular, biofilms are often sticky, and soils may adhere to surfaces containing such biofilms. Furthermore, biofilms may be the source of undesirable odors, for example, on washed parts or in the tubes of a washer, which may result from biofilm degradation. Biofilms may adversely affect the long-term functionality of cleaning equipment. Summary of the Invention [Problem to be solved by the invention]

[0004] One problem that the present invention aims to solve is to provide detergent formulations that have anti-biofilm activity, meaning that biofilm formation on the surfaces of the equipment and / or items to be cleaned is inhibited or reduced. [Means for solving the problem]

[0005] The present invention has solved this problem by providing a liquid composition comprising (a) at least one compound selected from the group consisting of chlorinated hydroxydiphenyl ethers, (b) at least one compound exhibiting DNAse activity, and (C) at least one compound selected from anionic surfactants and nonionic surfactants. DETAILED DESCRIPTION OF THE INVENTION

[0006] Throughout this specification, including the claims, the terms "comprising one" or "comprising a" shall be understood as synonymous with the term "comprising at least one" unless otherwise specified, and "between" shall be understood to include both limits.

[0007] The terms "a", "an" and "the" are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article.

[0008] The term "and / or" is inclusive, meaning "and" and "or."

[0009] The use of the phrase "can / could / may" within this specification refers to particular embodiments and not merely optional features.

[0010] It should be noted that when specifying any range of concentrations, weight ratios, or amounts, any particular upper concentration, weight ratio, or amount can be associated with any particular lower concentration, weight ratio, or amount, respectively.

[0011] In one aspect, the present invention provides Component (a): at least one compound selected from chlorinated hydroxydiphenyl ethers according to formula (I), Component (b): at least one compound exhibiting DNAse activity; and Component (c): at least one compound selected from anionic surfactants and nonionic surfactants; The present invention relates to a liquid composition comprising:

[0012] The compositions of the present invention are, in one embodiment, liquid at 20°C and 101.3 kPa. The liquid preferably has a viscosity of 1500 mPa at 25°C. * In embodiments, the viscosity at 25°C is less than 1000 mPa. * less than 800 mPa * is less than s.

[0013] Component (a) Component (a) is a compound of formula (I):

[0014] [ka] wherein R1, R2, and R3 are selected from H and Cl, and at least one of R1, R2, and R3 is Cl. The compound contains at least one compound selected from the group of chlorinated hydroxydiphenyl ethers according to

[0015] In one embodiment, the compound according to formula (I) is characterized in that at least two of R1, R2 and R3 are Cl.

[0016] In one preferred embodiment, R1 and R2 are Cl and R3 is H (4,4'-dichloro 2-hydroxydiphenyl ether; such compounds are also referred to herein as DCPP or Diclosan).

[0017] In yet another embodiment, all of R1, R2, and R3 are Cl (2,4,4'-trichloro-2'-hydroxydiphenyl ether, such compounds are also referred to herein as Triclosan).

[0018] Ingredient (b) The composition of the present invention comprises at least one compound exhibiting DNAse activity. Compounds exhibiting DNAse activity include DNAses that typically catalyze the hydrolytic cleavage of phosphodiester bonds in DNA. DNases are classified into, for example, EC3.1.11, EC3.1.12, EC3.1.15, EC3.1.16, EC3.1.21.X (wherein X = 1, 2, 3, 4, 5, 6, 7, 8, or 9), EC3.1.22.Y (wherein Y = 1, 2, 4, or 5), EC3.1.23, EC3.1.24, EC3.1.25, and EC3.1.30.Z (wherein Z = 1 or 2) and EC3.1.31.1. Preferably, the DNAse is selected from the classes EC 3.1.21.x (wherein x=1, 2, 3, 4, 5, 6, 7, 8 or 9), EC 3.1.22.y (wherein y=1, 2, 4 or 5), EC 3.1.30.z (wherein z=1 or 2), EC 3.1.31.1 and mixtures thereof: Nucleases of the class EC 3.1.21.x cleave at the 3' hydroxyl to release the 5' phosphomonoester. Those where x=1 are particularly preferred. Nucleases of class EC 3.1.22.y cleave at the 5' hydroxyl, liberating a 3' phosphomonoester. Enzymes of class EC 3.1.30.z may be preferred, as they act on both DNA and RNA and liberate a 5'-phosphomonoester. Suitable examples from class EC 3.1.31.2 are described in US 2012 / 0135498 A, e.g., SEQ ID NO: 3 therein. Nuclease enzymes from class EC 3.1.31.1 generate 3' phosphomonoesters.

[0019] DNAse activity or DNA degradation activity refers to the catalytic effect exerted by an enzyme, preferably a DNAse, in terms of molecules of substrate converted per minute per enzyme molecule (molecular activity) or, typically, units per milligram of enzyme (specific activity). DNAse activity is determined by test schemes known to those skilled in the art. For example, DNAse activity can be determined using DNase Test Agar with Methyl Green (BD, Franklin Lakes, NJ, USA), which must be prepared according to the supplier's instructions. Briefly, 21 g of agar is dissolved in 500 ml of water and then autoclaved at 121°C for 15 minutes. The autoclaved agar is then heated to 10-48°C in a water bath, and 20 ml of the agar is poured into a Petri dish and allowed to solidify by incubation at room temperature. 5 μl of enzyme solution is added to the solidified agar plate, and DNAse activity is observed as a colorless zone around the spotted enzyme solution. DNAse activity may alternatively be determined by using the DNAseAlert™ Kit (11-02-01-04, IDT Integrated DNA Technologies) according to the supplier's manual. Briefly, 95 μl of DNase sample is mixed with 5 μl of substrate in a microtiter plate, and fluorescence is immediately measured (excitation 536 nm, emission 556 nm) using a Clariostar® microtiter reader from BMG Labtech. Any suitable test scheme can be used to determine DNAse activity.

[0020] DNAses are enzymes. In the context of the present invention, enzymes are identified by their "polypeptide sequence" (also referred to herein as "amino acid sequence"). In the present invention, the term "enzyme" is used synonymously with the term "polypeptide" having enzymatic activity.

[0021] The peptide sequences refer to parent enzymes and / or their variant enzymes, both of which have enzymatic activity. As used herein, an enzyme is a polypeptide that is enzymatically active or that effects an enzymatic transformation, meaning that the enzyme acts on substrates and converts them into products.

[0022] The term "enzyme" as used herein preferably excludes inactive mutants of enzymes. Thus, mutant enzymes according to the present invention are preferably functional mutants that have enzymatic activity, where the enzymatic activity is of the same type as that exerted by the respective parent enzyme.

[0023] The polypeptide sequence specifies the three-dimensional structure of an enzyme, including its "active site," which in turn determines the enzyme's catalytic activity. The polypeptide sequence can be identified by a SEQ ID NO. In accordance with the World Intellectual Property Organization (WIPO) Standard ST.25 (1998), amino acids herein are represented using three-letter codes with the first letter capitalized, or their single-letter equivalents. Enzyme names are known to those skilled in the art based on recommendations from the Nomenclature Commission of the International Union of Biochemistry and Molecular Biology (IUBMB). The enzyme name includes the EC (Enzyme Commission) number, recommended name, synonyms (if any), catalytic activity, and other factors.

[0024] A "parent" sequence (of a parent protein or enzyme, also called a "parent enzyme") is a starting sequence for introducing modifications into the sequence (e.g., by introducing one or more amino acid substitutions, insertions, deletions, or a combination thereof) to result in a "variant" of the parent sequence. The term parent sequence includes sequences of wild-type enzymes and synthetically produced sequences that are used as starting sequences for introducing (further) modifications. As used herein, the terms "parent sequence" and "parent enzyme" are used interchangeably.

[0025] The term "enzyme variant" or "sequence variant" or "variant enzyme" refers to an enzyme that differs to some extent in amino acid sequence from the parent enzyme. Unless otherwise indicated, the variant enzyme "has enzymatic activity."

[0026] When describing sequence variants, the following nomenclature is used: Amino acid substitutions are described by indicating the original amino acid of the parent enzyme, followed by the position number in the amino acid sequence, followed by the substituted amino acid. Amino acid deletions are described by indicating the original amino acid of the parent enzyme, followed by the position number in the amino acid sequence, followed by an asterisk " * ". Amino acid insertions are described by indicating the original amino acid of the parent enzyme, followed by the position number in the amino acid sequence, followed by the original amino acid and the additional amino acid. It is clear that degeneracy in nomenclature occurs when an amino acid residue identical to an existing amino acid residue is inserted. When different changes can be introduced at a position, the different changes are separated by commas.

[0027] In one embodiment, an enzyme variant is defined by its sequence identity compared to the parent enzyme. Sequence identity is usually expressed as "% sequence identity" or "% identity." To calculate sequence identity, the first step is to create a sequence alignment. According to the present invention, a pairwise global alignment must be created, which means that the two sequences must be aligned over their entire length, and this is usually done by using a mathematical approach called an alignment algorithm.

[0028] According to the present invention, alignments are made by using the algorithm of Needleman and Wunsch (J. Mol. Biol. (1979) 48, pp. 443-453). Preferably, the program "NEEDLE" (The European Molecular Biology Open Software Suite (EMBOSS)) is used for the purposes of the present invention using the program's default parameters (gap open=10.0, gap extension=0.5 and matrix=EBLOSUM62).

[0029] According to the present invention, the following calculation of % identity is applied: % identity = (identical residues / length of the alignment region representing each sequence of the present invention over its full length) x 100.

[0030] In one embodiment, enzyme variants are described as amino acid sequences that are at least n% identical to the amino acid sequence of their respective parent enzymes, where "n" is an integer between 10 and 100. In one embodiment, the variant enzymes are at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical when compared to the full-length amino acid sequence of the parent enzyme, and the enzyme variants have enzymatic activity.

[0031] A variant enzyme herein refers to a variant enzyme that has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100% enzymatic activity compared to the enzymatic activity of the respective parent enzyme. In one embodiment, the variant enzyme has increased enzymatic activity compared to the respective parent enzyme. Increased means greater than 100% enzymatic activity compared to the enzymatic activity of the respective parent enzyme. In a preferred embodiment, increased enzymatic activity refers to increased enzymatic activity compared to the respective parent enzyme after storage of the enzyme at a particular temperature for a particular time.

[0032] Polypeptides with DNAse activity have been previously described, for example in WO 2015 / 155350 (Novozymes A / S) and WO 2015 / 155351 (Novozymes A / S), which describe fungal DNases. Vibrissea flavovirens, Penicillium reticulisporum, Acremonium dichromosporum, Preussia aemulans, Colletotrichum circinans, Clavicipitaceae, Trichurus spiralis, Pyrenochaetopsis sp., Aspergillus sydowii, Cladosporium cladosporioides, Rhinocladiella sp., Pyronema domestica It can be obtained from Aspergillus domesticum, Aspergillus niger, Phialophora geniculata, Paradendryphiella salina, Aspergillus insuetus, Purpureocillium lilacinum, Warcupiella spinulosa, Stenocarpella maydis, Acrophialophora fusispora, Chaetomium luteum, or Arthrinium arundinis.

[0033] The DNAse may, in one embodiment, be a Bacillus, such as Bacillus cibi, Bacillus horikoshii, Bacillus horneckiae, Bacillus idriensis, Bacillus algicola, Bacillus vietnamensis, Bacillus hwajinpoensis, Paenibacillus mucilanginosus, Bacillus indicus, Bacillus luciferensis, Bacillus marisflavii, Bacillus marisflavi) and their mutants.

[0034] In one embodiment, the at least one DNAse included in component (b) is selected from polypeptides having an amino acid sequence at least 80% identical to SEQ ID NOs: 1-24 and 27-28 of WO 2019 / 081724 or WO 2019 / 081721. The mutant DNAse comprises one or both of the motifs [D,M,L][S,T]GYSR[D,N] (SEQ ID NO: 25 of WO 2019 / 081724 or WO 2019 / 081721) and ASXNRSKG (SEQ ID NO: 26 of WO 2019 / 081724 or WO 2019 / 081721).

[0035] In one embodiment, the at least one DNAse is SEQ ID NO: 4 and variants thereof disclosed in U.S. Pat. No. 9,675,736; SEQ ID NOs: 1 to 3 and variants thereof disclosed in EP 3088502, and SEQ ID NOs: 1 to 6 and their variants disclosed in WO 2017 / 001471 is selected from.

[0036] In one embodiment, the at least one DNAse is selected from polypeptides comprising one or more of the motifs selected from [G,Y,W,F,A,H]NI[R,Q,D,E,V] (SEQ ID NO: 73 of WO 2017 / 060493), SDH[D,H,L]P[SEQ ID NO: 74 of WO 2017 / 060493) and GGNI[R,Q] (SEQ ID NO: 75 of WO 2017 / 060493). Preferably, the DNAse has an amino acid sequence that is at least 80%, at least 82%, at least 85%, at least 90%, at least 92%, at least 95%, or at least 99% identical when compared to a polypeptide selected from SEQ ID NOs: 6, 7, 10, 27, 30, 33, 36, 39, 42, 45, 48, 51, 57, 60, 63, 66, 69, and 72, all of which are disclosed in WO 2017 / 060493. In one embodiment, the at least one DNAse is selected from polypeptides having at least 85% sequence identity to the polypeptides of SEQ ID NOs: 10, 27, and 63, all of which are disclosed in WO 2017 / 060493. In one embodiment, the at least one DNAse is selected from polypeptides having at least 95% sequence identity to the polypeptides of SEQ ID NOs: 7, 39, 48, and 60, all of which are disclosed in WO 2017 / 060493. In one embodiment, the at least one DNAse is a polypeptide having at least 99% sequence identity to the polypeptide of SEQ ID NO: 42 disclosed in WO 2017 / 060493.

[0037] In one embodiment, the at least one DNAse comprised in component (b) is 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%, at least 96%, at least 97%, at least 98% or at least 99% identical to the full-length polypeptide sequence of SEQ ID NO: 1 according to the sequence listing of the present application. Preferably, the DNAse has the polypeptide sequence of SEQ ID NO: 1 according to the sequence listing of the present application.

[0038] SEQ ID NO: 1 is characterized by having the following amino acid sequence: MKKWMAGLFLAAAVLLCLLMVPQQIQGASLYDKVLYFPLSRYPETGDHIKDAIADGHSDICTIDRDGADKRRQESLKGIPTKPGYDRDEWPMAVCEEGGAGADVRYVTPSDNRGAGSWVGNQMSGYPDGTRVLFIVQ

[0039] Ingredient (c) Component (c) comprises at least one surfactant selected from the group consisting of anionic surfactants and nonionic surfactants.

[0040] Anionic surfactants (AIS) In one embodiment, the liquid compositions of the present invention comprise at least one anionic surfactant (AIS). Anionic surfactants include, but are not limited to, surface-active compounds that contain a hydrophobic group and at least one water-solubilizing anionic group, typically selected from sulfate, sulfonate, and carboxylate, to form water-soluble compounds.

[0041] In one embodiment, the liquid composition of the present invention comprises a compound of general formula (AIS I):

[0042] [ka] The composition comprises at least one anionic surfactant selected from the group consisting of:

[0043] The variables of general formula (AIS I) are defined as follows: R 1 is C1~C 23 -Alkyl (e.g., 1-, 2-, 3-, 4-C1-C 23 -alkyl) and C2-C 23 -alkenyl, where alkyl and / or alkenyl are linear or branched, and 2-, 3-, or 4-alkyl are, for example, n-CH 15 , n-C9H 19 , nC 11 H23 , nC 13 H 27 , nC 15 H 31 , nC 17 H 35 , i-C9H 19 ,I C 12 H 25 is. R 2 H, C1~C 20 -Alkyl and C2-C 20 -alkenyl, wherein alkyl and / or alkenyl are linear or branched. R 3 and R 4 are independent of each other, C1 to C 16 -alkyl, where alkyl is linear or branched, examples are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, 1,2-dimethylpropyl, isoamyl, n-hexyl, isohexyl, sec-hexyl, n-heptyl, n-octyl, 2-ethylhexyl, n-nonyl, n-decyl, isodecyl. A - -RCOO - , -SO3 - and RSO3 - and R is selected from linear or branched C1-C8 alkyl and C1-C4 hydroxyalkyl, and is alkyl. - is SO3 - In the case of (fatty) alcohol / alkyl (ethoxy / ether) sulfate [(F)A(E)S] and A - Ga-RCOO - In this case, it can be called (fatty) alcohol / alkyl (ethoxy / ether) carboxylate [(F)A(E)C]. M + is selected from H and salt-forming cations. The salt-forming cations may be monovalent or polyvalent, and therefore M + is 1 / v M v+Examples include, but are not limited to, sodium, potassium, magnesium, calcium, ammonium, and ammonium salts of mono-, di-, and triethanolamine.

[0044] m is in the range of 0 to 200, preferably 1 to 80, more preferably 3 to 20; n and o are each independently in the range of 0 to 100; n is preferably in the range of 1 to 10, more preferably 1 to 6; o is preferably in the range of 1 to 50, more preferably 4 to 25. The sum of m, n, and o is at least 1, and preferably the sum of m, n, and o is in the range of 5 to 100, more preferably 9 to 50.

[0045] The anionic surfactants of general formula (AIS I) can be of any structure, block copolymers or random copolymers.

[0046] In one embodiment, the liquid composition of the present invention comprises at least one anionic surfactant according to formula (AIS I), wherein R 1 nC 11 H 23 and R 2 is H and A - is SO3 - and m, n, and o are 0. M + is preferably NH4 + Such a compound may be referred to herein as ammonium lauryl sulfate (ALS).

[0047] In one embodiment, the liquid composition of the present invention comprises at least one anionic surfactant according to formula (AIS I), wherein R 1 nC 11 H 23 and R 2 is selected from H, and A - is SO3 - where m is 2 to 5, preferably 3, and n and o are 0. M + is preferably Na +Such compounds may be referred to herein as lauryl ether sulfates (LES), preferably sodium lauryl ether sulfate (SLES).

[0048] The liquid composition of the present invention, in one embodiment, comprises at least two anionic surfactants selected from compounds of general formula (AIS I), one of the anionic surfactants being R 1 C 11 and R 2 is H, m is 2, n and o=0, A - is SO3 - and M + Na + and the other surfactant is R 1 C 13 and R 2 is H, m is 2, n and o=0, A - is SO3 - and M + Na + It is characterized in that:

[0049] Further suitable anionic surfactants include C 12 ~C 18 Sulfofatty acid alkyl esters (e.g., C 12 ~C 18 Sulfo fatty acid methyl esters), C 10 ~C 18 -Alkylarylsulfonic acid (e.g., nC 10 ~C 18 -alkylbenzenesulfonic acid) and C 10 ~C 18 Salts of alkyl alkoxycarboxylates (M + ) are included. In all cases, M + is selected from salt-forming cations. The salt-forming cations may be monovalent or polyvalent, and therefore M + is 1 / v M v+ Examples include, but are not limited to, sodium, potassium, magnesium, calcium, ammonium, and ammonium salts of mono-, di-, and triethanolamine.

[0050] Non-limiting examples of further suitable anionic surfactants include branched alkylbenzenesulfonates (BABS), phenylalkanesulfonates, alpha-olefinsulfonates (AOS), olefinsulfonates, alkenesulfonates, alkane-2,3-diylbis(sulfates), hydroxyalkanesulfonates and disulfonates, secondary alkane sulfonates (SAS), paraffin sulfonates (PS), sulfonated fatty acid glycerol esters, alkyl- or alkenyl succinic acids, fatty acid derivatives of amino acids, diesters and monoesters of sulfosuccinic acid.

[0051] In one embodiment, the liquid composition comprises a compound of general formula (AIS II):

[0052] [ka] (Wherein R of formula (AIS II) 1 is C 10 ~C 13 alkyl) In one embodiment, the liquid composition of the present invention comprises a salt, preferably a sodium salt, of a compound according to formula (AIS II). In one embodiment, the liquid composition of the present invention comprises at least two anionic surfactants selected from compounds of general formula (AIS II), one of which is represented by R 1 C 10 and the other surfactant is R 1 C 13 In one embodiment, the liquid composition comprises at least two anionic surfactants selected from the sodium salts of compounds of general formula (AIS II), one of which is R 1 C 10 and the other surfactant is R 1 C 13Such compounds may also be referred to herein as LAS (linear alkylbenzene sulfonates).

[0053] In one embodiment, the liquid composition of the present invention comprises a compound of general formula (AIS III):

[0054] [ka] and at least one anionic surfactant selected from the group consisting of N-acyl amino acid surfactants.

[0055] The variables of general formula (AIS III) are defined as follows: R 19 is a straight chain or branched C6-C 22 -Alkyl and straight or branched C6-C 22 -alkenyl, for example oleyl. R 20 is selected from H and C1-C4-alkyl. R 21 is selected from H, methyl, —(CH2)3NHC(NH)NH2, —CH2C(O)NH2, —CH2C(O)OH, —(CH2)2C(O)NH2, —(CH2)2C(O)OH, (imidazol-4-yl)-methyl, —CH(CH3)C2H5, —CH2CH(CH3)2, —(CH2)4NH2, benzyl, hydroxymethyl, —CH(OH)CH3, (indol-3-yl)-methyl, (4-hydroxy-phenyl)-methyl, isopropyl, —(CH2)2SCH3, and —CH2SH. R 22 is selected from -COOX and -CH2SO3X, where X is Li + , Na + and K. + (selected from).

[0056] Non-limiting examples of suitable N-acyl amino acid surfactants include: Mono- and di-carboxylates of N-acylated glutamic acid (e.g., sodium, potassium, ammonium, and ammonium salts of mono-, di-, and triethanolamine), for example, sodium cocoyl glutamate, sodium lauroyl glutamate, sodium myristoyl glutamate, sodium palmitoyl glutamate, sodium stearoyl glutamate, disodium cocoyl glutamate, disodium stearoyl glutamate, potassium cocoyl glutamate, potassium lauroyl glutamate, and potassium myristoyl glutamate; carboxylates of N-acylated alanine (e.g., sodium, potassium, ammonium, and mono-, di-, and triethanolamine), for example, sodium cocoyl glutamate, sodium lauroyl glutamate, sodium myristoyl glutamate, sodium palmitoyl glutamate, sodium stearoyl glutamate, disodium cocoyl glutamate, disodium stearoyl glutamate, potassium cocoyl glutamate, potassium lauroyl glutamate, and potassium myristoyl glutamate; ammonium salts of amines), such as sodium cocoyl alanine and lauroyl alanine triethanolamine; carboxylic acid salts of N-acylated glycines (e.g., sodium, potassium, ammonium, and ammonium salts of mono-, di-, and triethanolamine), such as sodium cocoyl glycine and potassium cocoyl glycine; carboxylic acid salts of N-acylated sarcosines (e.g., sodium, potassium, ammonium, and ammonium salts of mono-, di-, and triethanolamine), such as sodium lauroyl sarcosine, sodium cocoyl sarcosine, sodium myristoyl sarcosine, sodium oleoyl sarcosine, and ammonium lauroyl sarcosine.

[0057] In one embodiment, the liquid composition of the present invention comprises at least one anionic surfactant selected from the group of soaps. Suitable are saturated and unsaturated C 12 ~C 18 Salts of fatty acids, such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, and (hydrated) erucic acid (M + ) is. M + is selected from salt-forming cations. The salt-forming cations may be monovalent or polyvalent, and therefore M + is 1 / v M v+Examples include, but are not limited to, sodium, potassium, magnesium, calcium, ammonium, and ammonium salts of mono-, di-, and triethanolamine.

[0058] Further non-limiting examples of suitable soaps include soap mixtures derived from natural fatty acids, such as tallow, coconut oil, palm kernel oil, bay oil, olive oil, or canola oil, which contain varying amounts of lauric and / or myristic and / or palmitic and / or stearic and / or oleic and / or linoleic acid soaps, depending on the natural fatty acid from which the soap is derived.

[0059] Further non-limiting examples of suitable anionic surfactants include sulfate, sulfonate, or carboxylate salts of natural fatty acids (e.g., derived from tallow, coconut oil, palm kernel oil, bay oil, olive oil, or canola oil) (M + Such anionic surfactants include sulfates, sulfonates or carboxylates of lauric acid and / or myristic acid and / or palmitic acid and / or stearic acid and / or oleic acid and / or linoleic acid in varying amounts, depending on the natural fatty acid from which the soap is derived.

[0060] In one embodiment, the liquid composition of the present invention comprises a mixture of two or more different anionic surfactants. In one embodiment, the liquid composition comprises at least two anionic surfactants selected from compounds of general formula (AIS I), wherein one of the anionic surfactants is R 1 C 11 and R 2 is H, m is 2, n and o=0, A - is SO3 - and M + Na + and the other surfactant is R 1 C 13 and R 2 is H, m is 2, n and o=0, A -is SO3 - and M + Na + In another embodiment, the liquid composition comprises at least two anionic surfactants selected from compounds of general formula (AIS II), one of which is R 1 C 10 and the other surfactant is R 1 C 13 It is characterized in that:

[0061] In one embodiment, the liquid composition of the present invention comprises at least one anionic surfactant, and the total amount of anionic surfactant may be in the range of 0 to 50% by weight, preferably in the range of 0.5 to 45% by weight, all relative to the total weight of the liquid composition.

[0062] In one embodiment, the liquid composition of the present invention comprises at least two anionic surfactants in an amount ranging from 0.5 to 25% by weight, in the range of 1 to 20% by weight, or in the range of 1.5 to 15% by weight, all relative to the total weight of the liquid composition, wherein the at least two anionic surfactants are selected from compounds of general formula (AIS I), and one of the anionic surfactants is R 1 C 11 and R 2 is H, m is 2, n and o=0, A - is SO3 - and M + Na + and the other surfactant is R 1 C 13 and R 2 is H, m is 2, n and o=0, A - is SO3 - and M + Na + It is characterized in that:

[0063] In one embodiment, the liquid composition of the present invention comprises at least two anionic surfactants in an amount ranging from 0.5 to 25% by weight, in the range of 1 to 20% by weight, or in the range of 1.5 to 15% by weight, all relative to the total weight of the liquid composition, wherein the at least two anionic surfactants are selected from compounds of general formula (AIS II), and one of the anionic surfactants is R 1 C 10 and the other surfactant is R 1 C 13 It is characterized in that:

[0064] Nonionic surfactants (NIS) In one embodiment, the liquid composition of the present invention comprises at least one non-ionic surfactant.

[0065] In one embodiment, the liquid composition of the present invention comprises compounds of general formula (NIS Ia) and (NIS Ib):

[0066] [ka] TIFF0007798425000006.tif45161.

[0067] The variables of general formulas (NIS Ia) and (NIS Ib) are defined as follows: R 1 is C1~C 23 Alkyl and C2-C 23 Alkenyl, wherein alkyl and / or alkenyl are linear or branched, examples of which include n-CH 15 , n-C9H 19 , nC 11 H 23 , nC 13 H 27 , nC 15 H 31 , nC 17 H 35 , i-C9H 19 ,I C 12 H 25 is. R 2 H, C1~C 20 Alkyl and C2-C 20 alkenyl, wherein the alkyl and / or alkenyl are linear or branched. R 3 and R 4 are independent of each other, C1 to C 16 It is selected from alkyl, which is linear or branched, examples being methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, 1,2-dimethylpropyl, isoamyl, n-hexyl, isohexyl, sec-hexyl, n-heptyl, n-octyl, 2-ethylhexyl, n-nonyl, n-decyl, isodecyl. R 5 is H and C1~C 18 alkyl, which is straight chain or branched.

[0068] The integers in the general formulae (NIS Ia) and (NIS Ib) are defined as follows: m is in the range of 0 to 200, preferably 1 to 80, more preferably 3 to 20; n and o are each independently in the range of 0 to 100; n is preferably in the range of 1 to 10, more preferably 1 to 6; o is preferably in the range of 1 to 50, more preferably 4 to 25. The sum of m, n, and o is at least 1, and preferably the sum of m, n, and o is in the range of 5 to 100, more preferably 9 to 50.

[0069] Compounds according to formula (NIS Ia) may also be referred to herein as alkyl polyethylene glycol ethers (AEOs). Compounds according to formula (NIS Ib) may also be referred to herein as alkylphenol polyethylene glycol ethers (APEOs).

[0070] In one embodiment, the liquid composition of the present invention comprises at least one nonionic surfactant selected from the general formula (NIS Ia), wherein m is in the range of 3 to 11, preferably 7 or less, n and o are 0, and R 1 is C 12 ~C 14 and R 2 and R 5 is H. Preferably, the liquid composition comprises at least two nonionic surfactants selected from compounds of general formula (NIS Ia), one of said nonionic surfactants being R 1 C 12 and R 2 and R 5 is H, m is 7, n and o=0, and the other surfactant is R 1 C 14 and R 2 and R 5 is H, m is 7, and n and o=0.

[0071] In one embodiment, the liquid composition of the present invention comprises at least two nonionic surfactants selected from compounds of general formula (NIS Ia), one of said nonionic surfactants being R 1 nC 11 H 23 and R 5 is H, m is 7, n and o=0, and the other surfactant is R 1 C 13 H 27 and R 5 is H, m is 7, and n and o=0.

[0072] The nonionic surfactants of general formulas (NIS Ia) and (NIS Ib) can be of any structure, be it a block structure or a random structure, and are not limited to the shown sequences of formulas (NIS Ia) and (NIS Ib).

[0073] In one embodiment, the liquid composition of the present invention comprises a compound of general formula (NIS II):

[0074] [ka] and at least one nonionic surfactant selected from the group consisting of alkyl polyglycosides (APGs).

[0075] The variables of general formula (NIS II) are defined as follows: R 1 is C1~C 17 Alkyl and C2-C 17 Alkenyl, wherein alkyl and / or alkenyl are linear or branched, examples of which include n-CH 15 , n-C9H 19 , nC 11 H 23 , nC 13 H 27 , nC 15 H 31 , nC 17 H 35 , i-C9H 19 ,I C 12 H 25 is. R 2 H, C1~C 17 Alkyl and C2-C 17 alkenyl, wherein the alkyl and / or alkenyl are linear or branched. G 1 is selected from monosaccharides having 4 to 6 carbon atoms, for example, glucose and xylose. The integer w in the general formula (NIS II) is in the range of 1.1 to 4, and w is an average number.

[0076] In one embodiment, the liquid composition of the present invention comprises a compound of general formula (NIS III):

[0077] [ka] The composition comprises at least one nonionic surfactant selected from the group consisting of:

[0078] The variables of general formula (NIS III) are defined as follows: AO is selected from ethylene oxide (EO), propylene oxide (PO), butylene oxide (BO), and mixtures thereof. R 6 is C5~C 17 Alkyl and C5-C 17 alkenyl, wherein the alkyl and / or alkenyl are linear or branched. R 7 H, C1~C 18 - alkyl, wherein alkyl is linear or branched. The integer y in the general formula (NIS III) is a number ranging from 1 to 70, preferably from 7 to 15.

[0079] In one embodiment, the liquid composition of the present invention comprises a compound of the general formula (NIS IV):

[0080] [ka] The composition comprises at least one nonionic surfactant.

[0081] where in formula (NIS IV) the following applies: AO are the same or different alkylene oxides selected from CH2-CH2-O, (CH2)3-O, (CH2)4-O, CH2CH(CH3)-O, CH(CH3)-CH2-O- and CH2CH(n-C3H7)-O. R 1 is a straight chain (linear, n-) or branched C4-C 30 - alkyl and linear or branched C4-C having at least one C-C double bond 30 -alkylene. 1 is preferably a linear or branched C4-C 30 -Alkyl, n-C4~C 30 -Alkyl, n-C7~C 15 Alkyl or nC 10 ~C 12 -alkyl. R 2 is linear (straight chain, n-) or branched C1-C30 - alkyl and linear or branched C2-C having at least one C-C double bond 30 -alkylene. 2 is preferably a linear or branched C6-C 20 -Alkyl, preferably linear or branched C8-C 12 -alkyl, more preferably linear or branched C 10 ~C 12 -alkyl. The integer x in the general formula (NIS IV) is preferably a number in the range of 5-70, 10-60, 15-50, or 20-40.

[0082] In a preferred embodiment, the detergent formulation of the present invention comprises at least one nonionic surfactant according to formula (NIS IV), R 1 n-C3~C 17 alkyl, and R 2 is straight chain or branched C8-C 14 Preferably, AO is -(CH2CH2O) x2 -(CH2CH(CH3)-O) x3 , -(CH2CH2O) x2 -(CH(CH3)CH2-O) x3 , and -(CH2CH2O) x4 where x2 and x4 are numbers in the range of 15 to 50, and x3 is a number in the range of 1 to 15.

[0083] In a preferred embodiment, the detergent formulation of the present invention comprises at least one nonionic surfactant according to formula (NIS IV), R 1 is n-C8 alkyl, and R 2 is branch C 11 alkyl, AO is CH2-CH2-O, and x is 22.

[0084] In a preferred embodiment, the detergent formulation of the present invention comprises at least one nonionic surfactant according to formula (NIS IV), R 1 is n-C8 alkyl, and R 2 n-C8~C 10alkyl, AO is CH2-CH2-O, and x is 40.

[0085] In a preferred embodiment, the detergent formulation of the present invention comprises at least one nonionic surfactant according to formula (NIS IV), R 1 is n-C8 alkyl, and R 2 nC 10 alkyl, and AO is -(CH2CH2O) x2 -(CH2CH(CH3)-O) x3 , -(CH2CH2O) x2 -(CH(CH3)CH2-O) x3 where x2 is 22 and x3 is 1.

[0086] In one embodiment, the at least one nonionic surfactant is selected from sorbitan esters and / or ethoxylated or propoxylated sorbitan esters. Non-limiting examples are products sold under the trade names SPAN and TWEEN.

[0087] The nonionic surfactant may further be selected from alkoxylated mono- or di-alkylamines, fatty acid monoethanolamides (FAMA), fatty acid di-ethanolamides (FADA), ethoxylated fatty acid monoethanolamides (EFAM), propoxylated fatty acid monoethanolamides (PFAM), polyhydroxyalkyl fatty acid amides, or N-acyl N-alkyl derivatives of glucosamine (glucamides, GA, or fatty acid glucamides, FAGA), and combinations thereof.

[0088] In one embodiment, the liquid composition of the present invention comprises at least one nonionic surfactant, and the total amount of nonionic surfactant may be in the range of 0% to 80% by weight, preferably in the range of 0.5% to 70% by weight, all based on the total weight of the liquid composition.

[0089] In one embodiment, the liquid composition of the present invention comprises at least one nonionic surfactant in the range of 0.3% to 30% by weight, in the range of 0.4% to 20% by weight, or in the range of 0.5% to 10% by weight, all based on the total weight of the liquid composition. In one embodiment, the at least one nonionic surfactant is selected from surfactants according to the general formula (NIS Ia), wherein m is 7, n and o are 0, and R 1 is C 12 ~C 14 and R 2 and R 5 is H. In one embodiment, the liquid composition of the present invention comprises two nonionic surfactants selected from compounds of general formula (NIS Ia), one of said nonionic surfactants being R 1 C 12 and R 2 and R 5 is H, m is 7, n and o=0, and the other surfactant is R 1 C 14 and R 2 and R 5 is H, m is 7, and n and o=0.

[0090] In one embodiment, the detergent formulation comprising at least one nonionic surfactant according to formula (NIS IV), preferably as disclosed above, is an automatic dishwashing detergent. Preferably, the automatic dishwashing detergent comprises at least one compound according to formula (NIS IV) in the range of about 0.3% to 10% by weight, about 0.5% to 5% by weight, or about 1% to 3% by weight, all based on the total weight of the detergent formulation. In one embodiment, the at least one nonionic surfactant is a compound according to formula (NIS IV), and R 1 is n-C8 alkyl, and R 2 is branch C 11 alkyl, AO is CH2-CH2-O, and x is 22.

[0091] In one embodiment, the liquid composition of the present invention comprises a mixture of two or more different nonionic surfactants.

[0092] In one embodiment of the present invention, component (c) comprises at least one anionic surfactant and at least one nonionic surfactant, and the weight ratio of the anionic surfactant to the nonionic surfactant is 5:1 to 1:5, preferably 2:1 to 1:4.

[0093] In another embodiment of the invention, the liquid composition has a weight ratio of anionic surfactant to nonionic surfactant of 4:1 to 1:4. Preferably, said liquid composition is a detergent for laundry and / or manual dishwashing and / or cleaning of medical equipment.

[0094] Ingredient (d) In one aspect, the liquid composition of the present invention includes a component (d) comprising at least one hydrolase other than DNAse. In one embodiment, the liquid composition of the present invention includes at least one protease and / or at least one amylase and / or at least one cellulase and / or at least one lipase and / or at least one mannanase. Preferably, the composition of the present invention includes at least one hydrolase other than DNAse that can degrade extracellular matrix components, preferably selected from extracellular polymeric substances. Examples of EPS include, but are not limited to, polysaccharides, proteins, and lipids.

[0095] In one embodiment, the hydrolase, which is different from the DNAse, is capable of hydrolyzing proteins.

[0096] In one embodiment, the hydrolase different from DNAse is capable of hydrolyzing β-1,4-glycosidic bonds and is preferably capable of degrading a substrate selected from amylopectin, amylose and cellulose.

[0097] In one embodiment, the hydrolase, which is different from DNAse, is capable of hydrolyzing β-1,6-glycosidic bonds and preferably capable of degrading poly-N-acetylglucosamine.

[0098] Proteases In one embodiment, the compositions of the invention comprise at least one protease. The protease is a member of class EC 3.4, including aminopeptidases (EC 3.4.11), dipeptidases (EC 3.4.13), dipeptidyl-peptidases and tripeptidyl-peptidases (EC 3.4.14), peptidyl-dipeptidases (EC 3.4.15), serine-type carboxypeptidases (EC 3.4.16), metallocarboxypeptidases (EC 3.4.17), cysteine-type carboxypeptidases (EC 3.4.18), and the like. endopeptidases (EC 3.4.18), omega peptidases (EC 3.4.19), serine endopeptidases (EC 3.4.21), cysteine ​​endopeptidases (EC 3.4.22), aspartic endopeptidases (EC 3.4.23), metallo-endopeptidases (EC 3.4.24), threonine endopeptidases (EC 3.4.25) or endopeptidases of unknown catalytic mechanism (EC 3.4.99).

[0099] In a preferred embodiment, the at least one protease is selected from serine proteases (EC 3.4.21). Serine proteases or serine peptidases are characterized by having a serine in the catalytic active site, which forms a covalent adduct with the substrate during catalysis. Serine proteases in the context of the present invention are selected from the group consisting of chymotrypsin (e.g., EC 3.4.21.1), elastase (e.g., EC 3.4.21.36), elastase (e.g., EC 3.4.21.37 or EC 3.4.21.71), granzymes (e.g., EC 3.4.21.78 or EC 3.4.21.79), kallikreins (e.g., EC 3.4.21.34, EC 3.4.21.35, EC 3.4.21.118 or EC 3.4.21.119), plasmin (e.g., EC 3.4.21.7), trypsin (e.g., EC 3.4.21.4), thrombin (e.g., EC 3.4.21.5) and subtilisin. Subtilisins are also known as subtilopeptidases, eg EC 3.4.21.62, which is also referred to below as "subtilisin".

[0100] A protease is an active protein that exerts "protease activity" or "proteolytic activity." Proteolytic activity relates to the rate of degradation of a protein by a protease or proteolytic enzyme over a defined time course.

[0101] Methods for analyzing proteolytic activity are well known in the literature (see, e.g., Gupta et al. (2002), Appl. Microbiol. Biotechnol. 60: 381-395). Proteolytic activity can be determined by using succinyl-Ala-Ala-Pro-Phe-p-nitroanilide as a substrate (Suc-AAPF-pNA, short AAPF; see, e.g., DelMar et al. (1979), Analytical Biochem 99, 316-320). pNA is cleaved from the substrate molecule by proteolytic cleavage, resulting in the release of a yellow color of free pNA, and an increase in OD 405 This can be quantified by measuring

[0102] In a preferred embodiment of the present invention, the at least one protease is selected from the following: subtilisin from Bacillus amyloliquefaciens BPN' (described by Vasantha et al. (1984) J. Bacteriol. Volume 159, pp. 811-819 and J.A. Wells et al. (1983) in Nucleic Acids Research, Volume 11, pp. 7911-7925), subtilisin from Bacillus licheniformis (subtilisin Carlsberg; E.L. Smith et al. (1968) in J. Biol. Chem., Volume 243, pp. 2184-2191 and Jacobs et al. (1985) in Nucl. Acids Res., Vol. 13, pp.8913-8926), subtilisin PB92 (the original sequence of alkaline protease PB92 is described in EP 283075), subtilisin 147 and / or 309 (Esperase® and Savinase®, respectively) disclosed in WO 89 / 06279, subtilisins from Bacillus lentus disclosed in WO 91 / 02792, such as subtilisin from Bacillus lentus DSM 5483 or variants of Bacillus lentus DSM 5483 described in WO 95 / 23221, subtilisins from Bacillus alkalophilus (DSM 11233) disclosed in German Patent No. 10064983, subtilisin from Bacillus gibsonii disclosed in WO 2003 / 054184, subtilisin from Bacillus sp. (DSM 14391) disclosed in WO 2003 / 056017, subtilisin from Bacillus sp. (DSM 14390) disclosed in WO 2003 / 055974, subtilisin from Bacillus sp. (DSM 14392) disclosed in WO 2003 / 054184, subtilisin having SEQ ID NO: 4 disclosed in WO 2005 / 063974, subtilisin having SEQ ID NO: 4 disclosed in WO 2005 / 103244, subtilisin having SEQ ID NO: 7 disclosed in WO 2005 / 103244, and subtilisin having SEQ ID NO: 2 disclosed in German Patent Application No. 102005028295.4.

[0103] In one embodiment, the composition of the invention comprises at least subtilisin 309 (which may be referred to herein as Savinase) as disclosed as sequence a) in Table I of WO 89 / 06279 or a variant thereof that is at least 80% identical and has proteolytic activity.

[0104] Suitable proteases include proteolytically active protease variants that are at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical when compared to the full-length polypeptide sequence of the parent enzymes disclosed above.

[0105] In one embodiment, the at least one protease has SEQ ID NO: 22 (which is the sequence of the mature alkaline protease from Bacillus lentus DSM 5483) described in EP 1921147, or a protease at least 80% identical thereto and having proteolytic activity. Preferred variants of the subtilisin protease derived from SEQ ID NO: 22 described in EP 1 921 147 are characterized by one or more amino acid substitutions at the following positions: 3, 4, 9, 15, 24, 27, 33, 36, 57, 68, 76, 77, 87, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 106, 118, 120, 123, 128, 129, 130, 131, 154, 160, 167, 170, 194, 195, 199, 205, 206, 217, 218, 222, 224, 232, 235, 236, 245, 248, 252 and 274 (according to BPN numbering) which have proteolytic activity. Most preferably, such proteases are not mutated at positions Asp32, His64 and Ser221 (according to BPN numbering).

[0106] In one embodiment, the protease is characterized by having the amino acid glutamic acid (E), or aspartic acid (D), or asparagine (N), or glutamine (Q), or alanine (A), or glycine (G), or serine (S) at position 101 (according to BPN numbering) and has proteolytic activity. In one embodiment, the protease comprises one or more additional substitutions: (a) threonine at position 3 (3T), (b) isoleucine at position 4 (4I), (c) alanine, threonine, or arginine at position 63 (63A, 63T, or 63R), (d) aspartic acid or glutamic acid at position 156 (156D or 156E), (e) proline at position 194 (194P), (f) methionine at position 199 (199M), (g) isoleucine at position 205 (205I), (h) aspartic acid, glutamic acid, or glycine at position 217 (217D, 217E, or 217G), or (i) a combination of two or more amino acids from (a)-(h). At least one protease may be at least 80% identical to SEQ ID NO: 22 described in EP 1921147 and characterized by comprising one amino acid (according to (a)-(h)) or a combination according to (i) together with amino acids 101E, 101D, 101N, 101Q, 101A, 101G, or 101S (according to BPN numbering) and having proteolytic activity. In one embodiment, the protease is characterized by comprising the mutations (according to BPN numbering) R101E, or S3T+V4I+V205I, or R101E and S3T, V4I and V205I, or S3T+V4I+V199M+V205I+L217D.

[0107] In one embodiment, the protease according to SEQ ID NO: 22 described in EP 1921147 is characterized by comprising the mutations (according to BPN numbering) S3T+V4I+S9R+A15T+V68A+D99S+R101S+A103S+I104V+N218D.

[0108] In accordance with the present invention, the liquid composition of the present invention comprises two or more proteases disclosed above.

[0109] Compositions comprising at least one protease also typically include at least one protease stabilizer selected from those disclosed herein.

[0110] amylase In one embodiment, the liquid composition of the present invention comprises at least one amylase. "Amylases" (alpha and / or beta) according to the present invention include those of bacterial or fungal origin (EC 3.2.1.1 and 3.2.1.2, respectively), including chemically modified or protein engineered mutants.

[0111] The amylases of the present invention have "amylolytic activity" or "amylase activity" which involves the (internal) hydrolysis of glycosidic bonds in polysaccharides. Alpha-amylase activity can be determined by assays that measure alpha-amylase activity known to those skilled in the art. Examples of assays that measure alpha-amylase activity include the following: Alpha-amylase activity can be determined by a method using Phadebas tablets as a substrate (Phadebas Amylase Test, supplied by Magle Life Science). Starch is hydrolyzed by alpha-amylase to yield a soluble blue fragment. The absorbance of the resulting blue solution, measured spectrophotometrically at 620 nm, is a function of alpha-amylase activity. The measured absorbance is directly proportional to the specific activity (activity / mg pure alpha-amylase protein) of the alpha-amylase in question under a given set of conditions.

[0112] In one embodiment, the at least one amylase is selected from: Amylase from Bacillus licheniformis having SEQ ID NO: 2 as described in WO 95 / 10603. Suitable variants having amylolytic activity and comprising substitutions at 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 are described in WO 95 / 10603. The variant is described in SEQ ID NO: 4 of WO 94 / 02597, WO 94 / 018314, WO 97 / 043424 and WO 99 / 019467. an amylase from B. stearothermophilus having SEQ ID NO: 6 as disclosed in WO 02 / 10355, or an amylase optionally having a C-terminal truncation relative to the wild-type sequence. Suitable variants of SEQ ID NO: 6 include those containing a deletion at positions 181 and / or 182 and / or a substitution at position 193. The amylase from Bacillus sp. 707 having SEQ ID NO: 6 as disclosed in WO 99 / 19467. Preferred variants of SEQ ID NO: 6 are those which have substitutions, deletions or insertions at one or more of the following positions: R181, G182, H183, G184, N195, I206, E212, E216 and K269. The amylase from Bacillus halmapalus having SEQ ID NO: 2 or SEQ ID NO: 7 as described in WO 96 / 23872, also referred to herein as SP-722. Preferred variants are described in WO 97 / 3296, WO 99 / 194671 and WO 2013 / 001078. The amylase from Bacillus sp. DSM12649 having SEQ ID NO: 4 as disclosed in WO 00 / 22103 and functional variants thereof. The amylase from Bacillus sp. strain TS-23 having SEQ ID NO: 2 as disclosed in WO 2009 / 061380 and functional variants thereof. - an amylase from Cytophaga sp. having SEQ ID NO: 1 as disclosed in WO 2013 / 184577. The amylase from Bacillus megaterium DSM90 having SEQ ID NO: 1 as disclosed in WO 2010 / 104675 and functional variants thereof. Amylase derived from Bacillus species comprising amino acids 1 to 485 of SEQ ID NO: 2 as described in WO 00 / 60060, and functional variants thereof. Amylases from Bacillus amyloliquefaciens, preferably selected from amylases according to SEQ ID NO: 3 described in WO 2016 / 092009, and functional variants thereof. an amylase having SEQ ID NO: 12 as described in WO 2006 / 002643 or an amylase variant comprising the substitutions Y295F and M202LITV in SEQ ID NO: 12. an amylase having SEQ ID NO: 6 as described in WO 2011 / 098531 or an amylase variant comprising a substitution in SEQ ID NO: 6 at one or more positions selected from the group consisting of 193 [G, A, S, T or M], 195 [F, W, Y, L, I or V], 197 [F, W, Y, L, I or V], 198 [Q or N], 200 [F, W, Y, L, I or V], 203 [F, W, Y, L, I or V], 206 [F, W, Y, N, L, I, V, H, Q, D or E], 210 [F, W, Y, L, I or V], 212 [F, W, Y, L, I or V], 213 [G, A, S, T or M] and 243 [F, W, Y, L, I or V]. an amylase having SEQ ID NO: 1 as described in WO 2013 / 001078 or an amylase variant comprising alterations at two or more (several) positions corresponding to positions G304, W140, W189, D134, E260, F262, W284, W347, W439, W469, G476 and G477 in SEQ ID NO: 1. an amylase having SEQ ID NO:2 as described in WO 2013 / 001087 or an amylase variant comprising a deletion of positions 181+182, or 182+183, or 183+184 in SEQ ID NO:2, and optionally comprising one or more modifications at any of the positions corresponding to W140, W159, W167, Q169, W189, E194, N260, F262, W284, F289, G304, G305, R320, W347, W439, W469, G476 and G477 in SEQ ID NO:2. amylases which are hybrid alpha-amylases derived from the above amylases and functional variants thereof, for example as described in WO 2006 / 066594; a hybrid amylase according to WO 2014 / 183920 having A and B domains with at least 90% identity to SEQ ID NO: 2 of WO 2014 / 183920 and a C domain with at least 90% identity to SEQ ID NO: 6 of WO 2014 / 183920, wherein the hybrid amylase has amylolytic activity, preferably a hybrid alpha-amylase with at least 95% identity to SEQ ID NO: 23 of WO 2014 / 183920, and wherein the hybrid amylase has amylolytic activity; A hybrid amylase according to WO2014 / 183921 having A and B domains having at least 75% identity to SEQ ID NO:2, SEQ ID NO:15, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:29, SEQ ID NO:26, SEQ ID NO:32 and SEQ ID NO:39 disclosed in WO2014 / 183921 and a C domain having at least 90% identity to SEQ ID NO:6 of WO2014 / 183921, wherein the hybrid amylase has amylolytic activity, preferably a hybrid alpha-amylase which is at least 95% identical to SEQ ID NO:30 disclosed in WO2014 / 183921 and has amylolytic activity.

[0113] In one embodiment, the liquid composition of the present invention comprises two or more amylases disclosed above.

[0114] cellulase In one embodiment, the liquid composition of the present invention comprises at least one cellulase selected from cellobiohydrolase (1,4-PD-glucan cellobiohydrolase, EC 3.2.1.91), endo-ss-1,4-glucanase (endo-1,4-PD-glucan 4-glucanohydrolase, EC 3.2.1.4), and ss-glucosidase (EC 3.2.1.21). Preferably, the composition comprises at least one cellulase from glycosyl hydrolase family 7 (GH7, pfam00840), preferably selected from endoglucanases (EC 3.2.1.4).

[0115] "Cellulase," "cellulase enzyme," or "cellulolytic enzyme" is an enzyme involved in the hydrolysis of cellulose. Assays for measuring "cellulase activity" or "cellulolytic activity" are known to those skilled in the art. For example, cellulolytic activity can be determined by the fact that cellulases hydrolyze carboxymethylcellulose to reducing carbohydrates, the reducing capacity of which is determined colorimetrically by the ferricyanide reaction according to Hoffman, WS, J. Biol. Chem. 120, 51 (1937).

[0116] The cellulases according to the present invention include those of bacterial or fungal origin. In one embodiment, at least one cellulase is selected from cellulases containing a cellulose-binding domain. In one embodiment, at least one cellulase is selected from cellulases containing only a catalytic domain, meaning that the cellulase lacks a cellulose-binding domain.

[0117] In one embodiment, the liquid composition comprises at least one cellulase that is 80% identical to the polypeptide sequence according to SEQ ID NO: 2 of WO 95 / 02675. In one embodiment, the composition comprises at least one cellulase that is 80% identical to the polypeptide sequence according to SEQ ID NO: 4 of WO 2004 / 053039. In one embodiment, the liquid composition comprises at least one cellulase that is 80% identical to the polypeptide sequence according to SEQ ID NO: 2 of WO 2002 / 99091.

[0118] In one embodiment, the liquid composition of the present invention comprises two or more cellulases, preferably endoglucanases (EC 3.2.1.4), as disclosed above.

[0119] Lipase In one embodiment, the liquid composition of the present invention comprises at least one lipase. The terms "lipase," "lipolytic enzyme," and "lipid esterase" all refer to enzymes in EC class 3.1.1 ("carboxylic acid ester hydrolase"). Lipase refers to an active protein having lipase activity (or lipolytic activity, triacylglycerol lipase, EC 3.1.1.3), cutinase activity (EC 3.1.1.74; enzymes having cutinase activity may be referred to herein as cutinases), sterol esterase activity (EC 3.1.1.13), and / or wax ester hydrolase activity (EC 3.1.1.50).

[0120] Methods for determining lipolytic activity are well known in the literature (see, e.g., Gupta et al. (2003), Biotechnol. Appl. Biochem. 37, pp. 63-71). For example, lipase activity can be measured by ester bond hydrolysis of the substrate para-nitrophenyl palmitate (palmitate-pNP, C:16), releasing pNP, which is yellow and can be detected at 405 nm.

[0121] Lipases include those of bacterial or fungal origin and known to those skilled in the art to be useful.

[0122] In one embodiment, the at least one lipase is selected from fungal triacylglycerol lipases (EC class 3.1.1.3). The fungal triacylglycerol lipase may be selected from lipases of Thermomyces lanuginosa. In one embodiment, the at least one Thermomyces lanuginosa lipase is selected from triacylglycerol lipases according to amino acids 1-269 of SEQ ID NO: 2 of U.S. Patent No. 5,869,438 and variants thereof having lipolytic activity. Preferably, the at least one Thermomyces lanuginosa lipase may be at least 80% identical to SEQ ID NO: 2 of U.S. Patent No. 5,869,438 and may be characterized by having amino acids T231R and N233R. The Thermomyces lanuginosa lipase variant preferably further comprises one or more of the following amino acid exchanges: Q4V, V60S, A150G, L227G, P256K.

[0123] In one embodiment, the liquid composition of the present invention comprises at least two lipases as disclosed above.

[0124] Mannan-degrading enzyme In one embodiment, the liquid composition of the present invention comprises at least one mannan-degrading enzyme selected from β-mannosidase (EC 3.2.1.25), endo-1,4-β-mannosidase (EC 3.2.1.78), and 1,4-β-mannobiosidase (EC 3.2.1.100). Preferably, the at least one mannan-degrading enzyme is selected from the group of endo-1,4-β-mannosidases (EC 3.2.1.78), a group of enzymes referred to herein as endo-β-1,4-D-mannanases, β-mannanases, or mannanases.

[0125] Polypeptides having mannanase activity can be tested for mannanase activity according to standard testing procedures known in the art, for example, by applying the solution to be tested to a 4 mm diameter hole punched into an agar plate containing 0.2% AZCL galactomannan (carob), a substrate for the assay of endo-1,4-beta-D-mannanase available from Megazyme, Inc., catalog number I-AZGMA (Megazyme's internet address: http: / / www.megazyme.com / Purchase / index.html).

[0126] In one embodiment, the liquid composition of the present invention comprises at least one mannanase selected from alkaline mannanases of family 5 or 26. The term "alkaline mannanase" is intended to encompass mannanases that have an enzymatic activity of at least 40% of its maximum activity at a given pH in the range of 7 to 12, preferably 7.5 to 10.5.

[0127] In one embodiment, the at least one mannanase contained in the composition of the present invention is, for example, a mannanase derived from a Bacillus species, as described in JP-0304706 [beta-mannanase derived from Bacillus species], JP-63056289 [alkaline, thermostable beta-mannanase], JP-63036774 [Bacillus microorganisms producing beta-mannanase and beta-mannosidase at alkaline pH FERM]. P-8856], JP-08051975 [alkaline beta-mannanase from alkaliphilic Bacillus sp. AM-001], WO 97 / 11164 [mannanase from Bacillus amyloliquefaciens], WO 91 / 18974 [mannanases active at extremes of pH and temperature], and WO 2014 / 100018 [endo-(3-mannanase 1 cloned from Bacillus circulans or Bacillus lentus strain CMG1240 (Bleman1; see U.S. Pat. No. 5,476,775)]. Suitable mannanases are described in WO 99 / 064619.

[0128] In one embodiment, the at least one mannanase included in the liquid composition of the present invention is a polypeptide that is 80% identical to SEQ ID NO: 2 of U.S. Patent No. 6,566,114. The at least one mannanase included in the liquid composition of the present invention is selected from mannanases derived from Trichoderma organisms, for example, as disclosed in WO 93 / 24622 or WO 2011 / 085747.

[0129] The mannanase variants of the present invention are 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%, at least 96%, at least 97%, at least 98% or at least 99% identical when compared to the full-length polypeptide sequence of the corresponding parent enzyme disclosed above.

[0130] In one embodiment, the liquid composition of the present invention comprises two or more mannan-degrading enzymes disclosed above, preferably endo-1,4-β-mannosidase (EC 3.2.1.78).

[0131] Further ingredients solvent In one embodiment, the liquid composition of the present invention comprises water in an amount ranging from 5% to 95% by weight, ranging from 5% to 70% by weight, ranging from 5% to 50% by weight, or ranging from 50% to 95% by weight, all of which weight percentages are based on the total weight of the liquid composition.

[0132] In one embodiment, the liquid composition of the present invention comprises at least one organic solvent, preferably selected from water-miscible organic solvents. Examples of such organic solvents include, but are not limited to, acetaldehyde, acetic acid, acetone, acetonitrile, butan-2-ol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, 2-butoxyethanol, 2-(2-butoxyethoxy)ethanol, butyric acid, 2-ethoxyethanol, diethanolamine, diethylene glycol, diethylenetriamine, dipropylene glycol, dimethylformamide, dimethoxyethane, dimethyl sulfoxide, 1,4-doxane, ethanol, ethylamine, ethylene glycol , formic acid, furfuryl alcohol, hexylene glycol, methanol, 2-methoxyethanol, methyldiethianolamine, methyl isocyanide, N-methyl-2-pyrrolidone, 1-propanol, 1,2-propanediol, 1,3-propanediol, 1,2,3-propanetriol, 1,2-pentanediol, 1,5-pentanediol, 2-propanol, propanoic acid, propylene glycol, 2-propoxyethanol, 2-(2-propoxyethoxy)ethanol, pyridine, tetrahydrofuran, and triethylene glycol.

[0133] Preferably, the liquid composition of the present invention comprises at least one organic solvent selected from ethanol, n-propanol, iso-propanol, n-butanol, iso-butanol, sec-butanol, ethylene glycol, 1,2-propylene glycol, 1,3-propanediol, 1,2,3-propanetriol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, diethylene glycol, 2-(2-butoxyethoxy)ethanol, 2-(2-propoxyethoxy)ethanol, hexylene glycol, 2-methoxyethanol, 2-ethoxyethanol, and 2-propoxyethanol, with ethanol, isopropanol, or 1,2-propylene glycol being more preferred.

[0134] The liquid composition of the present invention typically comprises an organic solvent in an amount ranging from 0% to 30% by weight, from 2% to 25% by weight, or from 5% to 20% by weight, based on the total weight of the liquid composition. In one embodiment, the liquid composition of the present invention comprises 5% to 10% of an organic solvent, preferably 1,2-propylene glycol.

[0135] A liquid composition that is intended to be enclosed in a water-soluble container, for example a pouch made of a water-soluble polymer film, preferably contains as little water as possible to ensure the shelf-life stability of the pouch, where shelf-life stability of the pouch means that the polymer film does not become leaky during the shelf-life period before the pouch is used in a washing process.

[0136] Compounds that stabilize the liquid composition itself "A compound that stabilizes the liquid composition itself" means any compound that can establish the storage stability of the liquid composition in an amount effective to ensure storage stability.

[0137] To those skilled in the art, shelf stability in relation to liquid compositions typically includes product appearance and dosage uniformity.

[0138] The appearance of the product is affected by the pH of the product and by the presence of compounds such as antioxidants, viscosity modifiers, emulsifiers, and the like.

[0139] Dose uniformity is usually related to product homogeneity.

[0140] The compositions according to the invention may be alkaline or may exhibit a neutral or slightly acidic pH value, for example, from 6 to 14, from 6.5 to 13, from 8 to 10.5, or from 8.5 to 9.0.

[0141] In one embodiment, the liquid composition of the present invention comprises at least one preservative selected from the group consisting of 2-phenoxyethanol, glutaraldehyde, 2-bromo-2-nitropropane-1,3-diol, and formic acid in acid form or as a salt thereof.

[0142] In one embodiment, the liquid composition comprises such a preservative, preferably 2-phenoxyethanol, at a concentration of 0.01% to 5% or 0.1% to 2% by weight relative to the total weight of the detergent formulation.

[0143] In another embodiment, the liquid composition comprises a preservative, preferably 2-bromo-2-nitropentane-1,3-diol, at a concentration of from 5 ppm to 5000 ppm or from 20 ppm to 1000 ppm by total weight of the detergent formulation.

[0144] In yet another embodiment of the invention, the liquid composition comprises glutaraldehyde in a concentration of from 2 ppm to 5000 ppm or from 10 ppm to 2000 ppm by total weight of the detergent formulation.

[0145] In yet another embodiment of the invention, the liquid composition comprises formic acid (as the acid or its salts) in a concentration of 0.01% to 3% or 0.05% to 0.5% by weight relative to the total weight of the detergent formulation.

[0146] In one embodiment, the liquid composition of the present invention comprises at least one enzyme stabilizer selected from boron-containing compounds, polyols, peptide aldehydes, other stabilizers, and mixtures thereof.

[0147] The boron-containing compound is selected from boric acid and its derivatives, boronic acid and its derivatives, such as arylboronic acids and their derivatives, salts thereof, and mixtures thereof. Boric acid may also be referred to herein as orthoboric acid.

[0148] In one embodiment, the at least one boron-containing compound is selected from the group consisting of arylboronic acids and their derivatives. In one embodiment, the boron-containing compound is selected from the group consisting of benzeneboronic acid (BBA), also known as phenylboronic acid (PBA), its derivatives, and mixtures thereof. In one embodiment, the phenylboronic acid derivative is a derivative of formula (1a) and formula (1b):

[0149] [ka] (In the formula, R1 is selected from the group consisting of hydrogen, hydroxy, unsubstituted or substituted C1-C6 alkyl, and unsubstituted or substituted C1-C6 alkenyl, and in a preferred embodiment, R is selected from the group consisting of hydroxy and unsubstituted C1 alkyl; R2 is selected from the group consisting of hydrogen, hydroxy, unsubstituted or substituted C1-C6 alkyl, and unsubstituted or substituted C1-C6 alkenyl, and in preferred embodiments, R is selected from the group consisting of H, hydroxy, and substituted C1 alkyl. is selected from the group consisting of:

[0150] In one embodiment, the phenylboronic acid derivative is selected from the group consisting of 4-formylphenylboronic acid (4-FPBA), 4-carboxyphenylboronic acid (4-CPBA), 4-(hydroxymethyl)phenylboronic acid (4-HMPBA), and p-tolylboronic acid (p-TBA).

[0151] In one embodiment, the liquid composition of the present invention comprises at least one polyol selected from polyols containing 2 to 6 hydroxyl groups, preferably selected from glycol, propylene glycol, 1,2-propanediol, 1,2-butanediol, 1,2-pentanediol, ethylene glycol, hexylene glycol, glycerol, sorbitol, mannitol, erythritol, glucose, fructose, lactose, and mixtures thereof.

[0152] In one embodiment, the liquid composition comprises at least one peptide aldehyde selected from di-, tri-, or tetrapeptide aldehydes and aldehyde analogues thereof (B1-BO-R, where R is H, CH3, CX3, CHX2, or CH2X (X = halogen), B1 is a single amino acid residue (in one embodiment, with an optionally substituted aliphatic or aromatic side chain), and B1 consists of one or more amino acid residues (in one embodiment, 1, 2, or 3) either in a form optionally containing an N-terminal protecting group or in a form as described in WO 09 / 118375 and WO 98 / 13459), or a protein-type protease inhibitor, for example, RASI, BASI, WASI (rice, barley, and wheat bifunctional alpha-amylase / subtilisin inhibitor), or CI2 or SSI.

[0153] In some embodiments, at least one peptide stabilizer is a compound of formula (2):

[0154] [ka] The compound is selected from salts thereof. R in equation (2) 1 , R 2 , R 3 , R 4 , R 5 and Z is defined as follows: R 1 is NH-CHR 1-CO is an L- or D-amino acid residue of Gly, Ala, Val, Leu, Ile, Met, Pro, Phe, Trp, Ser, Thr, Asp, Gln, Tyr, Cys, Lys, Arg, His, Asn, Glu, m-tyrosine, 3,4-dihydroxyphenylalanine, Nva, or Nle. 1 is NH-CHR 1 More preferably, R is a group such that -CO is an L- or D-amino acid residue of Ala, Val, Gly, Arg, Leu, Phe, Ile, His, or Thr. 1 is NH-CHR 1 The group -CO is an L- or D-amino acid residue of Ala, Val, Gly, Arg, Leu, Ile or His. R 2 is NH-CHR 2 -CO is an L- or D-amino acid residue of Gly, Ala, Val, Leu, Ile, Met, Pro, Phe, Trp, Ser, Thr, Asp, Gln, Tyr, Cys, Lys, Arg, His, Asn, Glu, m-tyrosine, 3,4-dihydroxyphenylalanine, Nva, or Nle. 2 is NH-CHR 2 More preferably, R is an L- or D-amino acid residue of Ala, Cys, Gly, Pro, Ser, Thr, Val, Nva, or Nle. 2 is NH-CHR 2 The group -CO is an L- or D-amino acid residue of Ala, Gly, Pro or Val. R 3 is NH-CHR 3 -CO is an L- or D-amino acid residue of Tyr, m-tyrosine, 3,4-dihydroxyphenylalanine, Phe, Val, Ala, Met, Nva, Leu, Ile, or Nle or other unnatural amino acids bearing an alkyl group. 3 is NH-CHR 3 The group -CO is an L- or D-amino acid residue of Tyr, Phe, Val, Ala or Leu.

[0155] In one embodiment, R 1 is NH-CHR 1 -CO is an L- or D-amino acid residue of Gly or Val, and R 2 is NH-CHR 2 -CO is an L- or D-amino acid residue of Ala, and R 3 is NH-CHR 3 The group is such that -CO is an L- or D-amino acid residue of Tyr, Ala or Leu.

[0156] In one embodiment, R 1 is NH-CHR 1 -CO is an L- or D-amino acid residue of Val, and R 2 is NH-CHR 2 -CO is an L- or D-amino acid residue of Ala, and R 3 is NH-CHR 3 The group is such that -CO is an L- or D-amino acid residue of Leu.

[0157] In one embodiment, R 4 and R 5 are each independently selected from methyl, ethyl, isopropyl, 2-butyl or 3-pentyl. More preferably, R 4 and R 5 are both methyl, ethyl, isopropyl, 2-butyl or 3-pentyl.

[0158] Z is selected from hydrogen, an N-terminal protecting group, and one or more amino acid residues optionally containing an N-terminal protecting group. Preferably, Z is an N-terminal protecting group.

[0159] When Z is one or more amino acid residues containing an N-terminal protecting group, the N-terminal protecting group is preferably a small aliphatic group such as formyl, acetyl, fluorenylmethyloxycarbonyl (Fmoc), tert-butyloxycarbonyl (Boc), methoxycarbonyl (Moc), methoxyacetyl (Mac), methyl carbamate, or methylaminocarbonyl / methylurea group. In the case of a tripeptide, the N-terminal protecting group is preferably a bulky aromatic group such as benzoyl (Bz), benzyloxycarbonyl (Cbz), p-methoxybenzylcarbonyl (MOZ), benzyl (Bn), p-methoxybenzyl (PMB), or p-methoxyphenyl (PMP).

[0160] Further suitable N-terminal protecting groups are described in Greene's Protective Groups in Organic Synthesis, 5th Edition, by Peter GMWuts, published by John Wiley & Sons, Inc. in 2014, and in Isidro-Llobet et al., Amino Acid-Protecting Groups, Chem. Rev. 2009 109(6), 2455-2504.

[0161] In a preferred embodiment, the peptide stabilizer is selected from a compound according to formula (2): ·R 1 and R 2 is NH-CHR 1 -CO and NH-CHR 2 -CO is a group such that each -CO is an L- or D-amino acid residue selected from Ala, Cys, Gly, Pro, Ser, Thr, Val, Nva, or Nle; 3 is NH-CHR 3 -CO is an L- or D-amino acid residue selected from Tyr, m-tyrosine, 3,4-dihydroxyphenylalanine, Phe, Val, Ala, Met, Nva, Leu, Ile or Nle, and, The N-terminal protecting group Z is selected from benzyloxycarbonyl (Cbz), p-methoxybenzylcarbonyl (MOZ), benzyl (Bn), benzoyl (Bz), p-methoxybenzyl (PMB), p-methoxyphenyl (PMP), formyl, acetyl (Ac), methyloxy, alkoxycarbonyl, methoxycarbonyl, fluorenylmethyloxycarbonyl (Fmoc), or tert-butyloxycarbonyl (Boc).

[0162] In one embodiment, the liquid composition comprises about 0.1-2 wt. % of at least one peptide stabilizer, based on the total weight of the liquid composition. Preferably, the liquid composition comprises about 0.15-1 wt. %, or 0.2-0.5 wt. %, or about 0.3 wt. % of at least one peptide stabilizer, based on the total weight of the liquid composition. More preferably, the liquid composition comprises about 0.3 wt. % of at least one peptide stabilizer, based on the total weight of the liquid composition. ·R 1 However, NH-CHR 1 -CO is an L- or D-amino acid residue of Val, and R 2 However, NH-CHR 2 -CO is an L- or D-amino acid residue of Ala, and R 3 However, NH-CHR 3 -CO is an L- or D-amino acid residue of Leu, and The N-terminal protecting group Z is benzyloxycarbonyl (Cbz). The peptide stabilizer according to formula (2) is characterized in that:

[0163] In one embodiment, the liquid composition of the present invention comprises at least one polyol as disclosed above and at least one peptide stabilizer as disclosed above.

[0164] In one embodiment, the liquid composition of the present invention comprises a salt preferably selected from the alkali salts of NaCl or KCl, lactic acid, formic acid, and mixtures thereof.

[0165] In one embodiment, the liquid compositions of the present invention comprise water-soluble sources of zinc(II), calcium(II) and / or magnesium(II) ions (E), as well as other metal ions (e.g., barium(II), scandium(II), iron(II), manganese(II), aluminum(III), tin(II), cobalt(II), copper(II), nickel(II), and oxovanadium(IV)) in the final composition that provide such ions to the enzyme.

[0166] As used herein, a compound that stabilizes the liquid composition itself means any compound other than enzyme stabilizers and antimicrobial compounds that is required to establish the storage stability of the liquid composition in an amount effective to ensure storage stability.

[0167] To those skilled in the art, shelf stability in relation to liquid compositions typically includes product appearance and dosage uniformity.

[0168] The appearance of the product is affected by the pH of the product and by the presence of compounds such as antioxidants, viscosity modifiers, emulsifiers, and the like.

[0169] Dose uniformity is usually related to product homogeneity.

[0170] The compositions of the present invention may be alkaline or may exhibit a neutral or slightly acidic pH value, for example, from 6 to 14, from 6.5 to 13, from 8 to 10.5, or from 8.5 to 9.0.

[0171] Use of component (a) to provide a homogeneous liquid composition comprising component (b) In one aspect, the present invention provides a homogeneous liquid enzyme-containing composition having anti-biofilm activity, where homogeneous means that the liquid is a uniform liquid without phase separation.

[0172] The present invention provides the use of component (a) as disclosed herein to provide a homogeneous liquid composition comprising component (b), wherein the liquid composition preferably has anti-biofilm activity.

[0173] The present invention relates to a method for providing a liquid antimicrobial composition by adding at least one chlorinated hydroxydiphenyl ether according to formula (I) to a liquid composition having DNAse activity, preferably the composition has anti-biofilm activity.

[0174] The liquid antimicrobial composition, in one embodiment, is stable on storage for at least 7 days at 4° C. In another embodiment, the liquid antimicrobial composition is stable on storage for at least 7 days at 30° C. or 37° C.

[0175] In one embodiment, storage stability refers to the homogeneity stability of the liquid antimicrobial composition. The homogeneity stability of the liquid antimicrobial composition preferably means that no phase separation occurs during storage time.

[0176] In another embodiment, storage stability refers to storage stability of DNAse activity. Storage stability of DNAse activity preferably means that the remaining DNAse activity after storage is at least 60% compared to the DNAse activity before storage. Preferably, the remaining DNAse activity after storage is at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% compared to the DNAse activity before storage.

[0177] Use of the liquid composition The liquid compositions of the present invention are referred to herein as "detergent formulations" or "cleaning formulations," which means formulations intended for cleaning soiled materials. Cleaning can refer to laundry or hard surface cleaning. Soiled materials according to the present invention include textiles and / or hard surfaces.

[0178] The term "laundry" refers to the process of treating textiles with a solution comprising the detergent formulation of the present invention, both in relation to domestic and industrial laundering. The laundering process may be carried out by using technical equipment, for example, domestic or industrial washing machines. Alternatively, the laundering process may be carried out by hand.

[0179] The term "textile" refers to any textile material, such as yarn (single yarns made of natural or synthetic fibers used in knitting and weaving), yarn intermediates, fibers, nonwoven materials, natural materials, synthetic materials, etc., as well as fabrics (textiles made by weaving, knitting, or felting fibers) made from these materials, such as garments (any clothing made from textiles), cloth, and other articles.

[0180] The term "fiber" includes natural fibers, synthetic fibers, and mixtures thereof. Examples of natural fibers are fibers of plants (e.g., flax, jute, cotton) or fibers of animal origin containing proteins such as collagen, keratin, and fibroin (e.g., silk, wool, angora, mohair, cashmere). Examples of fibers of synthetic origin are polyurethane fibers, such as Spandex® or Lycra®, polyester fibers, polyolefins, such as Elastofine, or polyamide fibers, such as nylon. A fiber can be a single filament or part of a textile, such as knitwear, woven fabric, or nonwoven fabric.

[0181] The term "hard surface cleaning" is defined herein as cleaning a hard surface, which may include any hard surface in the home, such as floors, furniture, walls, sanitary ware, glass, metal surfaces including cutlery, or tableware. Thus, the term "hard surface cleaning" also refers to "dishwashing," which refers to any form of dishwashing, such as washing by hand or automatic dishwasher (ADW). Dishwashing includes, but is not limited to, cleaning all forms of dining tableware sets, such as plates, cups, glasses, bowls, all forms of cutlery, such as spoons, knives, forks, and serving utensils, as well as ceramics, plastics, such as melamine, metal, porcelain, glass, and acrylic.

[0182] In one aspect, the present invention provides a method for inhibiting and / or reducing biofilm formation on hard and / or flexible surfaces present in washing and cleaning equipment and / or on textile surfaces, comprising: (1) Using the liquid composition of the present invention; (2) contacting the surface with a liquid composition; and (3) thereby preventing one or more microorganisms from adhering to and / or growing on and / or forming a biofilm on the surface; The present invention relates to a method for inhibiting and / or reducing the activity of a protein by

[0183] Preferably, the liquid composition of the present invention is diluted with water in the cleaning device or container before or during contacting the surface with the liquid composition of the present invention. Preferably, the liquid composition is diluted 1:100 to 1:3000 in water.

[0184] By reduced microbial growth is meant that biofilm formation on hard and flexible surfaces present in washing and cleaning implements and devices or on textile surfaces that come into contact with the detergent formulations of the present invention is reduced by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, in each case compared to biofilm formation on such surfaces that come into contact with a detergent formulation lacking component (a) and / or component (b). In one embodiment, biofilm formation is completely or nearly completely inhibited, meaning that "substantially no biofilm" is formed. "Substantially no biofilm" means that less than about 20%, less than about 18%, less than about 15%, or less than about 12% of biofilm is formed compared to biofilm formation on devices or textile surfaces that come into contact with a detergent formulation lacking component (a) and / or component (b).

[0185] In one embodiment, the biofilm according to the above embodiment comprises at least one microorganism selected from the group of bacteria and fungi.

[0186] The bacteria contained in biofilms according to the present invention can be divided into two major groups, Gram-positive and Gram-negative, based on their Gram-stain retention characteristics, which result from significant differences in the ultrastructure and chemical composition of the bacterial cell wall.

[0187] In one aspect of the present invention, at least one microorganism growing in the biofilm is selected from Gram-positive bacteria, preferably from cocci and bacilli. In one embodiment, the at least one microorganism is selected from catalase-positive cocci, for example, from the genus Staphylococcus and Micrococcus. In another embodiment, the at least one microorganism is selected from Gram-negative cocci, for example, from the genus Streptococcus. Staphylococci can be subdivided into coagulase-positive species (e.g., Staphylococcus aureus) and coagulase-negative species (e.g., Staphylococcus epidermidis, S. saprophyticus). In a preferred embodiment, the at least one microorganism is selected from coagulase-positive staphylococci. More preferably, the at least one microorganism is Staphylococcus aureus.

[0188] The Gram-positive bacteria may also be selected from the genera Aeromicrobium, Microbacterium, and Micrococcus.

[0189] In one aspect of the invention, the at least one microorganism growing in the biofilm according to the invention is selected from Gram-negative bacteria, preferably from Gram-negative Proteobacteria, which are usually subdivided into the following classes: Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Epsilonproteobacteria, Zetaproteobacteria, Acidithiobacilla, Hydrogenophilia, and Oligoflexia.

[0190] Alphaproteobacteria include, but are not limited to, Brucella, Rhizobium, Agrobacterium, Caulobacter, Rickettsia, and Wolbachia. Preferably, at least one microorganism growing in a biofilm according to the present invention is from the order Caulobacterales, more preferably from the genus Brevundimonas.

[0191] Betaproteobacteria include, but are not limited to, the following genera: Bordetella, Ralstonia, Neisseria, and Nitrosomonas.

[0192] Gammaproteobacteria include, but are not limited to, the following genera: Escherichia, Shigella, Salmonella, Stenotrophomonas, Yersinia, Buchnera, Haemophilus, Vibrio, and Pseudomonas. Preferably, the at least one microorganism is selected from the order Enterobacterales, Xanthomonadales, and Pseudomonadales. Preferably, the at least one microorganism is selected from the species Escherichia coli, Pseudomonas aeruginosa, and Pseudomonas putida.

[0193] The fungi may be selected from the class Microbotryomycetes, preferably from the genus Rhodotorula, for example from a species selected from Rhodotorula mucilaginosa, R. glutinis and R. minuta.

[0194] The fungus may also be selected from the class Saccharomycetes, preferably from the order Saccharomycetales, more preferably from the genus Candida, for example from the species Candida albicans.

[0195] The individual detergent components and amounts used in detergent formulations are known to those skilled in the art. Suitable detergent components include, inter alia, surfactants, builders, polymers, alkalis, bleaching systems, optical brighteners, foam suppressors and stabilizers, hydrotropes, and rust inhibitors. Further examples are described, for example, in "Complete Technology Book on Detergents with Formulations (Detergent Cake, Dishwashing Detergents, Liquid & Paste Detergents, Enzyme Detergents, Cleaning Powder & Spray Dried Washing Powder)", Engineers India Research Institute (EIRI), 6th Edition (2015). Another reference book for those skilled in the art is "Detergent Formulations Encyclopedia", Solverchem Publications, 2016.

[0196] It is understood that where ingredients are already provided in the composition by incorporating it into a liquid detergent formulation, detergent formulations containing the compositions of the present invention will need to be adjusted in concentration if these ingredients are to be effective for the desired purposes, which include stabilization of the product against deterioration and cleaning performance.

[0197] A detergent ingredient may serve more than one function in the end use of the detergent formulation, and therefore any detergent ingredient referred to herein with respect to a particular function may also serve another function in the end use of the detergent formulation. The function of a particular detergent ingredient in the end use of the detergent formulation typically depends on its amount in the detergent formulation, i.e., the effective amount of the detergent ingredient.

[0198] The term "effective amount" includes the amount of an individual ingredient that provides effective stain removal and / or effective cleaning conditions (e.g., pH, amount of sudsing), the amount of a particular ingredient to effectively provide an optical benefit (e.g., optical brightening, color transfer prevention), and / or the amount of a particular ingredient that effectively aids processing (those that maintain physical properties during processing, storage, and use, e.g., viscosity modifiers, hydrotropes, drying agents).

[0199] In one embodiment, the detergent formulation is a blend of more than two detergent ingredients, at least one ingredient effective in removing soils, at least one ingredient effective in providing optimal cleaning conditions, and at least one ingredient effective in maintaining the physical properties of the detergent.

[0200] The liquid detergent formulation of the present invention comprises component (a) in an amount of ...% by weight and component (b) in an amount of ...% by weight, all of said weight percentages being based on the total weight of the liquid detergent formulation.

[0201] In one embodiment, the detergent formulations of the present invention are liquid at 20° C. and 101.3 kPa.

[0202] In one embodiment of the present invention, the liquid detergent formulation includes one or more viscosity modifiers. Non-limiting examples of suitable viscosity modifiers include agar, carrageenan, tragacanth, gum arabic, xanthan gum, alginate, pectin, hydroxyethyl cellulose, hydroxypropyl cellulose, starch, gelatin, locust bean gum, cross-linked poly(meth)acrylates, such as polyacrylic acid cross-linked with bis-(meth)acrylamide, as well as silicic acid, clays, such as, but not limited to, montolilonite, zeolite, dextrin, and casein. The viscosity modifier may be included in an amount effective to provide the desired viscosity.

[0203] In one embodiment of the present invention, the liquid detergent formulation includes one or more hydrotropes, which can be organic solvents such as, but not limited to, ethanol, isopropanol, ethylene glycol, 1,2-propylene glycol, and additional organic solvents that are miscible with water under normal conditions. Further examples of suitable hydrotropes are the sodium salts of toluenesulfonic acid, xylenesulfonic acid, and cumenesulfonic acid. The hydrotrope is typically included in an amount that promotes or enables the dissolution of compounds that exhibit only slight solubility in water.

[0204] In one embodiment, the liquid detergent formulations of the present invention are bleach-free, e.g. free of inorganic peroxide compounds or chlorine bleaches, e.g. sodium hypochlorite, which means that the liquid detergent formulations according to the invention contain a total of not more than 0.01% by weight of inorganic peroxide compounds and chlorine bleaches, said % by weight being in each case relative to the total weight of the liquid detergent formulation.

[0205] laundry In one embodiment, the liquid composition of the present invention is a liquid laundry detergent. Typically, laundry detergents comprise a relatively high amount of surfactants, preferably selected from at least one nonionic surfactant according to formula (NIS-1) and / or at least one anionic surfactant according to formula (AS-I) and / or at least one anionic surfactant according to formula (AS-II).

[0206] In one embodiment, the liquid laundry detergent comprises anionic surfactants in an amount ranging from about 5% to 50% by weight, preferably in an amount ranging from about 10% to 40% by weight, all of said weight percentages being based on the total weight of the liquid laundry detergent composition.

[0207] In one embodiment, the liquid laundry detergent comprises nonionic surfactants in an amount ranging from about 5% to 50% by weight, preferably in an amount ranging from about 5% to 40% by weight, all of said weight percentages being based on the total weight of the liquid laundry detergent composition.

[0208] In one embodiment, the liquid laundry detergent of the present invention comprises at least one hydrolase disclosed herein. Preferably, at least one protease disclosed herein is combined with one or more detergent ingredients by addition to the liquid composition of the present invention. Preferably, the at least one protease is present in an amount of about 0.005% to 0.15% by weight, more preferably about 0.01% to 0.1% by weight, all of which are based on the total weight of the detergent formulation.

[0209] In one embodiment, the liquid laundry detergent further comprises at least one lipase disclosed herein, preferably in an amount of about 0.001% to 0.005%, more preferably 0.001% to 0.002%, by weight, all of which are based on the total weight of the detergent formulation. The at least one lipase is selected from a fungal triacylglycerol lipase selected from Thermomyces lanuginosa lipase and variants thereof, as disclosed herein.

[0210] In one embodiment, the liquid laundry detergent further comprises at least one alpha-amylase disclosed herein, preferably in an amount of about 0.001% to 0.015% by weight, more preferably 0.002% to 0.015% by weight, all of which are based on the total weight of the detergent formulation.

[0211] In one embodiment, the liquid laundry detergent further comprises at least one cellulase disclosed herein, preferably in an amount of about 0.001% to 0.01% by weight, more preferably 0.002% to 0.009% by weight, all of which are based on the total weight of the detergent formulation. The at least one cellulase is selected from endoglucanases (EC 3.2.1.4), preferably from those having the amino acid sequences disclosed in Figures 14A-E of WO 91 / 17244 and variants thereof, as disclosed herein.

[0212] In one embodiment, the liquid laundry detergent further comprises at least one mannanase disclosed herein, preferably in an amount of about 0.0005% to 0.005% by weight, more preferably 0.0005% to 0.002% by weight, all of which are based on the total weight of the detergent formulation. The at least one mannanase is selected from the endo-1,4-β-mannosidases (EC 3.2.1.78) disclosed herein.

[0213] In one embodiment, the liquid laundry detergent comprises at least one aminocarboxylate selected from ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), methylglycine diacetate (MGDA), and glutamic acid diacetate (GLDA) in an amount ranging from 1 wt. % to 10 wt. %, from 1.5 wt. % to 7 wt. %, or from 2 wt. % to 5 wt. %, all as disclosed above, all of which wt. % are based on the total weight of the liquid laundry detergent.

[0214] In one embodiment, the liquid laundry detergent comprises citric acid in the range of 0.1% to 10%, 0.5% to 8%, 1% to 5%, or 2% to 4% by weight, all based on the total weight of the detergent formulation. In one embodiment, the citric acid is provided as a mixture with a formate, e.g., sodium citrate:sodium formate=9:1.

[0215] In one embodiment, the liquid laundry detergent comprises at least one phosphonate disclosed herein, preferably selected from HEDP and DTPMP, in an amount ranging from 0.5 wt. % to 3.0 wt. %, or ranging from 1.0 wt. % to 2.5 wt. %, all of said wt. % being based on the total weight of the detergent formulation.

[0216] In one embodiment, the liquid laundry detergent comprises at least one ethoxylated polyethyleneimine polymer based on a polyethylene core and a polyethylene oxide shell. Preferably, the polyethyleneimine core molecule has an average molecular weight M in the range of 500 to 5000 g / mol. WMore preferred is a polyethyleneimine having a molecular weight of 500 to 1000 g / mol, and even more preferred is a polyethyleneimine having a molecular weight of 600 to 800 g / mol. W The ethoxylated polymer has an average of 5 to 50, preferably 10 to 30, and even more preferably 15 to 25 EO (ethoxy) groups per —NH group, and an average molecular weight M in the range of 5,000 to 20,0000, preferably 8,000 to 100,000, more preferably 8,000 to 50,000, even more preferably 10,000 to 30,000, and most preferably 10,000 to 20,000 g / mol. W Preferably, the at least one ethoxylated polyethyleneimine polymer is present in an amount ranging from about 0.5% to 5%, about 1% to 4%, or about 1.5% to 3% by weight, all of which weight percentages are based on the total weight of the liquid laundry detergent.

[0217] In one embodiment, the liquid laundry detergent comprises at least one ethoxylated hexamethylene diamine polymer, preferably having an average molecular weight Mw in the range of 2000-10,000 g / mol, more preferably 3,000-8,000, and most preferably 4,000-6,000, which is preferably quaternized and optionally sulfated. Preferably, the ethoxylated hexamethylene diamine polymer, preferably quaternized and optionally sulfated, contains an average of 10-50, preferably 15-40, and even more preferably 20-30 EO (ethoxy) groups per —NH group, and has an average molecular weight Mw in the range of 2,000-10,000 g / mol, more preferably 3,000-8,000, and most preferably 4,000-6,000. WIn a preferred embodiment, the ethoxylated hexamethylenediamine is quaternized and also sulfated, preferably having two cationic ammonium groups and two anionic sulfate groups. Preferably, the at least one ethoxylated hexamethylenediamine polymer is present in an amount ranging from about 0.5% to 5%, about 1% to 4%, or about 1.5% to 3% by weight, all of which are based on the total weight of the liquid laundry detergent.

[0218] Automatic dishwashing In one embodiment, the liquid composition of the present invention is a liquid automatic dishwashing detergent.

[0219] Typically, automatic dishwashing detergents do not contain significant amounts of anionic surfactants. In one aspect of the invention, the liquid automatic dishwashing detergent contains anionic surfactants in an amount of about 0% to 3% by weight, preferably less than 1% by weight, and more preferably no anionic surfactants, all of which weight percentages are based on the total weight of the liquid automatic dishwashing detergent composition.

[0220] In one aspect of the invention, the liquid automatic dishwashing detergent comprises nonionic surfactants in an amount of about 0% to 10% by weight, preferably less than 5% by weight, more preferably less than 3% by weight, all of said weight percentages being based on the total weight of the liquid automatic dishwashing detergent composition.

[0221] Preferably, the liquid automatic dishwashing detergent of the present invention comprises at least one nonionic surfactant according to formula (NIS-IV), more preferably R 1 is n-C8 alkyl, and R 2 is branch C 11 alkyl, AO is CH2-CH2-O, and x is 22. The automatic dishwashing detergent preferably contains such compounds in an amount ranging from about 0.3% to 10% by weight, from about 0.5% to 5% by weight, or from about 1% to 3% by weight, all of which weight percentages are based on the total weight of the liquid automatic dishwashing detergent.

[0222] In one embodiment, the liquid automatic dishwashing detergent comprises at least one aminocarboxylate as disclosed above in an amount of 5% to 15% by weight based on the total weight of the detergent formulation.

[0223] Preferably the automatic dishwashing detergent comprises: Ethylenediaminetetraacetic acid (EDTA) and / or diethylenetriaminepentaacetic acid (DTPA) and / or methylglycine diacetic acid (MGDA) and / or glutamic acid diacetic acid (GLDA), as disclosed above, in an amount ranging from 0.1% to 15% by weight, ranging from 1% to 10% by weight, ranging from 3% to 8% by weight, or ranging from 2.5% to 5% by weight (all said % by weight being relative to the total weight of the detergent formulation), optionally citric acid in an amount in the range of 0.1% to 10% by weight, in the range of 0.5% to 8% by weight, in the range of 1% to 5% by weight or in the range of 2% to 4% by weight (all said weight percentages relative to the total weight of the detergent formulation), in one embodiment provided as a mixture with formate, for example Na-citrate:Na-formate=9:1, optionally at least one phosphonate in an amount ranging from 0.1% to 5% by weight, from 0.5% to 3% by weight, or from 1% to 2% by weight (all said weight percentages being relative to the total weight of the detergent formulation), which phosphonate is preferably selected from derivatives of polyphosphonic acids, such as diphosphonic acids, for example sodium salts of HEDP, and derivatives of aminopolyphosphonic acids, such as aminoalkylenephosphonic acids, for example DTPMP, optionally at least one polycarboxylate in an amount ranging from 0% to 10%, 0.5% to 7%, 1% to 5%, or 2.5% to 5% by weight (all said weight percentages being relative to the total weight of the detergent formulation), selected from homopolymers having repeating monomers which are the same unsaturated carboxylic acid, such as polyacrylic acid (PAA), and copolymers having repeating monomers which are at least two different unsaturated carboxylic acids, such as copolymers of acrylic acid and methacrylic acid, copolymers of acrylic acid or methacrylic acid with maleic acid and / or fumaric acid, wherein the polyacrylic acid homopolymers may be partially neutralized or sulfonated, Includes builder series including.

[0224] In one embodiment, the liquid automatic dishwashing detergent of the present invention comprises at least one hydrolase disclosed herein selected from a protease, an amylase, and a lipase. The at least one protease disclosed herein is combined with one or more detergent ingredients by addition to the liquid composition of the present invention. Preferably, the at least one protease is present in an amount of about 0.10% to 0.25% by weight, more preferably about 0.12% to 0.21% by weight, all of which are based on the total weight of the liquid automatic dishwashing detergent.

[0225] In one embodiment, the liquid automatic dishwashing detergent further comprises at least one alpha-amylase disclosed herein, preferably in an amount of about 0.002% to 0.015% by weight, more preferably 0.004 to 0.01% by weight, all of which are based on the total weight of the detergent formulation.

[0226] In one embodiment, the liquid automatic dishwashing detergent comprises at least one zinc salt. The zinc salt is preferably selected from water-soluble and water-insoluble zinc salts. In this context, water-insoluble is used within the context of the present invention to refer to zinc salts that have a solubility of 0.1 g / l or less in distilled water at 25° C. Accordingly, zinc salts that have a higher solubility in water are referred to as water-soluble zinc salts within the context of the present invention.

[0227] The zinc salt may be selected from zinc benzoate, zinc gluconate, zinc lactate, zinc formate, ZnCl2, ZnSO4, zinc acetate, zinc citrate, Zn(NO3)2, Zn(CH3SO3)2 and zinc gallate, preferably ZnCl2, ZnSO4, zinc acetate, zinc citrate, Zn(NO3)2, Zn(CH3SO3)2 and zinc gallate.

[0228] In another embodiment of the invention, the zinc salt is selected from ZnO, ZnO·aq, Zn(OH) 2 and ZnCO 3 , with ZnO·aq being preferred.

[0229] In one embodiment of the present invention, the zinc salt is selected from zinc oxide having an average particle size (weight average) in the range of 10 nm to 100 μm.

[0230] The cations in zinc salts can be present in complexed form, for example, complexed with ammonium or water ligands, and in particular in hydrated form. For simplicity of notation, within the context of the present invention, the ligands are generally omitted when they are water ligands.

[0231] Depending on the method for adjusting the pH of the mixture according to the invention, the zinc salt can be changed. Thus, for example, it is possible to use zinc acetate or ZnCl2 to prepare the formulation according to the invention, which in an aqueous environment at a pH of 8 or 9 converts to ZnO, Zn(OH)2 or ZnO·aq, which can exist in uncomplexed or complexed form.

[0232] The zinc salt is preferably present in the liquid detergent formulation in dissolved or colloidal form.

[0233] In one embodiment of the invention, the automatic dishwashing formulation of the invention comprises a total of 0.05 to 0.4 wt. % of zinc salts, in each case based on the solids content of the formulation. Herein, the fraction of zinc salts is given as zinc or zinc ions. From this, the fraction of counterions can be calculated. [Example]

[0234] [Example 1] In general, biofilms were cultured in nutrient medium in microtiter plates in the presence of component (a) and / or component (b) disclosed above. After culturing, the biofilms were stained with a dye (safranin), which was then redissolved in a solvent. The absorbance of the dye solution at 540 nm is a measure of the amount of biofilm grown in the well.

[0235] Component (a) was 4,4'-dichloro-2-hydroxydiphenyl ether (CAS-No. 3380-30-1), commercial product Tinosan HP 100, added in the form of a 30% solution in 1,2 propylene glycol.

[0236] Component (b) is a protein sequence at a concentration of approximately 33%. MKKWMAGLFLAAAVLLCLLMVPQQIQGASLYDKVLYFPLSRYPETGDHIKDAIADGHSDICTIDRDGADKRRQESLKGIPTKPGYDRDEWPMAVCEEGGAGADVRYVTPSDNRGAGSWVGNQMSGYPDGTRVLFIVQ It was a lyophilized powder containing DNAse having the following structure:

[0237] Both test organisms, Pseudomonas aeruginosa DSM1117 and Staphylococcus aureus DSM20231, were cultured on trypticase soy agar for 24 hours at 35°C. Primary passages were stored at +4°C for 9 days.

[0238] Inoculum: 30% TSB diluted with water in a 200 mL shake flask at 35°C * + First passage in 200 ml of glucose 2.5 g / L, (i) Five single colonies for P. aeruginosa and (ii) Two single colonies for Staphylococcus aureus was prepared by suspending the overnight cultures on a shaker at approximately 160 rpm for 24 hours. The optical density (OD) of the overnight cultures was determined photometrically at 595 nm (OD595nm) and adjusted to OD595nm = 0.4 in 60% TSB + 5 g / L glucose. * TSB 100% is a trypticase soy broth medium: 1 L of aqueous medium containing 17 g of casein peptone, 3 g of soy peptone, 5 g of NaCl, and 2.5 g of K2HPO4.

[0239] Solutions of component (a), component (b), and component (a) + (b) were prepared in deionized water at twice the final test concentration, sterile filtered, and 75 μl was added to wells of a clear 96-well microtiter plate. Additionally, 75 μL of an OD595nm=0.4 cell suspension (inoculum) was transferred to the above wells of the microtiter plate, resulting in a total volume of 150 μl. Biofilms were then cultured in 30% TSB + 2.5 g / L glucose. The plates were incubated in a humidified chamber at 33°C and 40 rpm for 24 hours.

[0240] After incubation, the supernatant containing the floating cells was removed by pipette, and the wells were washed three times with 195 μL of 0.85% NaCl solution. After removing the NaCl solution, the plate was tapped on a tissue to minimize residual NaCl in the wells, and the empty plate was dried under laminar airflow.

[0241] Safranin (Gram Safranin, Sigma-Aldrich) was used for biofilm staining, 175 μL per well, and the plate was left at room temperature (RT) for 30 min.

[0242] The supernatant was removed by pipetting, and the wells were washed four times with 195 μL of 0.85% NaCl solution. Finally, the liquid was removed by pipetting, and the plate was tapped on a tissue to minimize residual NaCl in the wells. The empty plate was dried under laminar airflow. The wells were filled with 175 μL of 30% acetic acid in deionized H2O (dye solvent). The dye was further dissolved by pipetting up and down, and the liquid was transferred to a fresh microtiter plate.

[0243] The absorbance of the safranine solution was determined using a plate reader at 540 nm.

[0244] Each composition was tested in at least three wells in parallel. The average result of more than three was taken. From each of these average absorbance values, a blank background absorbance value (average of at least three wells containing growth medium but no bacteria) was subtracted. This net average absorbance is a measure of the amount of biofilm formed inside the well.

[0245] In the blank experiment, the highest amount of biofilm was observed and the biofilm inhibition effect was 0% (by definition). An active ingredient that did not affect biofilm formation at all (net absorbance = 0) would indicate a 100% biofilm inhibition effect.

[0246] Five systems were tested: (1) No antibiofilm agent was added to the biofilm growth medium, but instead, a saline solution was added ("blank experiment without agent"). (2) Addition of 10 ppm of component (b) to growth medium, where the ppm indicated refers to the weight of lyophilized powder in mg per L of liquid volume in each well of a microtiter plate. (3) Addition of 20 ppm of component (b) to the growth medium, where the ppm indicated refers to the weight of lyophilized powder in mg per L of liquid volume in each well of the microtiter plate. (4) Addition of 0.6 ppm of component (a) (i.e., 4,4'-dichloro-2-hydroxydiphenyl ether) to the growth medium, where the ppm indicated refers to the weight of component (a) in mg relative to the volume of liquid in L in each well of the microtiter plate. (5) Addition of a combination of 2 ppm component (a) and 10 ppm component (b)

[0247] [Table 1]

[0248] [Example 2] The following liquid laundry detergent formulations can be prepared:

[0249] [Table 2] AEO: Lutensol AO7 (BASF); two nonionic surfactants selected from compounds of general formula (Ia), one of which is R 1 C 12 and R 2 and R 5 is H, m is 7, n and o=0, and the other surfactant is R 1 C 14 and R 5 is H, m is 7, and n and o=0. AES: Texapon N70 (BASF); two anionic surfactants selected from compounds of general formula (AIS Ia), one of which is R 1 C 11 and R 2 is H, m is 2, n and o=0, A - is SO3 - and M + Na + and the other surfactant is R 1 C 13 and R 2 is H, m is 2, n and o=0, A - is SO3- and M + Na + It is characterized in that: LAS: Maranil DBS / LC (BASF); two anionic surfactants selected from compounds of general formula (AIS II), one of which is R 1 C 10 and the other surfactant is R 1 C 13 It is characterized in that: Palm Fatty Acids: Edenor K12-18 (Emery Oleochemicals) A protease having a polypeptide sequence according to SEQ ID NO: 22 as described in EP 1921147, optionally with a protease:R101E substitution.

[0250] The above formulations were prepared by first preparing a premix containing surfactant, solvent, fatty acid, citric acid, NaOH, and water up to 90%. This premix was prepared by adding all ingredients to the appropriate amount of water and stirring at room temperature. The pH was then set to pH=8.5 using NaOH. The final formulation was then prepared by stirring at room temperature: 90% of this premix, the appropriate concentration of the composition with or without protease, and water up to 100%. [Sequence List Free Text]

[0251] SEQ ID NO: 2: Protein: Artificial (motif of SEQ ID NO: 25 of WO 2019 / 081724) SEQ ID NO: 3: Protein: Artificial (motif of SEQ ID NO: 26 of WO 2019 / 081724) SEQ ID NO: 4: Protein: Artificial (motif of SEQ ID NO: 73 of WO 2017 / 060493) SEQ ID NO: 5: Protein: Artificial (motif of SEQ ID NO: 74 of WO 2017 / 060493) SEQ ID NO: 6: Protein: Artificial (motif of SEQ ID NO: 75 of WO 2017 / 060493)

Claims

1. (a) Formula (I) 【Chemistry 1】 wherein R1, R2, and R3 are selected from H and Cl, and at least one of R1, R2, and R3 is Cl. at least one chlorinated hydroxydiphenyl ether according to (b) at least one compound that exhibits DNAse activity, and (c) at least one compound selected from anionic surfactants and nonionic surfactants Including, A liquid composition, wherein the compound of (b) is at least 90% identical to the full-length polypeptide sequence of SEQ ID NO:

1.

2. 10. The composition of claim 1, wherein component (c) comprises at least one anionic surfactant and at least one nonionic surfactant.

3. 3. The composition of claim 1 or 2, wherein the liquid composition has anti-biofilm activity.

4. The composition of claim 3 , wherein the film comprises Pseudomonas aeruginosa and / or Staphylococcus aureus.

5. The composition of claim 1 or 2, further comprising at least one hydrolase.

6. 1. A method for providing a liquid anti-biofilm composition by adding at least one chlorinated hydroxydiphenyl ether according to formula (I) to a liquid composition having DNAse activity, comprising: Formula (I) is as follows: 【Chemistry 2】 wherein R1, R2, and R3 are selected from H and Cl, and at least one of R1, R2, and R3 is Cl. A method, wherein the liquid composition comprises a compound that is at least 90% identical to the full-length polypeptide sequence of SEQ ID NO:

1.

7. 4. A method for reducing and / or inhibiting biofilm formation on hard and / or flexible surfaces and / or textile surfaces present in washing and cleaning devices by using a liquid composition according to any one of claims 1 to 3 for washing and cleaning, wherein the reduction and / or inhibition of microbial growth is determined compared to microbial growth in the absence of components (a) and (b).

Images