Alkanolamine formate for enzyme stabilization in liquid formulations

Abstract

An enzyme preparation or detergent formulation comprising alkanolamine formate for stabilizing an enzyme, preferably a hydrolase, in a liquid environment. In a first aspect, the present invention relates to a liquid enzyme formulation comprising a) from 0.5% to 15% by weight of at least one hydrolase (EC 3) and b) from 2% to 70% by weight of at least one alkanolamine formate. In further aspects, the present invention provides a liquid detergent formulation comprising (A) 0.0005% to 0.4% by weight of at least one hydrolase (EC 3), (B) 4% to 20% by weight of an alkanolamine formate, and (C) at least 5% of at least one anionic compound.

Description

Technical Field

[0001] The present invention relates to the technical field of enzyme stabilization in liquid formulations. It is necessary to stabilize the enzymes contained in liquid formulations (LF), such as liquid enzyme preparations (LEP) or liquid detergent formulations (LDF), to avoid loss of function. Therefore, the present invention provides a compound that has been identified as stabilizing an enzyme, preferably a hydrolase, in a liquid environment, such as a liquid enzyme preparation (LEP) and / or a liquid detergent formulation (LDF). Further provided are methods for preparing such LEP or LDF and uses thereof. Background Art

[0002] In liquid formulations, enzymes tend to be unstable and are prone to losing activity when stored. Therefore, there is a continuous need to identify stable compounds that improve enzymes (especially hydrolases, such as proteases) in liquid formulations. Typically, liquid enzyme formulations contain stabilizing systems to improve enzyme stability. These enzyme stabilizers typically contain expensive enzyme inhibitors, particularly when there is protease. Therefore, it is necessary to identify stable alternative compounds that improve enzymes in liquid formulations to reduce or replace the demand for expensive enzyme inhibitors, particularly when there is protease.

[0003] A particular challenge for enzyme stabilization in LDF is the inclusion of anionic compounds in the LDF that tend to complex salts present in the formulation, such as complexing anionic compounds (also called builders) and / or surface anionic compounds (also called anionic surfactants). However, salts are required in the LDF to stabilize the enzymes, preferably proteases and / or lipases. Usually, the salt content cannot be increased arbitrarily, because, depending on the type of salt, saturation concentrations of salts can be achieved without sufficient enzyme stabilization. Common salts are salts selected from the group consisting of:

[0004] (a) Monovalent cations, such as Na + , K + and NH4 + ,as well as

[0005] (b) a monovalent organic anion of 1 to 6 carbons, which is preferably a small monocarboxylic acid of 1 to 6 carbons, such as formate, acetate, propionate or lactate.

[0006] Sodium formate is believed to increase the stability of subtilisin protease in LDF in amounts of about 0.1% to 5% by weight relative to the total weight of the detergent formulation. However, at concentrations of about ≥2.5% by weight relative to the total weight of the detergent formulation, sodium formate tends to precipitate in LDF or cause turbidity and phase separation. This is particularly true when the water content of the liquid detergent formulation is less than 50% by weight relative to the total weight of the detergent formulation. Therefore, further increasing the concentration of sodium formate does not further increase the stability of subtilisin protease.

[0007] Therefore, it is an additional object of the present invention to find a salt which, when used in a good stabilizing amount, not only improves the stability of enzymes such as hydrolases, preferably subtilisins and / or triacylglycerol lipases in LDF, but also does not precipitate in LDF. Summary of the invention

[0008] Therefore, in order to solve the above problems, the present invention provides a compound according to formula (I):

[0009]

[0010] Where R 1 and R 2 Selected from H and C2H4OH, R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, and m, n, o are each independently 0-2, preferably 0-1, more preferably 0,

[0011] It has been identified as stabilizing hydrolases in liquid environments such as liquid enzyme preparations (LEPs) and / or liquid detergent formulations (LDFs).

[0012] Compounds according to formula (I), such as alkanolamine formate (AAF) as described herein, have additionally and surprisingly been found to increase the stability of enzymes, preferably hydrolases, even more preferably subtilisin and / or triacylglycerol lipase in LDF when used in good stabilizing amounts, alone or in combination with a salt, preferably in combination with 0.5%-2.5% by weight of a salt relative to the total weight of the detergent formulation, without increasing the total salt concentration within the LDF to saturation concentrations and thus without precipitation in the LDF.

[0013] (a) Monovalent cations, such as Na + , K + and NH4 + ,as well as

[0014] (b) a monovalent organic anion of 1 to 6 carbons, which is preferably a small monocarboxylic acid of 1 to 6 carbons, such as formate, acetate, propionate or lactate.

[0015] Thus, in a first aspect, the present invention relates to a liquid enzyme preparation comprising a) 0.5 to 15% by weight of at least one enzyme, preferably a hydrolase (EC 3) and b) 2 to 70% by weight of at least one compound according to formula (I) as described herein, wherein the amount of hydrolase refers to 100% active hydrolase.

[0016] In a further aspect, the present invention provides a liquid detergent formulation comprising (A) 0.0005% to 0.4% by weight of at least one enzyme, preferably a hydrolase (EC 3), (B) 4% to 20% by weight of a compound according to formula (I) as described herein and (C) at least 5% of at least one anionic compound. DETAILED DESCRIPTION

[0017] According to the invention, a liquid formulation (LF) is understood to mean a product comprising at least one hydrolase (EC 3) and a compound according to formula (I), for example a liquid enzyme preparation (LEP) or a liquid detergent formulation (LDF). According to the invention, a liquid formulation contains at least one compound according to formula (I), resulting in a stabilization of the at least one hydrolase contained.

[0018] Therefore, in one embodiment, the present invention relates to a liquid enzyme preparation (LEP) comprising

[0019] a. 0.5 to 15% by weight of at least one enzyme, preferably a hydrolase (EC 3), and

[0020] b. 2 to 70% by weight of at least one compound according to formula (I)

[0021]

[0022] Where R 1 and R 2 Selected from H and C2H4OH,

[0023] R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, and m, n, o are each independently 0-2, preferably 0-1, more preferably 0;

[0024] The amount of hydrolase refers to 100% active hydrolase.

[0025] In another aspect, the present invention relates to a liquid detergent formulation (LDF) comprising:

[0026] (A) 0.0005% to 0.4% by weight of at least one enzyme, preferably a hydrolase (EC 3)

[0027] (B) 4 to 20% by weight of a compound according to formula (I)

[0028]

[0029] Where R 1 and R 2 Selected from H and C2H4OH, R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, and m, n, o are each independently 0-2, preferably 0-1, more preferably 0; and

[0030] (C) at least 5% of at least one anionic compound.

[0031] Component a. / (A) – Hydrolase (EC 3)

[0032] The liquid formulation (LF) of the invention comprises at least one enzyme, preferably a hydrolase (EC 3).

[0033] Hydrolase means an enzyme that exerts enzymatic activity. Enzyme activity is related to the ability of the hydrolase to degrade the corresponding substrate. The at least one hydrolase preferably originates from fermentation production.

[0034] " fermentation production " means the process of cultivating recombinant cells, which express the desired lytic enzyme in a suitable water-based nutrient medium that allows the recombinant host cell to grow and express the desired lytic enzyme. When the fermentation ends, the fermented liquid is usually collected, and the liquid fraction is separated from the solid fraction. Depends on whether the lytic enzyme has been secreted into the liquid fraction, the desired lytic enzyme can be reclaimed from the liquid fraction of the fermented liquid or from the cell lysate. The recovery of the desired lytic enzyme uses methods well known to those skilled in the art. Suitable methods for reclaiming lytic enzyme from the fermented liquid include but are not limited to collection, centrifugation, filtration, extraction and precipitation.

[0035] In one embodiment, the liquid formulation contains an "enzyme concentrate", meaning that the fermentation broth containing the hydrolytic enzymes has been purified and concentrated. The liquid enzyme concentrate typically contains hydrolytic enzymes in an amount of up to 40% by weight, or up to 30% by weight, or up to 25% by weight, all relative to the total weight of the enzyme concentrate.

[0036] The enzyme concentrate produced by fermentation contains water and potentially additional residual components, such as salts derived from the fermentation medium, cell debris derived from the production host cells, metabolites produced by the production host cells during fermentation. The residual components may be contained in the liquid enzyme concentrate in an amount of less than 20% by weight relative to the total weight of the enzyme concentrate. Preferably, the residual components are contained in an amount of less than 10% by weight, more preferably less than 5% by weight, all relative to the total weight of the enzyme concentrate.

[0037] In another aspect, the liquid formulation contains at least one solid hydrolase (EC 3) dissolved in at least one solvent selected from water and an organic solvent. Preferably, the liquid formulation comprises an amount of hydrolase below the saturation concentration of the hydrolase, meaning that the hydrolase is dissolved in the liquid formulation and no precipitation occurs.

[0038] Hydrolase can be a parent hydrolase or its variant." Parent hydrolase " or " parent sequence " (parent protein or polypeptide) is to be used for introducing the starting sequence of variation (for example, by introducing one or more amino acid replacements, insertions, deletions or its combination) to sequence, produces " variant " of parent sequence.Term parent enzyme (or parent sequence) comprises wild-type enzyme (sequence) and synthetic generation sequence (enzyme), and it is used as starting sequence for introducing (further) variation.Term " hydrolase variant " or " sequence variant " or " variant hydrolase " refers to the hydrolase whose amino acid sequence is different from parent hydrolase to a certain extent.If not otherwise specified, variant enzyme " having enzymatic activity " means that this variant enzyme has enzymatic activity of the same type as corresponding parent enzyme.

[0039] When describing the hydrolase variant, when compared with the parent sequence, usually replace, lack and insert.Nomenclature known to persons skilled in the art is used in this article.Amino acid substitution is usually described by providing the original amino acid, followed by the position number in the amino acid sequence, followed by the substituted amino acid.Amino acid deletion is usually described by providing the original amino acid, followed by the position number in the amino acid sequence, followed by *.Amino acid insertion is usually described by providing the original amino acid, followed by the position number in the amino acid sequence, followed by the original amino acid and extra amino acid.Under the situation that different changes can be introduced at a certain position, these different changes are separated by slashes.

[0040] When compared with the parent hydrolase, the hydrolase variant is usually defined by its sequence identity.Sequence identity is usually provided in the form of "sequence identity %" or "identity %".In order to calculate sequence identity, in the first step, a sequence alignment must be generated.According to the present invention, a paired global alignment must be generated, which means that two sequences must be compared on their full length, which is usually produced by using a mathematical method called an alignment algorithm.According to the present invention, an alignment is generated by using the algorithm of Needleman and Wunsch (J.Mol.Biol. [Journal of Molecular Biology] (1979) 48, 443-453 pages).Preferably, program "NEEDLE" (European Molecular Biology Open Software Suite (EMBOSS)) is used for the purpose of the present invention, wherein using program default parameters (gap open (gap open) = 10.0, gap extension (gap extend) = 0.5 and matrix = EBLOSUM62). According to the present invention, the following calculation of % identity applies: % identity = (same residues / length of the aligned region showing the corresponding sequences of the invention over the entire length)*100.

[0041] According to the present invention, a hydrolase variant is described as an amino acid sequence having at least n% identity with the amino acid sequence of the corresponding parent hydrolase, wherein "n" is an integer between 10 and 100. In one embodiment, it is increasingly preferred that the variant hydrolase has 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% identity when compared to the full-length amino acid sequence of the parent hydrolase, wherein the enzyme variant has enzymatic activity.

[0042] "Enzyme activity" is usually related to the degradation of the target substrate of the hydrolase. "Enzyme activity" means the catalytic effect exerted by the hydrolase, which is usually expressed in units per milligram of enzyme (specific activity), which is related to the substrate molecules converted per enzyme molecule per minute (molecular activity).

[0043] In a preferred embodiment, the hydrolase is selected from the group consisting of proteases, amylases, lipases, cellulases, hemicellulases, mannanases, xylanases, DNases, dispersins, pectinases and cutinases, preferably selected from the group consisting of subtilisin (EC 3.4.21.62), α-amylases (EC 3.2.1.1) and triacylglycerol lipases (EC 3.1.1.3).

[0044] Protease

[0045] Proteases (EPr) are members of enzyme class EC 3.4. Proteases include aminopeptidases (EC 3.4.11, EPr1), dipeptidases (EC 3.4.13, EPr2), dipeptidyl-peptidases and tripeptidyl-peptidases (EC 3.4.14, EPr3), peptidyl-dipeptidases (EC 3.4.15, EPr4), serine-type carboxypeptidases (EC 3.4.16, EPr5), metallocarboxypeptidases (EC 3.4.17, EPr6), cysteine-type carboxypeptidases (EC 3.4.18, EPr7), omega-peptidases (EC 3.4.19, EPr8), serine endopeptidases (EC 3.4.21, EPr9), cysteine ​​endopeptidases (EC 3.4.22, EPr10), aspartate endopeptidases (EC 3.4.23, EPr11), metalloendopeptidases (EC 3.4.24, EPr12), threonine endopeptidases (EC 3.4.25, EPr13), and oligopeptidases (EC 3.4.26, EPr14). 3.4.25, EPr13), or endopeptidases of unknown catalytic mechanism (EC 3.4.99, EPr14).

[0046] Protease means an enzyme that exerts proteolytic activity. Proteolytic activity is related to the ability of the protease to degrade proteins.

[0047] The protease may be a parent enzyme or a variant thereof, wherein the parent protease comprises a wild-type protease and a starting protease for further mutation. A variant protease means a mutated parent protease. The parent protease and the variant protease must have proteolytic activity to be a protease according to the present disclosure.

[0048] When compared to the parent protease, the protease variant can have a proteolytic activity less than, substantially equal to, or increased. The proteolytic activity of the variant is preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100% of the proteolytic activity of the corresponding parent protease. When compared to the corresponding parent protease, the increased proteolytic activity of the variant means a proteolytic activity greater than 100%, preferably at least 105%.

[0049] The LEP or LDF disclosed herein may comprise at least one protease selected from serine proteases (EC 3.4.21, EPr9). 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 the catalytic reaction. In one embodiment, the LEP or LDF comprises at least one EPr9 selected from the group consisting of chymotrypsin (EPr9a; EC 3.4.21.1), calcitonin (EPr9b; EC 3.4.21.2), elastase (EPr9c; EC 3.4.21.36, EC 3.4.21.37, EC 3.4.21.70, EC 3.4.21.71), granzyme (EPr9d; EC 3.4.21.78 or EC 3.4.21.79), kallikrein (EPr9e; EC 3.4.21.34, EC 3.4.21.35, EC 3.4.21.118, EC 3.4.21.119), plasmin (EPr9f; EC 3.4.21.7), trypsin (EPr9g; EC 3.4.21.36, EC 3.4.21.37, EC 3.4.21.70, EC 3.4.21.71), 3.4.21.4), thrombin (EPr9h, EC 3.4.21.5) and subtilisin (EPr9i). EPr9i is also known as subtilisin, such as EC 3.4.21.62, which is also referred to as "subtilisin" hereinafter.

[0050] Subtilisin (EPr9i) and chymotrypsin (EPr9a) are related serine proteases that both have a catalytic triad comprising aspartic acid, histidine and serine. In EPr9i, the relative order of these amino acids, read from the amino terminus to the carboxyl terminus, is aspartic acid-histidine-serine. In EPr9a, the relative order is histidine-aspartic acid-serine. A variety of EPr9i have been identified, and the amino acid sequences of many subtilases have been determined. For a more detailed description of such subtilases and their amino acid sequences, reference is made to Siezen et al. (1997), Protein Science [protein science] 6: 501-523.

[0051] In one embodiment, the LEP or LDF comprises at least one EPr9i which is a bacterial subtilisin. The bacterial protease may be a Gram-positive bacterial polypeptide such as Bacillus, Clostridium, Enterococcus, Geobacillus, Lactobacillus, Lactococcus, Oceanobacillus, Staphylococcus, Streptococcus, or Streptomyces. ces) proteases, or Gram-negative bacterial polypeptides, such as Campylobacter, Escherichia, Flavobacterium, Fusobacterium, Helicobacter, llyobacter, Neisseria, Pseudomonas, Salmonella, or Ureaplasma proteases. For example, R. Siezen's "Subtilases: Subtilisin-like Proteases" in "Subtilisin enzymes" edited by R. Bott and C. Betzel, New York, 1996, pp. 75-95, provides an overview of this family.

[0052] In one embodiment, the LEP or LDF comprises at least one EPr9i selected from subtilisin derived from Bacillus alcalophilus, Bacillus amyloliquefaciens, Bacillus brevis, Bacillus circulans, Bacillus clausii, Bacillus coagulans, Bacillus firmus, Bacillus gibsonii, Bacillus lautus, Bacillus lentus, Bacillus licheniformis, Bacillus megaterium, Bacillus pumilus, Bacillus sphaericus ... sphaericus), Bacillus stearothermophilus, Bacillus subtilis or Bacillus thuringiensis.

[0053] Specifically, at least one EPr9i can be selected from the following: subtilisin from Bacillus amyloliquefaciens BPN' (described by Vasantha et al. (1984) J. Bacteriol. [Journal of Bacteriology], Vol. 159, pp. 811-819 and Wells et al. (1983) in Nucleic Acids Research [Nucleic Acids Research], Vol. 11, pp. 7911-7925); subtilisin from Bacillus licheniformis (subtilisin Carlsberg [Bacillus-derived subtilisin]; disclosed in Smith et al. (1968) J. Biol Chem [Journal of Biological Chemistry], Vol. 243, pp. 2184-2191, and Jacobs et al. (1985) in Nucl. Acids Research [Nucleic Acids Research], Vol. 11, pp. 7911-7925); Res [Nucleic Acids Research], Vol. 13, pp. 8913-8926); subtilisin PB92 (the original sequence of alkaline protease PB92 is described in EP 283075A2); subtilisin 147 Subtilisin 309 ( See Table I of WO 89 / 06279); a subtilisin from Bacillus lentus as disclosed in WO 91 / 02792, such as from Bacillus lentus DSM 5483 or a variant of Bacillus lentus DSM 5483 as described in WO 95 / 23221; a subtilisin from an alkalophilic Bacillus (DSM 11233) as disclosed in DE 10064983; a subtilisin from Bacillus gibsonii (DSM 14391) as disclosed in WO 2003 / 054184; a subtilisin from Bacillus (DSM 14390) as disclosed in WO 2003 / 056017; a subtilisin from Bacillus (DSM 14392) as disclosed in WO 2003 / 055974; a subtilisin from WO subtilisin having SEQ ID NO: 4 as described in WO 2005 / 063974; subtilisin having SEQ ID NO: 4 as described in WO 2005 / 103244; subtilisin having SEQ ID NO: 7 as described in WO 2005 / 103244; and subtilisin having SEQ ID NO: 2 as described in application DE 102005028295.4.

[0054] Examples of subtilisin enzymes included in LEP or LDF include, but are not limited to, variants described in WO 92 / 19729, WO 95 / 23221, WO 96 / 34946, WO 98 / 20115, WO 98 / 20116, WO 99 / 11768, WO01 / 44452, WO 02 / 088340, WO 03 / 006602, WO 2004 / 03186, WO 2004 / 041979, WO 2007 / 006305, WO 2011 / 036263, WO 2011 / 036264 and WO 2011 / 072099.

[0055] In one embodiment, the LEP or LDF comprises at least one subtilisin having at least 80% identity with the polypeptide sequence according to SEQ ID NO: 22 as described in EP 1921147 (which is the sequence of the mature alkaline protease from Bacillus lentus DSM 5483; a sequence with 100% identity may be referred to herein as BLAP WT). Preferably, the subtilisin has no mutations at positions Asp32, His64 and Ser221 (according to BPN' numbering). A subtilisin having at least 80% identity with the polypeptide sequence according to SEQ ID NO: 22 as described in EP1921147 may be referred to herein as EPr9iA.

[0056] In one embodiment, EPr9iA has at least a substitution at position 101 (preferably selected from R101E, R101D and R101S) (according to BPN' numbering).

[0057] In one embodiment, EPr9iA has one or more substitutions selected from 3T, 4I, 63A / T / R, 156D / E, 194P, 199M, 205I and 217D / E / G, optionally together with a substitution at position 101 (selected from R101E, R101D and R101S), wherein numbering is according to BPN' numbering.

[0058] In one embodiment, EPr9iA has one or more selected from S156D, L262E, Q137H, S3T, R45E / D / Q, P55N, T58W,Y,L, Q59D / M / N / T, G61D / R, S87E, G97S, A98D / E / R, S106A / W, N117E, H120V / D / K / N, S125M, P129D, E1 101S, and wherein the numbering is according to BPN' numbering.

[0059] In one embodiment, the LF of the present invention comprises

[0060] (A) at least one serine protease EPr9iA having at least R101E or R101D or R101S, preferably R101E (according to BPN' numbering), and

[0061] (B) at least one alkanolamine formate according to formula (I) as described herein.

[0062] In one embodiment, the LEP of the present invention comprises

[0063] a. at least one serine protease EPr9iA having at least R101E or R101D or R101S, preferably R101E (according to BPN' numbering), and

[0064] b. At least one alkanolamine formate according to formula (I) as described herein.

[0065] In one embodiment, the LDF of the present invention comprises

[0066] (A) at least one serine protease EPr9iA having at least R101E or R101D or R101S, preferably R101E (according to BPN' numbering), and

[0067] (B) at least one alkanolamine formate according to formula (I) as described herein, and

[0068] (C) at least 5% of at least one anionic compound.

[0069] In one embodiment, component a. / (A) comprises at least one EPr9iA having one or more substitutions selected from 3T, 4I, 63A / T / R, 156D / E, 194P, 199M, 205I and 217D / E / G, and optionally further having the substitutions R101E or R101D or R101S, wherein the numbering is according to BPN' numbering.

[0070] In one embodiment, component a. / (A) comprises at least one EPr9iA having one or more selected from S156D, L262E, Q137H, S3T, R45E / D / Q, P55N, T58W / Y / L, Q59D / M / N / T, G61D / R, S87E, G97S, A98D,E,R, S106A / W, N117E, H120V / D / K / N, S 125M, P129D, E136Q, S144W, S161T, S163A / G, Y171L, A172S, N185Q, V199M, Y209W, M222Q, N238H, V244T, N261T / D and L262N / Q / D, and optionally further having the substitution R101E or R101D or R101S, wherein the numbering is according to BPN' numbering.

[0071] In one embodiment, component a. / (A) comprises at least one EPr9iA having mutations selected from S3T+V4I+V205I, S3T+V4I+R101E+V205I and S3T+V4I+V199M+V205I+L217D (according to BPN' numbering).

[0072] In one embodiment, component a. / (A) comprises at least one EPr9iA having the mutations S3T+V4I+S9R+A15T+V68A+D99S+R101S+A103S+I104V+N218D (numbering according to BPN').

[0073] EPr9, preferably EPr9i, more preferably EPr9iA may be stabilized by at least one enzyme stabilizer selected from the group consisting of boron-containing stabilizers and peptide stabilizers.

[0074] Thus, in one embodiment, in addition to at least one protease, the LEP or LDF further comprises at least one boron-containing stabilizer (PSB) selected from the group consisting of

[0075] (a) boric acid or its derivatives,

[0076] (b) boronic acid or a derivative thereof, such as an arylboronic acid or a derivative thereof,

[0077] (c) a salt of (a) or (b), and

[0078] (d) mixtures thereof.

[0079] Boric acid herein may be referred to as orthoboric acid. In one embodiment, the boron-containing stabilizer is selected from the group consisting of phenylboronic acid (BBA), which is also referred to as phenylboronic acid (PBA), derivatives thereof, and mixtures thereof.

[0080] In one embodiment, at least one phenyl-boronic acid derivative is selected from 4-formylphenylboronic acid (4-FPBA, PSB1), 4-carboxyphenylboronic acid (4-CPBA, PSB2), 4-(hydroxymethyl)phenylboronic acid (4-HMPBA, PSB3) and p-tolylboronic acid (p-TBA, PSB4), wherein PSB1 is preferred.

[0081] Therefore, in one embodiment, in addition to at least one protease, LEP or LDF also comprises at least one enzyme stabilizer that is a peptide stabilizer (PSP), which is preferably selected from a tripeptide compound comprising three amino acids selected from glycine, valine, alanine, tyrosine and leucine. The tripeptide stabilizer is preferably selected from peptide aldehydes, peptide acetals and peptide aldehyde bisulfite adducts. Typically, the tripeptide stabilizer carries an N-terminal protecting group. Preferably, the tripeptide stabilizer is selected from a compound comprising a combination of glycine-alanine-tyrosine (GAY, PSP1, preferably Z-GAY-H) and valine-alanine-leucine (VAL, PSP2, preferably Z-VAL-H) ​​with an N-terminal protecting group such as benzyloxycarbonyl (Cbz). The tripeptide stabilizer VAL with a CbZ protecting group may be referred to herein as Z-VAL.

[0082] In another embodiment, the LEP or LDF comprises, in addition to at least one protease, at least one enzyme stabilizer that is a peptide stabilizer (PSP) and at least one stabilizer that is a boron-containing stabilizer.

[0083] Specifically, LDF may contain one of the following combinations (C-PrPS):

[0084] id Actual combination C-PrPS1 EPr9iA+PSB1 C-PrPS2 EPr9iA+PSB2 C-PrPS3 EPr9iA+PSB3 C-PrPS4 EPr9iA+PSB4 C-PrPS5 EPr9iA+PSP1 C-PrPS6 EPr9iA+PSP2

[0085] Preferably, however, the peptide stabilizer is a peptide aldehyde, a peptide aldehyde bisulfite adduct, or a peptide acetal, preferably a peptide aldehyde, most preferably Z-GAY-H or Z-VAL-H. Preferably, EPr9iA in C-PrPS1 to C-PrPS6 is an EPr9iA having at least a substitution at position 101 (preferably selected from R101E, R101D and R101S, preferably R101E). In one embodiment, PSP2 in C-PrPS6 means Z-VAL, preferably Z-VAL-H.

[0086] In one embodiment, LEP or LDF comprises more than one protease. Specifically, it may comprise one of the following combinations:

[0087]

[0088] Amylase

[0089] Amylase means an enzyme that exerts "amylolytic activity" or "amylase activity". Amylolytic activity is related to the ability of the amylase to hydrolyze glycosidic bonds in polysaccharides, preferably at endo-positions. The amylase may be a parent amylase or a variant thereof. The parent amylase as well as the variant amylase must have amylolytic activity to be an amylase according to the present invention.

[0090] In one embodiment, the LEP or LDF comprises at least one α-amylase (EC 3.2.1.1; Amyα). Preferably, the LEP or LDF comprises at least one Amyα selected from the group consisting of:

[0091] An amylase (Amyα1) from Bacillus licheniformis having SEQ ID NO: 2 as described in WO 95 / 10603 and variants thereof having at least 95% identity. Suitable variants are described in WO 95 / 10603 comprising one or more substitutions at 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 having amylolytic activity. Variants are described in SEQ ID NO: 4 of WO 94 / 02597, WO 94 / 018314, WO 97 / 043424 and WO 99 / 019467.

[0092] An amylase (Amyα2) from Bacillus stearothermophilus having SEQ ID NO:6 as disclosed in WO 02 / 10355 and variants thereof having at least 95% identity. The amylase may be truncated at the C-terminus (Amyα2a). Suitable variants of SEQ ID NO:6 include those comprising a deletion at position 181 and / or 182 and / or a substitution at position 193.

[0093] An amylase (Amyα3) from Bacillus sp. 707 having SEQ ID NO: 6 as disclosed in WO 99 / 19467 and variants thereof having at least 95% identity. Preferred variants of SEQ NO: 6 are those having substitutions, deletions or insertions at one or more of the following positions: R181, G182, H183, G184, N195, I206, E212, E216 and K269.

[0094] • An amylase from Bacillus halmapalus having SEQ ID NO: 2 or SEQ ID NO: 7 as described in WO 96 / 23872, also described herein as SP-722 (Amyα4). Preferred variants are described in WO 97 / 3296, WO 99 / 194671 and WO 2013 / 001078.

[0095] • Amylase (Amyα5) from Bacillus sp. DSM 12649 having SEQ ID NO: 4 as disclosed in WO 00 / 22103 and variants having at least 95% identity thereto.

[0096] An amylase (Amyα6) from Bacillus sp. A 7-7 (DSM 12368) having an amino acid sequence at least 95% identical to SEQ ID NO: 2, particularly over the region from amino acids 32 to 516 according to SEQ ID NO: 2, as disclosed in WO 02 / 10356.

[0097] • Amylase (Amyα7) from Bacillus strain TS-23 having SEQ ID NO: 2 as disclosed in WO 2009 / 061380 and variants having at least 95% identity thereto.

[0098] • An amylase (Amyα8) from Cytophaga sp. having SEQ ID NO: 1 as disclosed in WO 2013 / 184577 and variants having at least 95% identity thereto.

[0099] • Amylase (Amyα9) from Bacillus megaterium DSM 90 having SEQ ID NO: 1 as disclosed in WO 2010 / 104675 and variants having at least 95% identity thereto.

[0100] • An amylase (Amyα10) from Bacillus sp. comprising amino acids 1 to 485 of SEQ ID NO: 2 as described in WO 00 / 60060 and variants having at least 95% identity thereto.

[0101] • An amylase from Bacillus amyloliquefaciens or a variant having at least 95% identity thereto (preferably selected from the amylase according to SEQ ID NO: 3 as described in WO 2016 / 092009) (Amyα11).

[0102] - An amylase having SEQ ID NO: 12 as described in WO 2006 / 002643, or an amylase variant having at least 95% identity thereto (Amyα12), preferably comprising the substitutions Y295F and M202L / I / T / V within said SEQ ID NO: 12.

[0103] an amylase having SEQ ID NO: 6 as described in WO 2011 / 098531, or an amylase variant having at least 95% identity thereto (Amyα13), preferably comprising one or more substitutions within said SEQ ID NO: 6 at a position selected from the group consisting of 193G / A / S / T / M, 195F / W / Y / L / I / V, 197F / W / Y / L / I / V, 198Q / N, 200F / W / Y / L / I / V, 203F / W / Y / L / I / V, 206F / W / Y / N / L / I / V / H / Q / D / E, 210F / W / Y / L / I / V, 212F / W / Y / L / I / V, 213G / A / S / T / M and 243F / W / Y / L / I / V.

[0104] An amylase having SEQ ID NO: 1 as described in WO 2013 / 001078, or an amylase variant (Amyα14) having at least 95% identity thereto, preferably comprising alterations at two or more (several) positions corresponding to positions G304, W140, W189, D134, E260, F262, W284, W347, W439, W469, G476 and G477 within said SEQ ID NO: 1.

[0105] An amylase having SEQ ID NO: 2 as described in WO 2013 / 001087, or an amylase variant (Amyα15) having at least 95% identity thereto, preferably comprising a deletion of positions 181+182, or 182+183, or 183+184 within said SEQ ID NO: 2; optionally the sequence comprises one or two or more modifications at any position selected from W140, W159, W167, Q169, W189, E194, N260, F262, W284, F289, G304, G305, R320, W347, W439, W469, G476, and G477 within said SEQ ID NO: 2.

[0106] - An amylase which is a hybrid alpha-amylase derived from the above mentioned amylases, as described for example in WO 2006 / 066594 (Amy alpha 16).

[0107] According to WO 2014 / 183920, a hybrid amylase having A and B domains that are at least 90% identical to SEQ ID NO: 2 of WO 2014 / 183920 and a C domain that is at least 90% identical to SEQ ID NO: 6 of WO 2014 / 183920; preferably, the hybrid alpha-amylase is at least 95% identical to SEQ ID NO: 23 of WO 2014 / 183920 (Amyα17).

[0108] According to WO 2014 / 183921, a hybrid amylase having A and B domains and a C domain, the A and B domains having at least 75% identity with 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 as disclosed in WO 2014 / 183921, and the C domain having at least 90% identity with SEQ ID NO: 6 of WO 2014 / 183921; preferably, the hybrid alpha-amylase has at least 95% identity with SEQ ID NO: 30 as disclosed in WO 2014 / 183921 (Amyα18).

[0109] · A hybrid amylase according to WO 2021 / 032881, comprising A and B domains derived from an alpha amylase (from Bacillus sp. A 7-7 (DSM 12368)) and a C domain derived from an alpha amylase (from Bacillus cereus), wherein the hybrid amylase has amylolytic activity; preferably, the A and B domains have at least 95% identity with the amino acid sequence of SEQ ID NO: 42, and the C domain has at least 95% identity with the amino acid sequence of SEQ ID NO: 44, both sequences are as disclosed in WO 2021 / 032881; more preferably, the hybrid amylase is SEQ ID NO: 54 (Amyα19) as disclosed in WO 2021 / 032881.

[0110] In one embodiment, the LEP or LDF comprises more than one Amyα. Preferably, the LEP comprises two or more Amyα selected from Amyα3, Amyα6, Amyα10, Amyα13, Amyα14, Amyα15, Amyα17, Amyα18 and Amyα19.

[0111] In one embodiment, LEP or LDF comprises more than one amylase. Specifically, it may comprise one of the following combinations:

[0112]

[0113]

[0114]

[0115] Lipase

[0116] "Lipase", "lipolytic enzyme" and "lipid esterase" all refer to enzymes of EC class 3.1.1 ("carboxylic ester hydrolase"). Lipase means an enzyme 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). The lipase may be a parent lipase or a variant thereof. The parent lipase as well as the variant lipase must have lipase activity to be a lipase according to the present invention.

[0117] In one embodiment, LEP or LDF comprises at least one lipase selected from triacylglycerol lipase (EC class 3.1.1.3). Preferably, the triacylglycerol lipase is selected from the lipase of Thermomyces lanuginosa. In one embodiment, LEP or LDF comprises at least one lipase selected from the triacylglycerol lipase of amino acids 1-269 of SEQ ID NO: 2 according to US5869438 and variants thereof, preferably a lipase (Lip1) having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity when compared to the full-length polypeptide sequence of amino acids 1-269 of SEQ ID NO: 2 of US5869438.

[0118] In one embodiment, LEP or LDF comprises at least one lipase selected from a lipase having a polypeptide sequence having at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to amino acids 1-269 of SEQ ID NO: 2 of US5869438, comprising at least amino acid substitutions T231R and N233R (Lip1a). When compared to amino acids 1-269 of SEQ ID NO: 2 of US5869438, the lipase variant may further comprise one or more of the following amino acid exchanges: Q4V, V60S, A150G, L227G, P256K (Lip1b).

[0119] In one embodiment, the LEP or LDF comprises at least one lipase (Lip2) having at least 95% identity to the full-length polypeptide sequence of amino acids 1-269 of SEQ ID NO: 1 of WO 2015 / 010009, preferably comprising at least the amino acid substitutions N11K+A18K+G23K+K24A+V77I+D130A+V154I+V187T+T189Q (Lip2a) or

[0120] N11K+A18K+G23K+K24A+L75R+V77I+D130A+V154I+V187T+T189Q(Lip2b).

[0121] Combination of protease, amylase and lipase

[0122] In one embodiment, the LEP or LDF comprises a combination of different types of hydrolases selected from proteases, amylases and lipases, preferably a combination of at least one protease and at least one amylase, or at least one protease and at least one lipase, or at least one protease, at least one amylase and at least one lipase.

[0123] In one embodiment, the LEP or LDF comprises at least one protease and at least one amylase, preferably at least one EPr9 and at least one amylase, more preferably at least one EPr9i and at least one Amyα.

[0124] In one embodiment, the LEP or LDF comprises at least one protease and at least one lipase, preferably at least one EPr9 and at least one Lip1, more preferably at least one EPr9i and at least one Lip1a.

[0125] In one embodiment, the LEP or LDF comprises at least one protease and at least one lipase, preferably at least one EPr9 and at least one Lip2, more preferably at least one EPr9i and at least one Lip2a.

[0126] In one embodiment, the LEP or LDF comprises at least one lipase and at least one amylase, preferably at least one Lip1 and at least one amylase, more preferably at least one Lip1a and at least one Amyα.

[0127] In one embodiment, the LEP or LDF comprises at least one lipase and at least one amylase, preferably at least one Lip2 and at least one amylase, more preferably at least one Lip2a and at least one Amyα.

[0128] Specifically, LEP or LDF may contain one of the following combinations:

[0129]

[0130]

[0131]

[0132]

[0133] In one embodiment, EPr9iA in the combination C-Hyd172 to C-Hyd190, C-Hyd200 and C-Hyd210 has at least a substitution at position 101 (preferably selected from R101E, R101D and R101S) (according to BPN' numbering).

[0134] In one embodiment, EPr9iA in the combination C-Hyd172 to C-Hyd190, C-Hyd200 and C-Hyd210 has one or more substitutions selected from 3T, 4I, 63A / T / R, 156D / E, 194P, 199M, 205I and 217D / E / G, optionally together with a substitution at position 101 (selected from R101E, R101D and R101S), where the numbering is according to BPN' numbering.

[0135] In one embodiment, EPr9iA in the combination C-Hyd172 to C-Hyd190, C-Hyd200 and C-Hyd210 has one or more selected from S156D, L262E, Q137H, S3T, R45E / D / Q, P55N, T58W,Y,L, Q59D / M / N / T, G61D / R, S87E, G97S, A98D / E / R, S106A / W, N117E, H120V / D / K / N, S125M, P129D, E136Q, S144W, S161T, S163A / G, Y171L, A172S, N185Q, V199M, Y209W, M222Q, N238H, V244T, N261T / D and L262N / Q / D, and optionally a substitution at position 101 selected from R101E, R101D and R101S, and wherein the numbering is according to BPN' numbering.

[0136] Component b. / (B)-Alkanolamine formate

[0137] The liquid formulation of the present invention (eg, LEP or LDF) comprises at least one compound according to formula (I), alkanolamine formate (AFF):

[0138]

[0139] Where R 1 and R 2 Selected from H and C2H4OH,

[0140] R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, and m, n, o are each independently 0-2, preferably 0-1, more preferably 0.

[0141] In one embodiment, R in the general formula (I) 1 and R 2 is H, which is referred to herein as AAF1. Preferably, m, n, o are 0 (ie AAF1a).

[0142] In one embodiment, R in the general formula (I) 1 , R 2 and R 3 is H, m and o are 0, and n is 1 (ie, AAF1b).

[0143] In one embodiment, R in the general formula (I) 1 and R 2 Yes H, R 3 is methyl, m and o are 0, and n is 1 (ie, AAF1c).

[0144] In one embodiment, R in the general formula (I) 1 is H and R 2 is C2H4OH, which is referred to herein as AAF2. Preferably, m, n, o are each independently 0 (ie AAF2a).

[0145] In one embodiment, R in the general formula (I) 1 and R 3 Yes H, R 2 is C2H4OH, m is 0, and n and o are 1 (ie AAF2b).

[0146] In one embodiment, R in the general formula (I) 1 Yes H, R 2 is C2H4OH, R 3 is methyl, m is 0, and n and o are 1 (ie, AAF2c).

[0147] In one embodiment, R in the general formula (I) 1 and R 2 is C2H4OH, which is referred to herein as AAF3. Preferably, m, n, o are 0 (ie AAF3a, also known as triethanolamine formate).

[0148] In one embodiment, R in the general formula (I) 1 and R 2 is C2H4OH, R 3 is H, and m, n, and o are all 1 (i.e. AAF3b).

[0149] In one embodiment, R in the general formula (I) 1 and R 2 is C2H4OH, R 3 is a methyl group, and m, n, and o are all 1 (ie, AAF3c).

[0150] Component a. / (A)+Component b. / (B)

[0151] The LEP or LDF according to the present invention may comprise one of the following combinations:

[0152] In one embodiment, the LEP or LDF of the present invention comprises at least one protease and at least one AAF selected from the group consisting of AAF1, AAF2 and AAF3, preferably selected from the group consisting of AAF1a, AAF2a and AAF3a.

[0153] In one embodiment, the LEP or LDF of the present invention comprises at least one protease and at least one AAF selected from the group consisting of AAF1b, AAF2b and AAF3b.

[0154] In one embodiment, the LEP or LDF of the present invention comprises at least one protease and at least one AAF selected from the group consisting of AAF1c, AAF2c and AAF3c.

[0155] Specifically, LEP or LDF may contain one of the following combinations:

[0156]

[0157]

[0158] In one embodiment, the LEP or LDF of the present invention comprises one of the combinations C-Pr1 to C-Pr44 and at least one AAF selected from AAF1a, AAF2a and AAF3a.

[0159] In one embodiment, the LEP or LDF of the present invention comprises one of the combinations C-Pr1 to C-Pr44 and at least one AAF selected from AAF1b, AAF2b and AAF3b.

[0160] In one embodiment, the LEP or LDF of the present invention comprises one of the combinations C-Pr1 to C-Pr44 and at least one AAF selected from AAF1c, AAF2c and AAF3c.

[0161] In one embodiment, the LEP or LDF comprises

[0162] (A) at least one serine protease EPr9iA having at least the substitution R101E or R101D or R101S (according to BPN' numbering), and

[0163] (B) AAF3a.

[0164] In one embodiment, the LEP or LDF comprises

[0165] (A) at least one EPr9iA having one or more substitutions selected from 3T, 4I, 63A / T / R, 156D / E, 194P, 199M, 205I and 217D / E / G, optionally together with the substitutions R101E or R101D or R101S (wherein the numbering is according to BPN' numbering), and

[0166] (B) AAF3a.

[0167] In one embodiment, the LEP or LDF comprises

[0168] (A) at least one EPr9iA having one or more selected from S156D, L262E, Q137H, S3T, R45E / D / Q, P55N, T58W / Y / L, Q59D,M,N,T, G61D / R, S87E, G97S, A98D / E / R, S106A / W, N117E, H120V / D / K / N, S125M, P12 9D, E136Q, S144W, S161T, S163A / G, Y171L, A172S, N185Q, V199M, Y209W, M222Q, N238H, V244T, N261T / D and L262N / Q / D, and optionally together with the substitutions R101E or R101D or R101S (wherein the numbering is according to BPN' numbering), and

[0169] (B) AAF3a.

[0170] In one embodiment, the LEP or LDF comprises

[0171] (A) at least one EPr9iA having a combination of mutations selected from S3T+V4I+V205I, S3T+V4I+R101E+V205I and S3T+V4I+V199M+V205I+L217D (according to BPN' numbering), and

[0172] (B) AAF3a.

[0173] In one embodiment, the LEP or LDF comprises

[0174] (A) at least one EPr9iA having the mutations S3T+V4I+S9R+A15T+V68A+D99S+R101S+A103S+I104V+N218D (numbering according to BPN'), and

[0175] (B) AAF3a.

[0176] In one embodiment, the LEP or LDF of the present invention comprises at least one amylase and at least one AAF selected from the group consisting of AAF1, AAF2 and AAF3, preferably selected from the group consisting of AAF1a, AAF2a and AAF3a.

[0177] In one embodiment, the LEP or LDF of the present invention comprises at least one amylase and at least one AAF selected from the group consisting of AAF1b, AAF2b and AAF3b.

[0178] In one embodiment, the LEP or LDF of the present invention comprises at least one amylase and at least one AAF selected from the group consisting of AAF1c, AAF2c and AAF3c.

[0179] Specifically, LEP or LDF may contain one of the following combinations:

[0180]

[0181]

[0182]

[0183] In one embodiment, the LEP or LDF of the present invention comprises one of the combinations C-Amyα1 to C-Amyα171 and at least one AAF selected from AAF1a, AAF2a, AAF3a, AAF1b, AAF2b, AAF3b, AAF1c, AAF2c and AAF3c.

[0184] In one embodiment, the LEP or LDF of the present invention comprises at least one lipase and at least one AAF selected from AAF1, AAF2 and AAF3 (preferably selected from AAF1a, AAF2a and AAF3a).

[0185] In one embodiment, the LEP or LDF of the present invention comprises at least one lipase and at least one AAF selected from the group consisting of AAF1b, AAF2b and AAF3b.

[0186] In one embodiment, the LEP or LDF of the present invention comprises at least one lipase and at least one AAF selected from the group consisting of AAF1c, AAF2c and AAF3c.

[0187] Specifically, LEP or LDF may contain one of the following combinations:

[0188]

[0189] In one embodiment, the LEP or LDF comprises a combination of different types of hydrolases (preferably according to one of the combinations C-Hyd1 to C-Hyd248) and at least one AAF selected from AAF1, AAF2 and AAF3 (preferably selected from AAF1a, AAF2a, AAF3a, AAF1b, AAF2b, AAF3b, AAF1c, AAF2c and AAF3c).

[0190] Preferably, LEP or LDF comprises one of the following combinations:

[0191]

[0192]

[0193]

[0194]

[0195] In one embodiment, the combination of LF280 to LF469 as disclosed above further comprises Lip1a.

[0196] In one embodiment, the combination of LF1 to LF527 as disclosed above comprises at least one additional hydrolase different from protease, amylase and lipase. The hydrolase different from protease, amylase and lipase is preferably selected from cellulase and mannanase.

[0197] In one embodiment LF1 to LF527 comprises at least one cellulase, preferably at least one β-1 ,4-glucanase (EC 3.2.1.4), also referred to herein as endoglucanase.

[0198] In one embodiment, LF1 to LF527 comprises at least one Humicola insolens DSM1800 endoglucanase having at least 80% identity to the amino acid sequence disclosed in Figures 14A-E of WO 91 / 17244, preferably to the sequence according to amino acids 20-434. Preferably, the endoglucanase has one or more substitutions at positions selected from 182, 223 and 231, most preferably selected from P182S, A223V and A231V. In one embodiment, LF1 to LF498 comprises at least one endoglucanase having at least 80% identity to the polypeptide according to SEQ ID NO: 2 of WO 95 / 02675.

[0199] In one embodiment LF1 to LF527 comprises at least one Bacillus sp. endoglucanase having at least 80% identity to the amino acid sequence from position 1 to position 773 of SEQ ID NO: 2 of WO 2004 / 053039.

[0200] In one embodiment LF1 to LF527 comprises at least one Thielavia terrestris endoglucanase having at least 80% identity to the amino acid sequence from position 1 to position 299 of SEQ ID NO: 4 of WO 2004 / 053039.

[0201] In one embodiment LF1 to LF527 comprises at least one mannanase, preferably at least one β-mannanase (EC 3.2.1.78).

[0202] In one embodiment, LF1 to LF527 comprises at least one β-mannanase selected from GH5 family mannanases. In one embodiment, LF1 to LF527 comprises at least one β-mannanase having at least 90% identity to SEQ ID NO: 12 of WO 2018 / 184767. In one embodiment, LF1 to LF527 comprises at least one β-mannanase having at least 90% identity to SEQ ID NO: 16 of WO 2018 / 184767. In one embodiment, LF1 to LF527 comprises at least one β-mannanase having at least 90% identity to SEQ ID NO: 20 of WO 2018 / 184767. Preferably, LF1 to LF527 comprises at least one mannanase having 95% identity to the polypeptide sequence of SEQ ID NO: 20 of WO 2018 / 184767, the at least one mannanase having at least one substitution selected from A101V, E405G and Y459F.

[0203] In one embodiment, LF1 to LF527 comprises at least one β-mannanase derived from a Trichoderma organism, such as those disclosed in WO 93 / 24622. Preferably, at least one β-mannanase has 80% identity with SEQ ID NO: 1 of WO 2008 / 009673. More preferably, the β-mannanase according to SEQ ID NO: 1 of WO 2008 / 009673 comprises at least one substitution selected from S3R, S66P, N113Y, V181H, L207F, A215T and F274L.

[0204] In one embodiment, LF1 to LF527 comprises at least one β-mannanase having a polypeptide sequence having at least 85% identity to SEQ ID NO: 16 as disclosed in WO 2018 / 185367.

[0205] In one embodiment, LF1 to LF527 comprises at least one β-mannanase having a polypeptide sequence at least 85% identical to SEQ ID NO: 12 as disclosed in WO 2018 / 185367.

[0206] In one embodiment, LF1 to LF527 comprises at least one β-mannanase having a polypeptide sequence at least 85% identical to SEQ ID NO: 20 as disclosed in WO 2018 / 185367.

[0207] In one embodiment LF1 to LF527 comprises at least one β-mannanase having a polypeptide sequence at least 85% identical to SEQ ID NO: 388 as disclosed in WO 2005 / 003319.

[0208] In one embodiment, the LEP according to the present invention is substantially free of surface-active anionic compounds and complexing anionic compounds. "Substantially free of surface-active anionic compounds and complexing anionic compounds" means that such compounds are not intentionally added, meaning that it contains preferably no more than 0.5%, more preferably no more than 0.01%, and most preferably 0% of surface-active anionic compounds and complexing anionic compounds. Surface-active anionic compounds preferably mean anionic surfactants.

[0209] Complex anionic compounds preferably mean citrates and aminocarboxylates.

[0210] In one embodiment, the LEP or LDF according to the present invention, preferably those comprising proteases such as LF1 to LF90 (preferably LF82 to LF90) and LF280 to LF489 (preferably LF451 to LF469, LF479 and LF489), comprises at least one enzyme stabilizer selected from boron-containing stabilizers and peptide stabilizers.

[0211] In one embodiment LF82 to LF90 comprises 4-formylphenylboronic acid (4-FPBA).Preferably, at least one serine protease EPr9iA of LF82 to LF90 has at least the substitution R101E or R101D or R101S, preferably R101E (according to BPN' numbering).

[0212] In one embodiment, LF451 to LF469, LF479 and LF489 comprise 4-formylphenylboronic acid (4-FPBA).Preferably, at least one serine protease EPr9iA of LF451 to LF469, LF479 and LF489 has at least the substitution R101E or R101D or R101S (according to BPN' numbering).

[0213] In one embodiment LF82 to LF90 comprises Z-VAL, preferably Z-VAL-H. Preferably, at least one serine protease EPr9iA in LF82 to LF90 has at least the substitution R101E or R101D or R101S (according to BPN' numbering).

[0214] In one embodiment, LF451 to LF469, LF479 and LF489 comprise Z-VAL, preferably Z-VAL-H. Preferably, at least one serine protease EPr9iA in LF451 to LF469, LF479 and LF489 has at least the substitution R101E or R101D or R101S, preferably R101E (according to BPN' numbering).

[0215] In a preferred embodiment, the LEP comprises a reduced amount of an enzyme stabilizer selected from boron-containing stabilizers and / or peptide stabilizers. In a preferred embodiment, the LEP is substantially free of an enzyme stabilizer selected from boron-containing stabilizers and peptide stabilizers. "Substantially free of enzyme stabilizers" means that such compounds are not intentionally added to the LEP, meaning that preferably no more than 0.5%, more preferably no more than 0.01%, and most preferably 0% of enzyme stabilizers are contained.

[0216] In preferred embodiments, the LEP comprises water in an amount increasingly preferably not exceeding 50%, 40%, 30%, 20%, or 15% by weight.

[0217] In one embodiment, the LEP or LDF of the present invention comprises at least one salt of a monovalent cation and a monovalent anion of 1-6 carbons, preferably C1-3 carbons. Preferably, the monovalent cation is selected from Na + (SALT1), K + (SALT2) and NH4 + (SALT3). Preferably, the monovalent anion is selected from the group consisting of formate (SALT#a), acetate (SALT#b), propionate (SALT#c) and lactate (SALT#d).

[0218] In one embodiment, the LEP or LDF comprises at least one salt selected from the group consisting of NaCl (SALT4), KCl (SALT5), CaCl2 (SALT6), and Na2SO4 (SALT7).

[0219] In one embodiment, LF1 to LF527 comprises sodium formate (SALT1a) or SALT6. Preferably, at least one serine protease EPr9iA in LF82 to LF90 has at least the substitution R101E or R101D or R101S (according to BPN' numbering). Preferably, at least one serine protease EPr9iA in LF451 to LF469, LF479 and LF489 has at least the substitution R101E or R101D or R101S (according to BPN' numbering).

[0220] In a preferred embodiment, the LDF comprises ≤ 3% by weight, preferably ≤ 2% by weight, more preferably ≤ 1% by weight of sodium formate.

[0221] In one embodiment, the LEP or LDF of the present invention comprises at least one solvent selected from water (SOL1) and an organic solvent. In one embodiment, the organic solvent is included in an amount of about 30% to 60% by weight relative to the total weight of the LEP.

[0222] In one embodiment, the LEP or LDF comprises at least one organic solvent selected from monohydric alcohols (SOL2), dihydric alcohols (also known as diols) (SOL3), trihydric alcohols (also known as triols) (SOL4), and sugar alcohols (SOL5).

[0223] At least one monohydric alcohol (SOL2) is selected from C2H6O, 1-propanol, propan-2-ol, 1-butanol, 2-methyl-1-propanol, butan-2-ol, ethylene glycol methyl ether, ethylene glycol ethyl ether, ethylene glycol propyl ether and ethylene glycol phenyl ether.

[0224] At least one diol (SOL3) is chosen from butane-1,3-diol, pentane-1,4-diol, pentane-1,5-diol, pentane-2,4-diol, hexane-2,5-diol, vicinal diols (OH-group at the ortho C position; SOL3a) and α-ω diols (OH-group at one of the ends of a linear molecule (HO-R-OH), SOL3b).

[0225] In one embodiment, the LEP or LDF comprises at least one vicinal diol (SOL3a), which is preferably selected from ethane-1,2-diol, propane-1,2-diol, butane-1,2-diol, butane-2,3-diol, pentane-1,2-diol, pentane-2,3-diol, hexane-2,3-diol, hexane-3,4-diol, heptane-1,2-diol, heptane-2,3-diol, heptane-3,4-diol, octane-1,2-diol, octane-2,3-diol, octane-3,4-diol and octane-4,5-diol.

[0226] In one embodiment, the LEP or LDF comprises at least one α-ω diol (SOL3b), which is preferably selected from butane-1,4-diol, hexane-1,6-diol, propane-1,3-diol, 2-(2-hydroxyethoxy)ethanol, 2-(2-propoxyethoxy)ethanol, 2-(2-butoxyethoxy)ethanol and 2-methyl-2,4-pentanediol.

[0227] As the triol (SOL4), propane-1,2,3-triol may be included.

[0228] In one embodiment, the LEP comprises at least one sugar alcohol (alditol, SOL5), such as sorbitol, mannitol, and erythritol, with sorbitol being preferred.

[0229] In one embodiment, LF1 to LF527 comprises SOL2 (preferably ethylene glycol phenyl ether), SOL3a (preferably propane-1,2-diol), SOL3b, SOL4 or SOL5 (preferably sorbitol). In one embodiment, LF1 to LF527 comprises a mixture of two solvents selected from SOL2 (preferably ethylene glycol phenyl ether), SOL3a (preferably propane-1,2-diol), SOL3b, SOL4 and SOL5 (preferably sorbitol). Preferably, at least one serine protease EPr9iA in LF82 to LF90 has at least the substitution R101E or R101D or R101S, preferably R101E (according to BPN' numbering). Preferably, at least one serine protease EPr9iA in LF451 to LF469, LF479 and LF489 has at least the substitution R101E or R101D or R101S, preferably R101E (according to BPN' numbering).

[0230] In one embodiment, the LEP comprises 0.5% to 15%, 1% to 15%, 2% to 15%, 0.5% to 10%, 0.5% to 8%, 0.5% to 6%, or 2% to 6% by weight of at least one hydrolase.

[0231] In one embodiment, the LEP increasingly preferably comprises 5% to 70%, 10% to 70%, 20% to 70%, 30% to 70%, 40% to 70%, 10% to 60%, 20% to 60%, 30% to 60%, 40% to 60%, 10% to 55%, 10% to 50%, 20% to 50%, 30% to 50% or 40% to 50% by weight of at least one compound according to formula (I) as described herein.

[0232] In one embodiment, the LEP comprises

[0233] a. 0.5% to 15% by weight of at least one hydrolase as disclosed above (EC 3)

[0234] b. 2 to 70% by weight, preferably 10 to 50% by weight, of at least one compound according to formula (I) as described herein, and

[0235] About 30% of at least one solvent selected from SOL3a (preferably propane-1,2-diol), SOL4 and SOL5 (preferably sorbitol).

[0236] In another embodiment, the LEP comprises

[0237] a. 0.5% to 15% by weight of at least one hydrolase as disclosed above (EC 3)

[0238] b. 2 to 70% by weight, preferably 10 to 50% by weight, of at least one compound according to formula (I) as described herein,

[0239] c. water in an amount not exceeding 15% by weight, and

[0240] d. About 30% of at least one solvent selected from SOL3a (preferably propane-1,2-diol), SOL4 and SOL5 (preferably sorbitol).

[0241] In one embodiment, the LEP may comprise at least two solvents selected from SOL3a (preferably propane-1,2-diol), SOL4 and SOL5 (preferably sorbitol), wherein the weight ratio of SOL3a:SOL4 or SOL3a:SOL5 is 2:1.

[0242] In one embodiment, the LEP comprises a component according to formula (I) as described herein as organic solvent, and no additional organic solvent as disclosed above.

[0243] Therefore, in a preferred embodiment, the present invention relates to a liquid enzyme preparation (LEP) comprising

[0244] a. 0.5 to 15% by weight of at least one hydrolase (EC 3), preferably selected from proteases, amylases and lipases, most preferably a protease as described herein, and

[0245] b. 2 to 70% by weight, preferably 20-60% by weight, of at least one compound according to formula (I)

[0246]

[0247] Where R 1 and R 2 Selected from H and C2H4OH,

[0248] R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, and m, n, o are each independently 0-2, preferably 0-1, more preferably 0; preferably wherein compound (b) is triethanolamine formate,

[0249] wherein the amount of hydrolase refers to 100% active hydrolase; and

[0250] water, preferably in an amount not exceeding 15% by weight, and

[0251] Preferably about 30% of at least one solvent selected from SOL3a (preferably propane-1,2-diol), SOL4 and SOL5 (preferably sorbitol);

[0252] and preferably further comprises at least one salt selected from monovalent cations and monovalent anions of 1 to 6 carbon atoms, NaCl, KCl, CaCl2 and Na2SO4

[0253] wherein the LEP is preferably free of surface active anionic compounds and complexing anionic compounds; and

[0254] Wherein the LEP preferably does not contain enzyme inhibitors, preferably does not contain boron-containing compounds and peptide stabilizers.

[0255] The LEP as described above may be prepared according to the following method: mixing at least one hydrolase (EC 3) with at least one compound according to formula (I) as described herein.

[0256] In a preferred embodiment, the method for preparing LEP comprises mixing the at least one hydrolase with at least one compound according to formula (I) as described herein, wherein the at least one hydrolase is contained in a liquid enzyme concentrate prior to mixing with the at least one compound according to formula (I) as described herein, wherein the liquid enzyme concentrate is preferably derived from fermentation enzyme production.

[0257] In a preferred embodiment, the at least one hydrolase is dissolved in a solvent selected from water and an organic solvent, preferably water, before mixing with the at least one compound according to formula (I) as described herein.

[0258] In a preferred embodiment, at least one compound according to formula (I) as described herein is used to provide a liquid enzyme preparation that is homogeneous in its appearance and has increased stability of at least one hydrolase when compared to a liquid enzyme preparation lacking the compound according to formula (I) as described herein.

[0259] In preferred embodiments, a compound according to formula (I) as described herein contained in the LEP or LDF is triethanolamine formate.

[0260] Component (C) – Anionic compound

[0261] Compared to LEP, LDF additionally comprises at least 5% of at least one anionic compound. Thus, the LDF according to the invention comprises:

[0262] (A) 0.0005% to 0.4% by weight of at least one hydrolase (EC 3)

[0263] (B) 4 to 20% by weight of a compound according to formula (I)

[0264]

[0265] Where R 1 and R 2 Selected from H and C2H4OH,

[0266] R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl,

[0267] and m, n, o are each independently 0-2, preferably 0-1, more preferably 0; and

[0268] (C) at least 5% of at least one anionic compound.

[0269] Preferably, the LDF comprises 0.001% to 0.4%, 0.01% to 0.2%, or 0.01% to 0.1% by weight of at least one hydrolase.

[0270] Preferably, the LDF comprises 5% to 20%, 6% to 20%, 5% to 18%, or 5% to 16% by weight of at least one compound according to formula (I) as described herein.

[0271] Preferably, the LDF comprises 5 to 80%, 10 to 70%, 20 to 60%, or 30 to 50% by weight of at least one anionic compound.

[0272] Preferably, the LDF comprises water in an amount of less than 80% by weight, preferably less than 60-70% by weight, more preferably less than 50% by weight, all relative to the total weight of the detergent formulation.

[0273] The LDF generally also comprises at least one compound selected from the group consisting of rheology modifiers, fragrances and colorants.

[0274] LDF can be prepared according to the following method: at least one hydrolase (EC 3) is mixed with at least one compound according to formula (I) as described herein and at least one anionic compound in one or more steps, wherein the at least one hydrolase is preferably contained in a liquid enzyme preparation before mixing with the at least one anionic compound.

[0275] In a preferred embodiment, at least one compound according to formula (I) as described herein is used to stabilize at least one hydrolase contained in a liquid detergent formulation comprising at least one anionic compound selected from surface-active anionic compounds and complexing anionic compounds, or to provide a liquid detergent formulation that is homogeneous in its appearance and has increased stability of at least one hydrolase when compared to a liquid detergent formulation lacking the compound according to formula (I).

[0276] In a preferred embodiment, a method for improving the detergency of a liquid detergent formulation by adding at least one compound according to formula (I) as described herein to a liquid detergent formulation containing a hydrolase is described, wherein the detergency is preferably improved for at least one stain selected from the group consisting of protease-sensitive stains, amylase-sensitive stains and lipase-sensitive stains.

[0277] The detergent composition may be a combination of liquid and solid detergent compositions.The liquid detergent composition may be a gel detergent composition.

[0278] The detergent composition may be a unit dose or multi-dose composition. The detergent composition may be in the form of a pouch (including a multi-compartment pouch). The detergent composition may be a clothing or hard surface cleaning composition suitable for home care and / or industrial and institutional (I&I) cleaning. Preferably, the hard surface cleaning composition may be a dishwashing detergent composition. The hard surface cleaning composition may be a medical cleaning composition, preferably a medical device cleaning composition. The hard surface cleaning composition may be an agrochemical device cleaning composition, preferably a spray can cleaning composition. Both clothing and dishwashing detergent compositions may be in the form of hand wash or automatic washing compositions.

[0279] The present invention therefore also relates to a method for cleaning, preferably laundry or hard surface cleaning, comprising the step of contacting a subject, preferably a textile or a hard surface, with a composition comprising a composition as described herein, preferably wherein the composition comprises at least one additional detergent component, preferably a surfactant and / or a builder.

[0280] Anionic compounds according to the present invention include, but are not limited to, surface-active anionic compounds (also called anionic surfactants, C1) and complexing anionic compounds (also called builders, C2).

[0281] Component (C1) – Surface-active anionic compound

[0282] Anionic surfactants herein include, but are not limited to, surface active compounds containing a hydrophobic group and at least one water-soluble anionic group (generally selected from sulfate, sulfonate and carboxylate to form water-soluble compounds).

[0283] AS1

[0284] In one embodiment, the LDF comprises at least one anionic surfactant selected from compounds of formula (AS1):

[0285]

[0286] The variables in the general formula (AS1) are defined as follows:

[0287] R 1 Selected from C1-C 23 -alkyl and C2-C 23 -alkenyl, wherein the alkyl and / or alkenyl groups are straight chain (linear; n-) or branched; an example is n-C7H 15 、n-C9H 19 、nC 11 H 23 、nC 12 H 25 、nC 13 H 27 、nC 14 H 29 、nC 15 H 31 、nC 16 H 33 、nC 17 H 35 、nC 18 H 37 、i-C9H 19 、iC 12 H 25 .

[0288] R 2 Selected from H, C1-C 20 -alkyl and C2-C 20 -alkenyl, wherein the alkyl and / or alkenyl groups are straight-chain (linear; n-) or branched.

[0289] R 3 and R 4 Each independently selected from C1-C 16-alkyl, where alkyl is linear (straight-chain; n-) 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, isopentyl, n-hexyl, isohexyl, sec-hexyl, n-heptyl, n-octyl, 2-ethylhexyl, n-nonyl, n-decyl, isodecyl.

[0290] A - Selected from-RCOO - 、-SO3 - and -RSO3 - , wherein R is selected from linear (straight chain; n-) or branched C1-C8-alkyl and C1-C4 hydroxyalkyl. - Yes-SO3 - When A - Yes - RCOO - When the compound is present, it can be referred to as a (fatty) alcohol / alkyl (ethoxy / ether) carboxylate [(F)A(E)C].

[0291] M + is selected from H and salt-forming cations. The salt-forming cations may be monovalent or polyvalent; thus, M + Equal to 1 / v M v + Examples include, but are not limited to, sodium, potassium, magnesium, calcium, ammonium, and ammonium salts of mono-, di-, and triethanolamine.

[0292] The integers in the general formula (AS1) are defined as follows:

[0293] m is in the range of zero to 200, preferably 1-80, more preferably 3-20; n and o are each independently in the range of zero to 100; n is preferably in the range of 0 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 one, preferably the sum of m, n and o is in the range of 5 to 100, more preferably in the range of from 9 to 50.

[0294] The anionic surfactant of the general formula (AS1) may have any structure, a block copolymer or a random copolymer.

[0295] In one embodiment, the LDF comprises at least one anionic surfactant according to formula (AS1), wherein R 1 Yes nC 10 To nC 18 , R 2 It is H, A - SO3 - , m, n and o are 0.+ Preferably Na + Such compounds may be referred to herein as secondary alkane sulfonates (SAS) or paraffin sulfonates.

[0296] In one embodiment, the LDF comprises at least one anionic surfactant according to formula (AS1), wherein R 1 Yes nC 11 H 23 , R 2 It is H, A - SO3 - , m, n and o are 0. + Preferably NH4 + Such compounds may be referred to herein as ammonium lauryl sulfate (ALS) or AS1a.

[0297] In one embodiment, the LDF comprises at least one anionic surfactant according to formula (AS1), wherein R 1 Yes nC 11 H 23 , R 2 Selected from H, A - SO3 - , m is 2-5, preferably 3, and n and o are 0. M + Preferably Na + Such compounds may be referred to herein as lauryl ether sulfate (LES) or sodium lauryl ether sulfate (SLES) or AS1b.

[0298] Additional suitable anionic surfactants include C 12 -C 18 Sulfonic fatty acid alkyl esters (such as C 12 -C 18 Sulfo fatty acid methyl ester), C 10 -C 18 -Alkyl aryl sulfonic acid (such as nC 10 -C 18 -alkylbenzenesulfonic acid) and C 10 -C 18 Salts of alkyl alkoxy carboxylates.

[0299] In one embodiment, the LDF comprises at least two anionic surfactants, both selected from compounds of formula (AS1a), wherein one of the anionic surfactants is characterized by R 1 It is C 11 , R 2 is H, m is 2, n and o = 0, A - SO3 - , salt forming cation (M + ) is Na+ , and the other surfactant is characterized by R 1 It is C 13 , R 2 is H, m is 2, n and o = 0, A - SO3 - , M + Yes + The anionic surfactant may be referred to herein as AS1c.

[0300] AS2

[0301] In one embodiment, the LDF comprises at least one anionic surfactant selected from compounds of general formula (AS2):

[0302]

[0303] Where R in formula (AS2) 1 It is C 10 -C 16 Alkyl. LDF may comprise a salt, preferably a sodium salt, of the compound according to formula (AS2).

[0304] In one embodiment, the compound according to formula (AS2) refers to a phenylalkanesulfonate, wherein R 1 It is C 12 -、C 13 -、C 14 -, or C 16 -alkyl.

[0305] In one embodiment, the LDF comprises an alkylbenzene sulfonate. In one aspect, this means a compound according to formula (AS2), wherein R 1 It is a branched chain C 12 Alkyl, which may be referred to herein as BABS. In one aspect, this means a compound according to formula (AS2), wherein R 1 It is a straight chain C 12 An alkyl group, which may be referred to herein as AS2a.

[0306] In one embodiment, the LDF comprises at least two anionic surfactants, both selected from compounds of formula (AS2), wherein one of the anionic surfactants is characterized by R 1 It is C 10 , and the other surfactant is characterized by R 1 It is C 13 Said combination may be referred to herein as AS2b. In one embodiment, said sodium salt of said compound is contained in LDF and is referred to herein as AS2c.

[0307] AS3

[0308] In one embodiment, the LDF comprises at least one anionic surfactant selected from compounds of the general formula (AS3), which may be referred to as N-acyl amino acid surfactants:

[0309]

[0310] The variables in the general formula (AS3) are defined as follows:

[0311] R 6 Selected from straight chain (straight chain; n-) or branched C6-C 22 -alkyl and straight chain (straight chain; n-) or branched C6-C 22 -alkenyl, such as oleyl.

[0312] R 7 is selected from H and C1-C4-alkyl.

[0313] R 8 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.

[0314] R 9 Selected from –COOX and –CH2SO3X, where X is selected from Li + 、Na + and K + .

[0315] Non-limiting examples of suitable N-acyl amino acid surfactants are mono- and di-carboxylates of N-acylated glutamic acids (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., carboxylates of N-acylated glycine (e.g., sodium, potassium, ammonium and ammonium salts of mono-, di- and triethanolamine), for example, sodium cocoyl alaninate and triethanolamine lauroyl alaninate; carboxylates of N-acylated glycine (e.g., sodium, potassium, ammonium and ammonium salts of mono-, di- and triethanolamine), for example, sodium cocoyl glycinate and potassium cocoyl glycinate; carboxylates of N-acylated sarcosine (e.g., sodium, potassium, ammonium and ammonium salts of mono-, di- and triethanolamine), for example, sodium lauroyl sarcosinate, sodium myristoyl sarcosinate, sodium oleoyl sarcosinate and ammonium lauroyl sarcosinate.

[0316] AS4

[0317] In one embodiment, the LDF comprises at least one anionic surfactant selected from the group of soaps (AS4). In one embodiment, the soaps are selected from 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, (hydrated) erucic acid. The salt-forming cation (M+) can be monovalent or polyvalent; thus, M + Equal to 1 / vM v+ Examples include, but are not limited to, sodium, potassium, magnesium, calcium, ammonium, and ammonium salts of mono-, di-, and triethanolamine.

[0318] Other non-limiting examples of suitable soaps include soap mixtures derived from natural fatty acids such as tallow, coconut oil, palm kernel oil, laurel oil, olive oil, or canola oil. Depending on the natural fatty acid from which the soap is derived, such soap mixtures contain varying amounts of lauric and / or myristic and / or palmitic and / or stearic and / or oleic and / or linoleic soaps.

[0319] Other non-limiting examples of suitable anionic surfactants include sulfates, sulfonates or carboxylates derived from natural fatty acids such as tallow, coconut oil, palm kernel oil, laurel oil, olive oil or canola oil. Depending on the natural fatty acid from which the soap is derived, 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.

[0320] A+B+C1

[0321] In one embodiment, the LDF of the present invention comprises

[0322] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0323] at least one AS1, preferably selected from AS1a, AS1b and AS1c, and

[0324] At least one AAF selected from AAF1, AAF2 and AAF3, preferably selected from AAF1a, AAF2a and AAF3a.

[0325] In one embodiment, the LDF of the present invention comprises

[0326] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0327] at least one AS2, which is preferably selected from AS2a, AS2b and AS2c, and

[0328] At least one AAF selected from AAF1, AAF2 and AAF3, preferably selected from AAF1a, AAF2a and AAF3a.

[0329] In one embodiment, the LDF of the present invention comprises

[0330] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0331] At least one AS3 and

[0332] At least one AAF selected from AAF1, AAF2 and AAF3, preferably selected from AAF1a, AAF2a and AAF3a.

[0333] In one embodiment, the LDF of the present invention comprises

[0334] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0335] At least one AS4 and

[0336] At least one AAF selected from AAF1, AAF2 and AAF3, selected from AAF1a, AAF2a and AAF3a.

[0337] In one embodiment, the LDF of the present invention comprises

[0338] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0339] At least two different anionic surfactants and

[0340] At least one AAF selected from AAF1, AAF2 and AAF3, selected from AAF1a, AAF2a and AAF3a.

[0341] Preferably, the at least two different anionic surfactants are selected from AS1c, AS2b and AS4.

[0342] In one embodiment, the LDF of the present invention comprises

[0343] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0344] at least one AS1, preferably selected from AS1a, AS1b and AS1c, and

[0345] At least one AAF selected from the group consisting of AAF1b, AAF2b, and AAF3b.

[0346] In one embodiment, the LDF of the present invention comprises

[0347] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0348] at least one AS2, which is preferably selected from AS2a, AS2b and AS2c, and

[0349] At least one AAF selected from the group consisting of AAF1b, AAF2b, and AAF3b.

[0350] In one embodiment, the LDF of the present invention comprises

[0351] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0352] At least one AS3 and

[0353] At least one AAF selected from the group consisting of AAF1b, AAF2b, and AAF3b.

[0354] In one embodiment, the LDF of the present invention comprises

[0355] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0356] At least one AS4 and

[0357] At least one AAF selected from the group consisting of AAF1b, AAF2b, and AAF3b.

[0358] In one embodiment, the LDF of the present invention comprises

[0359] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0360] At least two different anionic surfactants and

[0361] At least one AAF selected from the group consisting of AAF1b, AAF2b, and AAF3b.

[0362] Preferably, the at least two different anionic surfactants are selected from AS1c, AS2b and AS4.

[0363] In one embodiment, the LDF of the present invention comprises

[0364] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0365] at least one AS1, preferably selected from AS1a, AS1b and AS1c, and

[0366] At least one AAF selected from the group consisting of AAF1c, AAF2c, and AAF3c.

[0367] In one embodiment, the LDF of the present invention comprises

[0368] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0369] at least one AS2, which is preferably selected from AS2a, AS2b and AS2c, and

[0370] At least one AAF selected from the group consisting of AAF1c, AAF2c, and AAF3c.

[0371] In one embodiment, the LDF of the present invention comprises

[0372] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0373] At least one AS3 and

[0374] At least one AAF selected from the group consisting of AAF1c, AAF2c, and AAF3c.

[0375] In one embodiment, the LDF of the present invention comprises

[0376] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0377] At least one AS4 and

[0378] At least one AAF selected from the group consisting of AAF1c, AAF2c, and AAF3c.

[0379] In one embodiment, the LDF of the present invention comprises

[0380] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0381] At least two different anionic surfactants and

[0382] At least one AAF selected from the group consisting of AAF1c, AAF2c, and AAF3c.

[0383] Preferably, the at least two different anionic surfactants are selected from AS1c, AS2b and AS4.

[0384] Specifically, LDF may include one of the following combinations (LDF):

[0385]

[0386]

[0387] In one embodiment, EPr9i in LDF1 to LDF90 is EPr9iA having at least a substitution at position 101 (according to BPN' numbering) (preferably selected from R101E, R101D and R101S, preferably R101E).

[0388] In one embodiment, the LDF comprises one of the following combinations:

[0389]

[0390] In one embodiment, EPr9Ai in LDF91 to LDF126 has at least a substitution at position 101 (according to BPN' numbering) (preferably selected from R101E, R101D and R101S, preferably R101E).

[0391] In a preferred embodiment, the LDF is substantially free of enzyme stabilizers selected from boron-containing stabilizers and peptide stabilizers. "Substantially free of enzyme stabilizers" means that the molar ratio of enzyme to enzyme stabilizer selected from boron-containing stabilizers and peptide stabilizers in the liquid detergent formulation is at least higher than 1, more preferably higher than 5 and more preferably higher than 10.

[0392] Component (C2) – Complexing anionic compound

[0393] In one embodiment, the LDF of the present invention comprises

[0394] (A) 0.0005% to 0.4% by weight of at least one hydrolase (EC 3)

[0395] (B) 4 to 20% by weight of a compound according to formula (I)

[0396]

[0397] Where R 1 and R 2 Selected from H and C2H4OH,

[0398] R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, and m, n, o are each independently 0-2, preferably 0-1, more preferably 0, and

[0399] (C) at least one complexing anionic compound (C2).

[0400] Complexing anionic compounds herein (also referred to herein as "builders" or "building agents") are compounds selected from complexing agents (chelating agents, sequestrating agents), precipitants and ion exchange compounds, which can form water-soluble complexes with calcium and magnesium. The term is not intended to limit such compounds to the stated function in the final application of LDF.

[0401] Preferably, the LDF disclosed herein is substantially free of phosphate-based builders (PBBs). PBBs include, but are not limited to, sodium metaphosphate, sodium orthophosphate, sodium hydrogen phosphate, sodium pyrophosphate, trisodium phosphate, hexasodium metaphosphate, and polyphosphates, such as pentasodium tripolyphosphate (STP).

[0402] "Substantially free of PBB" means that no phosphate based builders are intentionally added to the LDF, preferably the PBB content in the LDF amounts to less than 0.2% by weight, preferably not more than 10 ppm, as determined gravimetrically. % by weight is relative to the total weight of the detergent formulation.

[0403] In one embodiment, the LDF comprises at least one non-phosphate based builder (NPB) including sodium gluconate, one or more sodium citrates, one or more sodium silicates, one or more sodium carbonates, one or more sodium phosphonates, one or more sodium aminocarboxylates, one or more sodium polycarboxylates, one or more sodium polysulfonates, and one or more sodium polyphosphonates.

[0404] Citrate (NPB1)

[0405] In one embodiment, the LDF comprises one or more citrates (NPB1). NPB1 includes monoalkali metal salts and dialkali metal salts, and in particular monosodium citric acid and preferably trisodium citric acid, ammonium or substituted ammonium citric acid and citric acid itself. Citrates can be used as anhydrous compounds or as hydrates, for example as sodium citrate dihydrate.

[0406] Carbonate (NPB2)

[0407] In one embodiment, the LDF comprises one or more carbonates (NPB2). NPB2 includes alkali metal carbonates and alkali metal bicarbonates, preferably sodium salts. Particularly suitable is sodium carbonate (Na2CO3).

[0408] Aminocarboxylate (NPB3)

[0409] In one embodiment, the LDF comprises one or more aminocarboxylates (NPB3 or NPBAC). NPB3 or NPBAC include, but are not limited to, diethanolglycine (DEG), dimethylglycine (DMG), nitrilotriacetic acid (NTA), N-hydroxyethylaminodiacetic acid, ethylenediaminetetraacetic acid (EDTA), N-(2-hydroxyethyl)iminodiacetic acid (HEIDA), hydroxyethylenediaminetriacetic acid, N-hydroxyethyl-ethylenediaminetriacetic acid (HEDTA), hydroxyethylenediaminetetraacetic acid, diethylenetriaminepentaacetic acid (DTPA) and methylglycinediacetic acid (MGDA), glutamic acid-diacetic acid (GLDA), iminodisuccinic acid (IDS), hydroxyiminodisuccinic acid, ethylenediaminedisuccinic acid (EDDS), aspartic acid-diacetic acid and alkali metal or ammonium salts thereof. Also suitable are aspartic acid-N-monoacetic acid (ASMA), aspartic acid-N,N-diacetic acid (ASDA), aspartic acid-N-monopropionic acid (ASMP), N-(2-sulfomethyl)aspartic acid (SMAS), N-(2-sulfoethyl)aspartic acid (SEAS), N-(2-sulfomethyl)glutamate (SMGL), N-(2-sulfoethyl)glutamate (SEGL), N-methyliminodiacetic acid (MIDA), α-alanine-N,N-diacetic acid (α-ALDA), serine-N,N-diacetic acid (SEDA), isoserine-N,N-diacetic acid (ISDA), phenylalanine-N,N-diacetic acid (PHDA), anthranilic acid-N,N-diacetic acid (ANDA), para-aminobenzenesulfonic acid-N,N-diacetic acid (SLDA), taurine-N,N-diacetic acid (TUDA) and sulfomethyl-N,N-diacetic acid (SMDA) and their alkali metal or ammonium salts. The term "ammonium salt" as used in this context refers to a salt having at least one cation carrying a nitrogen atom that is permanently or temporarily quaternized. Examples of cations carrying at least one nitrogen atom that is permanently quaternized include tetramethylammonium, tetraethylammonium, dimethyldiethylammonium and nC 10 -C 20 Examples of cations carrying at least one nitrogen atom that is temporarily quaternized include protonated amines and ammonia, such as monomethylammonium, dimethylammonium, trimethylammonium, monoethylammonium, diethylammonium, triethylammonium, nC 10 -C 20 - alkyldimethylammonium 2-hydroxyethylammonium, bis(2-hydroxyethyl)ammonium, tris(2-hydroxyethyl)ammonium, N-methyl 2-hydroxyethylammonium, N,N-dimethyl-2-hydroxyethylammonium, and especially NH4 + .

[0410] Preferably, the LDF contains less than 0.2% by weight, or from 0.01% to 0.1% by weight, of nitrilotriacetic acid (NTA), all relative to the total weight of the detergent formulation.

[0411] In one embodiment, the LDF comprises at least one compound selected from the group consisting of iminodisuccinic acid (IDS; NPBAC1), methylglycine diacetate (MGDA, NPBAC2), glutamic acid diacetate (GLDA, NPBAC3), and corresponding salts thereof.

[0412] In one embodiment, LDF comprises at least one aminocarboxylate selected from methylglycine diacetate (MGDA), glutamic acid diacetate (GLDA) and corresponding salts thereof, for example, alkali metal (such as sodium) salts thereof, in an amount in the range of 0.1% to 25.0% by weight, in the range of 1.0% to 18.0% by weight, in the range of 3.0% to 15.0% by weight, in the range of 3.0% to 10.0% by weight, or in the range of 5.0% to 8.0% by weight relative to the total weight of the detergent composition. Suitable salts of MGDA and GLDA include the tri-alkali metal salt of MGDA (formula NPBAC2) and the tetra-alkali metal salt of GLDA (formula NPBAC3):

[0413] [CH3-CH(COO)-N(CH2-COO)2]M3(NPBAC2)

[0414] [OOC-(CH2)2-CH(COO)-N(CH2-COO)2]M4(NPBAC3)

[0415] The variables in formulas (NPBAC2) and (NPBAC3) are defined as follows:

[0416] M is selected from the same or different alkali metal cations, such as cations of lithium, sodium, potassium, rubidium, cesium and a combination of at least two of the foregoing. Preferred examples of alkali metal cations are sodium and potassium and a combination of sodium and potassium. More preferred is the sodium salt.

[0417] In one embodiment, the alkali metal salt of MGDA is selected from [CH3-CH(COO)-N(CH2-COO)2]Na 3-y H y and [CH3-CH(COO)-N(CH2-COO)2]Na 3-x-y (NH4) x H y ,in:

[0418] x is selected from 0.0 to 1.0, preferably 0.1 to 0.5, more preferably 0.1 to 0.3;

[0419] y is selected from 0.0 to 1.0, preferably 0.0005 to 0.5.

[0420] Examples include Na 3-y H y 、[Na 0.7 (NH4) 0.3 ] 3-y H y 、[(NH4) 0.7 Na 0.3 ] 3-y H y The preferred embodiment is selected from Na 3-y H y .

[0421] In one embodiment, MGDA is chosen from at least one alkali metal salt of racemic MGDA and an alkali metal salt chosen from a mixture of L- and D-enantiomers according to formula (NPBAC2), said mixture containing predominantly the corresponding L-isomer with an enantiomeric excess (ee) ranging from 5% to 99%, preferably from 5% to 95%, more preferably from 10% to 75% and even more preferably from 10% to 66%.

[0422] In one embodiment, the total alkali neutralization degree of MGDA is in the range of from 0.80 mol-% to 0.98 mol-%, preferably 0.90% to 0.97%. The total alkali neutralization degree does not take into account any neutralization with ammonium.

[0423] In one embodiment, the alkali metal salt of GLDA is selected from

[0424] [OOC-(CH2)2-CH(COO)-N(CH2-COO)2]Na 4--y H y and [OOC-(CH2)2-CH(COO)-N(CH2-COO)2]M 4-x-y (NH4) y H x ,in:

[0425] x is selected from 0.0 to 2.0, preferably from 0.02 to 0.5, more preferably from 0.1 to 0.3, y is selected from 0.0 to 1.0, preferably from 0.0005 to 0.5.

[0426] In one embodiment, the alkali metal salt of GLDA can be selected from the alkali metal salts of the L- and D-enantiomers according to formula (NPBAC3), the mixture containing a racemic mixture or preferably mainly containing the corresponding L-isomer, for example with an enantiomeric excess (ee) in the range of from 5% to 99%, preferably 5% to 95%.

[0427] The enantiomeric excess can be determined, for example, by measuring polarization (polarimetry) or preferably by chromatography, for example by HPLC with a chiral column (e.g. with one or more cyclodextrins as stationary phase). Preferred is the determination of the enantiomeric excess by HPLC with a fixed optically active ammonium salt (such as D-penicillamine).

[0428] Typically, in the context of the present disclosure, small amounts of MGDA and / or GLDA may also carry cations other than alkali metals. Thus, it is possible that small amounts of the builder (such as 0.01% to 5 mol-% of the total builder) may carry alkaline earth metal cations (such as, for example, Mg 2+ or Ca 2+ ), or transition metal cations (such as, for example, Fe 2+ or Fe 3+ Cationic). The "small amount" of MGDA and / or GLDA herein means 0.1% to 1 w / w% in total relative to the corresponding builder.

[0429] In one embodiment, the LDF of the present invention comprises more than one aminocarboxylate (NPB3). Specifically, the LDF may comprise one of the following combinations (C-NPBAC):

[0430] id Actual combination C-NPBAC1 NPBAC1+NPBAC2 C-NPBAC2 NPBAC1+NPBAC3 C-NPBAC3 NPBAC2+NPBAC3 C-NPBAC4 NPBAC1+NPBAC2+NPBAC3

[0431] A+B+C2

[0432] In one embodiment, the LDF of the present invention comprises

[0433] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0434] at least one NPB1, preferably sodium citrate dihydrate, and

[0435] At least one AAF selected from AAF1, AAF2 and AAF3, preferably selected from AAF1a, AAF2a and AAF3a.

[0436] In one embodiment, the LDF of the present invention comprises

[0437] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0438] at least one NPB2, preferably Na2CO3, and

[0439] At least one AAF selected from AAF1, AAF2 and AAF3, preferably selected from AAF1a, AAF2a and AAF3a.

[0440] In one embodiment, the LDF of the present invention comprises

[0441] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0442] at least one NPB3, preferably selected from NPBAC1, NPBAC2 and NPBAC3, and

[0443] At least one AAF selected from AAF1, AAF2 and AAF3, preferably selected from AAF1a, AAF2a and AAF3a.

[0444] In one embodiment, the LDF of the present invention comprises

[0445] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0446] At least one selected from C-NPBAC1, C-NPBAC2, C-NPBAC3 and C-NPBAC4, and

[0447] At least one AAF selected from AAF1, AAF2 and AAF3, preferably selected from AAF1a, AAF2a and AAF3a.

[0448] In one embodiment, the LDF of the present invention comprises

[0449] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0450] at least one NPB1, preferably sodium citrate dihydrate, and

[0451] At least one AAF selected from the group consisting of AAF1b, AAF2b, and AAF3b.

[0452] In one embodiment, the LDF of the present invention comprises

[0453] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0454] at least one NPB2, preferably Na2CO3, and

[0455] At least one AAF selected from the group consisting of AAF1b, AAF2b, and AAF3b.

[0456] In one embodiment, the LDF of the present invention comprises

[0457] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0458] at least one NPB3, preferably selected from NPBAC1, NPBAC2 and NPBAC3, and

[0459] At least one AAF selected from the group consisting of AAF1b, AAF2b, and AAF3b.

[0460] In one embodiment, the LDF of the present invention comprises

[0461] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0462] At least one selected from C-NPBAC1, C-NPBAC2, C-NPBAC3 and C-NPBAC4, and

[0463] At least one AAF selected from the group consisting of AAF1b, AAF2b, and AAF3b.

[0464] In one embodiment, the LDF of the present invention comprises

[0465] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0466] at least one NPB1, preferably sodium citrate dihydrate, and

[0467] At least one AAF selected from the group consisting of AAF1c, AAF2c, and AAF3c.

[0468] In one embodiment, the LDF of the present invention comprises

[0469] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0470] at least one NPB2, preferably Na2CO3, and

[0471] At least one AAF selected from the group consisting of AAF1c, AAF2c, and AAF3c.

[0472] In one embodiment, the LDF of the present invention comprises

[0473] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0474] at least one NPB3, preferably selected from NPBAC1, NPBAC2 and NPBAC3, and

[0475] At least one AAF selected from the group consisting of AAF1c, AAF2c, and AAF3c.

[0476] In one embodiment, the LDF of the present invention comprises

[0477] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA,

[0478] At least one selected from C-NPBAC1, C-NPBAC2, C-NPBAC3 and C-NPBAC4, and

[0479] At least one AAF selected from the group consisting of AAF1c, AAF2c, and AAF3c.

[0480] In one embodiment, one of the following combinations of builders (C-NPB) may be included in the LDF of the present invention:

[0481]

[0482]

[0483] In one embodiment, the LDF comprises

[0484] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA, in combination with at least one of C-NPB1 to C-NPB22, and

[0485] At least one AAF selected from AAF1, AAF2 and AAF3, preferably selected from AAF1a, AAF2a and AAF3a.

[0486] In one embodiment, the LDF comprises

[0487] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA, in combination with at least one of C-NPB1 to C-NPB22, and

[0488] At least one AAF selected from the group consisting of AAF1b, AAF2b, and AAF3b.

[0489] In one embodiment, the LDF comprises

[0490] at least one EPr9, preferably at least one EPr9i, more preferably at least one EPr9iA, in combination with at least one of C-NPB1 to C-NPB22, and

[0491] At least one AAF selected from the group consisting of AAF1c, AAF2c, and AAF3c.

[0492] Specifically, LDF may include one of the following combinations (LDF):

[0493]

[0494]

[0495] In one embodiment, EPr9i in LDF127 to LDF207 is EPr9iA having at least a substitution at position 101 (according to BPN' numbering) (preferably selected from R101E, R101D and R101S, preferably R101E).

[0496] In one embodiment, the LDF comprises one of the following combinations:

[0497]

[0498]

[0499] In one embodiment, EPr9Ai in LDF208 to LDF261 has at least a substitution at position 101 (according to BPN' numbering) (preferably selected from R101E, R101D and R101S, preferably R101E).

[0500] A+B+C1+C2

[0501] In one embodiment, the LDF of the present invention comprises

[0502] (A) 0.0005% to 0.4% by weight of at least one hydrolase (EC 3)

[0503] (B) 4 to 20% by weight of a compound according to formula (I) as described herein, and

[0504] (C) at least 5% of anionic compounds, wherein the anionic compounds comprise at least one anionic surfactant (C1) and at least one builder (C2).

[0505] Components (A), (B), (C1) and (C2) have the preferences as disclosed above.

[0506] In one embodiment, the LDF of the present invention comprises at least one of the combinations LDF1 to LDF126, and at least one NPB1, preferably sodium citrate dihydrate.

[0507] In one embodiment, the LDF of the present invention comprises at least one of the combinations LDF1 to LDF126, and at least one NPB2, preferably Na2CO3.

[0508] In one embodiment, the LDF of the present invention comprises at least one of the combinations LDF1 to LDF126, and at least one NPB3, preferably selected from NPBAC1, NPBAC2 and NPBAC3.

[0509] In one embodiment, the LDF of the present invention comprises at least one of the combinations LDF1 to LDF126, and at least one of the combinations C-NPBAC1, C-NPBAC2, C-NPBAC3, and C-NPBAC4.

[0510] In one embodiment, the LDF includes at least one of the combinations LDF1 to LDF126 and at least one of the combinations C-NPB1-C-NPB22.

[0511] In one embodiment, the LDF of the present invention comprises LDF28 and at least one NPB1, preferably sodium citrate dihydrate.

[0512] In one embodiment, the LDF of the present invention comprises LDF58 and at least one NPB1, preferably sodium citrate dihydrate.

[0513] In one embodiment, the LDF of the present invention comprises LDF88 and at least one NPB1, preferably sodium citrate dihydrate.

[0514] In one embodiment, the LDF of the present invention comprises LDF28 and at least one NPB2, preferably Na2CO3.

[0515] In one embodiment, the LDF of the present invention comprises LDF58 and at least one NPB2, preferably Na2CO3.

[0516] In one embodiment, the LDF of the present invention comprises LDF88 and at least one NPB2, preferably Na2CO3.

[0517] In one embodiment, the LDF of the present invention comprises LDF28 and at least one NPB3, preferably selected from NPBAC1, NPBAC2 and NPBAC3.

[0518] In one embodiment, the LDF of the present invention comprises LDF58 and at least one NPB3, preferably selected from NPBAC1, NPBAC2 and NPBAC3.

[0519] In one embodiment, the LDF of the present invention comprises LDF88 and at least one NPB3, preferably selected from NPBAC1, NPBAC2 and NPBAC3.

[0520] In one embodiment, the LDF of the present invention comprises LDF28 and at least one of the combinations C-NPBAC1, C-NPBAC2, C-NPBAC3, and C-NPBAC4.

[0521] In one embodiment, the LDF of the present invention comprises LDF58 and at least one of the combinations C-NPBAC1, C-NPBAC2, C-NPBAC3, and C-NPBAC4.

[0522] In one embodiment, the LDF of the present invention comprises LDF88 and at least one of the combinations C-NPBAC1, C-NPBAC2, C-NPBAC3, and C-NPBAC4.

[0523] In one embodiment, the LDF comprises a surface active anionic compound preferably selected from LAS (linear alkylbenzene sulfonate) or AES (alkyl ether sulfate) and a complexing anionic compound preferably selected from citrate (NPB1) and aminocarboxylate (NPB3).

[0524] Therefore, preferably, the LDF according to the present invention comprises:

[0525] (A) 0.0005% to 0.4% by weight of at least one hydrolase (EC 3), preferably a protease as described herein,

[0526] (B) 4 to 20% by weight of a compound according to formula (I) as described herein, preferably triethanolamine formate; and

[0527] (C) at least 5% of at least one anionic compound, wherein the anionic compound is selected from surface-active anionic compounds and complexing anionic compounds, preferably wherein the surface-active anionic compound is selected from LAS or AES, and preferably wherein the complexing anionic compound is selected from citrate (NPB1) and aminocarboxylate (NPB3);

[0528] Therein preferably the liquid detergent formulation comprises ≤3% by weight, preferably ≤2% by weight, of sodium formate.

[0529] Component (D) – Additional detergent components

[0530] In one embodiment, the LDF of the present invention comprises

[0531] (A) 0.0005% to 0.4% by weight of at least one hydrolase (EC 3)

[0532] (B) 4 to 20% by weight of a compound according to formula (I) as described herein, and

[0533] (C) at least one anionic compound, wherein the anionic compound is selected from the group consisting of surface active anionic (C1) compounds and complexing anionic compounds (C2), and

[0534] (D) at least one additional detergent component.

[0535] Components (A), (B), (C1) and (C2) are as disclosed above, with the preferences as disclosed above.

[0536] At least one additional detergent component may be selected from:

[0537] At least one nonionic surfactant (D1)

[0538] At least one additional hydrolase selected from the group consisting of amylase, lipase, cellulase and mannanase (D2)

[0539] At least one solvent (D3)

[0540] At least one polymer (D4)

[0541] At least one antimicrobial agent (D5)

[0542] Component D1 – Nonionic surfactant

[0543] In one embodiment, the LDF of the present invention comprises

[0544] (A) 0.0005% to 0.4% by weight of at least one hydrolase (EC 3)

[0545] (B) 4 to 20% by weight of a compound according to formula (I) as described herein, and

[0546] (C) at least one anionic compound, wherein the anionic compound is selected from the group consisting of surface active anionic (C1) compounds and complexing anionic compounds (C2), and

[0547] (D) at least one nonionic surfactant (D1).

[0548] NIS1

[0549] In one embodiment, the LDF comprises at least one nonionic surfactant selected from compounds of formula (NIS1a) and (NIS1b):

[0550]

[0551] The variables of formula (NIS1a) and (NIS1b) are defined as follows:

[0552] R 1 Selected from C1-C23 Alkyl and C2-C 23 Alkenyl, wherein the alkyl and / or alkenyl is straight chain (linear; n-) or branched; an example is n-C7H 15 、n-C8H 17 、n-C9H 19 、nC 11 H 23 、nC 13 H 27 、nC 15 H 31 、nC 17 H 35 、i-C9H 19 、iC 12 H 25 .

[0553] R 2 Selected from H, C1-C 20 Alkyl and C2-C 20 Alkenyl, wherein the alkyl and / or alkenyl groups are straight chain (linear; n-) or branched.

[0554] R 3 and R 4 Each independently selected from C1-C 16 Alkyl, wherein the alkyl group is straight-chain (straight-chain; n-) 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, isopentyl, n-hexyl, isohexyl, sec-hexyl, n-heptyl, n-octyl, 2-ethylhexyl, n-nonyl, n-decyl, isodecyl.

[0555] R 5 Selected from H and C1-C 18 Alkyl, wherein the alkyl group is straight chain (linear; n-) or branched.

[0556] The integers of the general formula (NIS1a) and (NIS1b) are defined as follows:

[0557] m is in the range of zero to 200, preferably 1-80, more preferably 3-20; n and o are each independently in the range of zero 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 one, preferably the sum of m, n and o is in the range of 5 to 100, more preferably in the range of from 9 to 50.

[0558] Compounds according to formula (NIS1a) may be referred to herein as alkyl polyglycol ethers (AEO). Compounds according to formula (NIS1b) may be referred to herein as alkylphenol polyglycol ethers (APEO).

[0559] In one embodiment, the detergent formulation comprises at least one nonionic surfactant selected from compounds of general formula (NIS1a), wherein R 1 Yes nC 13 H 27 , R 2 and R 5 is H, m is 3-20, n and o=0.

[0560] In one embodiment, the detergent formulation comprises at least one nonionic surfactant selected from compounds of general formula (NIS1a), wherein R 1 Is it a straight chain or branched chain C 10 Alkyl, R 2 and R 5 is H, m is 3-14, n and o=0.

[0561] In one embodiment, the LDF comprises at least two nonionic surfactants, both selected from compounds of formula (NIS1a), wherein one of the nonionic surfactants is characterized by R 1 Yes nC 15 H 31 , R 2 and R 5 is H, m is 9-80, n and o=0, and another surfactant is characterized in that R 1 Yes nC 17 H 35 , R 2 and R 5 is H, m is 9-80, n and o = 0. In one embodiment, m in both nonionic surfactants is 9-80, preferably 25-80. The NIS may be referred to herein as NIS1a1.

[0562] In one embodiment, the detergent formulation comprises at least one nonionic surfactant selected from the general formula (NIS1a), wherein m is in the range of 3 to 11, preferably not more than 10, more preferably not more than 7; n and o are 0, R 1 It is a straight chain C9-C 17 Alkyl, R 2 and R 5 It's H.

[0563] In one embodiment, the detergent formulation comprises at least two nonionic surfactants, both selected from compounds of formula (NIS1a), wherein one of the nonionic surfactants is characterized by R 1 Yes nC 12 H 25 , R 2 and R 5 is H, m is 3-30, preferably 7, n and o=0, and the other surfactant is characterized in that R 1 Yes nC 14 H 29 , R 2 and R 5 is H, m is 3-30, preferably 7, n and o = 0. The NIS may be referred to herein as NIS1a2.

[0564] In one embodiment, the detergent formulation comprises at least two nonionic surfactants, both selected from compounds of formula (NIS1a), wherein one of the nonionic surfactants is characterized by R 1 Yes nC 11 H 23 , R 2 and R 5 is H, m is 4-10, n and o=0, and the other surfactant is characterized in that R 1 Selected from nC 11 H 23 and nC 17 H 35 , R 2 and R 5 is H, m is 4-10, n and o = 0. The NIS may be referred to herein as NIS1a3.

[0565] In one embodiment, the detergent formulation comprises at least two nonionic surfactants, both selected from compounds of formula (NIS1a), wherein one of the nonionic surfactants is characterized by R 1 It is n-C9H 19 , R 2 and R 5 is H, m is 5-7, n and o=0, and another surfactant is characterized in that R 1 Yes nC 17 H 35 , R 2 and R 5 is H, m is 5-7, n and o = 0. The NIS may be referred to herein as NIS1a4.

[0566] In one embodiment, the detergent formulation comprises at least two nonionic surfactants, both selected from compounds of formula (NIS1a), wherein one of the nonionic surfactants is characterized by R 1 Yes nC 11 H 23 , R 5 is H, m is 7, n and o=0, and the other surfactant is characterized in that R 1 It is C 13 H 27 , R 5 is H, m is 7, n and o = 0. The NIS may be referred to herein as NIS1a5.

[0567] In one embodiment, the detergent formulation comprises at least one nonionic surfactant according to the general formula (NIS1a), wherein R 1 C3 to C 18 Straight chain alkyl, R 2 Yes H, R 3 and R 4 Each independently selected from

[0568] ·Methyl, wherein n or o is 2-25, or

[0569] · ethyl, and n or o is 1-3, or

[0570] ·propyl, and n or o is 1-3,

[0571] And wherein m+n+o is equal to 5-50. In one embodiment, R 5 is H. In one embodiment, R 5 is selected from the group consisting of methyl, butyl, benzyl and tert-butyl. Said NIS may be referred to herein as NIS1a6.

[0572] The nonionic surfactants of the general formula (NIS1a) and (NIS1b) may have any structure, be it a block or random structure, and are not limited to the sequence of the shown formula (NIS1a) and (NIS1b).

[0573] In one embodiment, the detergent formulation comprises at least one compound according to formula (NIS1a), wherein R 2 is H, m is 10-50, R 3 Is a straight chain or branched C8-C 12 alkyl, n is 1 or 2, with 1 being preferred, o is 0 or 1 and R 5 It's H.

[0574] In one embodiment, the detergent formulation comprises at least one compound according to formula (NIS1a), wherein R 2is H, m is 10-50, R 3 Is a straight chain or branched C8-C 12 alkyl, n is 1 or 2, with 1 being preferred, o is 0 or 1 and R 5 It's H.

[0575] In one embodiment, the detergent formulation comprises at least one nonionic surfactant selected from compounds according to formula (NIS1a), wherein R 1 is n-C8 alkyl, R 2 Yes H, R 3 It is a branched chain C 11 Alkyl, R 5 is H, m is 22, n is 1 and o is 0. The NIS may be referred to herein as NIS1a7.

[0576] In one embodiment, the detergent formulation comprises at least one nonionic surfactant selected from compounds according to formula (NIS1a), wherein R 1 is n-C8 alkyl, R 2 Yes H, R 3 It is a branched chain C 11 Alkyl, R 5 is H, m is 19, n is 1 and o is 0. The NIS may be referred to herein as NIS1a8.

[0577] In one embodiment, the detergent formulation comprises at least one nonionic selected from compounds according to formula (NIS1a), wherein R 1 is n-C8 alkyl, R 2 Yes H, R 3 It is n-C8-C 10 Alkyl, R 5 is H, m is 40, n is 1 and o is 0. The NIS may be referred to herein as NIS1a9.

[0578] In one embodiment, the detergent formulation comprises at least one nonionic surfactant selected from compounds according to formula (NIS1a), wherein R 1 is n-C8 alkyl, R 2 Yes H, R 3 is methyl, R 4 Yes nC 10 Alkyl, R 5 is H, m is 22, n is 1 and o is 1. The NIS may be referred to herein as NIS1a10.

[0579] NIS2

[0580] In one embodiment, the detergent formulation comprises at least one nonionic surfactant selected from compounds of the general formula (NIS2), which may be referred to herein as alkyl-polyglycosides (APGs):

[0581]

[0582] R in the general formula (NIS2) 1 Selected from C1-C 17 Alkyl and C2-C 17 Alkenyl, wherein the alkyl and / or alkenyl is straight chain (linear; n-) or branched; an example is n-C7H 15 、n-C9H 19 、nC 11 H 23 、nC 13 H 27 、nC 15 H 31 、nC 17 H 35 、i-C9H 19 、iC 12 H 25 .

[0583] R in the general formula (NIS2) 2 Selected from H, C1-C 17 Alkyl and C2-C 17 Alkenyl, wherein the alkyl and / or alkenyl groups are straight chain (linear; n-) or branched.

[0584] G in the general formula (NIS2) 1 Selected from monosaccharides having 4 to 6 carbon atoms, such as glucose and xylose.

[0585] The integer w in the general formula (NIS2) is in the range from 1.1 to 4, w being an average number.

[0586] NIS3

[0587] In one embodiment, the detergent formulation comprises at least one nonionic surfactant selected from compounds of general formula (NIS3):

[0588]

[0589] The variables of the general formula (NIS3) are defined as follows:

[0590] AO is selected from the group consisting of ethylene oxide (EO), propylene oxide (PO), butylene oxide (BO) and mixtures thereof.

[0591] R 6 Selected from C5-C 17 Alkyl and C5-C17 Alkenyl, wherein the alkyl and / or alkenyl groups are straight chain (linear; n-) or branched.

[0592] R 7 Selected from H, C1-C 18 Alkyl, wherein the alkyl group is straight chain (linear; n-) or branched.

[0593] The integer y in the general formula (NIS3) is a number in the range of 1-70, preferably 7-15.

[0594] NIS4

[0595] In one embodiment, the detergent formulation comprises at least one nonionic surfactant selected from sorbitan esters (NIS4a) and / or ethoxylated (NIS4b) or propoxylated (NIS4c) sorbitan esters. Non-limiting examples are the products sold under the trade names SPAN and TWEEN.

[0596] NIS5

[0597] In one embodiment, the detergent formulation comprises at least one nonionic surfactant selected from alkoxylated mono- or di-alkylamines (NIS5a), fatty acid monoethanolamides (FAMA, NIS5b), fatty acid diethanolamides (FADA, NIS5c), ethoxylated fatty acid monoethanolamides (EFAM, NIS5d), propoxylated fatty acid monoethanolamides (PFAM, NIS5e), polyhydroxyalkyl fatty acid amides (NIS5f), or N-acyl N-alkyl derivatives of glucosamine (NIS5g), such as glucamide (GA), fatty acid glucamide (FAGA), and combinations thereof.

[0598] In one embodiment, the LDF of the present invention comprises

[0599] (A) at least one serine protease,

[0600] (B) at least one alkanolamine formate,

[0601] (C) at least one anionic surfactant (C1) and / or at least one builder (C2), and

[0602] (D) at least one nonionic surfactant (D1) selected from NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4 and NIS5.

[0603] Preferably, such LDF comprises C1 in an amount ranging from 10% to 30% by weight, preferably 12% to 25% by weight, relative to the total weight of LDF. C2 is preferably comprised in an amount ranging from 1% to 5% by weight, relative to the total weight of LDF. D1 is preferably comprised in an amount ranging from 5% to 15% by weight, relative to the total weight of LDF.

[0604] In one embodiment, LDF1-LDF261 includes NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5.

[0605] In one embodiment, LDF28 comprises at least one NPB1, preferably sodium citrate dihydrate, and NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5.

[0606] In one embodiment, LDF58 comprises at least one NPB1, preferably sodium citrate dihydrate, and NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5.

[0607] In one embodiment, LDF88 comprises at least one NPB1, preferably sodium citrate dihydrate, and NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5.

[0608] In one embodiment, LDF 28 comprises at least one NPB 2 , preferably Na 2 CO 3 , and NIS 1a2 , NIS 1a3 , NIS 1a4 , NIS 1a5 , NIS2 , NIS3 , NIS4 , or NIS5 .

[0609] In one embodiment, LDF58 comprises at least one NPB2, preferably Na2CO3, and NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5.

[0610] In one embodiment, LDF88 comprises at least one NPB2, preferably Na2CO3, and NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5.

[0611] In one embodiment, LDF28 comprises at least one NPB3, preferably selected from NPBAC1, NPBAC2 and NPBAC3, and NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5.

[0612] In one embodiment, LDF58 comprises at least one NPB3, preferably selected from NPBAC1, NPBAC2 and NPBAC3, and NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5.

[0613] In one embodiment, LDF88 comprises at least one NPB3, preferably selected from NPBAC1, NPBAC2 and NPBAC3.

[0614] In one embodiment, LDF88 comprises at least one NPB3, preferably selected from NPBAC1, NPBAC2 and NPBAC3, and NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5.

[0615] In one embodiment, the LDF of the present invention comprises

[0616] (A) at least one serine protease,

[0617] (B) at least one alkanolamine formate,

[0618] (C) at least one builder (C2), and

[0619] (D) at least one nonionic surfactant (D1) selected from NIS1a1, NIS1a6, NIS1a7, NIS1a8, NIS1a9 and NIS1a10.

[0620] Preferably, such LDF is substantially free of C1. C2 is preferably contained in an amount ranging from 5 to 20% by weight relative to the total weight of LDF. D1 is preferably contained in an amount lower than 5% by weight relative to the total weight of LDF.

[0621] In one embodiment, LDF127 to LDF261 comprises NIS1a1, NIS1a6, NIS1a7, NIS1a8, NIS1a9 or NIS1a10, wherein LDF127 to LDF261 are substantially free of C1.

[0622] Component D2 – Additional hydrolases

[0623] In one embodiment, the LDF of the present invention comprises

[0624] (A) 0.0005% to 0.4% by weight of at least one hydrolase (EC 3), preferably a protease as described herein,

[0625] (B) 4 to 20% by weight of a compound according to formula (I) as described herein, and

[0626] (C) at least one anionic compound, wherein the anionic compound is selected from the group consisting of surface-active anionic compounds (C1) and complexing anionic compounds (C2), and

[0627] (D) at least one hydrolase different from hydrolase EC 3, preferably wherein the hydrolase different from hydrolase EC 3 is selected from the group consisting of proteases, amylases, lipases, cellulases, mannanases as described herein, preferably wherein hydrolase EC 3 is a protease as described herein and the hydrolase different from hydrolase EC 3 is preferably an amylase, preferably at least one α-amylase (EC 3.2.1.1).

[0628] Preferably, the LDF comprises at least one alpha-amylase selected from hybrid amylases. In one embodiment, the LDF comprises at least one hybrid amylase having at least 95% identity to SEQ ID NO: 23 of WO 2014 / 183920 (Amy1). In one embodiment, the LDF comprises at least one hybrid amylase having at least 95% identity to SEQ ID NO: 30 of WO 2014 / 183921 (Amy2). In one embodiment, the LDF comprises at least one hybrid amylase having at least 95% identity to SEQ ID NO: 54 of WO 2021 / 032881 (Amy3).

[0629] In an embodiment, LDF1 to LDF126 comprises at least one hybrid amylase Amy1, Amy2 or Amy3, and optionally comprises NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5 and optionally C2 as disclosed herein.

[0630] In one embodiment, LDF127 to LDF261 comprises at least one hybrid amylase Amy1, Amy2, or Amy3. The LDF is preferably substantially free of C1. In one embodiment, the LDF comprises NIS1a1, NIS1a6, NIS1a7, NIS1a8, NIS1a9, or NIS1a10.

[0631] In one embodiment, the LDF comprises at least one lipase, preferably at least one triacylglycerol lipase (EC 3.1.1.3) as described herein.

[0632] Preferably, the LDF comprises at least one triacylglycerol lipase having at least 80% identity to amino acids 1-269 of SEQ ID NO: 2 of US5869438. In one embodiment, the lipase comprises at least the amino acid substitutions T231R and N233R (Lip1). In one embodiment, the LDF comprises at least one lipase comprising T231R and N233R and one or more of the following amino acid exchanges when compared to amino acids 1-269 of SEQ ID NO: 2 of US5869438: Q4V, V60S, A150G, L227G, P256K.

[0633] In one embodiment, the LDF comprises at least one lipase having at least 95% identity to the full-length polypeptide sequence of amino acids 1-269 of SEQ ID NO: 1 of WO 2015 / 010009, preferably comprising at least the amino acid substitutions N11K+A18K+G23K+K24A+V77I+D130A+V154I+V187T+T189Q (Lip2a) or

[0634] N11K+A18K+G23K+K24A+L75R+V77I+D130A+V154I+V187T+T189Q(Lip2b).

[0635] In one embodiment, LDF1 to LDF126 comprises at least one triacylglycerol lipase Lip1 and optionally comprises NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5 and optionally C2 as disclosed herein.

[0636] In one embodiment, LDF1 to LDF126 comprises at least one triacylglycerol lipase Lip2a and optionally comprises NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5 and optionally C2 as disclosed herein.

[0637] In one embodiment the LDF comprises at least one cellulase (D2c), preferably at least one β-1 ,4-glucanase (EC 3.2.1.4), also referred to herein as endoglucanase.

[0638] In one embodiment, the LDF comprises at least one Humicola insolens DSM 1800 endoglucanase having at least 80% identity (Cel1) to the amino acid sequence disclosed in Figures 14A-E of WO 91 / 17244, preferably to the sequence according to amino acids 20-434. Preferably, the endoglucanase has one or more substitutions at positions selected from 182, 223 and 231, most preferably selected from P182S, A223V and A231V. In one embodiment, the LDF comprises at least one endoglucanase having at least 80% identity to the polypeptide according to SEQ ID NO: 2 of WO 95 / 02675.

[0639] In one embodiment, the LDF comprises at least one Bacillus sp. endoglucanase having at least 80% identity to the amino acid sequence from position 1 to position 773 of SEQ ID NO: 2 of WO 2004 / 053039.

[0640] In one embodiment, the LDF comprises at least one Thielavia terrestris endoglucanase having at least 80% identity to the amino acid sequence from position 1 to position 299 of SEQ ID NO: 4 of WO 2004 / 053039.

[0641] In one embodiment, LDF1 to LDF126 comprises at least one endoglucanase Cel1 and optionally comprises NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5 and optionally C2 as disclosed herein.

[0642] In one aspect, the LDF comprises at least one mannanase, preferably at least one β-mannanase (EC 3.2.1.78).

[0643] In one embodiment, LDF comprises at least one β-mannanase selected from the GH5 mannanase family. In one embodiment, LDF comprises at least one β-mannanase (Man1) with at least 90% identity to SEQ ID NO: 12 of WO 2018 / 184767. In one embodiment, LDF comprises at least one β-mannanase (Man2) with at least 90% identity to SEQ ID NO: 16 of WO 2018 / 184767. In one embodiment, LDF comprises at least one β-mannanase (Man3) with at least 90% identity to SEQ ID NO: 20 of WO 2018 / 184767. Preferably, LDF comprises at least one mannanase with 95% identity to the polypeptide sequence of SEQ ID NO: 20 of WO 2018 / 184767, and the at least one mannanase has at least one substitution selected from A101V, E405G and Y459F.

[0644] In one embodiment, the LDF comprises at least one β-mannanase (Man4) having a polypeptide sequence at least 90% identical to amino acids 29-324 of SEQ ID NO: 1 of WO 2021 / 058452.

[0645] In one embodiment, the LDF comprises at least one β-mannanase derived from a Trichoderma organism, such as those disclosed in WO 93 / 24622. Preferably, the at least one β-mannanase has 80% identity (Man5) to SEQ ID NO: 1 of WO 2008 / 009673. More preferably, the β-mannanase according to SEQ ID NO: 1 of WO 2008 / 009673 comprises at least one substitution selected from S3R, S66P, N113Y, V181H, L207F, A215T and F274L.

[0646] In one embodiment, LDF1 to LDF126 comprises at least one β-mannanase Man1, Man2, Man3, Man4, or Man5 and optionally comprises NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5 and optionally C2 as disclosed herein.

[0647] In one aspect, LDF1 to LDF126 comprise at least one DNase.

[0648] In one embodiment, the LDF comprises at least one DNA enzyme having at least 80% identity to SEQ ID NOs: 1-24 and SEQ ID NOs: 27-28 of WO 2019 / 081724 and WO 2019 / 081721. Preferably, the LDF comprises at least one DNA enzyme comprising one or two motifs selected from SEQ ID NOs: 25 and SEQ ID NOs: 26 of WO 2019 / 081724.

[0649] In one embodiment, the LDF comprises at least one DNA enzyme comprising one or more motifs selected from SEQ ID NO: 73, SEQ ID NO: 74 and SEQ ID NO: 75 of WO 2017 / 060493.

[0650] In one embodiment, LDF1 to LDF126 comprise at least one DNase and optionally comprise NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4, or NIS5 and optionally C2 as disclosed herein.

[0651] Component D3 - Solvent

[0652] In one embodiment, the LDF of the present invention comprises

[0653] (A) 0.0005% to 0.4% by weight of at least one hydrolase (EC 3)

[0654] (B) 4 to 20% by weight of a compound according to formula (I) as described herein, and

[0655] (C) at least one anionic surfactant (C1) and / or at least one builder (C2), and

[0656] (D) at least one solvent (D3) selected from water (SOL1) and organic solvents.

[0657] Preferably, at least one organic solvent is selected from water-miscible organic solvents. The "water miscibility" of an organic solvent means the property of the organic solvent to mix with water to form a homogeneous solution. Thus, a "solution" in this context means a homogeneous mixture of two or more organic solvents in water. When one of the solvents in the solution is water, the solution may be referred to as an "aqueous solution". "Homogeneity" generally refers to a uniform formulation of two or more components within a solution or mixture.

[0658] In one embodiment, the LDF comprises at least one organic solvent selected from the group consisting of monohydric alcohol (SOL2), dihydric alcohol (SOL3), trihydric alcohol (SOL4) and sugar alcohol (SOL5).

[0659] At least one monohydric alcohol (SOL2) is selected from C2H6O, 1-propanol, propan-2-ol, 1-butanol, 2-methyl-1-propanol, butan-2-ol, ethylene glycol methyl ether, ethylene glycol ethyl ether, ethylene glycol propyl ether and ethylene glycol phenyl ether.

[0660] The water-miscible SOL2 is preferably selected from C2H6O and propan-2-ol.

[0661] At least one diol (SOL3) is chosen from butane-1,3-diol, pentane-1,4-diol, pentane-1,5-diol, pentane-2,4-diol, hexane-2,5-diol, vicinal diols (OH-group at the ortho C position; SOL3a) and α-ω diols (OH-group at one of the ends of a linear molecule (HO-R-OH), SOL3b).

[0662] In one embodiment, the LDF comprises at least one vicinal diol (SOL3a), which is preferably selected from ethane-1,2-diol, propane-1,2-diol, butane-1,2-diol, butane-2,3-diol, pentane-1,2-diol, pentane-2,3-diol, hexane-2,3-diol, hexane-3,4-diol, heptane-1,2-diol, heptane-2,3-diol, heptane-3,4-diol, octane-1,2-diol, octane-2,3-diol, octane-3,4-diol and octane-4,5-diol.

[0663] In one embodiment, the LDF comprises at least one α-ω diol (SOL3b), which is preferably selected from butane-1,4-diol, hexane-1,6-diol, propane-1,3-diol, 2-(2-hydroxyethoxy)ethanol, 2-(2-propoxyethoxy)ethanol, 2-(2-butoxyethoxy)ethanol and 2-methyl-2,4-pentanediol.

[0664] The water-miscible SOL3 is preferably selected from butane-1,3-diol, propane-1,2-diol, pentane-1,4-diol, pentane-1,5-diol, pentane-2,4-diol, butane-1,4-diol and 1,6-hexanediol.

[0665] As the triol (SOL4), propane-1,2,3-triol may be included.

[0666] In one embodiment, the LDF comprises at least one sugar alcohol (alditol, SOL5), such as sorbitol, mannitol and erythritol, with sorbitol being preferred.

[0667] Furthermore, the LDF may include at least one organic solvent selected from compounds such as 2-butoxyethanol, isopropyl alcohol, and d-limonene.

[0668] In one embodiment, LDF1 to LDF126 contain SOL2, preferably ethylene glycol phenyl ether.

[0669] In one embodiment, LDF1 to LDF126 contain SOL3a, preferably propane-1,2-diol.

[0670] In one embodiment, LDF1 to LDF126 contain SOL4, preferably propane-1,2,3-triol.

[0671] In one embodiment, LDF1 to LDF126 comprise SOL5, preferably sorbitol.

[0672] In one embodiment, said LDF1 to LDF126 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS1a2 and optionally C2 as disclosed herein.

[0673] In one embodiment, said LDF1 to LDF126 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS1a3 and optionally C2 as disclosed herein.

[0674] In one embodiment, said LDF1 to LDF126 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS1a4 and optionally C2 as disclosed herein.

[0675] In one embodiment, said LDF1 to LDF126 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS1a5 and optionally C2 as disclosed herein.

[0676] In one embodiment, said LDF1 to LDF126 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS2 and optionally C2 as disclosed herein.

[0677] In one embodiment, said LDF1 to LDF126 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS3 and optionally C2 as disclosed herein.

[0678] In one embodiment, said LDF1 to LDF126 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS4 and optionally C2 as disclosed herein.

[0679] In one embodiment, said LDF1 to LDF126 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS5 and optionally C2 as disclosed herein.

[0680] In one embodiment, the LDF further comprises SOL1.

[0681] In one embodiment, LDF127 to LDF261 contain SOL2, preferably ethylene glycol phenyl ether. The LDF is preferably substantially free of C1.

[0682] In one embodiment, LDF127 to LDF261 contain SOL3a, preferably propane-1,2-diol. The LDF is preferably substantially free of C1.

[0683] In one embodiment, LDF127 to LDF261 contain SOL4, preferably propane-1,2,3-triol. The LDF is preferably substantially free of C1.

[0684] In one embodiment, LDF127 to LDF261 comprise SOL5, preferably sorbitol. The LDF is preferably substantially free of C1.

[0685] In one embodiment, the LDF127 to LDF261 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS1a1.

[0686] In one embodiment, the LDF127 to LDF261 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS1a6.

[0687] In one embodiment, the LDF127 to LDF261 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS1a7.

[0688] In one embodiment, the LDF127 to LDF261 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS1a8.

[0689] In one embodiment, the LDF127 to LDF261 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS1a9.

[0690] In one embodiment, the LDF127 to LDF261 comprising SOL2, SOL3a, SOL4, or SOL5 comprises NIS1a10.

[0691] In one embodiment, the LDF further comprises SOL1.

[0692] Component D4 - polymer

[0693] In one embodiment, the LDF of the present invention comprises

[0694] (A) 0.0005% to 0.4% by weight of at least one hydrolase (EC 3),

[0695] (B) 4 to 20% by weight of a compound according to formula (I) as described herein,

[0696] (C) at least one anionic surfactant (C1) and / or at least one builder (C2), and

[0697] (D) at least one polymer (D4).

[0698] In one embodiment, at least one polymer is selected from the group of "polycarboxylates" (which includes salts of polycarboxylates). The salt-forming cations may be monovalent or polyvalent. Suitable examples include, but are not limited to, sodium, potassium, magnesium, calcium, ammonium, and ammonium salts of mono-, di-, and triethanolamine.

[0699] Polycarboxylates include compounds containing monomers selected from unsaturated carboxylic acids of the general formula (CoC):

[0700]

[0701] The variables in the general formula (CoC) are defined as follows:

[0702] R 1 , R 2 and R 3 Independently selected from H; straight or branched C1-C 12 Alkyl, straight chain or branched C2-C 12 Alkenyl, wherein the alkyl and / or alkenyl may be substituted by: -NH2, -OH, or -COOH; -COOH; and -COOR 5 , where R 5 Selected from straight or branched C1-C 12 Alkyl and straight or branched C2-C 12 Alkenyl.

[0703] R 4 may be a spacer group, which is optionally selected from -(CH2) n -(wherein n is in the range of 0 to 4), -COO-(CH2) k - (wherein k is in the range of 1 to 6), -C(O)-NH- and -C(O)-NR 6 -, where R 6 Selected from straight or branched C1-C 22 Alkyl, straight chain or branched C2-C 22 Alkenyl and C6-C 22 Aryl.

[0704] Non-limiting examples of suitable unsaturated carboxylic acids include acrylic acid, methacrylic acid, 2-ethylacrylic acid, 2-phenylacrylic acid, malonic acid, crotonic acid, maleic acid (or maleic anhydride), fumaric acid, itaconic acid, aconitic acid, mesaconic acid, citraconic acid, sorbic acid, cinnamic acid, methylenemalonic acid, unsaturated C4-C 10 Dicarboxylic acids and mixtures thereof.

[0705] The polycarboxylates can be characterized by having a K-value, i.e. a molecular weight determined according to Fikentscher's K-value, which is a value measured via the viscosity of an aqueous solution at a defined polymer content and defined viscosity measurement conditions and thus relates to the molecular weight of the polymer of a given polymer class. The measurement is preferably carried out according to ISO 1628-1.

[0706] Polycarboxylates can be characterized by their weight average molecular weight (Mw). Preferably, Mw is determined by gel permeation chromatography using standard methods. Examples include polycarboxylates with a weight average molecular weight (Mw) in the range of about 500 g / mol to about 500,000 g / mol, in the range of about 1,000 g / mol to about 100,000 g / mol, or in the range of about 3,000 g / mol to about 80,000 g / mol.

[0707] The polycarboxylate may be a homopolymer having repeating monomers as the same unsaturated carboxylic acid according to formula CoC. Such homopolymers may be referred to herein as "HP".

[0708] In one embodiment, the LDF comprises at least one homopolymer of acrylic acid, which may be referred to herein as "HP1". HP1 herein includes salts of polyacrylic acid. The salt-forming cations may be monovalent or polyvalent, including sodium, potassium, magnesium, calcium, ammonium, and ammonium salts of mono-, di-, and triethanolamine, with sodium being preferred. Thus, HP1 may be completely or only partially neutralized by salt formation.

[0709] In one embodiment, the LDF comprises at least one HP1, preferably a sodium salt of polyacrylic acid, having a weight average molecular weight (Mw; measured by gel permeation chromatography) in the range of about 800 g / mol to about 12,000 g / mol, preferably in the range of about 900 g / mol to about 10,000 g / mol, more preferably in the range of about 1,000 g / mol to about 9,000 g / mol. The at least one CoHP1 may have an average molar mass selected from about 1,200 g / mol (HP1a), about 2,500 g / mol (HP1b), about 4,000 g / mol (HP1c), and about 8,500 g / mol (HP1d).

[0710] In one embodiment, the LDF comprises at least one HP1, preferably a sodium salt of polyacrylic acid, having a K-value in the range of about 10 to about 50, preferably in the range of about 15 to about 30, wherein the K-value is determined using about 1% dry matter in distilled water. The at least one HP1 may have a K-value selected from about 15 (HP1a), about 20 (HP1b), about 25 (HP1c), and about 30 (HP1d).

[0711] In one embodiment, LDF comprises at least one HP1. Specifically, it may comprise one of the following combinations:

[0712]

[0713]

[0714] In one embodiment, EPr9iA in LDF262 to LDF343 is EPr9iA with R101E.

[0715] In one embodiment, LDF262-LDF343 further comprises PSB1, PBS2, Amy1, Amy2, Amy3, Cel1, Lip1, or Lip2a.

[0716] In one embodiment, LDF262 to LDF343 further comprise Man1 or Man2 or Man3 or Man4 or Man5, preferably Man4.

[0717] In one embodiment, LDF262 to LDF343 further comprise at least one NIS selected from NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4 and NIS5.

[0718] In one embodiment, the LDF comprises one of the following combinations:

[0719]

[0720] In one embodiment, LDF344 to LDF432 are substantially free of anionic surfactants.

[0721] In one embodiment, EPr9iA in LDF344 to LDF432 is EPr9iA with R101E.

[0722] In one embodiment, LDF344 to LDF432 further comprise PSB1, PSP2, Amy1, Amy2, or Amy3.

[0723] In one embodiment, LDF344 to LDF432 further comprise at least one NIS selected from NIS1a1, NIS1a6, NIS1a7, NIS1a8, NIS1a9 and NIS1a10.

[0724] In one embodiment, the LDF comprises at least one copolymer of acrylic acid and maleic acid, which may be referred to herein as "CP1". CP1 herein includes salts of copolymers of acrylic acid and maleic acid. The salt-forming cations may be monovalent or polyvalent, including sodium, potassium, magnesium, calcium, ammonium, and ammonium salts of mono-, di-, and triethanolamine, with sodium being preferred. Thus, CP1 may be completely or only partially neutralized by salt formation.

[0725] In one embodiment, CP1 comprises 50 to 90% by weight acrylic acid and 50 to 10% by weight maleic acid.

[0726] In one embodiment, the LDF comprises at least one CP1, preferably a sodium salt of a copolymer of acrylic acid and maleic acid, having a weight average molecular weight (Mw; measured by gel permeation chromatography) in the range of about 30,000 g / mol to about 100,000 g / mol, preferably in the range of about 50,000 g / mol to about 90,000 g / mol, more preferably in the range of about 70,000 g / mol to about 85,000 g / mol. The at least one CP1 may have an average molar mass of about 70,000 g / mol.

[0727] In one embodiment, the LDF comprises at least one CP1, preferably a sodium salt of a copolymer of acrylic acid and maleic acid, having a K-value in the range of about 10 to about 100, preferably in the range of about 30 to about 80, more preferably in the range of about 45 to about 60, wherein the K-value is determined using about 1% dry matter in distilled water. The at least one CP1 may have a K-value of about 55.

[0728] In one embodiment, the LDF comprises at least one copolymer consisting of acrylic acid and at least one hydrophobic monomer, which may be referred to herein as "CP2a". Suitable hydrophobic monomers are, for example, isobutylene, diisobutylene, butene, pentene, hexene and styrene, olefins having 10 or more carbon atoms or mixtures thereof, such as, for example, 1-decene, 1-dodecene, 1-tetradecene, 1-hexadecene, 1-octadecene, 1-eicosene, 1-docosene, 1-tetracosene and 1-hexacosene, C 22 -α-olefins, C 20 -C 24-a mixture of alpha-olefins and polyisobutylene having an average of 12 to 100 carbon atoms per molecule. Such copolymers may be referred to herein as CoCP2 and include partially or fully neutralized forms thereof. Neutralization is preferably achieved by using a suitable base such as NaOH or KOH to form an alkali metal salt of such a polymer.

[0729] In one embodiment, the LDF comprises at least one copolymer consisting of maleic acid and at least one hydrophobic monomer as disclosed above. The hydrophobic monomer is preferably selected from the group consisting of isobutylene, diisobutylene, butene, or a mixture thereof. Neutralization is preferably achieved by using a suitable base such as NaOH or KOH to form an alkali metal salt of such a polymer.

[0730] In one embodiment, the copolymer comprises maleic acid and a hydrophobic monomer, preferably diisobutylene, in a ratio of 1:1, which may be referred to herein as "CP2b". Preferably, the CP2b is a sodium salt of a copolymer of maleic acid and diisobutylene. CP2b may have a K-value in the range of about 20 to about 80, preferably in the range of about 0 to about 50, more preferably in the range of about 35 to about 45, wherein the K-value is determined using about 1% dry matter in distilled water.

[0731] In one embodiment, the LDF comprises at least one copolymer consisting of at least one monomer according to formula CoC and at least one monomer having at least one sulfonate group. The copolymer may be referred to herein as "CP3" and is preferably composed of acrylic acid and AMPS. CP3 herein includes salts of the copolymer. The salt-forming cation may be monovalent or polyvalent, including sodium, potassium, magnesium, calcium, ammonium and ammonium salts of mono-, di- and triethanolamine, with sodium being preferred. Thus, CP3 may be completely or only partially neutralized by salt formation.

[0732] In one embodiment, CP3 comprises about 60% to 80% by weight acrylic acid and 20% to 40% by weight AMPS. Preferably, the weight ratio acrylic acid:AMPS is about 70:30. CP3 may have a K-value of about 40.

[0733] In one embodiment, LDF comprises at least one copolymer comprising at least one monomer from the group consisting of unsaturated carboxylic acids as defined in formula CoC and at least one hydrophilic monomer selected from nonionic monomers having hydroxyl functional groups or alkylene oxide groups. Such copolymers may be referred to herein as "CP4" and include salts of the polymer. The salt-forming cations may be monovalent or polyvalent, including sodium, potassium, magnesium, calcium, ammonium and mono-, di- and triethanolamine ammonium salts, of which sodium is preferred. Therefore, CP4 may be completely or only partially neutralized by salt formation.

[0734] Examples of such nonionic monomers having hydroxyl functional groups or alkylene oxide groups include, but are not limited to, allyl alcohol, isoprene alcohol, methoxy polyethylene glycol (meth) acrylate, methoxy polypropylene glycol (meth) acrylate, methoxy polybutylene glycol (meth) acrylate, methoxy poly(propylene oxide-co-ethylene oxide) (meth) acrylate, ethoxy polyethylene glycol (meth) acrylate, ethoxy polypropylene glycol (meth) acrylate, ethoxy polybutylene glycol (meth) acrylate, and ethoxy poly(propylene oxide-co-ethylene oxide) (meth) acrylate. The polyalkylene glycol herein may contain 3 alkylene oxide units (AO) to 50 AO / molecules, 5 AO to 40 AO / molecules, or 10 AO to 30 AO / molecules.

[0735] In one embodiment, the LDF comprises at least one polycarboxylate which is derived by alkoxylation such as ethoxylation and / or propoxylation. Such polycarboxylates may be referred to herein as "CP5".

[0736] Alkoxylated polycarboxylates include polyacrylates with one ethoxy side chain per 2 to 8 acrylate units. In one embodiment, the alkoxylated polycarboxylates include polyacrylates with one ethoxy side chain per 7 to 8 acrylate units. The side chains are connected to the polyacrylate "backbone" through an ester to provide a "comb-like" polymeric structure. The molecular weight can be in the range of about 2,000 g / mol to about 50,000 g / mol.

[0737] In one embodiment, the LDF comprises at least one amphoteric polymer comprising

[0738] (a) at least one ethylenically unsaturated carboxylic acid selected from acrylic acid and methacrylic acid,

[0739] (b) at least one amide selected from N-C1-C 10 - alkyl(meth)acrylamides, acrylamide and methacrylamide, and

[0740] (c) at least one comonomer selected from the group consisting of DADMAC (polydiallyldimethylammonium chloride), MAPTAC (3-methacrylamido-N,N,N-trimethylpropane-1-ammonium chloride) and APTAC ((3-acrylamidopropyl)-trimethylammonium chloride).

[0741] Such amphiphilic polymers may be referred to herein as "CP6".

[0742] In one embodiment, the LDF comprises one or more compounds selected from the group of polyaspartic acid and its salts, which may be referred to herein as "CP7". The salt-forming cation may be monovalent or polyvalent. Suitable examples include, but are not limited to, sodium, potassium, magnesium, calcium, ammonium, and ammonium salts of mono-, di- and triethanolamine.

[0743] CP7 can be prepared from aspartic acid, maleic acid or maleic anhydride or fumaric acid, which are reacted to obtain a first polysuccinimide (which in turn can be hydrolyzed to polyaspartic acid or directly to a salt of polyaspartic acid) or directly to polyaspartic acid or directly to a salt thereof. It is preferred to use L-aspartic acid as the only starting material for the preparation of thermal polyaspartic acid.

[0744] In one aspect, CP7 includes compounds produced by the polycondensation of aspartic acid and other carboxylic acids such as amino acids. In another aspect, CP7 includes compounds produced by the polycondensation of aspartic acid, diamines and amino alcohols. Therefore, CP7 includes copolymers of aspartic acid and salts thereof.

[0745] In one embodiment, the LDF comprises one or more compounds selected from the group of alkoxylated polyalkyleneimines or alkoxylated polyamines, which may be referred to herein as "CP8". "CP8" also includes the structures disclosed in WO 2021 / 165468, in particular in claim 1 and on pages 2 to 4 of WO 2021 / 165468, and the structures obtained by the methods described in WO 2022 / 136408 and WO 2022 / 136409, in particular in claim 1 and on page 3 of WO 2022 / 136408 and WO 2022 / 136409, respectively. The structure of the alkoxylated polyalkyleneimines or alkoxylated polyamines may be further described by the general formula (CP8a):

[0746]

[0747] The variables are defined as follows:

[0748] R means same or different

[0749] i) Straight or branched C2-C 12- An alkylene group or

[0750] ii) an ether alkyl unit of the following formula (CP8b):

[0751]

[0752] The variables are defined as follows:

[0753] R 10 , R 11 , R12 represents identical or different straight-chain or branched C2-C6-alkylene groups, and

[0754] d is an integer having a value in the range of 0 to 50, or

[0755] iii) C5-C5-C5-alkyl optionally substituted by at least one C1-C3-alkyl 10 -cycloalkylene;

[0756] B represents the additional moiety of the alkoxylated polyalkyleneimine by branching

[0757] y and z are each an integer having a value in the range of 0 to 150, with the proviso that, in the case where R is a C5-C3-alkyl group optionally substituted with at least one C1-C3-alkyl group, 10 In the case of -cycloalkylene, both z and y are 0

[0758] E1, E2, E3, E4, E5 are hydrogen or represent identical or different residues according to formula (CP8c),

[0759] wherein the residue according to formula (CP8c) is an alkyleneoxy unit as defined below

[0760]

[0761] The variables are defined as follows:

[0762] R 1 Represents C2-C 22 -(1,2-alkylene) group;

[0763] R 2 represents hydrogen and / or C1-C 22 -alkyl and / or C7-C 22 - aralkyl, if z in the general formula (CP8b) is an integer ≥ 1, or

[0764] R 2 represents hydrogen and / or C1-C4-alkyl and / or C7-C 22 - aralkyl, if z is 0 in the general formula (CP8b);

[0765] n is an integer having a value of at least 5 to 100;

[0766] Wherein 20% to 100% of the total amount of E1, E2, E3, E4 and E5 in general formula (CP8a) is a residue according to formula (CP8c).

[0767] In one embodiment, the nitrogen atoms present in CP8 are quaternized in order to tailor the alkoxylated polyalkyleneimine or alkoxylated polyamine to a specific formulation to achieve better compatibility and / or phase stability of the formulation.

[0768] The LDF may contain CP8 in an amount ranging from 0.1% to 10% by weight, preferably from about 0.25% to 5% by weight, more preferably from about 0.5% to 3% by weight and most preferably from about 1% to 3% by weight, relative to the total weight of the detergent formulation.

[0769] In one embodiment, the LDF comprises at least one of the following combinations:

[0770]

[0771]

[0772] In one embodiment, EPr9iA in LDF433 to LDF497 is EPr9iA with R101E.

[0773] In one embodiment, LDF433 to LDF497 further comprises PSBl, PSP2, Amy1, Amy2, Amy3, Cel1, Lip1, or Lip2a.

[0774] In one embodiment, LDF433 to LDF497 further comprise Man1 or Man2 or Man3 or Man4 or Man5, preferably Man4.

[0775] In one embodiment, LDF433 to LDF497 further comprise at least one NIS selected from NIS1a2, NIS1a3, NIS1a4, NIS1a5, NIS2, NIS3, NIS4 and NIS5.

[0776] In one embodiment, the LDF comprises one of the following combinations:

[0777]

[0778] In one embodiment, LDF498 to LDF608 are substantially free of anionic surfactants.

[0779] In one embodiment, EPr9iA in LDF498 to LDF608 is R101E.

[0780] In one embodiment, LDF498 to LDF608 further include PSB1.

[0781] In one embodiment, LDF498 to LDF608 further include PSP2.

[0782] In one embodiment, LDF498 to LDF608 further comprises Amy1.

[0783] In one embodiment, LDF498 to LDF608 further comprises Amy2.

[0784] In one embodiment, LDF498 to LDF608 further comprises Amy3.

[0785] In one embodiment, LDF498 to LDF608 further comprise at least one NIS selected from NIS1a1, NIS1a6, NIS1a7, NIS1a8, NIS1a9 and NIS1a10.

[0786] Component D5 - Antimicrobial Agent

[0787] In one embodiment, the LDF of the present invention comprises

[0788] (A) 0.0005% to 0.4% by weight of at least one hydrolase (EC 3),

[0789] (B) 4 to 20% by weight of a compound according to formula (I) as described herein,

[0790] (C) at least one anionic surfactant (C1) and / or at least one builder (C2), and

[0791] (D) at least one antimicrobial agent (D5).

[0792] In one embodiment, the LDF of the present invention comprises at least one D5 selected from 2-phenoxyethanol (AMic1; CAS-No. 122-99-6), 2-bromo-2-nitropropane-1,3-diol (AMic2), formic acid in acid form or as a salt thereof (AMic3), 4,4'-dichloro-2-hydroxydiphenyl ether (AMic4; CAS-No. 3380-30-1) and isothiazol-3-one (AMic5).

[0793] In one embodiment, LDF1 to LDF608 contain AMic1 in an amount ranging from 0.1% to 2% by weight relative to the total weight of the liquid formulation.

[0794] In one embodiment, LDF1 to LDF608 contain AMic2 in an amount ranging from 20 ppm to 1000 ppm.

[0795] In one embodiment, LDF1 to LDF608 contain AMic3 in an amount ranging from 0.05% to 0.5% by weight relative to the total weight of the liquid formulation.

[0796] In one embodiment, LDF1 to LDF608 contain AMic4 in an amount ranging from 0.001% to 3% by weight, 0.002% to 1% by weight, or 0.01% to 0.6% by weight, all relative to the total weight of the liquid formulation.

[0797] Component D6 – Other ingredients

[0798] In one embodiment, the LDF disclosed herein further comprises at least one amphoteric surfactant selected from the group consisting of AMS1, AMS2, AMS3 and AMS4.

[0799] In one embodiment, the detergent formulation comprises at least one amphoteric surfactant selected from compounds of the general formula (AMS1), which may be referred to as amino acids (proteinogenic and non-proteinogenic):

[0800]

[0801] The variables in the general formula (AMS1) are defined as follows:

[0802] R 8 Selected from H, C1-C4 alkyl, C2-C4 alkenyl, wherein the alkyl and / or is linear (straight chain; n-) or branched.

[0803] R 9 Selected from C1-C 22 Alkyl, C2-C 22 Alkenyl, C 10 -C 22 Alkylcarbonyl and C 10 -C 22 Alkenylcarbonyl.

[0804] R 10 Selected from H, methyl, -(CH2)3NHC(NH)NH 2、 -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.

[0805] R x is selected from H and C1-C4-alkyl.

[0806] AMS2

[0807] In one embodiment, the detergent formulation comprises at least one amphoteric surfactant (which may be referred to as betaine and / or sulfobetaine) (AMS2) selected from compounds of general formula (AMS2a), (AMS2b), or (AMS2c):

[0808]

[0809] The variables in formula (AMS2a), (AMS2b) and (AMS2c) are defined as follows:

[0810] R 11 Selected from straight chain (straight chain; n-) or branched C7-C 22 Alkyl and straight chain (straight chain; n-) or branched C7-C 22 Alkenyl.

[0811] R 12 Each is independently selected from straight chain (straight chain; n-) C1-C4 alkyl.

[0812] R 13 Selected from C1-C5 alkyl and hydroxy C1-C5 alkyl, for example, 2-hydroxypropyl.

[0813] A - Selected from carboxylate and sulfonate.

[0814] The integer r in the general formulae (AMS2a), (AMS2b) and (AMS2c) is in the range of 2-6.

[0815] AMS3

[0816] In one embodiment, the detergent formulation comprises at least one amphoteric surfactant (which may be referred to as an alkyl-amphocarboxylate) selected from compounds of the general formula (AMS3):

[0817]

[0818] The variables in the general formula (AMS3) are defined as follows:

[0819] R 11 Selected from C7-C 22 Alkyl and C7-C 22 Alkenyl, wherein the alkyl and / or alkenyl is straight-chain (linear; n-) or branched, preferably straight-chain.

[0820] R 14 Selected from -CH2C(O)O - M + 、-CH2CH2C(O)O- M + and -CH2CH(OH)CH2SO3 - M + .

[0821] R 15 Selected from H and -CH2C(O)O -

[0822] The integer r in the general formula (AMS3) is in the range of 2-6.

[0823] M+ is selected from salt-forming cations. Salt-forming cations can be monovalent or polyvalent; therefore, M+ is equal to 1 / v Mv+. Examples include, but are not limited to, sodium, potassium, magnesium, calcium, ammonium, and ammonium salts of mono-, di-, and triethanolamine.

[0824] AMS4

[0825] In one embodiment, the detergent formulation comprises at least one amphoteric surfactant (which may be referred to as an amine oxide (AO)) selected from compounds of the general formula (AMS4):

[0826]

[0827] The variables in the general formula (AMS4) are defined as follows:

[0828] R 16 Selected from C8-C 18 Alkyl, hydroxyl C8-C 18 Alkyl, acylamidopropionyl and C8-C 18 Alkylphenyl; wherein the alkyl and / or alkenyl groups are straight chain (straight chain; n-) or branched

[0829] R 17 Selected from C2-C3 alkylene, hydroxy C2-C3 alkylene and mixtures thereof

[0830] R 18 Selected from C1-C3 alkyl and hydroxy C1-C3 alkyl

[0831] The integer x in the general formula (AMS4) is in the range of 0 to 5, preferably from 0 to 3, most preferably 0.

[0832] In one embodiment, the LDF containing anionic surfactant (C1) contains at least one graft polymer "GP1" containing a polyether as a grafting matrix and a copolymer as a grafted side chain, the copolymer containing at least one comonomer selected from the following

[0833] CH2=CH-OC(O)-R 3 (Comonomer Ia)

[0834] CH2=CH-CH2-OC(O)-R 3 (Comonomer Ib)

[0835] CH2=CZ-CO-OR 4 (Comonomer Ic)

[0836] Where R 3 Selected from C1-C 21 Alkyl, for example, methyl, n-propyl, n-pentyl, n-heptyl, n-nonyl, isononyl, n-undecyl, n-tridecyl, n-pentadecyl, n-heptadecyl, or n-nonadecyl.

[0837] R 4 Selected from C2-C 20 Alkyl, preferably with an even number of carbon atoms, for example ethyl, n-propyl and isopropyl, n-hexyl, isohexyl, sec-hexyl, n-heptyl, n-octyl, 2-ethylhexyl, n-nonyl, n-decyl or isodecyl, nC 12 H 25 、nC 14 H 29 、nC 16 H 33 or nC 18 H 37 ,

[0838] And Z is selected from hydrogen and methyl, hydrogen being preferred.

[0839] In one embodiment, the polyether is, for example, having an average molecular weight M in the range of from 500 to 25,000 g / mol, preferably 1,000 to 15,000 g / mol, and even more preferably 1,500 to 10,000 g / mol. n of polyethylene glycol.

[0840] In one embodiment, the polyether is, for example, having an average molecular weight M in the range of from 500 to 20,000 g / mol, preferably 2,000 to 10,000 g / mol, and even more preferably 4,000 to 9,000 g / mol. n of polypropylene glycol.

[0841] In one embodiment, the polyether is a copolymer of ethylene glycol and propylene glycol units, such as a random copolymer and preferably a block copolymer, such as a diblock copolymer and a triblock copolymer.

[0842] In one embodiment, the grafting matrix of GP1 is selected from polyethylene glycol, polypropylene glycol and EO-PO block copolymer, each of which is non-terminated or with C1-C 20 Alkyl or C3-C 20- 2-Hydroxyalkyl terminated. The polyether is preferably non-terminated.

[0843] Comonomer Ia may in particular be chosen from vinyl acetate, vinyl propionate, vinyl butyrate, vinyl n-hexanoate, vinyl n-octanoate, vinyl-2-ethyl hexanoate, vinyl laurate, vinyl stearate, vinyl myristate and vinyl palmitate.

[0844] The comonomer Ib may in particular be chosen from allyl propionate, allyl butyrate, allyl n-hexanoate, allyl n-octanoate, allyl-2-ethyl hexanoate, allyl laurate, vinyl stearate, allyl myristate and allyl palmitate.

[0845] Comonomer Ic may in particular be chosen from 2-ethylhexyl (meth)acrylate, 2-n-propylheptyl (meth)acrylate, stearyl (meth)acrylate, lauryl (meth)acrylate, palmityl (meth)acrylate and myristyl (meth)acrylate.

[0846] In one embodiment, GP1 comprises side chains in copolymerized form of comonomers preferably selected from the group consisting of (comonomer la) and (comonomer lc).

[0847] In one embodiment, GP1 has an average molecular weight M in the range of from 2,250 to 200,000 g / mol. n , preferably 2,250 to 25,000 g / mol, even more preferably 2,500 to 10,000 g / mol. The average molecular weight M n This can be determined by gel permeation chromatography using polyethylene glycol as a comparative standard. The grafting itself can be confirmed by HPLC (high pressure liquid chromatography).

[0848] In one embodiment, the essentially anionic surfactant (C1 )-free LDF comprises 0.05 to 0.4% by weight, relative to the total weight of the detergent formulation, of at least one compound selected from the group of non-alkoxylated polyalkyleneimines (PEI).

[0849] In one embodiment, LDF comprises a branched homopolymer of 15-20 ethyleneimine units. The homopolymer preferably has an Mw of about 250 g / mol to 1,500 g / mol. 13 The ratio of primary amine:secondary amine:tertiary amine is preferably about 1:0.9:x, where x is 0.5-0.6, as measured by C-NMR spectroscopy.Such a homopolymer may be referred to herein as PEI1.

[0850] In one embodiment, the LDF comprises a branched homopolymer having a Mw of about 500 g / mol to 10,000 g / mol.13 The ratio of primary amine:secondary amine:tertiary amine is preferably about 1:1:x, where x is 0.7-0.9, as measured by C-NMR spectroscopy.Such homopolymers may be referred to herein as PEI2.

[0851] In one embodiment, the LDF comprises a branched homopolymer having a Mw of about 25,000 g / mol and when used 13 The ratio of primary amine:secondary amine:tertiary amine is preferably about 1:1.1:0.7 when measured by C-NMR spectroscopy. The homopolymer may be referred to herein as PEI3.

[0852] In one embodiment, the LDF comprises a branched homopolymer having a Mw of about 70,000 g / mol and when used 13 The ratio of primary amine:secondary amine:tertiary amine is preferably about 0.5:1:0.5 as measured by C-NMR spectroscopy. The homopolymer may be referred to herein as PEI4.

[0853] In one embodiment, the LDF comprises a branched homopolymer having a Mw of about 750,000 g / mol and when used 13 The ratio of primary amine:secondary amine:tertiary amine is preferably about 1:1:0.7 as measured by C-NMR spectroscopy. The homopolymer may be referred to herein as PEI5.

[0854] In one embodiment, the LDF substantially free of anionic surfactant (C1) comprises 0.05% to 0.4% by weight, relative to the total weight of the detergent formulation, of at least one zinc salt (ZS) selected from water-soluble and water-insoluble zinc salts. In this context, water-insoluble means a zinc salt having a solubility of 0.1 g / l or less in distilled water at 25°C. Therefore, zinc salts having a higher solubility in water are correspondingly referred to as water-soluble zinc salts.

[0855] The zinc salt can 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, among which ZnCl2 (ZS1), ZnSO4 (ZS2), zinc acetate (ZS3), zinc citrate (ZS4), Zn(NO3)2 (ZS5), Zn(CH3SO3)2 (ZS6) and zinc gallate (ZS7) are preferred.

[0856] In one embodiment, the zinc salt is selected from ZnO (ZS8), ZnO·aq (ZS9), Zn(OH)2 (ZS10) and ZnCO3 (ZS11). ZnO·aq is preferred.

[0857] In one embodiment, the LDF substantially free of anionic surfactant (C1) comprises at least one graft polymer "GP2" consisting of

[0858] GP2-A: at least one grafting substrate selected from nonionic monosaccharides, disaccharides, oligosaccharides and polysaccharides, and side chains obtainable by grafting on

[0859] GP2-B: at least one ethylenically unsaturated monocarboxylic acid or dicarboxylic acid, referred to as monocarboxylic acid or dicarboxylic acid, and

[0860] GP2-C: at least one compound selected from the following

[0861]

[0862] GP2-A is preferably selected from nonionic polysaccharides, in particular from starches, which are preferably not chemically modified, for example their hydroxyl groups are preferably neither esterified nor etherified. Examples are corn starch, rice starch, potato starch and wheat starch.

[0863] GP2-B is preferably selected from monocarboxylic acids, more preferably selected from ethylenically unsaturated C3-C 10 - monocarboxylic acids and their alkali metal or ammonium salts, in particular potassium and sodium salts. Preferred monocarboxylic acids are acrylic acid and methacrylic acid, and also sodium (meth)acrylate. 10 Mixtures of monocarboxylic acids and in particular mixtures of acrylic acid and methacrylic acid are also preferred.

[0864] Examples

[0865] Example 1: Liquid enzyme formulation with and without initial stabilizer (% means % by weight relative to the total weight of LEP)

[0866] Table 1: Composition of liquid enzyme preparations with and without initial stabilizer (% means % by weight relative to the total weight of LEP)

[0867]

[0868] Enzyme concentrate: EPr9iA with R101E; amount used in LEP was adjusted to provide 6% protease with 100% proteolytic activity

[0869] Initial enzyme stabilizer: Z-VAL

[0870] Table 2: Stability of proteases determined by residual enzyme activity after storage

[0871]

[0872] *When compared to the initial protease activity at t=0, which had been set to 100%.

[0873] The liquid enzyme preparation is stored at a temperature of 38°C for up to 10 weeks. 4 weeks of storage is considered equivalent to approximately 9 months at room temperature or >15 months of storage at 8°C. Protease activity is analyzed by measuring the reactivity to the peptide substrate Suc-AAPF-pNA. Here, pNA is cleaved from the substrate molecule using 100mM TRIS buffer at 30°C, pH 8.6. The cleavage rate, which is proportional to the protease activity, can be determined by measuring the OD405 (optical density at 405nm) by the increase in the yellow color of free pNA in the solution. Before and after storage, the proteolytic activity is determined.

[0874] Example 2: Liquid detergent formulations tested for proteases and amylases (% means % by weight relative to the total weight of the detergent formulation)

[0875] Table 3: Composition of the LDF tested for proteases and amylases

[0876]

[0877] LAS: anionic surfactant; linear dodecylbenzene sulfonic acid (CAS27176-87-0): Maranil DBS / LC

[0878] AES (alkyl ether sulfate); anionic surfactant; poly(oxy-1,2-ethanediyl), α.-sulfo-.ω.-hydroxy-, C 12-14 -Alkyl ether, sodium salt (CAS 68891-38-3): Texapon N70

[0879] NIS1: nonionic surfactant; C 13 C 15 Oxo alcohol ethoxylate (7EO): Lutensol AO 7

[0880] NIS2: nonionic surfactant; C 12 C 14 Oxo alcohol ethoxylate (7EO): Lutensol A 7N

[0881] NIS3: nonionic surfactant; C 12 C 18 Oxo alcohol ethoxylate (7EO): Dehydol LT 7

[0882] MEA: Monoethanolamine

[0883] TEA: triethanolamine

[0884] Protease product: A product containing about 4% EPr9iA with R101E (with 100% proteolytic activity) in about 50% diol blend

[0885] Amylase products: prime 100L (Novozymes)

[0886] Table 4: Storage stability of the enzymes determined by the residual activity of proteases and amylases after storage for a certain period of time at 38° C. in the detergent formulations Det 1 and Det 2.

[0887]

[0888] The liquid detergent formulations were stored for up to 6 weeks at a temperature of 38°C. 4 weeks of storage is considered equivalent to approximately 9 months at room temperature or >15 months at 8°C.

[0889] Protease activity was analyzed by measuring reactivity to the peptide substrate Suc-AAPF-pNA. Here, pNA was cleaved from the substrate molecule using 100 mM TRIS buffer at 30°C, pH 8.6. The cleavage rate, which is proportional to the protease activity, can be measured by OD 405 (optical density at 405 nm) was determined by the increase in the yellow color of free pNA in solution. Before and after storage, the proteolytic activity was determined.

[0890] Amylase activity after storage was quantitatively measured by releasing the chromophore p-nitrophenol (pNP) from the substrate Ethyliden-capped-pNPG7 (Roche Applied Science, Material No. 10880078103). Alpha-amylase degrades the substrate into smaller molecules, and alpha-glucosidase (Roche Applied Science, Material No. 11626329103) added in excess compared to alpha-amylase processes these smaller products until pNP is released; the release of pNP measured by the increase in absorption at 405nm is proportional to the alpha-amylase activity of the sample. Amylase standard: Termamyl 120L (Sigma 3403). Before and after storage, amylolytic activity was determined.

[0891] The residual enzyme activity corresponds to the enzyme activity remaining when compared to the initial enzyme activity available at time 0 before storage.

[0892] Example 3: Tested liquid detergent formulations with protease, amylase and lipase

[0893] Table 5: Composition of the tested liquid detergent formulations for proteases, amylases and lipases (% means % by weight relative to the total weight of the detergent formulation)

[0894]

[0895] LAS: anionic surfactant; linear dodecylbenzene sulfonic acid (CAS27176-87-0): Maranil DBS / LC

[0896] AES (alkyl ether sulfate); anionic surfactant; poly(oxy-1,2-ethanediyl), α.-sulfo-.ω.-hydroxy-, C 12-14 -Alkyl ether, sodium salt (CAS 68891-38-3): Texapon N70

[0897] NIS1a-I: nonionic surfactant; C 13 C 15 Oxo alcohol ethoxylate (7EO): Lutensol AO 7

[0898] NIS1a-II: nonionic surfactant; C 12 C 14 Oxo alcohol ethoxylate (7EO): Lutensol A7N

[0899] NIS1a-II: nonionic surfactant; C 12 C 18 Oxo alcohol ethoxylate (7EO): Dehydol LT 7

[0900] MEA: monoethanolamine; TEA: triethanolamine

[0901] * Enzymes: % refers to the % by weight of the enzyme product formulated into the detergent formulation relative to the total weight of the detergent formulation

[0902] Protease I: Enzyme formulation with 4% EPr9iA with R101E (with 100% proteolytic activity) + about 0.3% Z-VAL

[0903] Protease II: Enzyme formulation with 4% EPr9iA with R101E (with 100% proteolytic activity), lacking enzyme stabilizers (such as boron-containing compounds and peptide stabilizers)

[0904] Amylase: prime 100L (Novozymes)

[0905] Lipase: purchased from Sigma-Aldrich 100L (CAS-No. 9001-62-1, EC-No. 232-619-9).

[0906] Table 6: Stability of detergent formulations containing additives

[0907]

[0908] Table 7: Residual activity of proteases, amylases and lipases after storage at 38°C

[0909]

[0910]

[0911]

[0912] * The preparation is turbid and unstable

[0913] Detergent formulations Det 1 and Det 3 as described above, except for the enzyme components, were as follows:

[0914]

[0915] Table 8: Residual activity of proteases and amylases after storage at 38°C

[0916]

[0917] The liquid detergent formulations were stored at a temperature of 38°C for up to 6 or 8 weeks, respectively. 4 weeks of storage is considered equivalent to approximately 9 months at room temperature or >15 months at 8°C.

[0918] Protease activity was analyzed by measuring reactivity to the peptide substrate Suc-AAPF-pNA. Here, pNA was cleaved from the substrate molecule using 100 mM TRIS buffer at 30°C, pH 8.6. The cleavage rate, which is proportional to the protease activity, can be measured by OD 405 (optical density at 405 nm) was determined by the increase in the yellow color of free pNA in solution. Before and after storage, the proteolytic activity was determined.

[0919] Amylase activity after storage was quantitatively measured by releasing the chromophore p-nitrophenol (pNP) from the substrate Ethyliden-capped-pNPG7 (Roche Applied Science, Material No. 10880078103). Alpha-amylase degrades the substrate into smaller molecules, and alpha-glucosidase (Roche Applied Science, Material No. 11626329103) added in excess compared to alpha-amylase processes these smaller products until pNP is released; the release of pNP measured by the increase in absorption at 405nm is proportional to the alpha-amylase activity of the sample. Amylase standard: Termamyl 120L (Sigma 3403). Before and after storage, amylolytic activity was determined.

[0920] Lipase activity was determined by using p-nitrophenol-valerate (2.4 mM pNP-C5 in 100 mM Tris, pH 8.0, 0.01% Triton X100) as a substrate. The absorption at 405 nm was measured every 30 seconds at 20° C. for 5 minutes. The slope of the time-dependent absorption curve (increase in absorbance per minute at 405 nm) is proportional to the activity of the lipase.

[0921] The residual enzyme activity corresponds to the enzyme activity remaining when compared to the initial enzyme activity available at time 0 before storage.

Claims

1. A liquid enzyme preparation comprising a. 0.5% to 15% by weight of at least one hydrolase (EC 3), and b. 2 to 70% by weight of at least one compound according to formula (I) Where R 1 and R 2 Selected from H and C2H4OH, R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, and m, n, o are each independently 0-2, preferably 0-1, more preferably 0; The amount of hydrolase refers to 100% active hydrolase.

2. The liquid enzyme preparation according to claim 1, wherein The liquid enzyme preparation comprises water in an amount not exceeding 15% by weight.

3. A liquid enzyme preparation according to any one of the preceding claims, wherein The liquid enzyme preparation is substantially free of surface active anionic compounds and complexing anionic compounds.

4. A liquid enzyme preparation according to any one of the preceding claims, wherein The liquid enzyme preparation is substantially free of enzyme stabilizers selected from boron-containing compounds and peptide stabilizers.

5. A liquid enzyme preparation according to any one of the preceding claims, wherein The liquid enzyme preparation further comprises at least one solvent selected from diols, triols and sugar alcohols and / or further comprises at least one salt selected from the group consisting of monovalent cations and monovalent anions of 1-6 carbons, NaCl, KCl, CaCl2 and Na2SO4.

6. A method for preparing a liquid enzyme preparation comprising the step of mixing at least one hydrolase (EC 3) with at least one compound according to formula (I) Where R 1 and R 2 Selected from H and C2H4OH, R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, And m, n, o are each independently 0-2, preferably 0-1, more preferably 0.

7. The method according to claim 6, wherein: The at least one hydrolase is contained in a liquid enzyme concentrate before mixing with the at least one compound according to formula (I), wherein the liquid enzyme concentrate preferably originates from a fermentative enzyme production.

8. The method according to claim 6, wherein: Prior to mixing with at least one compound according to formula (I), the at least one hydrolase is dissolved in a solvent selected from water and an organic solvent, preferably water.

9. At least one compound according to formula (I) Where R 1 and R 2 Selected from H and C2H4OH, R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, and m, n, o are each independently 0-2, preferably 0-1, more preferably 0; Use for providing a liquid enzyme preparation which is homogeneous in its appearance and has increased stability of at least one hydrolase when compared to a liquid enzyme preparation lacking the compound according to formula (I).

10. A liquid detergent formulation comprising (A) 0.0005% to 0.4% by weight of at least one hydrolase (EC 3) (B) 4 to 20% by weight of a compound according to formula (I) Where R 1 and R 2 Selected from H and C2H4OH, R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, and m, n, o are each independently 0-2, preferably 0-1, more preferably 0; and (C) at least 5% of at least one anionic compound.

11. The liquid detergent formulation according to claim 10, wherein The anionic compound is selected from surface active anionic compounds and complex anionic compounds, wherein the surface active anionic compound is preferably selected from LAS or AES, and wherein the complex anionic compound is preferably selected from citrate (NPB1) and aminocarboxylate (NPB3).

12. A liquid detergent formulation according to any one of claims 10 or 11, wherein The liquid detergent formulation comprises ≤ 3% by weight, preferably ≤ 2% by weight, of sodium formate.

13. A method for preparing a liquid detergent formulation comprising reacting at least one hydrolase, at least one compound according to formula (I) Where R 1 and R 2 Selected from H and C2H4OH, R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, and m, n, o are each independently 0-2, preferably 0-1, more preferably 0; and at least one anionic compound in one or more steps, wherein the at least one hydrolase is preferably comprised in the liquid enzyme preparation according to claim 1 before mixing with the at least one anionic compound.

14. At least one compound according to formula (I) Where R 1 and R 2 Selected from H and C2H4OH, R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, and m, n, o are each independently 0-2, preferably 0-1, more preferably 0; Use of a method for stabilizing at least one hydrolase contained in a liquid detergent formulation comprising at least one anionic compound selected from surface-active anionic compounds and complexing anionic compounds or providing a liquid detergent formulation which is homogeneous in its appearance and has an increased stability of at least one hydrolase when compared to a liquid detergent formulation lacking the compound according to formula (I).

15. A method for improving the detergency of a liquid detergent formulation by adding at least one compound according to formula (I) to a liquid detergent formulation containing a hydrolase Where R 1 and R 2 Selected from H and C2H4OH, R 3 Each of R is independently selected from H, methyl and ethyl, preferably all R 3 is H or methyl, and m, n, o are each independently 0-2, preferably 0-1, more preferably 0; The stain removal is preferably improved on at least one stain selected from the group consisting of protease-sensitive stains, amylase-sensitive stains and lipase-sensitive stains.

16. The liquid enzyme preparation according to any one of claims 1 to 5 or the liquid detergent formulation according to any one of claims 10 to 12, wherein The hydrolase is selected from the group consisting of proteases, amylases, lipases, cellulases, hemicellulases, mannanases, xylanases, DNases, dispersins, pectinases and cutinases, preferably selected from the group consisting of subtilisin (EC 3.4.21.62), α-amylases (EC 3.2.1.1) and triacylglycerol lipases (EC 3.1.1.3).

17. The liquid enzyme preparation according to any one of claims 1-5 or 16 or the liquid detergent formulation according to any one of claims 10-12 or 16, wherein The at least one compound according to formula (I) is triethanolamine formate.

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