[go: up one dir, main page]

WO2025093368A1 - Stabilisation d'enzymes dans des compositions contenant un inhibiteur de protéase - Google Patents

Stabilisation d'enzymes dans des compositions contenant un inhibiteur de protéase Download PDF

Info

Publication number
WO2025093368A1
WO2025093368A1 PCT/EP2024/079772 EP2024079772W WO2025093368A1 WO 2025093368 A1 WO2025093368 A1 WO 2025093368A1 EP 2024079772 W EP2024079772 W EP 2024079772W WO 2025093368 A1 WO2025093368 A1 WO 2025093368A1
Authority
WO
WIPO (PCT)
Prior art keywords
weight
composition
protease
enzyme
liquid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/EP2024/079772
Other languages
English (en)
Inventor
Sonja KUEBELBECK
Oliver Spangenberg
Claudia Esper
Gabriele BOENEMANN
Hauke Rohwer
Priya ANAND
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BASF SE
Original Assignee
BASF SE
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by BASF SE filed Critical BASF SE
Publication of WO2025093368A1 publication Critical patent/WO2025093368A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/38—Products with no well-defined composition, e.g. natural products
    • C11D3/386—Preparations containing enzymes, e.g. protease or amylase
    • C11D3/38663—Stabilised liquid enzyme compositions
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/20—Organic compounds containing oxygen
    • C11D3/2003—Alcohols; Phenols
    • C11D3/2041—Dihydric alcohols
    • C11D3/2048—Dihydric alcohols branched
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/20—Organic compounds containing oxygen
    • C11D3/2068—Ethers
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/20—Organic compounds containing oxygen
    • C11D3/2096—Heterocyclic compounds
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/38—Products with no well-defined composition, e.g. natural products
    • C11D3/386—Preparations containing enzymes, e.g. protease or amylase
    • C11D3/38618—Protease or amylase in liquid compositions only
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/38—Products with no well-defined composition, e.g. natural products
    • C11D3/386—Preparations containing enzymes, e.g. protease or amylase
    • C11D3/3869—Enzyme enhancers or mediators

Definitions

  • the present invention relates to the field of enzyme stabilization in compositions containing a non-polar protease inhibitor.
  • the present invention provides liquid compositions comprising solubilizing agents, namely at least one neopentyl glycol, gamma-valerolactone, and diethylene glycol that have been identified to efficiently solubilize non-polar protease inhibitors, while additionally stabilizing enzymes, preferably hydrolases, in the liquid environment.
  • Enzymes are increasingly used in various applications, such as in the detergent industry, as sustainable alternative to petrochemistry. Particularly proteases are nowadays routinely used in cleaning compositions. Furthermore, enzyme blends including a protease and one or more further enzyme such as an amylase, a cellulase, and/or a mannanase are used to further improve the performance of cleaning compositions.
  • enzyme blends including a protease and one or more further enzyme such as an amylase, a cellulase, and/or a mannanase are used to further improve the performance of cleaning compositions.
  • the use of enzymes is hampered by their instability in different environments, especially in detergent compositions but also more generally in hydrophobic solvents.
  • proteases degrade themselves and also further enzymes present in the composition. To overcome the latter, protease inhibitors can be used as enzyme stabilizers.
  • protease inhibitors only temporarily inhibit the enzyme activity under storage conditions but are released in the final application of the enzyme, therefore allowing the enzyme to exert its catalytic activity.
  • protease inhibitors are peptide aldehydes and peptide aldehyde hydrosulfite adducts as e.g. described in W02009/118375 or WO2013/004636.
  • the types of amino acid side chains of the peptide aldehyde or the hydrosulfite adducts determine the polarity of the protease inhibitor. It remains a challenge to provide water-containing products containing enzymes, such as cleaning compositions, comprising non-polar protease inhibitors with limited solubility in water and/or water-miscible organic solvents, which need to be solubilized in hydrophobic solvents.
  • One part of this challenge is the negative effect of such hydrophobic solvents on the stability of enzymes also contained in the compositions. Therefore, there is a need to identify solubilizing agents, which efficiently solubilize the non-polar protease inhibitor without negatively impacting enzyme stability.
  • WO2022/063698 discloses solvent systems comprising at least two organic solvents, wherein one of them is 1 ,2 propane diol (monopropylene glycol, MPG) and the other one is selected from diols other than 1 ,2-propane diol, preferably 1 ,6-hexane diol.
  • MPG monopropylene glycol
  • the disclosed solvent systems allow dissolution of enzyme stabilizers having limited solubility in water and/or water-miscible organic solvents in a water-containing protease product.
  • WO2022/063698 is silent on alternative solubilizing agents, such as neopentyl glycol or gamma-valerolactone to enable solubilization of such enzyme inhibitors.
  • WO2022/063698 does not show any beneficial effect on further enzymes (meaning other than proteases) when formulating enzymes. Therefore, there is still a need for improved solvent systems with less impact on protease stability and stability of further enzymes, such as amylases, mannanases and cellulases that nevertheless allow solubilization of non-polar protease inhibitors in water-containing products.
  • compositions comprising solubilizing agents that have been identified to efficiently solubilize non-polar protease inhibitors and additionally stabilize enzymes, preferably hydrolases, in a liquid environment.
  • solubilizing agents selected from the group consisting of neopentyl glycol, gamma-valerolactone, and diethylene glycol both allow dissolution of a protease inhibitor having limited solubility in water and/or water-miscible organic solvents, meaning a non-polar protease inhibitor, while also ensuring enzyme stability of proteases but additionally also of further enzymes, such as e.g. amylases, cellulases, or mannanases.
  • compositions including liquid enzyme compositions and liquid cleaning compositions, comprising at least one protease, a protease inhibitor and at least one solubilizing agent selected from the group consisting of neopentyl glycol, gamma-valerolactone, and diethylene glycol.
  • solubilizing agent selected from the group consisting of neopentyl glycol, gamma-valerolactone, and diethylene glycol.
  • the terms "about” and “approximately” denote an interval of accuracy that a person skilled in the art will understand to still ensure the technical effect of the feature in question.
  • the term typically indicates a deviation from the indicated numerical value of ⁇ 20%, preferably ⁇ 15%, more preferably ⁇ 10%, and even more preferably ⁇ 5%.
  • first”, “second”, “third” or “(a)”, “(b)”, “(c)”, “(d)” etc. are used for distinguishing between elements and not necessarily for describing a sequential or chronological order. It is to be understood that the terms so used are interchangeable under appropriate circumstances and that the embodiments of the invention described herein are capable of operation in other sequences than described or illustrated herein.
  • first, “second”, “third” or “(a)”, “(b)”, “(c)”, “(d)”, “i”, “ii” etc. relate to steps of a method or use or assay, there is no time or time interval coherence between the steps, i.e. the steps may be carried out simultaneously or there may be time intervals of seconds, minutes, hours, days, weeks, months or even years between such steps, unless otherwise indicated in the application as set forth herein above or below.
  • compositions are a mixture of components (also called ingredients) prepared according to a specific formula.
  • Compositions can be liquid or solid.
  • Liquid compositions of the invention include solutions, emulsions and dispersions, gels etc. as long as the liquid is fluid and pourable.
  • Liquid compositions according to the present invention preferably have a dynamic viscosity in the range of about 500 to about 20,000 mPa*s, determined at 25°C according to Brookfield, for example spindle 3 at 20 rpm with a Brookfield viscosimeter LVT-II.
  • Gel means a shear thinning, lamellar gel, with a pouring viscosity in the range of from 100 to 5,000 mPa*s (milli Pascal seconds), more preferably less than 3,000 mPa*s, most preferably less than 1 ,500 mPa*s.
  • Compositions include enzyme compositions, e.g. liquid enzyme compositions as described herein, and cleaning compositions, e.g. liquid cleaning compositions as described herein.
  • the components of the composition can be separated by formulating them in different compartments, such as different compartments of multi-chamber-pouches or bottles having different chambers, from which the liquids are poured out at the same time in a predefined amount to assure the application of the right amount per individual point of use of each component from each chamber.
  • multi-compartment-pouches and bottles etc. are known to a person of skill as well.
  • compositions of the invention may be delivered in dual- or multi-compartment containers, in single-phase or multi-phase unit doses, in a spray or foam detergent, as premoistened wipes (i.e., the cleaning composition in combination with a nonwoven material such as that discussed in US 6,121 , 165, Mackey, et al.), and other homogeneous, non-homogeneous or single-phase or multiphase cleaning product forms.
  • a composition "essentially devoid” of a compound shall mean herein that the respective compound is not added to the composition (on purpose), preferably that at most non-effective amounts are present, most preferably 0% of the compound are contained in the composition.
  • compositions including liquid enzyme compositions and liquid cleaning compositions, and methods of the present invention comprises a protease as described herein.
  • proteases also called “peptidases” and protease variants described herein have “protease activity” (also called “proteolytic activity” herein). This property is related to the hydrolytic activity of a protease (proteolysis, which means hydrolysis of peptide bonds linking amino acids together in a polypeptide chain) on protein containing substrates, e.g. casein, haemoglobin and BSA. Quantitatively, proteolytic activity is related to the rate of degradation of proteins by a protease or proteolytic enzyme in a defined course of time. The methods for analyzing proteolytic activity and thereby the effect of an inhibitor on the proteolytic activity are well-known in the literature (see e.g. Gupta et al. (2002), Appl. Microbiol. Biotechnol. 60: 381 -395).
  • proteolytic activity can be determined by using Succinyl-Ala-Ala-Pro-Phe-p-nitroanilide (Suc-AAPF- pNA, short AAPF; see e.g. DelMar et al. (1979), Analytical Biochem 99, 316-320) as substrate.
  • pNA is cleaved from the substrate molecule by proteolytic cleavage, resulting in release of free pNA which has a yellow color that can be quantified by measuring the optical density at 405 nm (OD405).
  • Variant enzymes as generally used herein, meaning not solely applicable to proteases, differ from “parent” enzymes by certain amino acid alterations, preferably amino acid substitutions at one or more amino acid positions.
  • the abbreviations for single amino acids used are according to the accepted IUPAC single letter or three letter amino acid abbreviation.
  • amino acid alteration refers to amino acid substitution, deletion, or insertion.
  • substitutions are generally described herein by providing the original amino acid followed by the number of the position within the amino acid sequence followed by the amino acid, which substitutes the original amino acid. Substitutions can also be described by merely naming the resulting amino acid without specifying the initial amino acid at this position. Positions of substitutions can be described by merely providing the number of the position within the amino acid sequence.
  • “Insertions” are generally described herein by providing the original amino acid followed by the number of the position within the amino acid sequence followed by the original amino acid and the additional amino acid. Ways of introducing amino acid alterations, e.g., substitutions, into protein sequence are well known in the art.
  • Variant polynucleotide and variant polypeptide sequences may be generally defined herein by their sequence identity when compared to another sequence. Sequence identity is usually provided as "% sequence identity” or "% identity”. For calculation of sequence identities, in a first step a sequence alignment is produced. According to this invention, a pairwise global alignment is produced, meaning that two sequences are aligned over their complete length. The alignment is usually produced by using a mathematical approach called alignment algorithm.
  • the alignment is generated by using the algorithm of Needleman and Wunsch (J. Mol. Biol. (1979) 48, p. 443-453).
  • the program "NEEDLE” The European Molecular Biology Open Software Suite (EMBOSS)
  • EMBOSS European Molecular Biology Open Software Suite
  • %-identity (identical residues I length of the alignment region, which is showing the sequence of the invention from start to stop codon excluding introns over its complete length) *100.
  • Amino acid A is similar to amino acids S
  • Amino acid D is similar to amino acids E; N
  • Amino acid E is similar to amino acids D; K; Q
  • Amino acid F is similar to amino acids W; Y
  • Amino acid H is similar to amino acids N; Y
  • Amino acid I is similar to amino acids L; M; V;
  • Amino acid K is similar to amino acids E; Q; R
  • Amino acid L is similar to amino acids I; M; V
  • Amino acid M is similar to amino acids I; L; V
  • Amino acid N is similar to amino acids D; H; S;
  • Amino acid Q is similar to amino acids E; K; R
  • Amino acid R is similar to amino acids K; Q
  • Amino acid S is similar to amino acids A; N; T
  • Amino acid T is similar to amino acids S
  • Amino acid V is similar to amino acids I; L; M
  • Amino acid W is similar to amino acids F; Y Amino acid Y is similar to amino acids F; H; W
  • Non-polar proteinogenic amino acids are alanine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, and valine.
  • Enzyme properties generally include herein, but are not limited to, catalytic activity, substrate/cofactor specificity, product specificity, stability in the course of time, thermostability, pH stability, and chemical stability.
  • Enzymatic activity or catalytic activity generally means the catalytic effect exerted by an enzyme, expressed as units per milligram of enzyme (specific activity) or molecules of substrate transformed per minute per molecule of enzyme (molecular activity).
  • Enzymatic activity can be specified by the enzymes actual function, e.g., protease exert proteolytic activity by catalyzing the hydrolytic cleavage of peptide bonds, lipases exert lipolytic activity by hydrolytic cleavage of ester bonds, amylase activity involves hydrolysis of glycosidic linkages in polysaccharides, mannanase activity involves hydrolysis of the internal glycoside bonds in the mannan backbone etc.
  • enzyme stability relates to the retention of enzymatic activity as a function of time during storage or operation. Retention of enzymatic activity as a function of time during storage is called “storage stability” and is preferred within the context of the invention.
  • Storage-stability relating to hydrolase activity, preferably protease activity, amylase activity, mannanase activity, or cellulase activity, within a composition preferably means that the residual hydrolytic activity after storage for up to 14 days or up to 3 months at elevated temperatures of about 37°C or 45°C is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, or at least 85% when compared to the hydrolytic activity of a sample stored at -80°C.
  • Suitable proteases according to the invention have at least 70%, more preferably at least 80%, more preferably at least 85%, most preferably at least 90% residual protease activity after storage for 28 days in a composition according to the invention at 45 °C.
  • Suitable proteases according to the invention have proteolytic activity, preferably when determined according to the essays described above, they exhibit at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150% of the proteolytic activity of the protease according to SEQ ID NO: 1 comprising a R101 E substitution (according to BPN' (SEQ ID NO: 2) numbering).
  • Proteases are members of class EC 3.4.
  • Proteases include aminopeptidases (EC 3.4.11), dipeptidases (EC 3.4.13), dipeptidyl-peptidases and tripeptidyl-peptidases (EC 3.4.14), peptidyl-dipeptidases (EC 3.4.15), serine-type carboxypeptidases (EC 3.4.16), metallocarboxypeptidases (EC 3.4.17), cysteine-type carboxypeptidases (EC 3.4.18), omega peptidases (EC 3.4.19), serine endopeptidases (EC 3.4.21), cysteine endopeptidases (EC 3.4.22), aspartic endopeptidases (EC 3.4.23), metallo-endopeptidases (EC 3.4.24), threonine endopeptidases (EC 3.4.25), or endopeptidases of unknown catalytic mechanism (EC 3.4.99).
  • At least one protease may be selected from metal lo-endoproteases (EC 3.4.24).
  • a metallopro-tease may for example be a thermolysin from, e.g., family M4 or another metalloprotease such as those from M5, M7 or M8 families.
  • a metal loprotease may especially be derived from Bacillus amyloliquefaciens described in WO 07/044993A2, from Bacillus, Brevibacillus, Thermoactinomyces, Geobacillus, Paenibacillus, Lysinibacillus or Streptomyces spp.
  • At least one protease may be selected from serine proteases (EC 3.4.21). Serine proteases or serine peptidases are characterized by having a serine in the catalytically active site, which forms a covalent adduct with the substrate during the catalytic reaction.
  • a serine protease may be selected from the group consisting of chymotrypsin (e.g., EC 3.4.21.1), elastase (e.g., EC 3.4.21.36, EC 3.4.21.37, or EC 3.4.21.71), granzyme (e.g., EC 3.4.21 .78 or EC 3.4.21 .79), kalli krein (e.g., EC 3.4.21 .34, EC 3.4.21 .35, EC 3.4.21 .118, or EC 3.4.21 .119,) plasmin (e.g., EC 3.4.21.7), trypsin (e.g., EC 3.4.21.4), thrombin (e.g., EC 3.4.21.5), and subtilisin.
  • subtilisin is also known as subtilopeptidase, e.g., EC 3.4.21.62, the latter hereinafter also being referred to as "subtilisin”.
  • subtilisins and chymotrypsin-related serine proteases both have a catalytic triad comprising aspartate, histidine and serine.
  • subtilisin related proteases the relative order of these amino acids, reading from the amino to carboxy-terminus, is aspartate-histidine-serine.
  • the chymotrypsin-related proteases the relative order is histidine-aspartate-serine.
  • subtilisin herein refers to a serine protease having the catalytic triad of subtilisin related proteases.
  • subtilisins as described in WO 89/06276 and EP 0283075, WO 89/06279, WO 89/09830, WO 89/09819, WO 91/06637 and WO 91/02792.
  • Parent proteases of the subtilisin type (EC 3.4.21.62) and variants may be bacterial proteases.
  • Said bacterial protease may derived from gram-positive bacteria such as a Bacillus, Clostridium, Enterococcus, Geobacillus, Lactobacillus, Lactococcus, Oceanobacillus, Staphylococcus, Streptococcus, or Streptomyces protease, or gramnegative bacteria such as a Campylobacter, Escherichia (e.g. E. coll), Flavobacterium, Fusobacterium, Helicobacter, llyobacter, Neisseria, Pseudomonas, Salmonella, or Ureaplasma protease.
  • the parent proteases and variants may be derived from Bacillus alcalophil us, 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, Bacillus stearothermophilus, Bacillus subtilis, or Bacillus thuringiensis.
  • At least one protease may be selected from the following: subtilisin from Bacillus amyloliquefaciens BPN' (SEQ ID NO: 2 herein, described by Vasantha et al. (1984) J. Bacteriol. Volume 159, p. 811-819 and JA Wells et al. (1983) in Nucleic Acids Research, Volume 11 , p. 7911 -7925); subtilisin from Bacillus licheniformis (subtilisin Carlsberg; disclosed in EL Smith et al. (1968) in J. Biol Chem, Volume 243, pp. 2184-2191 , and Jacobs et al. (1985) in Nucl.
  • subtilisin PB92 original sequence of the alkaline protease PB92 is described in EP 283075 A2; subtilisin 147 and/or 309 (Esperase®, Savinase®, respectively) as disclosed in WO 89/06279; subtilisin from Bacillus lentus as disclosed in WO 91/02792, such as from Bacillus lentus DSM 5483 or the variants of Bacillus lentus DSM 5483 as described in WO 95/23221 ; subtilisin from Bacillus alcalophilus (DSM 11233) disclosed in DE 10064983; subtilisin from Bacillus gibsonii (DSM 14391) as disclosed in WO 2003/054184; subtilisin from Bacillus sp.
  • DSM 11233 subtilisin from Bacillus alcalophilus
  • DSM 14391 subtilisin from Bacillus gibsonii
  • component a comprises at least subtilisin 309 (which might be called Savinase herein) as disclosed as sequence a) in Table I of WO 89/06279 or a variant thereof which is at least 80% similar and/or identical thereto and has proteolytic activity.
  • subtilisin 309 which might be called Savinase herein
  • Examples of useful proteases in accordance with the present invention comprise the variants of subtilisin protease derived from SEQ ID NO: 22 as described in EP 1921147 (disclosed in SEQ ID NO: 1 herein, which is the sequence of mature alkaline protease from Bacillus lentus DSM 5483) with amino acid substitutions in one or more of the following positions: 3, 4, 9, 15, 24, 27, 33, 36, 57, 68, 76, 77, 87, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 106, 118, 120, 123, 128, 129, 130, 131 , 154, 160, 167, 170, 194, 195, 199, 205, 206, 217, 218, 222, 224, 232, 235, 236, 245, 248, 252 and 274 (according to the BPN' numbering), which have proteolytic activity.
  • such a protease is not mutated
  • the protease of component a has SEQ ID NO: 22 as described in EP 1921147 (SEQ ID NO: 1 herein), or a protease which is at least 80% identical thereto and has proteolytic activity.
  • said protease is characterized by having at least amino acid glutamic acid (E), or aspartic acid (D), or asparagine (N), or glutamine (Q), or alanine (A), or glycine (G), or serine (S) at position 101 (according to BPN' numbering) and has proteolytic activity.
  • the at least one protease of component a is at least 80% identical to SEQ ID NO: 22 as described in EP 1921147 (SEQ ID NO: 1 herein) and is characterized by comprising one amino acid (according to (a)-(h) above) or combinations according to (I) above together with the amino acid 101 E, 101D, 101 N, 101 Q, 101A, 101 G, or 101 S (according to BPN' numbering, SEQ ID NO: 2 herein) and having proteolytic activity.
  • said protease is characterized by comprising the substitution (according to BPN' numbering) R101 E, or S3T + V4I + V205I, or R101 E and S3T, V4I, and V205I, or S3T + V4I + V199M + V205I + L217D, and having proteolytic activity.
  • the at least one protease comprised in component a is at least 80% identical to SEQ ID NO: 22 as described in EP 1921147 (SEQ ID NO: 1 herein) and has at least the substitutions S3T+V4I+R101 E+V205I or
  • the at least one protease comprised in component a. is a protease according to SEQ ID NO: 22 as described in EP 1921147 (SEQ ID NO: 1 herein) is characterized by comprising the substitutions (according to BPN' numbering) S3T + V4I + S9R + A15T + V68A + D99S + R101 S + A103S + 1104V + N218D and having proteolytic activity.
  • the at least one protease comprised in component a is a protease 80% identical to SEQ ID NO: 22 as described in EP 1921147 (SEQ ID NO: 1 herein) having the substitution R101 E (according to BPN' numbering, SEQ ID NO: 2 herein).
  • the at least one protease comprised in component a is a protease 80% identical to SEQ ID NO: 22 as described in EP 1921147 (SEQ ID NO: 1 herein) having the substitution R101 E (according to BPN' numbering, SEQ ID NO: 2 herein).
  • SEQ ID NO: 22 is a protease 82%, 84%, 85%, 86%, 88%, 90%, 92%, 94%, 95%, 96%, 98% or 99% but less than 100% identical to SEQ ID NO: 22 as described in EP 1921147 (SEQ ID NO: 1 herein) having the substitution R101 E (according to BPN' numbering, SEQ ID NO: 2 herein).
  • the at least one protease comprised in component a is a protease identical to SEQ ID NO: 22 as described in EP 1921147 (SEQ ID NO: 1 herein) having the substitution R101 E according to BPN' numbering (SEQ ID NO: 2 herein).
  • component a in one embodiment, comprises a combination of at least two proteases, preferably selected from the group of serine endopeptidases (EC 3.4.21), more preferably selected from the group of subtilisin type proteases (EC 3.4.21 .62) - all as disclosed above.
  • proteases preferably selected from the group of serine endopeptidases (EC 3.4.21), more preferably selected from the group of subtilisin type proteases (EC 3.4.21 .62) - all as disclosed above.
  • At least one protease may be selected from those commercially available including but not limited to those sold under the trade names Alcalase®, Blaze®, DuralaseTM, DurazymTM, Relase®, Relase® Ultra, Savinase®, Savinase® Ultra, Primase®, Polarzyme®, Kannase®, Liquanase®, Liquanase® Ultra, Ovozyme®, Coronase®, Coronase® Ultra, Neutrase®, Everlase® and Esperase® (Novozymes A/S), those sold under the tradename Maxatase®, Maxacai®, Maxapem®, Purafect®, Purafect® Prime, Purafect MA®, Purafect Ox®, Purafect OxP®, Puramax®, Properase®, FN2®, FN3®, FN4®, Excellase®, Eraser®, Ultimase®, Opticlean®, Effectenz®, Preferenz® and
  • proteases include also those, which are variants of the above-described proteases, which have proteolytic activity.
  • protease variants include variants with at least 40 to 100% identity to the full-length polypeptide sequence of the parent enzyme as disclosed above.
  • protease variants having proteolytic activity are at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the full-length polypeptide sequence of the parent enzyme as disclosed above.
  • the invention relates to protease variants comprising conservative substitutions not pertaining the functional domain of the respective protease.
  • protease variants have proteolytic activity according to the present invention when said protease variants exhibit increased proteolytic activity when compared to the parent protease.
  • the liquid composition of the invention is a liquid enzyme composition that comprises amounts of protease (component a.) in the range of about 0.05% to 10% by weight relative to the total weight of the liquid composition, wherein the amount relates to 100% active enzyme.
  • the protease is comprised in amounts in the range of about 2% to about 12% by weight, about 2% to about 10% by weight, about 2% to about 8% by weight, about 2% to about 6% by weight, or more preferably of about 2% to about 5% by weight, all relative to the total weight of the liquid composition.
  • the liquid composition of the invention is a liquid cleaning composition that comprises amounts of the protease (component a.) in the range of about 0.0005% to 0.1 % by weight relative to the total weight of the liquid composition, wherein the amount relates to 100% active enzyme.
  • the protease is comprised in amounts in the range of about 0.02% to 0.06% by weight, more preferably of about 0.02% to 0.05% by weight, all relative to the total weight of the liquid composition.
  • protease inhibitors as used herein are compounds that slow down or halt enzymatic activity of proteases. Protease inhibitors frequently also stabilize the enzyme in its three-dimensional structure. Hence, protease inhibitors usually also act as protease stabilizers.
  • the protease inhibitor in the compositions of the present invention is a peptide aldehyde, or a salt or a derivative thereof, preferably the protease inhibitor in the composition of the present invention is a peptide aldehyde, a peptide aldehyde hydrosulfite adduct, or a combination thereof.
  • a “derivative” of a compound as used herein, e. g. a peptide aldehyde, is a compound that is derived from the compound by a chemical reaction, e. g. a peptide aldehyde hydrosulfite adduct.
  • salt as used herein is a compound consisting of an assembly of positively charged cations and negatively charged anions resulting in a compound with no net electric charge.
  • component b. comprises at least one peptide stabilizer, meaning peptide aldehyde herein.
  • the peptide aldehyde is a tripeptide aldehyde, preferably selected from a compound of formula (I) or a salt thereof or hydrosulfite adduct thereof:
  • R1 , R2, R3 and Z within formula (I) are defined as follows:
  • R1 is an amino acid side chain of an L or D amino acid selected from the group consisting of Gly, Ala, Vai, Leu, lie, Met, Pro, Phe, Trp, Ser, Thr, Asp, Gin, Tyr, Cys, Lys, Arg, His, Asn, Glu, m-tyrosine, 3,4-dihydroxyphenylalanine, Nva, or Nle.
  • R1 is an amino acid side chain of an L or D amino acid selected from the group consisting of Ala, Vai, Gly, Arg, Leu, Phe, lie, His or Thr. More preferably, R1 is an amino acid side chain of an L or D amino acid selected from the group consisting of Ala, Vai, Gly, Arg, Leu, lie or His.
  • R2 is an amino acid side chain of an L or D amino acid selected from the group consisting of Gly, Ala, Vai, Leu, lie, Met, Pro, Phe, Trp, Ser, Thr, Asp, Gin, Tyr, Cys, Lys, Arg, His, Asn, Glu, m-tyrosine, 3,4-dihydroxyphenylalanine, Nva, or Nle.
  • R2 is an amino acid side chain of an L or D amino acid selected from the group consisting of Ala, Cys, Gly, Pro, Ser, Thr, Vai, Nva or Nle. More preferably, R2 is an amino acid side chain of an L or D amino acid selected from the group consisting of Ala, Gly, Pro or Vai.
  • R3 is an amino acid side chain of an L or D amino acid selected from the group consisting of Tyr, m-tyrosine, 3,4- dihydroxyphenylalanine, Phe, Vai, Ala, Met, Nva, Leu, lie or Nle or other non-natural amino acids carrying alkyl groups.
  • R3 is an amino acid side chain of an L or D amino acid selected from the group consisting of Tyr, Phe, Vai, Ala or Leu.
  • R1 and R2 may be amino acid side chains of non-polar L or D amino acids which herein include amino acids comprising aliphatic or aromatic R groups.
  • Non-polar amino acids herein specifically include Gly, Ala, Vai, Leu, lie, Met, Pro, Phe and Trp.
  • R1 is an amino acid side chain of an L or D-amino acid residue of Gly or Vai
  • R2 is an amino acid side chain of an L or D-amino acid residue of Ala
  • R3 is an amino acid side chain of an L or D-amino acid residue of Tyr, Ala, or Leu.
  • At least two of R1 , R2 and R3 are amino acid side chains of non-polar amino acid residues, preferably independently from each other selected from an L or D-amino acid residue of Ala, Vai, Gly and Leu.
  • R1 , R2 and R3 are amino acid side chains of non-polar amino acid residues.
  • R1 is an amino acid side chain of an L or D-amino acid residue of Vai
  • R2 is an amino acid side chain of an L or D-amino acid residue of Ala
  • R3 is an amino acid side chain of an L or D-amino acid residue of Leu.
  • the more non-polar amino acids are comprised in the peptide aldehyde, the more challenges exist to solubilize the peptide stabilizer in water and/or water-miscible solvents.
  • Z in formula (I) selected from hydrogen, an N-terminal protection group, and one or more amino acid residues optionally comprising an N-terminal protection group.
  • Z is an N-terminal protection group.
  • the N-terminal protection group is preferably a small aliphatic group, e.g., formyl, acetyl, fluorenylmethyloxycarbonyl (Fmoc), tert-butyloxycarbonyl (Boc), methoxycarbonyl (Moc), methoxyacetyl (Mac), methyl carbamate or a methylamino carbonyl/methyl urea group.
  • a small aliphatic group e.g., formyl, acetyl, fluorenylmethyloxycarbonyl (Fmoc), tert-butyloxycarbonyl (Boc), methoxycarbonyl (Moc), methoxyacetyl (Mac), methyl carbamate or a methylamino carbonyl/methyl urea group.
  • the N-terminal protection group is preferably a bulky aromatic group such as benzoyl (Bz), benzyloxycarbonyl (Cbz), p-methoxybenzyl carbonyl (MOZ), benzyl (Bn), p-methoxybenzyl (PMB) or p-methoxyphenyl (PMP).
  • Bz benzoyl
  • Cbz benzyloxycarbonyl
  • MOZ p-methoxybenzyl carbonyl
  • Bn benzyl
  • PMB p-methoxybenzyl
  • PMP p-methoxyphenyl
  • N-terminal protection groups are described in Greene's Protective Groups in Organic Synthesis, Fifth Edition by Peter G. M. Wuts, published in 2014 by John Wiley & Sons, Inc and in Isidro-Llobet et al., Amino Acid-Protecting Groups, Chem. Rev. 2009 109(6), 2455-2504.
  • compositions of the invention comprise at least one peptide aldehyde (component b.) selected from compounds according to formula (I), wherein
  • R1 is an amino acid side chain of an L or D-amino acid residue of Vai
  • R2 is an amino acid side chain of an L or D- amino acid residue of Ala
  • R3 is an amino acid side chain of an L or D-amino acid residue of Leu and the N-terminal protection group Z is selected from benzyloxycarbonyl (Cbz), p-methoxybenzyl carbonyl (MOZ), benzyl (Bn), benzoyl (Bz), p-methoxybenzyl (PMB), p-methoxyphenyl (PMP), formyl, acetyl (Ac), methyloxy, alkoxycarbonyl, methoxycarbonyl, fluorenylmethyloxycarbonyl (Fmoc), or tert-butyloxycarbonyl (Boc).
  • Z-VAL-H This compound is referred to as Z-VAL-H herein.
  • R1 is an amino acid side chain of an L or D-amino acid residue of Vai
  • R2 is an amino acid side chain of an L or D- amino acid residue of Ala
  • R3 is an amino acid side chain of an L or D-amino acid residue of Leu and the N-terminal protection group Z of the peptide aldehyde is benzyloxycarbonyl (Cbz).
  • This compound is referred to as Cbz-VAL-H herein, wherein -H represents the aldehyde group.
  • the liquid composition of the invention is a liquid enzyme composition that comprises amounts of the peptide aldehyde (component b.) in the range of about 0.05% to 0.8% by weight relative to the total weight of the liquid composition, wherein the amount relates to 100% active content.
  • the peptide aldehyde is comprised in amounts in the range of about 0.05% to about 0.6% by weight, of about 0.05% to about 0.5% by weight, of about 0.05% to about 0.4%, of about 0.05% to about 0.25%, of about 0.15% to about 0.2% or of about 0.1 % to about 0.25% by weight, all relative to the total weight of the liquid composition.
  • the liquid composition of the invention is a liquid cleaning composition that comprises amounts of the peptide aldehyde (component b.) in the range of about 0.00005% to 0.06% by weight relative to the total weight of the liquid composition, wherein the amount relates to 100% active content.
  • the peptide aldehyde is comprised in amounts in the range of about 0.00005% to 0.04% by weight, of about 0.00005% to 0.004% by weight, of about 0.0005% to 0.004%, or of about 0.001 % to 0.003% by weight, all relative to the total weight of the liquid composition.
  • Component c. Solubilizing agents according to the invention
  • Component c. comprises at least on solubilizing agent selected from the group consisting of
  • component c. comprises neopentyl glycol (NPG; IUPAC name: 2,2-dimethylpropane-l ,3-diol, sometimes also referred to as 2,2-dimethyl-1 ,3-propanediol), having the formula (II):
  • the liquid composition of the invention comprises amounts of neopentyl glycol as component c. ranging from about 0.01% to 25%.
  • the liquid composition of the invention is a liquid enzyme composition that comprises amounts of neopentyl glycol as component c. ranging from about 5% to 25% by weight, preferably from about 10% to 20% by weight.
  • the total amount of neopentyl glycol as component c. in the liquid enzyme composition of the invention may be at least 5% by weight, at least 6% by weight, at least 7% by weight, at least 8% by weight, at least 9% by weight, or at least 10% by weight.
  • liquid enzyme composition in the liquid enzyme composition may not be more than 25% by weight, not more than 24% by weight, not more than 23% by weight, not more than 22% by weight, not more than 21% by weight, not more than 20% by weight, not more than 19% by weight, not more than 18% by weight, not more than 17% by weight, not more than 16% by weight, not more than 15% by weight, not more than 14% by weight, not more than 13% by weight, or not more than 12% by weight.
  • the liquid composition of the invention is a liquid cleaning composition that comprises amounts of neopentyl glycol as component c. ranging from about 0.01% to 1% by weight, preferably from about 0.05% to 0.2% by weight.
  • the total amount of neopentyl glycol as component c. in the liquid cleaning composition of the invention may be at least 0.01% by weight, at least 0.02% by weight, at least 0.03% by weight, at least 0.04% by weight, or at least 0.05% by weight.
  • the total amount of neopentyl glycol as component c. in the liquid cleaning composition may not be more than 1% by weight, not more than 0.8% by weight, not more than 0.6% by weight, not more than 0.4% by weight, not more than 0.2% by weight, or not more than 0.15% by weight.
  • component c. comprises gamma-valerolactone (GVL; IUPAC name: 5-methyloxolan-2- one, sometimes also referred to as y-valerolactone, 5-methyldihydrofuran-2(3H)-one, 4-pentanolide, 4- valerolactone, 4-pentalactone, 4-hydroxypentanoic acid lactone), having the formula (III):
  • the liquid composition of the invention comprises amounts of gamma-valerolactone as component c. ranging from about 0.005% to 10%.
  • the liquid composition of the invention is a liquid enzyme composition that comprises amounts of gamma-valerolactone as component c. ranging from about 1% to 10% by weight, preferably from about 2% to 8% by weight, more preferably from about 2% to 5% by weight.
  • the total amount of gamma- valerolactone as component c. in the liquid enzyme compositions of the invention may be at least 1% by weight, at least 2% by weight, at least 3% by weight, at least 4% by weight, or at least 4.5% by weight.
  • liquid enzyme compositions of the invention may not be more than 10% by weight, not more than 9% by weight, not more than 8% by weight, not more than 7% by weight, not more than 6% by weight, not more than 5.5% by weight or not more than 5% by weight, all relative to the total weight of the liquid composition.
  • the liquid composition of the invention is a liquid cleaning composition that comprises amounts of gamma-valerolacone as component c. ranging from about 0.005% to 0.5% by weight, preferably from about 0.01% to 0.1% by weight.
  • the total amount of gamma-valerolactone as component c. in the liquid cleaning composition of the invention may be at least 0.005% by weight, at least 0.01% by weight, at least 0.02% by weight, at least 0.03% by weight, at least 0.04% by weight, or at least 0.05% by weight.
  • the total amount of gamma-valerolactone as component c. in the liquid cleaning composition may not be more than 0.5% by weight, not more than 0.4% by weight, not more than 0.3% by weight, not more than 0.2% by weight, not more than 0.15% by weight, or not more than 0.1% by weight.
  • component c. comprises diethylene glycol (DEG; IUPAC name: 2, 2' -oxydiethanol, sometimes also referred to as 2,2'-oxybis(ethan-1-ol), 2-(2-hydroxyethoxy)ethan-1-ol, ethylene diglycol, diglycol, 2,2'-oxybisethanol, 2,2'-oxydiethanol, 3-oxa-1,5-pentanediol, dihydroxy diethyl ether, digenos, digol), having the formula (IV):
  • DEG diethylene glycol
  • IUPAC name: 2,2' -oxydiethanol sometimes also referred to as 2,2'-oxybis(ethan-1-ol), 2-(2-hydroxyethoxy)ethan-1-ol, ethylene diglycol, diglycol, 2,2'-oxybisethanol, 2,2'-oxydiethanol, 3-oxa-1,5-pentanediol, dihydroxy diethyl ether, digenos, digol
  • the liquid composition of the invention comprises amounts of diethylene glycol as component c. ranging from about 0.01% to 15%.
  • the liquid composition of the invention is a liquid enzyme composition that comprises amounts of diethylene glycol as component c. ranging from about 5% to about 15% by weight, preferably from about 8% to about 12% by weight.
  • the total amount of diethylene glycol as component c. in the liquid enzyme compositions of the invention may be at least 5% by weight, at least 6% by weight, at least 7% by weight, at least 8% by weight, at least 9% by weight, at least 9.5% by weight, or at least 10% by weight, all relative to the total weight of the liquid composition.
  • liquid enzyme compositions of the invention may not be more than 15% by weight, not more than 14% by weight, not more than 13% by weight, not more than 12% by weight, not more than 11 % by weight, or not more than 10.5% by weight all relative to the total weight of the liquid composition.
  • the liquid composition of the invention is a liquid cleaning composition that comprises amounts of diethylene glycol as component c. ranging from about 0.01 % to 1 % by weight, preferably from about 0.05% to 0.2% by weight.
  • the total amount of diethylene glycol as component c. in the liquid cleaning composition of the invention may be at least 0.01 % by weight, at least 0.02% by weight, at least 0.03% by weight, at least 0.04% by weight, or at least 0.05% by weight.
  • the total amount of neopentyl glycol as component c. in the liquid cleaning composition may not be more than 1 % by weight, not more than 0.8% by weight, not more than 0.6% by weight, not more than 0.4% by weight, not more than 0.2% by weight, or not more than 0.15% by weight.
  • composition according to the invention including liquid enzyme compositions and liquid cleaning compositions, may comprise further components.
  • compositions including the liquid enzyme compositions and the liquid cleaning compositions, according to the invention are provided in a liquid form, they contain preferably water as the main solvent.
  • Nonaqueous solvents can also or additionally be used. Suitable non-aqueous solvents encompass mono- or polyhydric alcohols, alkanolamines or glycol ethers, provided they are miscible with water in the specified concentration range.
  • the solvents are preferably selected from ethanol, n-propanol, iso-propanol, butanols, glycol, propanediol, butanediol, glycerol, diglycol, propyl diglycol, butyl diglycol, hexylene glycol, ethylene glycol methyl ether, ethylene glycol ethyl ether, ethylene glycol propyl ether, ethylene glycol mono-n-butyl ether, diethylene glycol methyl ether, diethylene glycol ethyl ether, propylene glycol methyl ether, propylene glycol ethyl ether, propylene glycol propyl ether, dipropylene glycol monomethyl ether, dipropylene glycol monoethyl ether, diisopropylene glycol monomethyl ether, diisopropylene glycol monoethyl ether, methoxytriglycol, ethoxytriglycol, but
  • the composition may contain at least one polyol as additional non-aqueous solvent.
  • the polyol can in particular encompass glycerol, 1,2 propanediol (mono propylene glycol, MPG), 1 ,3-propanediol, ethylene glycol, and/or dipropylene glycol.
  • the composition preferably contains a solvent mixture of glycerol and MPG.
  • the composition according to the invention comprises MPG as additional solvent.
  • the liquid composition of the invention is a liquid enzyme composition that comprises amounts of MPG as additional solvent ranging from about 1 % to about 35% by weight, preferably from about 5% to about 30% by weight.
  • the total amount of MPG in the liquid enzyme composition of the invention may be at least 1 % by weight, at least 2% by weight, at least 4% by weight, at least 5% by weight, at least 6% by weight, at least 8% by weight, or at least 9% by weight.
  • the total amount of MPG in the liquid enzyme composition may not be more than 40% by weight, not more than 35% by weight, not more than 30% by weight, not more than 28% by weight, not more than 26% by weight, not more than 24% by weight, not more than 22% by weight, not more than 20% by weight, not more than 18% by weight, not more than 16% by weight, not more than 14% by weight, not more than 12% by weight, or not more than 10% by weight.
  • the liquid composition of the invention is a liquid cleaning composition that comprises amounts of MPG as additional solvent ranging from about 0.01 % to about 3% by weight, preferably from about 0.05 to about 1 % by weight, more preferably from about 0.05 to about 0.5% by weight.
  • the composition according to the invention comprises glycerol as additional solvent.
  • the liquid composition of the invention is a liquid enzyme composition that comprises amounts of glycerol as additional solvent ranging from about 10% to about 50% by weight, preferably from about 15% to about 45% by weight.
  • the total amount of glycerol in the liquid enzyme composition of the invention may be at least 10% by weight, at least 12% by weight, at least 14% by weight, at least 15% by weight, at least 16% by weight, at least 18% by weight, or at least 19% by weight.
  • the total amount of glycerol in the liquid enzyme composition may not be more than 45% by weight, not more than 42% by weight, not more than 40% by weight, not more than 38% by weight, not more than 36% by weight, not more than 34% by weight, not more than 32% by weight, not more than 30% by weight, not more than 28% by weight, not more than 26% by weight, not more than 24% by weight, not more than 22% by weight, or not more than 20% by weight.
  • the liquid composition of the invention is a liquid cleaning composition that comprises amounts of glycerol as additional solvent ranging from about 0.01 % to about 5% by weight, preferably from about 0.05 to about 1 % by weight, more preferably from about 0.05 to about 0.5% by weight.
  • the cleaning composition comprises at least one preservative.
  • a preservative is an antimicrobial agent which may be added to aqueous products and compositions to maintain the original performance, characteristics and integrity of the products and compositions by killing contaminating microorganisms or inhibiting their growth.
  • An antimicrobial agent is a chemical compound that kills microorganisms or inhibits their growth or reproduction. Microorganisms can be bacteria, yeasts or molds.
  • composition may contain one or more antimicrobial agents and/or preservatives as listed in patent WO2021/115912 A1 ("Formulations comprising a hydrophobically modified polyethylene-imine and one or more enzymes”) on pages 35 to 39.
  • Especially of interest for the cleaning compositions and fabric and home care products and specifically in laundry formulations are any of the following antimicrobial agents and/or preservatives: 4,4'-dichloro 2-hydroxydiphenyl ether (further names: 5-chloro-2-(4-chlorophenoxy) phenol, Diclosan, DCPP), Tinosan® HP 100 (30wt.% of DCPP in 1 ,2-propylene glycol); 2-phenoxyethanol (further names: phenoxyethanol, methylphenylglycol, phenoxetol, ethylene glycol phenyl ether, ethylene glycol monophenyl ether, 2-(phenoxy) ethanol, 2-phenoxy-1 -ethanol); 2-bromo-2-nitropropane-1,3-diol (further names: 2-bromo-2-nitro-1,3-propanediol, bronopol); glutaraldehyde (further names: 1 -5-pentandial, pent
  • the composition contains 2-phenoxyethanol in a concentration of 0.1 to 2%, preferably ⁇ 0.9%, or 4,4'- dichloro 2-hydroxydiphenyl ether (DCPP) in a concentration of 0.005 to 0.6%.
  • 2-phenoxyethanol in a concentration of 0.1 to 2%, preferably ⁇ 0.9%, or 4,4'- dichloro 2-hydroxydiphenyl ether (DCPP) in a concentration of 0.005 to 0.6%.
  • DCPP 4,4'- dichloro 2-hydroxydiphenyl ether
  • the composition of the invention e.g. a liquid enzyme composition, comprises 0.8-1% 2- phenoxyethanol.
  • the invention also encompasses a method of preserving an aqueous composition according to the invention against microbial contamination or growth, which method comprises addition of at least one antimicrobial agent or preservative, preferably 2-phenoxyethanol.
  • the invention also encompasses a method of providing an antimicrobial effect on textiles after treatment with a solid laundry detergent (e.g. powders, granulates, capsules, tablets, bars etc.), a liquid laundry detergent, a softener or an after-rinse containing 4, 4' -dichloro 2-hydroxy-diphenyl ether (DCPP).
  • a solid laundry detergent e.g. powders, granulates, capsules, tablets, bars etc.
  • DCPP after-rinse containing 4, 4' -dichloro 2-hydroxy-diphenyl ether
  • compositions of the invention e.g. a liquid enzyme composition
  • compositions of the present invention may comprise at least one further enzyme in addition to the protease as described herein.
  • the at least one further enzyme is a detergent enzyme.
  • the further enzyme is classified as an oxidoreductase (EC 1), a transferase (EC 2), a hydrolase (EC 3), a lyase (EC 4), an isomerase (EC 5), or a Ligase (EC 6) (EC-numbering according to Enzyme Nomenclature, Recommendations (1992) of the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology including its supplements published 1993-1999).
  • the further enzyme is a hydrolase (EC 3), in one embodiment the further enzyme is a glycosidase (EC 3.2) or a peptidase (EC 3.4). In one embodiment, the further enzyme is selected from the group consisting of an amylase (in particular an alpha-amylase (EC 3.2.1.1)), a cellulase (EC 3.2.1.4), a lactase (EC 3.2.1.108), a mannanase (EC 3.2.1.25), a lipase (EC3.1.1.3), a phytase (EC 3.1.3.8), a nuclease (EC 3.1.11 to EC 3.1.31), and a protease.
  • an amylase in particular an alpha-amylase (EC 3.2.1.1)
  • a cellulase EC 3.2.1.4
  • a lactase EC 3.2.1.108
  • a mannanase EC 3.2.1.25
  • lipase
  • the further enzyme is selected from the group consisting of oxidoreductases, transferases, hydrolases, lyases, isomerases, ligases, aminopeptidases, amylases, asparaginases, carbohydrases, carboxypeptidases, catalases, cellulases, chitinases, cutinases, cyclodextrin glycosyltransferases, deoxyribonucleases, esterases, alpha-galactosidases, betagalactosidases, glucoamylases, alpha-glucosidases, beta-glucosidases, hyaluronic acid synthases, invertases, laccases, lipases, mannanases, mannosidases, mutanases, oxidases, pectinolytic enzymes, peroxidases, phytases, polyphenoloxidases, proteases, rib
  • the further enzyme is selected from the group consisting of mannanases, amylases, cellulases, lipases, dispersins, and DNases.
  • the further enzyme is selected from the group consisting of mannanases, amylases, and cellulases.
  • the further enzyme is a mannanase.
  • the compositions of the present invention comprise a mannanase.
  • Mannanases as described herein are enzymes selected from the group of mannan degrading enzymes that according to the invention have “mannanase activity”.
  • the mannanase may be selected from p-mannosidase (EC 3.2.1.25), endo-1 ,4-p-mannosidase (EC 3.2.1.78), and 1,4- p-mannobiosidase (EC 3.2.1.100).
  • the mannanase is selected from the group of endo-1,4-p- mannosidase (EC 3.2.1.78), a group of enzymes which may be called endo-p-1 ,4-D-mannanase, p-mannanase, or mannanase herein.
  • Endo-1 , 4-mannanases randomly hydrolyze the 1 ,4-linkages within the mannan backbone releasing manno-oligosaccharides (MOS).
  • Mannanase activity may be determined by assays for measurement of mannanase activity, which are known to those skilled in the art. For instance, a mannanase to be tested may be applied to 4 mm diameter holes punched out in agar plates comprising 0.2% AZCL galactomannan (carob), i.e. a substrate for the assay of endo-1 ,4-beta- D-mannanase. Carob is e.g. available as l-AZGMA from the company Megazyme. Mannan degrading activity may be tested in a liquid assay using carob galactomannan dyed with Remazol Brilliant Blue as described in McCleary, B. V.10 (1978).
  • carob AZCL galactomannan
  • Suitable mannanases according to the invention have mannanase activity, preferably when determined according to the essays described above, they exhibit at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150% of the mannanase activity of the mannanase variant comprising or consisting of SEC ID NO: 3 with the amino acid substitutions G59V, N66D, A89H, E234Q, W259M, N282Y, T318N, E319G, and S322G according to the numbering of SEC ID NO: 3.
  • the mannanase comprised in the composition of the invention may be selected from alkaline mannanase of Family 5 or 26 (i.e., GH5 or GH26).
  • alkaline mannanase is meant to encompass mannanases having an enzymatic activity of at least 40% of its maximum activity at a given pH ranging from 7 to 12, preferably 7.5 to 10.5.
  • the mannanase comprised in the composition of the invention may be selected from mannanases originating from Bacillus organisms, such as described in JP-0304706 [beta-mannanase from Bacillus sp.], JP-63056289 [alkaline, thermostable beta-mannanase], JP-63036774 [Bacillus microorganism FERM P-8856 producing beta-mannanase and beta-mannosidase at an alkaline pH], JP-08051975 [alkaline beta-mannanases from alkalophilic Bacillus sp.
  • Suitable mannanases are described in WO 99/064619],
  • the mannanase comprised in the composition of the invention may be selected from mannanases originating from Trichoderma organisms, such as disclosed in WO 93/24622.
  • the mannanase comprised in the composition of the invention maybe a sequence disclosed in W02005003319 as SEQ ID NO: 388.
  • Suitable mannanases include also those, which are variants of the above described mannanases which have mannan degrading activity.
  • the mannanase of the present invention is a mannanase that comprises an amino acid sequence which is at least 65% identical to SEQ ID NO: 3.
  • the mannanase is a variant of a parent mannanase, wherein the mannanase variant comprises one or more amino acid substitutions at positions selected from the group consisting of 59, 66, 89, 234, 259, 282, 318, 319, and 322, according to the numbering of SEQ ID NO: 3, and an amino acid sequence which is at least 65%, preferably at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, or even more preferably at least 98% but less than 100% identical SEQ ID NO: 3.
  • the mannanase is a variant of a parent mannanase that comprises at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight or all amino acid substitutions selected from the group consisting of X59V, X66D, X89H, X234Q, X259M, X282Y, X318N, X319G, and X322G according to the numbering of SEQ ID NO: 3 and an amino acid sequence which is at least 65%, preferably at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, or even more preferably at least 98% but less than 100% identical SEQ ID NO: 3.
  • the mannanase is a variant of a parent mannanase comprising or consisting of SEQ ID NO: 3 with the amino acid substitutions Q59V, N66D, A89H, E234Q, W259M, N282Y, T318N, E319G, and S322G according to the numbering of SEQ ID NO: 3 and optionally additionally comprises one or more, preferably 1 -10, more preferably 1-5 conservative amino acid substitutions.
  • mannanase variants include variants with at least 40 to 100% similarity and/or identity when compared to the full-length polypeptide sequence of the parent enzyme as disclosed above.
  • mannanase variants having mannan degrading activity are at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% similar and/or identical to the full-length polypeptide sequence of the parent enzyme as disclosed above.
  • mannanase variants have mannan degrading activity according to the pre-sent invention when said mannanase variants exhibit at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% of the mannan degrading activity of the respective parent mannanase.
  • At least one mannanase may be selected from a commercially available mannanase such as Mannaway® (Novozymes A/S).
  • compositions of the present invention including liquid enzyme compositions and liquid cleaning compositions, comprise an amylase.
  • Amylases according to the invention (alpha and/or beta) include amylases of bacterial or fungal origin (EC 3.2.1.1 and 3.2.1.2, respectively), preferably selected from the group of alpha-amylases (EC 3.2.1.1). Chemically modified or protein engineered variants are included.
  • Amylases according to the invention have "amylolytic activity” or “amylase activity” involving (en-do)hydrolysis of glucosidic linkages in polysaccharides.
  • Alpha-amylase activity may be determined by assays for measurement of alpha-amylase activity which are known to those skilled in the art.
  • Alpha-amylase activity can for example be determined by a method employing Phadebas tablets as substrate (Phadebas Amylase Test, supplied by Magle Life Science).
  • Starch is hydrolyzed by the alpha-amylase giving soluble blue fragments.
  • the absorbance of the resulting blue solution measured spectrophotometrically at 620 nm, is a function of the alpha-amylase activity. The measured absorbance is directly proportional to the specific activity (activity/mg of pure alpha-amylase protein) of the alpha-amylase in question under the given set of conditions.
  • Alpha-amylase activity can also be determined by a method employing the ethyliden-4-nitrophenyl-alpha-D- maltoheptaosid (EPS).
  • D-maltoheptaoside is a blocked oligosaccharide, which can be cleaved by an endo-amylase.
  • the alpha-glucosidase included in the kit digests the substrate to liberate a free PNP molecule, which has a yellow color and thus can be measured by visible spectrophotometry at 405nm.
  • Kits containing EPS substrate and alpha-glucosidase are manufactured by Roche Costum Biotech (cat. No. 10880078103). The slope of the time dependent absorption-curve is directly proportional to the specific activity (activity per mg enzyme) of the alpha-amylase in question under the given set of conditions.
  • Amylolytic activity may be provided in units per gram enzyme.
  • 1 unit alpha-amylase may liberate 1.0 mg of maltose from starch in 3 min at pH 6.9 at 20°C.
  • Suitable amylases according to the invention have amylase activity, preferably when determined according to the essays described above, they exhibit at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150% of the amylase activity of the amylase according to SEQ ID NO: 4, having the amino acid substitutions N25H, G4Q, R176K, G186E, T251 E, L405M, and Y482W according to the numbering of SEQ ID NO: 5.
  • Suitable amylases according to the invention have at least 40% residual activity after storage for 3 months at 45 °C in compositions according to the invention including liquid enzyme and liquid detergent compositions.
  • At least one amylase is selected from:
  • Suitable variants of SEQ ID NO: 6 include those comprising a deletion in positions 181 and/or 182 and/or a substitution in position 193.
  • Preferred variants of SEQ NO: 6 are those having a substitution, a deletion or an insertion in one or more of the following positions: R181 , G182, H183, G184, N195, 1206, E212, E216 and K269.
  • amylases from Bacillus sp. comprising amino acids 1 to 485 of SEQ ID NO: 2 as described in WO 00/60060.
  • amylases from Bacillus amyloliquefaciens or variants thereof, preferably selected from amylases according to SEQ ID NO: 3 as described in WO 2016/092009.
  • amylases having SEQ ID NO: 12 as described in WO 2006/002643 or amylase variants comprising the substitutions Y295F and M202LITV within said SEQ ID NO: 12.
  • amylases having SEQ ID NO: 6 as described in WO 2011/098531 or amylase variants comprising a substitution at one or more positions selected from the group consisting of 193 [G,A,S,T or M], 195 [F, W, Y, L, I or V], 197 [F, W,Y,L, I or V], 198 [Q or N], 200 [F, W, Y,L, I or V], 203 [F, W, Y,L, I or V], 206 [F,W,Y,N,L,I,V,H,Q,D or E], 210 [F,W,Y,L,I or V], 212 [F,W,Y,L,I or V], 213 [G,A,S,T or M] and 243 [F, W,Y,L, I or V] within said SEQ ID NO: 6.
  • amylases having SEQ ID NO: 1 as described in WO 2013/001078 or amylase variants comprising an alteration 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.
  • amylases having SEQ ID NO: 2 as described in WO 2013/001087 or amylase variants comprising a deletion of positions 181 +182, or 182+183, or 183+184, within said SEQ ID NO: 2, optionally comprising one or two or more modifications in any of positions corresponding to 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.
  • amylases which are hybrid alpha-amylases from above mentioned amylases as for example as described in WO 2006/066594.
  • hybrid amylases according to WO 2014/183920 with A and B domains having at least 90% identity to SEQ ID NO: 2 of WO 2014/183920 and a C domain having at least 90% identity to SEQ ID NO: 6 of WO 2014/183920, wherein the hybrid amylase has amylolytic activity; preferably the hybrid alpha-amylase is at least 95% identical to SEQ ID NO: 23 of WO 2014/183920, having amylolytic activity.
  • hybrid amylase according to WO 2014/183921 with A and B domains having at least 75% identity to SEQ ID NO: 2, SEQ ID NO: 15, SEQ ID NO: 20, SEQ ID NO: 23, SEQ ID NO: 29, SEQ ID NO: 26, SEQ ID NO: 32, or SEQ ID NO: 39 as disclosed in WO 2014/183921 and a C domain having at least 90% identity to SEQ ID NO: 6 of WO 2014/183921, wherein the hybrid amylase has amylolytic activity; preferably, the hybrid alpha-amylase is at least 95% identical to SEQ ID NO: 30 as disclosed in WO 2014/183921 , having amylolytic activity.
  • the amylase is a sequence having at least 80%, preferably at least 85%, more preferably at least 90%, more preferably at least 95%, even more preferably at least 98% sequence identity to SEQ ID NO: 4, having amylolytic activity.
  • said amylase is characterized by comprising the substitution N25H, G4Q, R176K, G186E, T251 E, L405M, and Y482W according to the numbering of SEQ ID NO: 5.
  • the amylase is a sequence according to SEQ ID NO: 4 comprising the substitutions N25H, G4Q, R176K, G186E, T251 E, L405M, and Y482W according to the numbering of SEQ ID NO: 5.
  • Suitable amylases also include those, which are variants of the above-described amylases, which have amylolytic activity.
  • amylase variants include variants with at least 40 to 100% identity when compared to the full-length polypeptide sequence of the parent enzyme as disclosed above.
  • amylase variants having amylolytic activity are at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the full-length polypeptide sequence of the parent enzyme as disclosed above.
  • the invention relates to amylase variants comprising conservative substitutions not pertaining the functional domain of the respective amylase.
  • Amylase variants of this embodiment having amylolytic activity may be at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% similar to the full- length polypeptide sequence of the parent enzyme.
  • amylase variants have amylolytic activity according to the present invention when said amylase variants exhibit increased amylolytic activity when compared to the parent amylase.
  • amylase variants have amylolytic activity according to the present invention when said amylase variants exhibit increased amylolytic activity when compared to the parent cellulase.
  • amylase variants have amylolytic activity according to the present invention when said amylase variants exhibit at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% of the amylolytic activity of the respective parent amylase.
  • At least one amylase is selected from commercially available amylases which include but are not limited to products sold under the trade names DuramylTM, TermamylTM, FungamylTM, StainzymeTM, Stainzyme PlusTM, NatalaseTM, Liquozyme X and BANTM, AmplifyTM, Amplify PrimeTM (from Novozymes A/S), and RapidaseTM, PurastarTM, PoweraseTM, EffectenzTM (MIOO from DuPont), PreferenzTM (S1000, S110 and F1000; from DuPont), PrimaGreenTM (ALL; DuPont), OptisizeTM (DuPont).
  • commercially available amylases which include but are not limited to products sold under the trade names DuramylTM, TermamylTM, FungamylTM, StainzymeTM, Stainzyme PlusTM, NatalaseTM, Liquozyme X and BANTM, AmplifyTM, Amplify PrimeTM
  • compositions of the present invention comprising a cellulase.
  • Cellulases according to the invention have cellulase activity and include those of bacterial or fungal origin.
  • Cellulases are enzymes involved in hydrolysis of cellulose. Assays for measurement of "cellulase activity” or “cellulolytic activity” are known to those skilled in the art. For example, cellulolytic activity may be determined using a calorimetric assay, where the cellulase hydrolyses carboxymethyl cellulose to reducing carboxyd rates, which can be determined calorimetrically via the ferricyanide reaction according to Hoffman, W. S., J. Biol. Chem. 120, 51 (1937). Suitable cellulases according to the invention have at least 50% residual activity after 14-day storage at 45°C in compositions according to the invention, e.g. liquid enzyme and liquid detergent compositions, including a protease and cellularlase blend.
  • Cellulolytic activity may be provided in units per gram enzyme. For example, 1 unit may liberate 1.0 pmole of glucose from cellulose in one hour at pH 5.0 at 37°C (2 hour incubation time).
  • At least one cellulase comprised in the composition of the invention may be selected from cellobiohydrolase (1,4- P-D-glucan cellobiohydrolase, EC 3.2.1.91), endo-ss-1 ,4-glucanase (EC 3.2.1.4) and ss-glucosidase (EC 3.2.1.21).
  • Endoglucanases of EC class 3.2.1.4 may be named endoglucanase, endo-1 ,4-ss-D-glucan 4-glucano hydrolase, endo-1 ,4-beta-glucanase, carboxymethyl cellulase, and beta-1 , 4-glucanase.
  • Endoglucanases may be classified by amino acid sequence similarities (Henrissat, B. Accessed at UniProt 10/26/2011) under family 5 containing more than 20 endoglucanases of EC 3.2.1.4. Reference is also made to T.-M. Enveri, "Microbial Cellulases” in W.M. Fogarty, Microbial Enzymes and Biotechnology, Applied Science Publishers, p. 183-224 (1983); Methods in Enzymology, (1988) Vol. 160, p. 200-391 (edited by Wood, W.A. and Kellogg, S.T.); Beguin, P., "Molecular Biology of Cellulose Degradation", Annu. Rev. Microbiol. (1990), Vol. 44, pp.
  • At least one cellulase comprised in the composition of the invention is selected of the glycosyl hydrolase family 7 (GH7, pfam00840), preferably selected from endoglucanases (EC 3.2.1.4).
  • GH7, pfam00840 glycosyl hydrolase family 7
  • endoglucanases EC 3.2.1.4
  • At least one cellulase comprised in the composition of the invention is selected from cellulases comprising a cellulose binding domain. In one embodiment, at least one cellulase is selected from cellulases comprising a catalytic domain only, meaning that the cellulase lacks cellulose binding domain.
  • Bacillus such as Bacillus sp. CBS 670.93 and CBS 669.93
  • Melanocarpus such as Melanocarpus albomyces as disclosed in WO 97/14804
  • Clostridium e.g. Clostridium thermocellum
  • Humicola such as Humicola insolens (DSM1800) as disclosed in EP 0495257, EP 0531315, EP 0531372, US 4435307, US 5648263, US 5776757, WO 89/09259, WO 91/17244, WO 94/07998 (sequence displayed in figure 1 "43kd human variants thereof), WO 95/24471 , WO 96/11262 and WO 98/12307.
  • DSM1800 Humicola insolens
  • Fusarium such as Fusarium oxysporum e.g. strain J79 (DSM2672) as disclosed in EP 0495257, EP 0531315, EP 0531372, US 5648263, US 5776757, WO 89/09259, WO 91/17244, WO 95/24471 and WO 96/11262
  • Thielavia such as Thielavia terrestris or Myceliophthora thermophila strain CBS 11765 as disclosed in EP 0531315, US 5648263, US 5776757, WO 89/09259, WO 91/17244, WO 95/24471 , WO 96/11262, WO 96/29397 (SEQ ID NO: 9 and variants thereof), and WO 98/12307.
  • Trichoderma such as Trichoderma reesei, Trichoderma longibrachiatum or Trichoderma harzianum as disclosed in EP 1305432, EP 1240525, WO 92/06165, WO 94/21801 , WO 94/26880, WO 95/02043, WO 95/24471 and WO 02/099091.
  • Aspergillus such as Aspergillus aculeatus as disclosed in WO 93/17244
  • Acremonium such as Acremonium sp., Acremoniu persicinum, Acremonium acremonium, Acremonium brachypenium, Acremonum dichromosporum, Acremonium obclavatum, Acremonium pinkertoniae, Acremonium roseogriseum, Acremonium incoloratum, and Acremonium furatum as disclosed in WO 96/11262 and WO 96/29397 (SEQ ID NO: 5 and variants thereof).
  • Cellvibrio such as Cellvibrio mixtus DSM 11683, Cellvibrio mixtus DSM 11684, Cellvibrio mixtus DSM 11685, Cellvibrio mixtus ACM 2601 , Cellvibrio mixtus DSM 1523, and Cellvibrio gilvus DSM 11686, as disclosed in WO 98/08940.
  • Cephalosporium such as Cephalosporium sp. RYM-202 as disclosed in WO 96/11262.
  • Suitable cellulases include also those, which are variants of the above described cellulases which have cellulolytic activity.
  • cellulase variants include variants with at least 40 to 100% identity when compared to the full length polypeptide sequence of the parent enzyme as disclosed above.
  • cellulase variants having cellulolytic activity are at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% similar and/or identical to the full length polypeptide sequence of the parent enzyme as disclosed above.
  • the composition of the invention comprises a Humicola insolens DSM 1800 cellulase complex having endoglucanase, cellobiohydrolase and beta-glucosidase activity.
  • the cellulase is a sequence having at least 80%, preferably 85%, more preferably 90%, more preferably 92%, more preferably 95%, more preferably 96% more preferably 98%, more preferably 99%, or 100% sequence identity to SEQ ID NO: 6, having cellulase activity.
  • the composition of the invention comprises at least one Humicola insolens DSM 1800 endoglucanase (EC 3.2.1.4) having the amino acid sequence disclosed in Fig. 14A-E of WO 91/17244, preferably amino acids 20-434 according said sequence, more preferably having one or more substitutions at positions selected from 182, 223, and 231 , most preferably selected from P182S, A223V, and A231V.
  • the endoglucanase is at least 80% similar and/or identical to a polypeptide according to SEQ ID NO: 2 of WO 95/02675.
  • the composition of the invention comprises at least a Bacillus sp. Cellulase (EC 3.2.1.4) selected from a polypeptide at least 80% similar and/or identical to the amino acid sequence of position 1 to position 773 of SEQ ID NO: 2 of WO 2004/053039 or a catalytically active fragment thereof.
  • the composition of the invention comprises at least a Thielavia terrestris cellulase (EC 3.2.1.4) having a polypeptide at least 80% similar and/or identical to the amino acid sequence of position 1 to position 299 of SEQ ID NO: 4 of WO 2004/053039 or a catalytically active fragment thereof.
  • cellulase variants have cellulolytic activity according to the present invention when said cellulase variants exhibit increased cellulolytic activity when compared to the parent cellulase.
  • cellulase variants have cellulolytic activity according to the present invention when said cellulase variants exhibit at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% of the cellulolytic activity of the respective parent cellulase.
  • At least one cellulase may be selected from Renozyme®, Celluzyme®, Celluclean®, Endolase® and Carezyme® (Novozymes A/S), ClazinaseTM, and Puradax HATM (Genencor Int. Inc.), and KAC-500(B)TM (Kao Corporation).
  • At least one peroxidases may be selected from GuardzymeTM (Novozymes A/S).
  • compositions of the present invention comprising enzyme blends of enzymes as described herein (meaning a combination of different enzymes).
  • such blends are blends of two enzymes, e.g. protease and mannanase, protease and amylase, or protease and cellulase.
  • such blends are blends of three enzymes, e.g. protease, mannanase and amylase, or protease, amylase and cellulase or protease, mannanase and cellulase.
  • enzymes e.g. protease, mannanase and amylase, or protease, amylase and cellulase or protease, mannanase and cellulase.
  • such blends are blends of four enzymes, e.g. protease, mannanase, amylase and cellulase.
  • enzymes e.g. protease, mannanase, amylase and cellulase.
  • such blends are blends of more than four enzymes including any enzyme combination of any enzymes as described herein.
  • compositions according to the invention can comprise additional components such as buffer, preservative, dyestuff or fragrance.
  • the invention relates to liquid enzyme compositions, characterized by a concentration of the protease of 0.05-12%, 0.05-10%, preferably 2-6%, a concentration of the protease inhibitor of 0.05-0.4%, preferably 0.1 - 0.25%, and a concentration of the at least on solubilizing agent, selected from the group consisting of, of: i. neopentyl glycol: 5-25%, preferably 8-12%,
  • the liquid enzyme composition is essentially devoid of anionic surfactants, builders, polymers, alkalis, bleaching systems, fluorescent whitening agents, suds suppressors and stabilizers, hydrotropes, and/or corrosion inhibitors.
  • the liquid enzyme composition is essentially devoid of anionic surfactants and builders.
  • the liquid enzyme composition according to the invention comprises components supporting enzyme stability, such as e.g. calcium chloride dihydrate (CaCl2*2H2O), preferably 0.1 -0.5% by weight.
  • enzyme stability such as e.g. calcium chloride dihydrate (CaCl2*2H2O), preferably 0.1 -0.5% by weight.
  • liquid enzyme composition according to the invention comprises "further components” as described herein.
  • the composition contains further solvents.
  • the liquid enzyme composition comprises monopropylene glycol (MPG) as additional solvent.
  • MPG monopropylene glycol
  • the liquid composition of the invention is a liquid enzyme composition that comprises amounts of MPG as additional solvent ranging from about 1% to about 35% by weight, preferably from about 5% to about 30% by weight.
  • the total amount of MPG in the liquid enzyme composition of the invention may be at least 1% by weight, at least 2% by weight, at least 4% by weight, at least 5% by weight, at least 6% by weight, at least 8% by weight, or at least 9% by weight.
  • the total amount of MPG in the liquid enzyme composition may not be more than 40% by weight, not more than 35% by weight, not more than 30% by weight, not more than 28% by weight, not more than 26% by weight, not more than 24% by weight, not more than 22% by weight, not more than 20% by weight, not more than 18% by weight, not more than 16% by weight, not more than 14% by weight, not more than 12% by weight, or not more than 10% by weight.
  • the liquid enzyme composition comprises glycerol as additional solvent.
  • the liquid composition of the invention is a liquid enzyme composition that comprises amounts of glycerol as additional solvent ranging from about 10% to about 50% by weight, preferably from about 15% to about 45% by weight.
  • the total amount of glycerol in the liquid enzyme composition of the invention may be at least 10% by weight, at least 12% by weight, at least 14% by weight, at least 15% by weight, at least 16% by weight, at least 18% by weight, or at least 19% by weight.
  • the total amount of glycerol in the liquid enzyme composition may not be more than 45% by weight, not more than 42% by weight, not more than 40% by weight, not more than 38% by weight, not more than 36% by weight, not more than 34% by weight, not more than 32% by weight, not more than 30% by weight, not more than 28% by weight, not more than 26% by weight, not more than 24% by weight, not more than 22% by weight, or not more than 20% by weight.
  • the liquid enzyme composition contains a solvent mixture of glycerol and MPG.
  • liquid enzyme compositions according to the invention comprise specific ratios of component c. and the additional solvents as described herein.
  • the liquid enzyme composition according to the invention comprises different amounts of and different ratios of component c., MPG and glycerol, for example those listed in tables 1 and 2 below (by weight means relative to the total weight of the liquid composition):
  • Table 1 Example formulations of component c., MPG and glycerol in amounts % by weight
  • the liquid enzyme composition comprises neopentyl glycol as component c. and MPG and glycerol as additional components.
  • neopentyl glycol, MPG and glycerol are formulated according to formulation a:
  • Formulation a neopentyl glycol:MPG:glycerol in ratios of 1 : ⁇ 3: ⁇ 6, or 1 : ⁇ 3: ⁇ 5, or 1 : ⁇ 3: ⁇ 4, or 1 : ⁇ 3: ⁇ 3.5 wherein the total amount of neopentyl glycol, MPG and glycerol in % by weight contained in formulation a is set to be more than 45%, preferably at least 48%, most preferably at least 50% and optionally at most 70%, preferably at most 65%, more preferably at most 60%, most preferably at most 55%.
  • the liquid enzyme composition comprises gamma-valerolactone as component c. and MPG and glycerol as additional components.
  • gamma-valerolactone, MPG and glycerol are formulated according to formulation b:
  • Formulation b gamma-valerolactone:glycerol in a ratio of 1 : ⁇ 35, preferably 1 : ⁇ 30, more preferably 1 : ⁇ 25, even more preferably 1 : ⁇ 20 or most preferred 1 : ⁇ 18 wherein the total amount of gamma-valerolactone, MPG and glycerol in % by weight contained in formulation b is set to be at least 50% and optionally at most 70%.
  • the liquid enzyme composition comprises diethylene glycol as component c. and MPG and glycerol as additional components.
  • diethylene glycol, MPG and/or glycerol are formulated according to formulation c:
  • Formulation c diethylene glycol: MPG:glycerol in a ratio of 1 :2:2 optionally wherein the total amount of diethylene glycol, MPG and glycerol in % by weight contained in formulation c is set to be at least 50% and optionally at most 70%, preferably at most 65%, more preferably at most 60%, most preferably at most 55%.
  • the liquid enzyme composition comprises a preservative as described herein. In a preferred embodiment, the liquid enzyme composition comprises 0.1 to 2%, preferably 0.8-1 % 2-phenoxythanol.
  • the liquid enzyme composition is essentially devoid of preservatives as described herein.
  • the liquid enzyme composition comprises at least one further enzyme, I. e. an enzyme blend as described herein.
  • the liquid enzyme composition additionally comprises a mannanase as described herein.
  • the mannanase is comprised in a concentration range of 0.01 to 0.5%, more preferably 0.05 to 0.15%.
  • the liquid enzyme composition additionally comprises an amylase as described herein.
  • the amylase is comprised in a concentration range of 0.1 to 3%, more preferably 0.3 to 1 .0%.
  • the liquid enzyme composition additionally comprises a cellulase as described herein.
  • the cellulase is comprised in a concentration range of 0.05 to 1 .0%, more preferably 0.1 to 0.6%.
  • the liquid enzyme composition comprises at least two further enzymes, i.e. an enzyme blend as described herein.
  • the invention relates to liquid enzyme compositions, characterized by a concentration of the protease of O.05-12%, 0.05-10%, preferably 2-6%, a concentration of the protease inhibitor of 0.05-0.4%, preferably 0.15-0.25%, and a concentration of neopentyl glycol of 5-25%, preferably 8-16%, most preferably 8-12%, wherein the composition additionally comprises 5-30% MPG, preferably 8-20% MPG, more preferably 8-12% MPG and 10-40% glycerol, preferably 20-35% glycerol, more preferably 28-32% glycerol, and wherein the composition optionally additionally comprises an amylase and/or cellulase and/or mannanase, and wherein the composition optionally additionally comprises 0.1 -0.5% CaCI2, and optionally wherein the pH of the composition is set to 6.
  • the invention relates to liquid enzyme compositions, characterized by a concentration of the protease of 0.05-12%, 0.05-10%, preferably 2-6%, a concentration of the protease inhibitor of 0.05-0.4%, preferably 0.1 -0.25%, and a concentration of neopentyl glycol of 5-25%, preferably 8-16%, most preferably 8-12%, wherein the composition additionally comprises 10-40% MPG, preferably 20-35% MPG, more preferably 28-34% MPG and 10-40% glycerol, preferably 12-30% glycerol, more preferably 14-20% glycerol and wherein the composition optionally additionally comprises an amylase and/or cellulase and/or mannanase, and wherein the composition optionally additionally comprises 0.1 -0.5% CaCI2, and optionally wherein the pH of the composition is set to 6.
  • the invention relates to liquid enzyme compositions, characterized by a concentration of the protease of 0.05-12%, 0.05-10%, preferably 2-6%, a concentration of the protease inhibitor of 0.05-0.4%, preferably 0.1 -0.25%, and a concentration of neopentyl glycol of 5-25%, preferably 8-16%, more preferably 8-12%, wherein the composition additionally comprises 10-30% MPG, preferably 15-25% MPG, more preferably 18-22% MPG and 10-40% glycerol, preferably 15-30% glycerol, more preferably 18-22% glycerol, and wherein the composition optionally additionally comprises an amylase and/or cellulase and/or mannanase, and wherein the composition optionally additionally comprises 0.1 -0.5% CaCI2, and optionally wherein the pH of the composition is set to 6.
  • the invention relates to liquid enzyme compositions, characterized by a concentration of the protease of 0.05-12%, 0.05-10%, preferably 2-6%, a concentration of the protease inhibitor of 0.05-0.4%, preferably 0.1 -0.25%, and a concentration of gamma-valerolactone of 1 -15%, preferably 2-10%, more preferably 2-6%, wherein the composition additionally comprises 2-25% MPG, preferably 2-15% MPG, more preferably 4-8% MPG and 30-50% glycerol, preferably 35-48% glycerol, more preferably 40-45% glycerol and wherein the composition optionally additionally comprises an amylase and/or cellulase and/or mannanase, and wherein the composition optionally additionally comprises 0.1 -0.5% CaCI2, and optionally wherein the pH of the composition is set to 6.
  • the invention also relates to a method for preparation of a liquid enzyme composition according to the invention, comprising the steps of mixing at least components a. and b. and at least one of c. - all as disclosed herein - in one or more steps in any order.
  • the invention also relates to a method for preparation of a liquid enzyme composition according to the invention, comprising the steps of mixing the solubilizing agent (component c.) with MPG in a first step and subsequently adding the protease inhibitor (component b.), the protease (component a.), glycerol and optionally additional enzymes.
  • the invention also relates to a method for preparation of a liquid enzyme composition according to the invention, comprising the steps of mixing the protease inhibitor (component b.) and a solubilizing agent (component c.) optionally with MPG in a first step and subsequently adding the protease (component a.), glycerol and optionally additional enzymes.
  • the invention also relates to a method for stabilizing and solubilizing a protease and protease inhibitor in a liquid enzyme composition, wherein the method comprises bringing together the protease (component a.), protease inhibitor (component b.) and a solubilizing agent (component c.) selected from the group consisting of:
  • neopentyl glycol ii. gamma-valerolactone, and ill. diethylene glycol.
  • the invention also relates to the use of component c. as described herein for the solubilization of a protease inhibitor (component b.) as described herein.
  • the invention relates to compositions comprising a. at least one protease and b. a protease inhibitor and c. at least one solubilizing agent selected from the group consisting of:
  • compositions designated for cleaning soiled material, such as textiles or dishware. Cleaning compositions according to the invention include compositions for different applications such as laundry and hard surface cleaning.
  • compositions of the invention can for example be used in detergents, in hard surface cleaning compositions, in cosmetic and pharmaceutical formulations, in crop protection formulations, in coating agents, in adhesives, in leather treatment agents, in formulations for the textile industry, in fiber processing, in metal processing, in the food or feed industry, in water treatments, in the paper industry, or for fermentation or mineral processing. Therefore, the present invention also includes detergents, cleaning agents, wetting agents, coating agents, adhesives, leather treatment or textile treatment agents, or cosmetic, pharmaceutical, or crop protection formulations.
  • detergent component is defined herein to mean a type of chemical, which can be used in detergent compositions. Typical detergent components are surfactants and builders.
  • a detergent component includes amounts of certain components to provide effective stain removal and/or effective cleaning conditions (e. g. pH, temperature, water hardness, quantity of foaming), amounts of certain components to effectively provide optical benefits (e. g. optical brightening, dye transfer inhibition, color care), and amounts of certain components to effectively aid the processing (maintain physical characteristics during processing, storage and use; e.g. rheology modifiers, hydrotropes, desiccants).
  • textile means any textile material including yarns (thread made of natural or synthetic fibers used for knitting or weaving), yarn intermediates, fibers, non-woven materials, natural materials, synthetic materials, as well as fabrics made of these materials such as garments, cloths, and other articles.
  • fabric a textile made by weaving, knitting or felting fibers
  • garment any article of clothing made of textile
  • hard surface cleaning relates to both household hard surface cleaning and industrial hard surface cleaning and means the process of treating hard surfaces with a solution containing a cleaning composition of the present invention.
  • Hard surfaces may include any hard surfaces in a household or industry, such as floors, furnishing, walls, sanitary ceramics, glass, metallic surfaces including medical devices, cutlery, and dishes (also called “dishware”).
  • a particular form of hard surface cleaning is dishwashing, including manual dish washing (MDW) or automatic dishwashing (ADW).
  • wash refers to all forms of washing dishes or dishware, e. g. by hand or automatic dish washing.
  • Washing dishware includes, but is not limited to, the cleaning of all forms of crockery such as plates, cups, glasses, bowls, all forms of cutlery such as spoons, knives, forks and serving utensils as well as ceramics, plastics such as melamine, metals, china, glass, and acrylics.
  • stains In the field of detergency, usually the term “stains” is used with reference to laundry, e.g., cleaning for textiles, fabric, or fibers, whereas the term “soils” is usually used with reference to hard surface cleaning, e.g., cleaning of dishes and cutlery.
  • stain and “soil” are used interchangeably.
  • Detergent components vary in type and/or amount in a cleaning composition, e.g. a detergent formulation, depending on the desired application such as laundering white textiles, colored textiles, and wool.
  • the component(s) chosen further depend on physical form of a cleaning composition, e.g. a detergent formulation (liquid, solid, gel, provided in pouches or as a tablet, etc).
  • the component(s) chosen e.g. for laundering formulations further depend on regional conventions which themselves are related to aspects like washing temperatures used, mechanics of laundry machine (vertical vs. horizontal axis machines), water consumption per wash cycle etc. and geographical characteristics like average hardness of water.
  • detergent components and usage in cleaning compositions are known to those skilled in the art.
  • Suitable detergent components comprise inter alia surfactants, builders, polymers, alkalis, bleaching systems, fluorescent whitening agents, suds suppressors and stabilizers, hydrotropes, and corrosion inhibitors. Further examples are described e.g. in "complete Technology Book on Detergents with Formulations (Detergent Cake, Dishwashing Detergents, Liquid & Paste Detergents, Enzyme Detergents, Cleaning Powder & Spray Dried Washing Powder)”, Engineers India Research Institute (EIRI), 6 th edition (2015). Another reference book for those skilled in the art may be “Detergent Formulations Encyclopedia”, Solverchem Publications, 2016.
  • one or more of the detergent components, preferably the surfactant and/or the builder is bio-degradable and/or bio-based.
  • detergent components are in addition to the components comprised in the compositions of the invention. If a component comprised in the liquid compositions of the invention is also a detergent component, it might be that its concentrations need to be adjusted for the component to be effective for the purpose desired in the cleaning composition, e.g. detergent formulation.
  • Detergent components may have more than one function in the final application of a cleaning composition, e.g. a detergent formulation, therefore any detergent component mentioned in the context of a specific function herein, may also have another function in the final application of a cleaning composition, e.g. a detergent formulation.
  • the function of a specific detergent component in the final application of a cleaning composition, e.g. a detergent formulation usually depends on its amount within the detergent formulation, i.e. the effective amount of a detergent component.
  • a cleaning composition e.g. a detergent formulation
  • a cleaning composition is a formulation of more than two detergent components, wherein at least one component is effective in stain-removal, at least one component is effective in providing the optimal cleaning conditions, and at least one component is effective in maintaining the physical characteristics of the cleaning composition.
  • Cleaning compositions of the invention can be liquid or solid, preferably they are liquid. In one embodiment, at least a part of the cleaning composition of the invention is provided as a liquid. Depending on whether a water-soluble package is enclosing the liquid cleaning composition, e.g. detergent formulation, the liquid cleaning composition, e.g. detergent formulation comprises water or is essentially free from water.
  • liquid cleaning compositions comprise enzyme blends as described herein.
  • Cleaning compositions according to the invention comprise at least one compound selected from surfactants, builders, polymers, fragrances and dyestuffs.
  • the cleaning compositions of the invention comprise at least one surfactant selected from anionic surfactants, non-ionic surfactants, amphoteric surfactants, and cationic surfactants.
  • surfactant (synonymously used herein with “surface active agent”) means an organic chemical that, when added to a liquid, changes the properties of that liquid at an interface. According to its ionic charge, a surfactant is called non-ionic, anionic, cationic, or amphoteric.
  • the cleaning compositions e.g. detergent formulations, in one embodiment, comprise 2% to 30% by weight of anionic surfactants and/or 2% to 30% by weight of non-ionic surfactants, all relative to the total weight of the cleaning compositions, e.g. detergent formulation.
  • laundry detergents comprise higher amounts of surfactants than detergents for automated dish washing.
  • the cleaning compositions e.g.
  • detergent formulations of the invention comprise at least one non-ionic surfactant selected from alkoxylated alcohols, di- and multiblock copolymers of ethylene oxide and propylene oxide and reaction products of sorbitan with ethylene oxide or propylene oxide, alkyl polyglycosides (APG), hydroxyalkyl mixed ethers and amine oxides.
  • non-ionic surfactant selected from alkoxylated alcohols, di- and multiblock copolymers of ethylene oxide and propylene oxide and reaction products of sorbitan with ethylene oxide or propylene oxide, alkyl polyglycosides (APG), hydroxyalkyl mixed ethers and amine oxides.
  • the cleaning compositions e.g. detergent formulations, of the invention comprise at least one anionic surfactant selected from alkali metal and ammonium salts of Cs-Cis-alkyl sulfates, of Cs-C -fatty alcohol polyether sulfates, of sulfuric acid half-esters of ethoxylated C4-Ci2-alkylphenols (ethoxylation: 1 to 50 mol of ethylene oxide/mol), C12-C18 sulfo fatty acid alkyl esters, for example of C12-C18 sulfo fatty acid methyl esters, furthermore of Ci2-Ci8-alkylsulfonic acids and of Cio-Cis-alkylarylsulfonic acids.
  • anionic surfactant selected from alkali metal and ammonium salts of Cs-Cis-alkyl sulfates, of Cs-C -fatty alcohol polyether sulfates, of sulfuric acid half-esters of ethoxy
  • alkali metal salts of the aforementioned compounds particularly preferably the sodium salts.
  • suitable anionic surfactants include branched alkylbenzenesulfonates (BABS), phenylalkanesulfonates, alpha-olefinsulfonates (AOS), olefin sulfonates, alkene sulfonates, alkane-2,3-diylbis(sulfates), hydroxyalkanesulfonates and disulfonates, secondary alkanesulfonates (SAS), paraffin sulfonates (PS), sulfonated fatty acid glycerol esters, alkyl- or alkenylsuccinic acid, fatty acid derivatives of amino acids, diesters and monoesters of sulfo-succinic acid.
  • BABS branched alkylbenzenesulfonates
  • AOS alpha-olefinsulfonates
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least one anionic surfactant of general formula (AS-I):
  • R 1 is selected from Ci-C23-alkyl (such as 1-, 2-, 3-, 4- Ci-C23-alkyl) and C2-C23-alkenyl, wherein alkyl and/or alkenyl are linear (straight-chain; n-) or branched, and wherein 2-, 3-, or 4-alkyl; examples are n-CzHis, n- C9H19, n-CnH23, n-Ci3H27, n-CisHsi, n-CizFhe, i-CgH , i-Cigfe.
  • Ci-C23-alkyl such as 1-, 2-, 3-, 4- Ci-C23-alkyl
  • C2-C23-alkenyl wherein alkyl and/or alkenyl are linear (straight-chain; n-) or branched, and wherein 2-, 3-, or 4-alkyl
  • examples are n-CzHis, n- C9H19,
  • R 2 is selected from H, Ci-Cgo-alkyl and Cg-Cgo-alkenyl, wherein alkyl and/or alkenyl are linear (straightchain; n-) or branched.
  • R 3 and R 4 each independently selected from Ci-Ci6-alkyl, wherein 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, isoamyl, n-hexyl, isohexyl, sec-hexyl, n-heptyl, n- octyl, 2-ethylhexyl, n-nonyl, n-decyl, isodecyl.
  • A’ is selected from -RCOQ-, -SOr and RSOr, wherein R is selected from linear (straight-chain; n-) or branched Ci-Cs-alkyl, wherein alkyl for example is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, secbutyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, 1 ,2-dimethylpropyl, isoamyl, n-hexyl, isohexyl, sec-hexyl, n-heptyl, n-octyl, 2-ethylhexyl, or any other isomer of Ci-Cs-hydroxyalkyl and C1-C4- hydroxyalkyl is selected from hydroxymethyl, 1 -hydroxyethyl, 2-hydroxyethyl, 1-
  • M + is selected from H and salt forming cations.
  • Salt forming cations usually are monovalent or multivalent; hence M + equals 1/v M v+ . Examples include but are not limited to sodium, potassium, magnesium, calcium, ammonium, and the ammonium salt of mono-, di, and triethanolamine.
  • m is in the range of zero to 200, preferably 1 -80, more preferably 3-20; n and o, each independently in the range of zero to 100; n preferably is in the range of 1 to 10, more preferably 1 to 6; o preferably is 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.
  • Anionic surfactants of the general formula (AS-I) can be of any structure, block copolymers or random copolymers.
  • the cleaning compositions, e.g. detergent formulations of the invention comprise at least one anionic surfactant according to formula (AS-I), wherein R 1 is Ce-Cis-alkyl, R 2 is H, A’ is SOr, m is 2-30, and n and o are 0.
  • M + preferably is sodium, potassium, magnesium, calcium, ammonium, and the ammonium salt of mono-, di, and triethanolamine.
  • Such compounds may be called alcohol ethoxy sulphates (AES) herein.
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least one anionic surfactant according to formula (AS-I), wherein R 1 is n-CuFte, R 2 is H, A- is SOy, m, n and o being 0. M + preferably is NI .
  • Such compounds may be called ammonium lauryl sulfate (ALS) herein.
  • the cleaning compositions, e.g. detergent formulations of the invention comprise at least one anionic surfactant according to formula (AS-I), wherein R 1 is n-CuFte, R 2 is selected from H, A- is SOy, m being 2-5, preferably 3, and n and o being 0. M + preferably is Na + .
  • Such compounds may be called laurylethersulfates (LES) herein, preferably sodium laurylethersulfates (SLES).
  • laundry detergent formulations comprise compounds according to formula AS-I.
  • the cleaning compositions e.g. detergent formulations comprise at least one anionic surfactant selected from compounds of general formula (AS-II):
  • Cleaning compositions e.g. detergent formulations of the invention, in one embodiment, comprise salts of compounds according to formula (AS-II), preferably sodium salts.
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least two anionic surfactants, selected from compounds of general formula (AS-II), wherein one of said anionic surfactants is characterized in R 1 being C10, and the other surfactant is characterized in R 1 being C13.
  • AS-II general formula
  • Compounds like this may be called LAS (linear alkylbenzene sulfonates) herein.
  • laundry detergent formulations comprise compounds according to formula AS-II.
  • cleaning compositions e.g. detergent formulations comprise at least one anionic surfactant selected from AS-I and AS-II and at least one soap.
  • soaps are selected from salts (M + ) of saturated and unsaturated C12-C18 fatty acids, such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, (hydrated) erucic acid.
  • M + is selected from salt forming cations. Salt forming cations may be monovalent or multivalent; hence M + equals 1/v M v+ . Examples include but are not limited to sodium, potassium, magnesium, calcium, ammonium, and the ammonium salt of mono-, di, and triethanolamine.
  • 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.
  • Such soap mixtures comprise soaps of lauric acid and/or myristic acid and/or palmitic acid and/or stearic acid and/or oleic acid and/or linoleic acid in different amounts, depending on the natural fatty acids from which the soaps are derived.
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least one non-ionic surfactant of the general formula (NIS-I): wherein in formula (NIS-I) the following applies:
  • R 1 is selected from C1-C23 alkyl and C2-C23 alkenyl, wherein alkyl and/or alkenyl are linear (straight-chain; n-) or branched; examples are n-CzHis, n-CgHig, n-CnFte, n-Ci3H27, n-CisHsi, n-CizHgs, i-CgHig, i-Ci2H25.
  • R 2 is selected from H, C1-C20 alkyl and C2-C20 alkenyl, wherein alkyl and/or alkenyl are linear (straightchain; n-) or branched.
  • R 3 and R 4 each independently selected from C1-C16 alkyl, wherein 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, isoamyl, n-hexyl, isohexyl, sec-hexyl, n-heptyl, n- octyl, 2 -ethylhexyl, n-nonyl, n-decyl, isodecyl.
  • R 5 is selected from H and C1-C18 alkyl, wherein alkyl is linear (straight-chain; n-) or branched.
  • NIS-I The integers of the general formula (NIS-I) are defined as follows: m is in the range of zero to 200, preferably 1 -80, more preferably 3-20; n and 0, each independently in the range of zero to 100; n preferably is in the range of 1 to 10, more preferably 1 to 6; 0 preferably is in the range of 1 to 50, more preferably 4 to 25.
  • the sum of m, n and 0 is at least one, preferably the sum of m, n and 0 is in the range of 5 to 100, more preferably in the range of from 9 to 50.
  • the non-ionic surfactants of the general formula (NIS-I) can be of any structure, is it block or random structure, and is not limited to the displayed sequence of formula (NIS-I).
  • NIS-I general formula
  • NIS-I general formula
  • NIS-I non-ionic surfactants
  • NIS-I non-ionic surfactants
  • NIS-I non-ionic surfactants
  • NIS-I non-ionic surfactants
  • NIS-I non-ionic surfactants
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least one non-ionic surfactant of the general formula (NIS-I I):
  • - AO being identical or different alkylene oxides, selected from CH2-CH2-O, (CH2)3-O,
  • R 1 is selected from linear (straight-chain; n-) or branched C4-C3o-alkyl, and from straight-chain or branched
  • R 1 preferably is selected from straight-chain or branched C ⁇ Cso-alkyl, n-C ⁇ Cso-alkyl, n-C7-Ci5 alkyl, or n-Cio-Ci2-alkyl.
  • R 2 is selected from linear (straight-chain; n-) or branched Ci-Cso-alkyl, and from straight-chain or branched
  • R 2 preferably is selected from straight-chain or branched C6-C2o-alkyl, preferably straight-chain or branched C8-Ci2-alkyl, more preferably straight-chain or branched Cio-Ci2-alkyl.
  • the integer x of the general formula (NIS-II) preferably is a number in the range of 5 to 70, 10 to 60, 15 to
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least one non-ionic surfactant according to formula (NIS-II), wherein R 1 is n-C 3 -Ci7 alkyl, R 2 is linear or branched C 8 -Ci4 alkyl.
  • NIS-II non-ionic surfactant according to formula (NIS-II), wherein R 1 is n-C 3 -Ci7 alkyl, R 2 is linear or branched C 8 -Ci4 alkyl.
  • AO is selected from -(CH2CH2O) X 2-(CH 2 CH(CH3)-O) X 3, -(CH2CH 2 O) X 2-(CH(CH3)CH2-O) X 3, and -(CH 2 CH 2 O) X 4, wherein x2 and x4 is a number in the range of 15-50 and x3 is a number in the range of 1 to 15.
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least one non-ionic surfactant according to formula (NIS-II), wherein R 1 is n-Cs alkyl, R 2 is branched On alkyl, AO is CH2-CH2-O, and x is 22.
  • NIS-II non-ionic surfactant according to formula (NIS-II), wherein R 1 is n-Cs alkyl, R 2 is branched On alkyl, AO is CH2-CH2-O, and x is 22.
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least one non-ionic surfactant according to formula (NIS-II), wherein R 1 is n-Cs alkyl, R 2 is n-Cs-Cw alkyl, AO is CH2-CH2-O, and x is 40.
  • NIS-II non-ionic surfactant according to formula (NIS-II), wherein R 1 is n-Cs alkyl, R 2 is n-Cs-Cw alkyl, AO is CH2-CH2-O, and x is 40.
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least one non-ionic surfactant according to formula (NIS-II), wherein R 1 is n-Cs alkyl, R 2 is n-Cw alkyl, AO is selected from -(CH 2 CH2O)X2-(CH 2 CH(CH3)-O)X3, -(CH 2 CH 2 O)X 2 -(CH(CH 3 )CH 2 -O)X3, wherein x2 is 22 and x3 is 1.
  • NIS-II non-ionic surfactant according to formula (NIS-II)
  • cleaning compositions e.g. detergent formulations comprising at least one non-ionic surfactant according to formula (NIS-II), preferably those as disclosed above, are automated dishwashing detergents.
  • the automated dishwashing detergents comprise at least one compound according to formula (NIS-II) in the range of about 0.3% to 10% by weight, in the range of about 0.5% to 5% by weight, or in the range of about 1 % to 3%, all relative to the total weight of a cleaning composition.
  • At least one non-ionic surfactant is a compound according to formula (NIS-II), wherein R 1 is n-Cs alkyl, R 2 is branched Cn alkyl, AO is CH 2 -CH 2 -O, and x is 22.
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least one amphoteric surfactant according to general formula (AMS-I), which might be called modified amino acids (proteinogenic as well as non-proteinogenic):
  • AMS-I amphoteric surfactant according to general formula
  • AMS-I The variables in general formula (AMS-I) are defined as follows:
  • R 8 is selected from H, C1-C4 alkyl, C 2 -C4 alkenyl, wherein alkyl and/or are linear (straight-chain; n-) or branched.
  • R 9 is selected from C1-C22- alkyl, C 2 -C 22 - alkenyl, Cio-C 22 alkylcarbonyl, and C10-C22 alkenylcarbonyl.
  • - R 10 is selected from H, methyl, -(CH 2 ) 3 NHC(NH)NH 2 , -CH 2 C(O)NH 2 , -CH 2 C(O)OH, -(CH 2 ) 2 C(O)NH 2 , - (CH 2 ) 2 C(O)OH, (imidazole-4-yl)-methyl, -CH(CH 3 )C 2 H 5 , -CH 2 CH(CH 3 ) 2 , -(CH 2 ) 4 NH 2 , benzyl, hydroxymethyl, -CH(OH)CH3, (indole-3-yl)-methyl, (4-hydroxy-phenyl)-methyl, isopropyl, -(CH 2 ) 2 SCH3, and -CH 2 SH.
  • R x is selected from H and Ci-C4-alkyl.
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least one amphoteric surfactant of general formulae (AMS-lla), (AMS-llb), or (AMS-llc), which might be called betaines and/or sulfobetaines:
  • AMS-lla The variables in general formulae (AMS-lla), (AMS-llb) and (AMS-llc) are defined as follows:
  • R 11 is selected from linear (straight-chain; n-) or branched C7-C22 alkyl and linear (straight-chain; n-) or branched C7-C22 alkenyl.
  • R 12 are each independently selected from linear (straight-chain; n-) C1-C4 alkyl.
  • R 13 is selected from C1-C5 alkyl and hydroxy C1-C5 alkyl; for example 2-hydroxypropyl.
  • A’ is selected from carboxylate and sulfonate.
  • AMS-lla The integer r in general formulae (AMS-lla), (AMS-llb), and (AMS-llc) is in the range of 2 to 6.
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least one amphoteric surfactant of general formula (AMS-III), which might be called alkyl-amphocarboxylates:
  • AMS-III amphoteric surfactant of general formula
  • R 11 is selected from C7-C22 alkyl and C7-C22 alkenyl, wherein alkyl and/or alkenyl are linear (straight-chain; n-) or branched, preferably linear.
  • - R 14 is selected from -CH 2 C(O)O-M + , -CH 2 CH 2 C(O)O-M + and -CH 2 CH(OH)CH 2 SO3-M + .
  • R 15 is selected from H and -CH2C(O)O-
  • AMS-III The integer r in general formula (AMS-III) is in the range of 2 to 6.
  • Non-limiting examples of further suitable alkyl-amphocarboxylates include sodium cocoamphoacetate, sodium lauroamphoacetate, sodium capryloamphoacetate, disodium cocoamphodiacetate, disodium lauroamphodiacetate, disodium caprylamphodiacetate, disodium capryloamphodiacetate, disodium cocoamphodipropionate, disodium lauroamphodipropionate, disodium caprylamphodipropionate, and disodium capryloamphodipropionate.
  • the cleaning compositions e.g. detergent formulations of the invention comprise at least one amphoteric surfactant according to general formula (AMS-IV), which might be called amine oxides (AO):
  • R 16 is selected from Cs-Cis alkyl, hydroxy Cs-C alkyl, acylamidopropoyl and Cs-C alkyl phenyl group; wherein alkyl and/or alkenyl are linear (straight-chain; n-) or branched.
  • R 17 is selected from C2-C3 alkylene, hydroxy C2-C3 alkylene, and mixtures thereof.
  • each residue can be independently selected from C1-C3 alkyl and hydroxy C1-C3;
  • R 15 groups can be attached to each other, e.g., through an oxygen or nitrogen atom, to form a ring structure.
  • the integer x in general formula (AMS-IV) is in the range of 0 to 5, preferably from 0 to 3, most preferably 0.
  • Non-limiting examples of further suitable amine oxides include C10-C18 alkyl dimethyl amine oxides and Cs-Cis alkoxy ethyl dihydroxyethyl amine oxides.
  • Examples of such materials include dimethyloctyl amine oxide, diethyldecyl amine oxide, bis-(2-hydroxyethyl)dodecyl amine oxide, dimethyldodecylamine oxide, dipropyltetradecyl amine oxide, methylethylhexadecyl amine oxide, dodecylamidopropyl dimethyl amine oxide, cetyl dimethyl amine oxide, stearyl dimethyl amine oxide, tallow dimethyl amine oxide and dimethyl-2- hydroxyoctadecyl amine oxide.
  • a further example of a suitable amine oxide is cocamidylpropyl dimethylaminoxide, sometimes also called cocamidopropylamine oxide.
  • compositions e.g. detergent formulations according to the present invention.
  • manual dishwashing detergents comprise at least one amphoteric surfactant.
  • liquid cleaning compositions according to the invention comprise at least one amphoteric surfactant, wherein the total amount of amphoteric surfactant preferably is in the range from 0.01 % to 10%, in the range from 0.1 to 5%, or in the range from 0.5 to 1 % by weight, all relative to the total weight of the cleaning composition, e.g. detergent formulation.
  • at least one amphoteric surfactant is selected from compound according to formulae AMS-lla, AMS-llb, AMS-llc and AMS-IV.
  • cleaning compositions e.g. detergent formulations, of the invention comprise one or more complexing agents (chelating agents, sequestrating agents), precipitating agents, and/or ion exchange compounds, which usually form water-soluble complexes with calcium and magnesium.
  • complexing agents chelating agents, sequestrating agents
  • precipitating agents precipitating agents
  • ion exchange compounds which usually form water-soluble complexes with calcium and magnesium.
  • builders or building agents” herein, without meaning to limit such compounds to this function in the final application of a cleaning composition, e.g. detergent formulation.
  • Non-phosphate-based builders according to the invention include sodium gluconate, citrate(s), silicate(s), carbonate(s), phosphonate(s), amino carboxylate(s), polycarboxylate(s), polysulfonate(s), and polyphosphonate(s).
  • cleaning compositions e.g. detergent formulations, of the invention comprise one or more citrates.
  • citrate(s) includes the mono- and the dialkali metal salts and in particular the mono- and preferably the trisodium salt of citric acid, ammonium or substituted ammonium salts of citric acid as well as citric acid as such.
  • Citrate can be used as the anhydrous compound or as the hydrate, for example as sodium citrate dihydrate.
  • the cleaning compositions e.g. detergent formulations, of the invention, in one embodiment, comprise at least one phosphonate preferably selected from derivatives of polyphosphonic acids such as of diphosphonic acid such as sodium salt of HEDP, derivatives of aminopolyphosphonic acid such as aminoalkylene phosphonic acids such as DTPMP.
  • Said phosphonates preferably are comprised in amounts in the range of 0.1 % to 5.0% by weight, in the range of 0.5% to 3.0% by weight, or in the range of 1.0% to 2.0% by weight, all relative to the total weight of the cleaning composition.
  • Cleaning compositions of the invention may comprise one or more amino carboxylates.
  • suitable "amino carboxylates” include, but are not limited to: diethanol glycine (DEG), dimethylglycine (DMG), nitrilitriacetic acid (NTA), N-hydroxyethylaminodiacetic acid, eth-ylenediaminetetraacetic acid (EDTA), N- (2hydroxyethyl)iminodiacetic acid (HEIDA), hydroxyeth-ylenediaminetriacetic acid, N-hydroxyethyl- ethylenediaminetriacetic acid (HEDTA), hydroxyeth-ylenediaminetetraacetic acid, diethylenetriaminepentaacetic acid (DTPA), and methylglycinedi-acetic acid (MGDA), glutamic acid-diacetic acid (GLDA), iminodisuccinic acid (IDS), hydroxyimi-nodisuccinic acid, ethylenediaminedis
  • MGDA aspartic acid-N-monoacetic acid
  • ASDA aspartic acid-N, N-diacetic acid
  • ASMP aspartic acid-N-monopropionic acid
  • SMAS N-(2-sulfomethyl) aspartic acid
  • SEAS N-(2-sulfoethyl) aspartic acid
  • SMGL SGL
  • SEGL N-methyliminodiacetic acid
  • MIDA alpha-alanine-N, N-diacetic acid
  • SEDA isoserine-N, N-diacetic acid (ISDA), phenylalanine-N, N-diacetic acid
  • ISDA isoserine-N, N-diacetic acid
  • ISDA phenylalanine-N, N-diacetic acid
  • ammonium salts refers to salts with at least one cation that bears a nitrogen atom that is permanently or temporarily quaternized.
  • cations that bear at least one nitrogen atom that is permanently quaternized include tetramethylammonium, tetraethylammonium, dimethyldiethyl ammonium, and n-C10-C20-alkyl tri-methyl ammonium.
  • Examples of cations that bear at least one nitrogen atom that is temporarily quaternized include protonated amines and ammonia, such as monomethyl ammonium, dimethyl ammonium, trimethyl ammonium, monoethyl ammonium, diethyl ammonium, triethyl ammonium, n-C10-C20-alkyl dimethyl ammonium 2-hydroxyethylammonium, bis(2-hydroxyethyl) ammonium, tris(2-hydroxyethyl)ammonium, N-methyl 2- hydroxyethyl ammonium, N,N-dimethyl-2-hydroxyethylammonium, and especially NH4+.
  • protonated amines and ammonia such as monomethyl ammonium, dimethyl ammonium, trimethyl ammonium, monoethyl ammonium, diethyl ammonium, triethyl ammonium, n-C10-C20-alkyl dimethyl ammonium 2-hydroxyethylammoni
  • Said amino carboxylates preferably are comprised in amounts in the range of 0% to 30.0% by weight, in the range of 0.5% to 25.0% by weight, in the range of 1 % to 20% by weight, in the range of 2% to 15%, in the range of 2.5% to 10% by weight, in the range of 3% to 8% by weight, or in the range of 2.5 to 5% by weight, all relative to the total weight of the cleaning composition.
  • the compositions according to the invention are free from phosphates and polyphosphates, with hydrogenphosphates being subsumed, for example free from trisodiumphosphate, pentasodiumtripolyphosphate and hexasodiummetaphosphate.
  • hydrogenphosphates being subsumed, for example free from trisodiumphosphate, pentasodiumtripolyphosphate and hexasodiummetaphosphate.
  • "free from” is to be understood as meaning that the content of phosphate and polyphosphate is in total in the range from 10 ppm to 0.2% by weight, determined by gravimetry and relative to the total weight of the cleaning composition.
  • the composition of the present invention is a liquid cleaning composition additionally comprising at least one detergent component, wherein the detergent component is a compound selected from an anionic surfactant and a builder, wherein the anionic surfactant is preferably selected from linear alkylbenzene sulphonate (LAS) or alcohol ethoxy sulphates (AES) and wherein the builder preferably is a non-phosphate based builder, preferably selected from citrates, phosphonates and amino carboxylates.
  • the detergent component is a compound selected from an anionic surfactant and a builder
  • the anionic surfactant is preferably selected from linear alkylbenzene sulphonate (LAS) or alcohol ethoxy sulphates (AES)
  • the builder preferably is a non-phosphate based builder, preferably selected from citrates, phosphonates and amino carboxylates.
  • liquid cleaning compositions e.g. detergent formulations
  • liquid cleaning compositions e.g. detergent formulations
  • liquid cleaning compositions, e.g. detergent formulations of the invention usually comprise one or more rheology modifiers, which are also known as thickeners in the art.
  • the cleaning composition, e.g. detergent formulation of the invention comprises at least one naturally derived polymeric structurants, preferably selected from polysaccharide derivatives such as xanthan gum in amounts in the range of 0.1 % to about 5% by weight, or even from about 0.2% to about 0.5% by weight, relative to the total weight of the cleaning composition.
  • liquid cleaning compositions e.g. detergent formulations, of the invention are free from bleaches, for example free from inorganic peroxide compounds or chlorine bleaches such as sodium hypochlorite, meaning that liquid cleaning compositions, e.g. detergent formulations according to the invention comprise in total 0.8%, 0.5%, 0.1 % or 0.01 % by weight or less of inorganic peroxide compound and chlorine bleach, relative in each case on total weight of the liquid cleaning composition.
  • liquid cleaning compositions of the invention are preferably selected from liquid laundry detergents, liquid manual dishwashing detergents and liquid automated dishwashing detergents.
  • liquid enzyme compositions as described herein are added to the liquid cleaning compositions.
  • liquid enzyme compositions of the invention to a liquid cleaning composition, preferably a liquid detergent formulation, usually occurs in a weight ratio liquid composition:detergent formulation of about 1 :1000, 1 :500, 1 :100, 1 :50, 1 :30, 1 :25, 1 :20, or 1 : 10.
  • Components a. and b. and c., further components and detergent component(s) are such that their amounts in the liquid cleaning compositions comply with their ratios, amounts and total amounts as defined above in the chapters on the individual components or the chapter on the liquid enzyme composition, and thus the cleaning composition comprises those components in such amounts and ratios which result from the use of such liquid composition in such a cleaning composition, e.g. detergent formulation.
  • the liquid cleaning composition is characterized by a concentration of the protease of 0.0005-0.1%, preferably 0.02-0.06%, a concentration of the protease inhibitor of 0.00005-0.06%, preferably 0.0005-0.004%, and a concentration of the solubilizing agent of:
  • neopentyl glycol 0.01-1%, preferably 0.05-0.2%
  • ii. gamma-valerolactone 0.005-0.5%, preferably 0.01-0.1%
  • diethylene glycol 0.01-1%, preferably 0.05-0.2%
  • the invention also relates to a method for preparation of a cleaning composition of the invention, comprising the steps of mixing at least components a. and b. and at least one of c. - all as disclosed herein - with at least one detergent component in one or more steps in any order.
  • the method of preparation of a cleaning composition comprises the steps of mixing the liquid enzyme composition of the invention with at least one detergent component in one or more steps in any order.
  • the invention in one aspect relates to the use of the liquid enzyme compositions of the invention as a detergent component for the cleaning compositions such as l&l and homecare formulations for laundry and hard surface cleaning.
  • the invention also relates to a method for cleaning a soiled textile or dishware, wherein the method comprises providing a solid textile or dishware and bringing the soiled fabric or dishware in contact with a liquid cleaning composition according as described herein.
  • a composition comprising: a. at least one protease and b. a protease inhibitor and c. at least one solubilizing agent selected from the group consisting of:
  • neopentyl glycol ii. gamma-valerolactone, and ill. diethylene glycol.
  • a composition comprising: b. a protease inhibitor and c. at least one solubilizing agent selected from the group consisting of:
  • the composition according to any of embodiments 1 to 3, wherein the solubilizing agent is gammavalerolactone.
  • the protease inhibitor is selected from the group consisting of peptide aldehydes, salts or derivates thereof, preferably peptide aldehydes or peptide aldehyde hydrosulfite adducts and combinations thereof, more preferably a peptide aldehyde or peptide aldehyde hydrosulfite adduct comprising non-polar amino acid side chains.
  • composition according to embodiment 10 wherein the protease inhibitor is a peptide aldehyde, salt, or hydrosulfite adduct thereof, preferably a tripeptide aldehyde.
  • R1, R2, R3 are amino acid side chains of non-polar amino acids
  • Z is an N-terminal protection group, preferably selected from benzyloxycarbonyl (Cbz), p-methoxybenzyl carbonyl (MOZ), benzyl (Bn), benzoyl (Bz), p-methoxybenzyl (PMB), p-methoxyphenyl (PMP), formyl, acetyl (Ac), methyloxy, alkoxycarbonyl, methoxycarbonyl, fluorenylmethyloxycarbonyl (Fmoc), and tertbutyloxycarbonyl (Boc).
  • composition according to embodiment 12 wherein R1, R2 and R3 are amino acid side chains selected from the group consisting of Gly, Ala, Vai, Leu, lie, Met, Pro, Phe and Trp.
  • the composition according to embodiment 13 wherein R1 is an amino acid side chain of Vai, R2 is an amino acid side chain of Ala, and R3 is an amino acid side chain of Leu.
  • the composition according to embodiment 14, wherein the protease inhibitor is Z-VAL-H.
  • the composition according to any of embodiments 1 and 3 to 16 wherein the at least one protease comprised in component a.
  • the composition according to embodiment 18, wherein the at least one protease comprised in component a. is a protease identical to SEQ ID NO: 1 having the substitution R101 E according to the numbering of SEQ ID NO: 2.
  • the composition according to embodiment 21 wherein the at least one further enzyme is an amylase.
  • composition according to embodiment 21 wherein the at least one further enzyme is a cellulase.
  • the composition according to embodiment 21 wherein the at least one further enzyme is a mannanase.
  • the composition according to embodiment 21, wherein the composition comprises an amylase and a cellulase.
  • the composition according to any of the preceding embodiments wherein the composition further comprises glycerol and/or further comprises MPG.
  • the composition according to embodiment 29, wherein the composition comprises 0.01 % to 35% MPG.
  • liquid enzyme composition preferably 8-12%, ii. gamma-valerolactone: 1 -6%, preferably 4.5-5.5%, and/or ill. diethylene glycol: 8-12%, preferably 9.5-10.5%.
  • the liquid enzyme composition according to any of embodiments 33 or 34 wherein the liquid enzyme composition is essentially devoid of anionic surfactants and builders.
  • the liquid enzyme composition according to any of embodiments 32 to 35 wherein the composition additionally comprises calcium chloride dihydrate, preferably 0.1 -0.5% by weight, more preferably 0.2%- 0.4% by weight.
  • the liquid enzyme composition according to any of embodiments 32 to 36 wherein the pH of the composition is set to 6.
  • the liquid enzyme composition according to embodiment 38 wherein the composition comprises glycerol and MPG.
  • liquid enzyme composition according to any of embodiments 38 or 39, wherein the composition comprises glycerol from about 10% to 50% by weight, preferably from about 15% to 45% by weight.
  • the liquid enzyme composition according to embodiment 39 wherein the composition comprises neopentyl glycol: MPG:glycerol in ratios of 1 : ⁇ 3: ⁇ 6, or 1 : ⁇ 3: ⁇ 5, or 1 : ⁇ 3: ⁇ 4, or 1 : ⁇ 3: ⁇ 3.5, preferably 1 : ⁇ 3: ⁇ 4 or 1 : ⁇ 3: ⁇ 3.5, most preferably 1 : ⁇ 3: ⁇ 3.5, wherein the total amount of neopentyl glycol, MPG and glycerol in % by weight contained in the composition is set to be more than 45%, preferably at least 48%, most preferably at least 50% and optionally at most 70%.
  • the liquid enzyme composition according to embodiment 39 wherein the composition comprises gamma- valerolactone:glycerol in a ratio of 1 : ⁇ 35, preferably 1 : ⁇ 30, more preferably 1 : ⁇ 25, even more preferably 1 : ⁇ 20 or most preferred 1 : ⁇ 18, wherein the total amount of gamma-valerolactone, MPG and glycerol in % by weight contained in the composition is set to be at least 50% and optionally at most 70%.
  • the liquid enzyme composition according to embodiment 44 wherein the total amount of diethylene glycol, MPG and glycerol in % by weight contained in the composition is set to be at least 50% and optionally at most 70%.
  • the liquid enzyme composition according to any of embodiments 32 to 45, wherein the composition comprises: a. 0.05-12%, 0.05-10%, preferably 2% to 6% by weight protease, b. 0.05-0.4%, preferably 0.1 % to 0.25% by weight protease inhibitor, c. 5-25%, preferably 8-16%, most preferably 8-12% by weight neopentyl glycol, d. 10-40%, preferably 12-30%, more preferably 14-20% glycerol, e. 10-40%, preferably 20-35%, more preferably 28-34% MPG, and f. 0.1 -0.5% CaCI 2 *2H 2 O wherein optionally the pH of the composition is set to 6.
  • the liquid enzyme composition according to any of embodiments 32 to 45, wherein the composition comprises: a. 0.05-12%, 0.05-10%, preferably 2-6% by weight protease, b. 0.05-0.4%, preferably 0.1 -0.25% by weight protease inhibitor, c. 5-25%, preferably 8-16%, more preferably 8-12% by weight neopentyl glycol, d. 10-40%, preferably 15-30%, more preferably 18-22% glycerol, e. 10-30%, preferably 15-25%, more preferably 18-22% MPG, and f. 0.1 -0.5% CaCI 2 *2H 2 O wherein optionally the pH of the composition is set to 6.
  • the liquid enzyme composition according to any of embodiments 32 to 45, wherein the composition comprises: a. 0.05-12%, 0.05-10%, preferably 2-6% by weight protease, b. 0.05-0.4%, preferably 0.1 -0.25% by weight protease inhibitor, c. 1-15%, preferably 2-10%, more preferably 2-6% by weight gamma-valerolactone, d. 30-50%, preferably 35-48%, more preferably 40-45% glycerol, e. 2-25%, preferably 2-15%, more preferably 4-8% MPG, and f. 0.1 -0.5% CaCI 2 *2H 2 O wherein optionally the pH of the composition is set to 6.
  • liquid enzyme composition according to any of embodiments 46 to 49, wherein the composition additionally comprises 0.1-2% amylase.
  • the liquid cleaning composition according to embodiment 52 wherein the detergent component is a compound selected from an anionic surfactant and a builder, wherein the anionic surfactant is preferably selected from linear alkylbenzene sulphonate (LAS) or alcohol ethoxy sulphates (AES) and wherein the builder preferably is a non-phosphate-based builder, preferably selected from citrates, phosphonates and amino carboxylates.
  • the concentration of the protease is 0.0005-0.1 %, preferably 0.02-0.06%
  • the concentration of the protease inhibitor is 0.00005- 0.06%, preferably 0.0005-0.004%
  • concentration of the solubilizing agent is:
  • composition II. gamma-valerolactone: 0.005-0.5%, preferably 0.01-0.1 %, and/or ill. diethylene glycol: 0.01-1 %, preferably 0.05-0.2%.
  • the composition according to any of embodiments 1 to 33 or 35 to 54, wherein the composition further comprises 2-phenoxyethanol, preferably in an amount ranging from 2 ppm to 5%, more preferably 0.1 to 2% by weight of the composition.
  • a method for cleaning a soiled textile or dishware comprising providing a soiled textile or dishware and bringing the soiled fabric or dishware in contact with a liquid cleaning composition according to any of embodiments 52 to 55.
  • a method for stabilizing and solubilizing a protease and a protease inhibitor in a liquid enzyme composition comprising bringing together the protease, protease inhibitor and a solubilizing agent selected from the group consisting of:
  • neopentyl glycol ii. gamma-valerolactone, and ill. diethylene glycol.
  • Condition 1 100% solubilizing agent*
  • Condition 2 50% solubilizing agent, 50% water
  • Condition 3 40% solubilizing agent, 10% glycerol, 50% water
  • Condition 4 10% solubilizing agent, 40% glycerol, 50% water
  • solubilizing agent was present as solid material (EC, NPG and sorbitol), the solubilizing agent was tested as 70% solution in water
  • the stabilizer was dissolved in the respective solubilizing agent followed by subsequent addition of glycerol and water.
  • the resulting solution was visually inspected to a) see if the dilution with glycerol and water led to precipitation of the inhibitor and b) to see if the inhibitor was completely soluble, meaning whether the solid material dissolved without leaving any residue within 24 h.
  • the results are displayed in table 3.
  • Enzyme stability was tested by determining enzyme activity after storage of the enzyme in different aqueous formulations containing the respective solubilizing agent over a course of 14 days.
  • the reference formulation was 50% glycerol aqueous solution. The results are displayed in tables 4 and 5.
  • the protease according SEQ ID NO: 1 having the substitution R101 E according to the numbering of SEQ ID NO: 2 (abbreviated as P herein, 2.5% active enzyme) was stored for 14 days in the respective solubilizing agent at pH 6 at 45°C without inhibitor.
  • the protease has been used as a pre-formulated concentrate (11 % (w/w) P in 20% (w/w) MPG).
  • the activity after storage was determined with a standard assay using dimethyl-casein (DMC, 0.3% (w/v)) as substrate.
  • DMC dimethyl-casein
  • the release of free amino groups of DMC by proteolytic cleavage was detected using 2,4,6- trinitrobenzenesulfonic acid (TNBS, 0.2% (w/v)), measuring CD at 405 nm.
  • CHES N-cyclohexyl-2-aminoethanesulfonic acid
  • the cellulase according to SEC ID NO: 6 (abbreviated as C herein, 0.3% active enzyme) was stored for 14 days in the respective solubilizing agent at pH 6 at 45°C without inhibitor.
  • the cellulase has been used as a pre-formulated concentrate (10% (w/w) C in 30% (w/w) glycerol).
  • the activity after storage was determined with a standard assay using 4-nitrophenyl-p-D-lactopyranoside (pNPL) as substrate.
  • 4-ni trophenol pNP, yellow color
  • pNP can be quantified by measuring the OD at 405 nm.
  • the amylase according SEQ ID NO: 4 comprising the substitutions N25H, G4Q, R176K, G186E, T251 E, L405M, and Y482W according to the numbering of SEQ ID NO: 5 (abbreviated as A herein, 0.4% active enzyme) was stored for 14 days in the respective solubilizing agent at pH 6 at 45°C without inhibitor.
  • the amylase has been used as pre-formulated concentrate (3% (w/w) A in 40% (w/w) glycerol).
  • the activity after storage was determined with a standard assay using ethylidene-4-nitrophenoyl-o-D- maltoheptasoide (EPS) as substrate and o-glucosidase as second enzyme.
  • EPS ethylidene-4-nitrophenoyl-o-D- maltoheptasoide
  • o-glucosidase Upon cleavage of the glucose chain by the amylase, the o-glucosidase present in excess can hydrolyze and release 4-nitrophenol (pNP). The resulting yellow color can be quantified by measuring the OD at 405 nm.
  • the mannanase according to SEQ ID NO: 3 comprising the amino acid substitutions Q59V, N66D, A89H, E234Q, W259M, N282Y, T318N, E319G, and S322G according to the numbering of SEQ ID NO: 3 (abbreviated as M herein, 0,05% active enzyme) was stored for 14 days in the respective solubilizing agent at pH 6 at 37°C without inhibitor.
  • the mannanase has been used as a pre-formulated concentrate (2% (w/w) M in 30% (w/w) glycerol).
  • the activity after storage was determined with a standard assay using azurine-crosslinked (AZCL) galactomannan as substrate.
  • Azurin blue color
  • Azurin can be quantified by measuring the OD at 590 nm.
  • Enzyme activity (protease P, cellulase C, amylase A) after storage in aqueous compositions comprising different solubilizing agents in comparison to storage in 50% glycerol aqueous solution.
  • Mannanase activity (mannanase M) after storage in aqueous compositions comprising different solubilizing agents in comparison to storage in 50% glycerol aqueous solution.
  • solubilizing agents DEG, GVL and NPG overall showed a stabilizing effect on different enzymes and were considered suitable solubilizing agents.
  • Example 3 Solubility of Cbz-VAL-H in different formulations comprising a combination of the respective solubilizing agent, glycerol, and MPG
  • the solubility of the protease inhibitor Cbz-VAL-H in different aqueous formulations including the respective suitable solubilizing agent, glycerol and monopropylene glycol (MPG) was analyzed.
  • the inhibitor was first dissolved in a mixture of the respective solubilizing agent and MPG (NPG/MPG, GVL/MPG or DEG/MPG). Glycerol and water were added subsequently.
  • the resulting solution was visually inspected to a) see if the dilution with glycerol and water led to precipitation of the inhibitor and b) to see if the inhibitor was completely soluble, meaning whether the solid material dissolved without leaving any residue within 24 h.
  • Example 4 Storage stability of protease and Cbz-VAL-H in suitable formulations comprising a combination of the respective solubilizing agent, glycerol, and MPG
  • Protease storage stability was tested by determining enzyme activity after storage of the enzyme together with Cbz- VAL-H in different aqueous formulations containing the respective solubilizing agent, glycerol and MPG. Enzyme activity was determined according to example 2.
  • Preformulated protease concentrate formulations were formulated with water, glycerol, MPG and the four solubilizing agents to be compared (DEG, NPG, GVL, HDO (as control)). Enzyme concentrations were set to 2.5-3% w/w active protease P. The concentration of the protease inhibitor Cbz-VAL-H was set to 0.15% w/w. The compositions additionally contain 0.3% w/w CaCl2*2H2O. The solutions were stored in closed vessels at -80 °C or at 45 °C for 14 or 28 days. Enzyme activity after storage was measured and the residual activity was normalized to the activity of the -80 °C samples. Formulations containing hexane diol (HDO) were used as comparison. The results are displayed in table 9.
  • HDO hexane diol
  • Example 5 Storage stability of enzymes in blends after storage in suitable formulations comprising a combination of the respective solubilizing agent, Cbz-VAL-H, glycerol, and MPG
  • Enzyme storage stability in blends was tested by determining enzyme activity after storage of the enzymes together with Cbz-VAL-H in different aqueous formulations containing the respective solubilizing agent, glycerol and MPG. Enzyme activity was determined according to example 2.
  • Protease/cellulase blend in formulations comprising DEG, NPG, or GVL as solubilizing agent
  • Preformulated enzyme concentrate formulations (Protease P, Cellulase C - see Example 2 for description) were formulated with water, glycerol, MPG and the three solubilizing agents to be compared (DEG, NPG, GVL).
  • Enzyme concentrations were set to 2-2.5% w/w active protease P and 0.3-0.6% w/w active cellulase C.
  • the concentration of the protease inhibitor Cbz-VAL-H was set to 0.15% w/w. All compositions additionally contain 0.3% w/w CaCl2*2H2O.
  • the pH value was adjusted to 6.0.
  • Preformulated enzyme concentrate formulations (protease P, amylase A, see Example 2 for description) were formulated with water, glycerol, MPG and the two solubilizing agents to be compared (NPG, GVL). Enzyme concentrations were set to 2.5-5% w/w active protease P and 0.3-1% w/w active amylase A. The concentration of the protease inhibitor Cbz-VAL-H was set to 0.15% w/w. All compositions additionally contain 0.3% w/w CaCl2*2H2O. The pH value was adjusted to 6.0. The solutions were stored in closed vessels at -80 °C and 45 °C for 14 days or 3 months.
  • Enzyme activity after storage was measured and the residual activity was normalized to the activity of the -80 °C samples.
  • Formulations containing hexane diol (HDO) were used as comparison. After 14 days, no significant reduction in activity was detected (results not displayed). The results of the 3-month storage experiment are displayed in table 11 .
  • Table 11 Residual activity of protease P and amylase A after three months storage. All percentages are given in w/w.
  • Preformulated enzyme concentrate formulations (protease P, amylase A, cellulase C, see Example 2 for description) were formulated with water, glycerol, MPG and the two solubilizing agents to be compared (NPG, GVL). Enzyme concentrations were set to 1 .8-4.5 active w/w protease P, 0.3-1 .0% w/w active amylase A and 0.2- 0.6 % w/w active cellulase C. The concentration of the protease inhibitor Cbz-VAL-H was set to 0.15% w/w. All compositions additionally contain 0.3% w/w CaCl2*2H2O. The pH value was adjusted to 6.0.
  • Table 12 Residual activity of cellulase C, amylase A and protease P after 14 days or 3 months storage. All percentages are given in w/w. It is further noted that aqueous formulations containing 10% w/w NPG, 10% w/w MPG, 30% w/w glycerol, 0.3% w/w CaCl2*2H20, 0.15% w/w Cbz-VAL-H, 1 .8% active w/w protease, 0.3% w/w active amylase and 0.6% w/w active cellulase that were stored for 14 days at 45°C showed 79% residual cellulase activity, whereas formulations containing 10% w/w NPG, 20% w/w MPG, 20% w/w glycerol, 0.3% w/w CaCl2*2H2O, 0.15% w/w Cbz-VAL-H 2.5% w/w active protease, 1 % w/w
  • formulations containing 5% w/w HDO, 45% w/w MPG, 10% w/w glycerol, 0.3% w/w CaCl2*2H2O, 0.15% w/w Cbz-VAL-H, 1 .8% w/w active protease, 0.3% w/w active amylase and 0.6% w/w active cellulase that were stored for 14 days at 45°C showed 0% residual cellulase activity.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Emergency Medicine (AREA)
  • Detergent Compositions (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

La présente invention concerne des agents de solubilisation qui ont été identifiés pour solubiliser efficacement des inhibiteurs de protéases non polaires, tout en stabilisant en outre des enzymes, de préférence des hydrolases, dans un environnement liquide. L'invention concerne en outre des procédés de stabilisation et de solubilisation d'une protéase et d'un inhibiteur de protéase dans une composition liquide comprenant de tels agents de solubilisation et des procédés de nettoyage d'un tissu ou d'une vaisselle souillés à l'aide d'une composition liquide comprenant de tels agents de solubilisation.
PCT/EP2024/079772 2023-11-02 2024-10-22 Stabilisation d'enzymes dans des compositions contenant un inhibiteur de protéase Pending WO2025093368A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP23207476 2023-11-02
EP23207476.5 2023-11-02

Publications (1)

Publication Number Publication Date
WO2025093368A1 true WO2025093368A1 (fr) 2025-05-08

Family

ID=88690422

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2024/079772 Pending WO2025093368A1 (fr) 2023-11-02 2024-10-22 Stabilisation d'enzymes dans des compositions contenant un inhibiteur de protéase

Country Status (1)

Country Link
WO (1) WO2025093368A1 (fr)

Citations (85)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4435307A (en) 1980-04-30 1984-03-06 Novo Industri A/S Detergent cellulase
JPS6336774B2 (fr) 1979-05-25 1988-07-21 Tokyo Shibaura Electric Co
EP0283075A2 (fr) 1987-02-27 1988-09-21 Gist-Brocades N.V. Clonage moléculaire et expression de gènes codant pour des enzymes protéolytiques
JPS6356289B2 (fr) 1979-11-15 1988-11-08 Kansai Denryoku Kk
WO1989006279A1 (fr) 1988-01-07 1989-07-13 Novo-Nordisk A/S Genes de subtilisine mutes
WO1989006276A2 (fr) 1988-01-08 1989-07-13 Dpz Deutsches Primatenzentrum Gesellschaft Mbh Retrovirus de primates du type hiv-2, vaccins, compositions diagnostiques et pharmaceutiques
WO1989009259A1 (fr) 1988-03-24 1989-10-05 Novo-Nordisk A/S Preparation de cellulase
WO1989009819A1 (fr) 1988-04-12 1989-10-19 Genex Corporation Mutations combinantes de stabilisation de subtilisine
WO1989009830A1 (fr) 1988-04-12 1989-10-19 Genex Corporation Mutations de subtilisine
JPH034706A (ja) 1989-05-31 1991-01-10 Kubota Corp 人工種子作成方法
WO1991002792A1 (fr) 1989-08-25 1991-03-07 Henkel Research Corporation Enzyme proteolytique alcaline et procede de production
WO1991006637A1 (fr) 1989-10-31 1991-05-16 Genencor International, Inc. Mutants de subtilisine
WO1991017244A1 (fr) 1990-05-09 1991-11-14 Novo Nordisk A/S Enzyme capable de degrader la cellulose ou l'hemicellulose
WO1991018974A1 (fr) 1990-05-29 1991-12-12 Chemgen Corporation HEMICELLULASE ACTIVE A DES VALEURS EXTREMES DE pH ET DE TEMPERATURE AINSI QUE SES MOYENS DE PRODUCTION
WO1992006165A1 (fr) 1991-06-11 1992-04-16 Genencor International, Inc. Compositions de detergent contenant des compositions de cellulase manquant de constituants de type cbh i
EP0495257A1 (fr) 1991-01-16 1992-07-22 The Procter & Gamble Company Compositions de détergent compactes contenant de la cellulase de haute activité
EP0531372A1 (fr) 1990-05-09 1993-03-17 Novo Nordisk As Preparation de cellulase comprenant un enzyme d'endoglucanase.
WO1993017244A1 (fr) 1992-02-28 1993-09-02 Institut Teplofiziki Pompe a vide du type a jet
WO1993024622A1 (fr) 1992-05-22 1993-12-09 Alko Ltd. Mannanases, genes les codant, procede d'isolement de ces genes, et procedes de blanchiment de pates de lignocellulose
WO1994002597A1 (fr) 1992-07-23 1994-02-03 Novo Nordisk A/S Alpha-amylase mutante, detergent, agent de lavage de vaisselle et de liquefaction
WO1994007998A1 (fr) 1992-10-06 1994-04-14 Novo Nordisk A/S Variantes de cellulase
WO1994018314A1 (fr) 1993-02-11 1994-08-18 Genencor International, Inc. Alpha-amylase stable a l'oxydation
WO1994021801A2 (fr) 1993-03-17 1994-09-29 Genencor International, Inc. Purification et clonage moleculaire de la cellulase eg iii
WO1994026880A1 (fr) 1993-05-10 1994-11-24 Gist-Brocades N.V. Action combinee des endoglucanases et des cellobiohydrolases
WO1995002043A1 (fr) 1993-07-06 1995-01-19 Novo Nordisk A/S ADN CODANT UNE ENZYME A ACTIVITE D'ENDOGLUCANASE ET ETANT ISOLEE A PARTIR DE $i(TRICHODERMA HARZIANUM)
WO1995002675A1 (fr) 1993-07-12 1995-01-26 Novo Nordisk A/S Composition detersive comprenant deux cellulases
WO1995010603A1 (fr) 1993-10-08 1995-04-20 Novo Nordisk A/S Variants d'amylase
WO1995023221A1 (fr) 1994-02-24 1995-08-31 Cognis, Inc. Enzymes ameliorees et detergents les contenant
WO1995024471A1 (fr) 1994-03-08 1995-09-14 Novo Nordisk A/S Nouvelles cellulases alcalines
JPH0851975A (ja) 1991-10-09 1996-02-27 Res Dev Corp Of Japan 新規なβ−マンナナーゼとその製造方法
WO1996011262A1 (fr) 1994-10-06 1996-04-18 Novo Nordisk A/S Enzyme et preparation enzymatique presentant une activite endoglucanase
WO1996023872A1 (fr) 1995-02-02 1996-08-08 Stichting Centraal Laboratorium Van De Bloedtransfusiedienst Van Het Nederlandse Rode Kruis Enrichissement de cellules souches hematopoietiques provenant du sang ou de la moelle osseuse
WO1996029397A1 (fr) 1995-03-17 1996-09-26 Novo Nordisk A/S Nouvelles endoglucanases
WO1997003296A1 (fr) 1995-07-08 1997-01-30 Hohmann Joerg Dispositif de mesure de l'allongement d'un boulon filete ou d'une vis
WO1997011164A1 (fr) 1995-09-20 1997-03-27 Genencor International, Inc. Mannanase purifiee provenant de bacillus amyloliquefaciens et procede de preparation
WO1997014804A1 (fr) 1995-10-17 1997-04-24 Röhn Enzyme Finland OY Cellulases, genes les codant et utilisation de ces cellulases
US5648263A (en) 1988-03-24 1997-07-15 Novo Nordisk A/S Methods for reducing the harshness of a cotton-containing fabric
WO1997043424A1 (fr) 1996-05-14 1997-11-20 Genencor International, Inc. α-AMYLASES MODIFIEES POSSEDANT DES PROPRIETES MODIFIEES DE FIXATION DU CALCIUM
WO1998008940A1 (fr) 1996-08-26 1998-03-05 Novo Nordisk A/S Nouvelle endoglucanase
WO1998012307A1 (fr) 1996-09-17 1998-03-26 Novo Nordisk A/S Variants de cellulase
WO1998013459A1 (fr) 1996-09-24 1998-04-02 The Procter & Gamble Company Detergents liquides contenant un enzyme proteolytique, un aldehyde peptidique et des ions calcium
WO1999019467A1 (fr) 1997-10-13 1999-04-22 Novo Nordisk A/S MUTANTS D'α-AMYLASE
WO1999064619A2 (fr) 1998-06-10 1999-12-16 Novozymes A/S Nouvelles mannanases
WO2000022103A1 (fr) 1998-10-13 2000-04-20 Novozymes A/S Polypeptide modifie a reponse immunitaire reduite
US6121165A (en) 1997-07-31 2000-09-19 The Procter & Gamble Company Wet-like cleaning articles
WO2000060060A2 (fr) 1999-03-31 2000-10-12 Novozymes A/S Polypeptides presentant une activite alcaline alpha-amylase et acides nucleiques les codant
WO2002010355A2 (fr) 2000-08-01 2002-02-07 Novozymes A/S Mutants d'alpha-amylase a proprietes modifiees
DE10064983A1 (de) 2000-12-23 2002-07-18 Henkel Kgaa Neue Alkalische Protease aus Bacillus alcalophilus (DSM 11233)
EP1240525A2 (fr) 1999-12-23 2002-09-18 PHARMACIA & UPJOHN COMPANY Titrages et methodes diagnostiques et therapeutiques bases sur l'utilisation des canaux sodiques comme cibles de proteine beta-amyloide ou d'agregats de celle-ci
WO2002099091A2 (fr) 2001-06-06 2002-12-12 Novozymes A/S Endo-beta-1,4-glucanase
EP1305432A2 (fr) 2000-08-04 2003-05-02 Genencor International, Inc. Cellulase trichoderma reesei egiii mutante, adn codant pour de telles compositions d'egiii et methodes d'obtention
WO2003054184A1 (fr) 2001-12-20 2003-07-03 Henkel Kommanditgesellschaft Auf Aktien Nouvelle protease alcaline extraite de bacillus gibsonii (dsm 14393) et agents de lavage et de nettoyage contenant cette nouvelle protease alcaline
WO2003055974A2 (fr) 2001-12-22 2003-07-10 Henkel Kommanditgesellschaft Auf Aktien Nouvelle protease alcaline obtenue a partir de bacillus sp. (dsm 14392) et agents de lavage et de nettoyage contenant cette protease alcaline
WO2003056017A2 (fr) 2001-12-22 2003-07-10 Henkel Kommanditgesellschaft Auf Aktien Nouvelle protease alcaline obtenue a partir de bacillus sp (dsm 14390) et agents de lavage et de nettoyage contenant cette protease alcaline
WO2004053039A2 (fr) 2002-12-11 2004-06-24 Novozymes A/S Composition detergente
WO2005003319A2 (fr) 2003-07-02 2005-01-13 Diversa Corporation Glucanases, acides nucleiques les codant et procedes de fabrication et d'utilisation associes
WO2005063974A1 (fr) 2003-12-23 2005-07-14 Henkel Kommanditgesellschaft Auf Aktien Nouvelle protease alcaline et produits detergents et nettoyants contenant cette nouvelle protease alcaline
WO2005103244A1 (fr) 2004-04-23 2005-11-03 Henkel Kommanditgesellschaft Auf Aktien Nouvelles proteases alcalines et agents de lavage et detergents renfermant celles-ci
WO2006002643A2 (fr) 2004-07-05 2006-01-12 Novozymes A/S Variants d'alpha-amylases presentant des proprietes modifiees
WO2006066594A2 (fr) 2004-12-23 2006-06-29 Novozymes A/S Variantes de l'alpha-amylase
WO2007044993A2 (fr) 2005-10-12 2007-04-19 Genencor International, Inc. Utilisation et production d'une metalloprotease neutre stable au stockage
WO2009061380A2 (fr) 2007-11-05 2009-05-14 Danisco Us Inc., Genencor Division Variants de bacillus sp. ts-23 alpha-amylase à propriétés modifiées
WO2009118375A2 (fr) 2008-03-26 2009-10-01 Novozymes A/S Compositions stabilisées d’enzymes liquides
WO2010104675A1 (fr) 2009-03-10 2010-09-16 Danisco Us Inc. Alpha-amylases associées à la souche bacillus megaterium dsm90, et leurs procédés d'utilisation
WO2011098531A1 (fr) 2010-02-10 2011-08-18 Novozymes A/S Variants et compositions contenant des variants à stabilité élevée en présence d'un agent chélateur
WO2013001087A2 (fr) 2011-06-30 2013-01-03 Novozymes A/S Procédé de criblage d'alpha-amylases
WO2013001078A1 (fr) 2011-06-30 2013-01-03 Novozymes A/S Variants d'alpha-amylase
WO2013004636A1 (fr) 2011-07-01 2013-01-10 Novozymes A/S Composition de subtilisine stabilisée
US20130025073A1 (en) * 2011-07-25 2013-01-31 Philip Frank Souter Detergent compositions
WO2013184577A1 (fr) 2012-06-08 2013-12-12 Danisco Us Inc. Variants d'alpha-amylase dérivés de l'alpha-amylase de cytophaga sp. amylase/ (cspamy2)
WO2014100018A1 (fr) 2012-12-19 2014-06-26 Danisco Us Inc. Nouvelle mannanase, compositions et procédés pour les utiliser
WO2014183920A1 (fr) 2013-05-17 2014-11-20 Novozymes A/S Polypeptides présentant une activité alpha-amylase
WO2014183921A1 (fr) 2013-05-17 2014-11-20 Novozymes A/S Polypeptides présentant une activité alpha-amylase
WO2014194032A1 (fr) 2013-05-29 2014-12-04 Danisco Us Inc. Métalloprotéases inédites
WO2014194054A1 (fr) 2013-05-29 2014-12-04 Danisco Us Inc. Métalloprotéases inédites
WO2014194117A2 (fr) 2013-05-29 2014-12-04 Danisco Us Inc. Métalloprotéases inédites
WO2015193488A1 (fr) 2014-06-20 2015-12-23 Novozymes A/S Métalloprotéase issue de kribbella aluminosa et compositions détergentes comprenant cette métalloprotéase
WO2016075078A2 (fr) 2014-11-10 2016-05-19 Novozymes A/S Métalloprotéases et leurs utilisations
WO2016092009A1 (fr) 2014-12-10 2016-06-16 Henkel Ag & Co. Kgaa Agent de lavage et de nettoyage solide comprenant de l'amylase
EP3677676A1 (fr) * 2019-01-03 2020-07-08 Basf Se Composés de stabilisation d'amylases dans des liquides
US20210009927A1 (en) * 2018-04-17 2021-01-14 Novozymes A/S Polypeptides Comprising Carbohydrate Binding Activity in Detergent Compositions And Their use in Reducing Wrinkles in Textile or Fabrics
WO2021115912A1 (fr) 2019-12-09 2021-06-17 Basf Se Formulations comprenant une polyéthylèneimine modifiée de manière hydrophobe et une ou plusieurs enzymes
WO2022063698A1 (fr) 2020-09-22 2022-03-31 Basf Se Composition liquide comprenant un aldéhyde peptidique
US20220333039A1 (en) * 2019-12-23 2022-10-20 The Procter & Gamble Company Compositions comprising enzymes
CN116574565A (zh) * 2023-04-24 2023-08-11 大连奥首科技有限公司 一种芯片封装用助焊剂清洗剂、其制备方法及用途

Patent Citations (90)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6336774B2 (fr) 1979-05-25 1988-07-21 Tokyo Shibaura Electric Co
JPS6356289B2 (fr) 1979-11-15 1988-11-08 Kansai Denryoku Kk
US4435307A (en) 1980-04-30 1984-03-06 Novo Industri A/S Detergent cellulase
EP0283075A2 (fr) 1987-02-27 1988-09-21 Gist-Brocades N.V. Clonage moléculaire et expression de gènes codant pour des enzymes protéolytiques
WO1989006279A1 (fr) 1988-01-07 1989-07-13 Novo-Nordisk A/S Genes de subtilisine mutes
WO1989006276A2 (fr) 1988-01-08 1989-07-13 Dpz Deutsches Primatenzentrum Gesellschaft Mbh Retrovirus de primates du type hiv-2, vaccins, compositions diagnostiques et pharmaceutiques
WO1989009259A1 (fr) 1988-03-24 1989-10-05 Novo-Nordisk A/S Preparation de cellulase
US5648263A (en) 1988-03-24 1997-07-15 Novo Nordisk A/S Methods for reducing the harshness of a cotton-containing fabric
US5776757A (en) 1988-03-24 1998-07-07 Novo Nordisk A/S Fungal cellulase composition containing alkaline CMC-endoglucanase and essentially no cellobiohydrolase and method of making thereof
WO1989009819A1 (fr) 1988-04-12 1989-10-19 Genex Corporation Mutations combinantes de stabilisation de subtilisine
WO1989009830A1 (fr) 1988-04-12 1989-10-19 Genex Corporation Mutations de subtilisine
JPH034706A (ja) 1989-05-31 1991-01-10 Kubota Corp 人工種子作成方法
WO1991002792A1 (fr) 1989-08-25 1991-03-07 Henkel Research Corporation Enzyme proteolytique alcaline et procede de production
US5352604A (en) 1989-08-25 1994-10-04 Henkel Research Corporation Alkaline proteolytic enzyme and method of production
WO1991006637A1 (fr) 1989-10-31 1991-05-16 Genencor International, Inc. Mutants de subtilisine
WO1991017244A1 (fr) 1990-05-09 1991-11-14 Novo Nordisk A/S Enzyme capable de degrader la cellulose ou l'hemicellulose
EP0531372A1 (fr) 1990-05-09 1993-03-17 Novo Nordisk As Preparation de cellulase comprenant un enzyme d'endoglucanase.
EP0531315A1 (fr) 1990-05-09 1993-03-17 Novo Nordisk As Enzyme capable de degrader la cellulose ou l"hemicellulose.
WO1991018974A1 (fr) 1990-05-29 1991-12-12 Chemgen Corporation HEMICELLULASE ACTIVE A DES VALEURS EXTREMES DE pH ET DE TEMPERATURE AINSI QUE SES MOYENS DE PRODUCTION
US5476775A (en) 1990-05-29 1995-12-19 Chemgen Corporation Hemicellulase active at PH and temperature extremes
EP0495257A1 (fr) 1991-01-16 1992-07-22 The Procter & Gamble Company Compositions de détergent compactes contenant de la cellulase de haute activité
WO1992006165A1 (fr) 1991-06-11 1992-04-16 Genencor International, Inc. Compositions de detergent contenant des compositions de cellulase manquant de constituants de type cbh i
JPH0851975A (ja) 1991-10-09 1996-02-27 Res Dev Corp Of Japan 新規なβ−マンナナーゼとその製造方法
WO1993017244A1 (fr) 1992-02-28 1993-09-02 Institut Teplofiziki Pompe a vide du type a jet
WO1993024622A1 (fr) 1992-05-22 1993-12-09 Alko Ltd. Mannanases, genes les codant, procede d'isolement de ces genes, et procedes de blanchiment de pates de lignocellulose
WO1994002597A1 (fr) 1992-07-23 1994-02-03 Novo Nordisk A/S Alpha-amylase mutante, detergent, agent de lavage de vaisselle et de liquefaction
WO1994007998A1 (fr) 1992-10-06 1994-04-14 Novo Nordisk A/S Variantes de cellulase
WO1994018314A1 (fr) 1993-02-11 1994-08-18 Genencor International, Inc. Alpha-amylase stable a l'oxydation
WO1994021801A2 (fr) 1993-03-17 1994-09-29 Genencor International, Inc. Purification et clonage moleculaire de la cellulase eg iii
WO1994026880A1 (fr) 1993-05-10 1994-11-24 Gist-Brocades N.V. Action combinee des endoglucanases et des cellobiohydrolases
WO1995002043A1 (fr) 1993-07-06 1995-01-19 Novo Nordisk A/S ADN CODANT UNE ENZYME A ACTIVITE D'ENDOGLUCANASE ET ETANT ISOLEE A PARTIR DE $i(TRICHODERMA HARZIANUM)
WO1995002675A1 (fr) 1993-07-12 1995-01-26 Novo Nordisk A/S Composition detersive comprenant deux cellulases
WO1995010603A1 (fr) 1993-10-08 1995-04-20 Novo Nordisk A/S Variants d'amylase
WO1995023221A1 (fr) 1994-02-24 1995-08-31 Cognis, Inc. Enzymes ameliorees et detergents les contenant
EP1921147A2 (fr) 1994-02-24 2008-05-14 Henkel Kommanditgesellschaft auf Aktien Enzymes améliorées et détergents les contenant
WO1995024471A1 (fr) 1994-03-08 1995-09-14 Novo Nordisk A/S Nouvelles cellulases alcalines
WO1996011262A1 (fr) 1994-10-06 1996-04-18 Novo Nordisk A/S Enzyme et preparation enzymatique presentant une activite endoglucanase
WO1996023872A1 (fr) 1995-02-02 1996-08-08 Stichting Centraal Laboratorium Van De Bloedtransfusiedienst Van Het Nederlandse Rode Kruis Enrichissement de cellules souches hematopoietiques provenant du sang ou de la moelle osseuse
WO1996029397A1 (fr) 1995-03-17 1996-09-26 Novo Nordisk A/S Nouvelles endoglucanases
WO1997003296A1 (fr) 1995-07-08 1997-01-30 Hohmann Joerg Dispositif de mesure de l'allongement d'un boulon filete ou d'une vis
WO1997011164A1 (fr) 1995-09-20 1997-03-27 Genencor International, Inc. Mannanase purifiee provenant de bacillus amyloliquefaciens et procede de preparation
WO1997014804A1 (fr) 1995-10-17 1997-04-24 Röhn Enzyme Finland OY Cellulases, genes les codant et utilisation de ces cellulases
WO1997043424A1 (fr) 1996-05-14 1997-11-20 Genencor International, Inc. α-AMYLASES MODIFIEES POSSEDANT DES PROPRIETES MODIFIEES DE FIXATION DU CALCIUM
WO1998008940A1 (fr) 1996-08-26 1998-03-05 Novo Nordisk A/S Nouvelle endoglucanase
WO1998012307A1 (fr) 1996-09-17 1998-03-26 Novo Nordisk A/S Variants de cellulase
WO1998013459A1 (fr) 1996-09-24 1998-04-02 The Procter & Gamble Company Detergents liquides contenant un enzyme proteolytique, un aldehyde peptidique et des ions calcium
US6121165A (en) 1997-07-31 2000-09-19 The Procter & Gamble Company Wet-like cleaning articles
WO1999019467A1 (fr) 1997-10-13 1999-04-22 Novo Nordisk A/S MUTANTS D'α-AMYLASE
WO1999064619A2 (fr) 1998-06-10 1999-12-16 Novozymes A/S Nouvelles mannanases
WO2000022103A1 (fr) 1998-10-13 2000-04-20 Novozymes A/S Polypeptide modifie a reponse immunitaire reduite
WO2000060060A2 (fr) 1999-03-31 2000-10-12 Novozymes A/S Polypeptides presentant une activite alcaline alpha-amylase et acides nucleiques les codant
EP1240525A2 (fr) 1999-12-23 2002-09-18 PHARMACIA & UPJOHN COMPANY Titrages et methodes diagnostiques et therapeutiques bases sur l'utilisation des canaux sodiques comme cibles de proteine beta-amyloide ou d'agregats de celle-ci
WO2002010355A2 (fr) 2000-08-01 2002-02-07 Novozymes A/S Mutants d'alpha-amylase a proprietes modifiees
EP1305432A2 (fr) 2000-08-04 2003-05-02 Genencor International, Inc. Cellulase trichoderma reesei egiii mutante, adn codant pour de telles compositions d'egiii et methodes d'obtention
DE10064983A1 (de) 2000-12-23 2002-07-18 Henkel Kgaa Neue Alkalische Protease aus Bacillus alcalophilus (DSM 11233)
WO2002099091A2 (fr) 2001-06-06 2002-12-12 Novozymes A/S Endo-beta-1,4-glucanase
WO2003054184A1 (fr) 2001-12-20 2003-07-03 Henkel Kommanditgesellschaft Auf Aktien Nouvelle protease alcaline extraite de bacillus gibsonii (dsm 14393) et agents de lavage et de nettoyage contenant cette nouvelle protease alcaline
WO2003055974A2 (fr) 2001-12-22 2003-07-10 Henkel Kommanditgesellschaft Auf Aktien Nouvelle protease alcaline obtenue a partir de bacillus sp. (dsm 14392) et agents de lavage et de nettoyage contenant cette protease alcaline
WO2003056017A2 (fr) 2001-12-22 2003-07-10 Henkel Kommanditgesellschaft Auf Aktien Nouvelle protease alcaline obtenue a partir de bacillus sp (dsm 14390) et agents de lavage et de nettoyage contenant cette protease alcaline
WO2004053039A2 (fr) 2002-12-11 2004-06-24 Novozymes A/S Composition detergente
WO2005003319A2 (fr) 2003-07-02 2005-01-13 Diversa Corporation Glucanases, acides nucleiques les codant et procedes de fabrication et d'utilisation associes
WO2005063974A1 (fr) 2003-12-23 2005-07-14 Henkel Kommanditgesellschaft Auf Aktien Nouvelle protease alcaline et produits detergents et nettoyants contenant cette nouvelle protease alcaline
WO2005103244A1 (fr) 2004-04-23 2005-11-03 Henkel Kommanditgesellschaft Auf Aktien Nouvelles proteases alcalines et agents de lavage et detergents renfermant celles-ci
WO2006002643A2 (fr) 2004-07-05 2006-01-12 Novozymes A/S Variants d'alpha-amylases presentant des proprietes modifiees
WO2006066594A2 (fr) 2004-12-23 2006-06-29 Novozymes A/S Variantes de l'alpha-amylase
WO2007044993A2 (fr) 2005-10-12 2007-04-19 Genencor International, Inc. Utilisation et production d'une metalloprotease neutre stable au stockage
WO2009061380A2 (fr) 2007-11-05 2009-05-14 Danisco Us Inc., Genencor Division Variants de bacillus sp. ts-23 alpha-amylase à propriétés modifiées
WO2009118375A2 (fr) 2008-03-26 2009-10-01 Novozymes A/S Compositions stabilisées d’enzymes liquides
WO2010104675A1 (fr) 2009-03-10 2010-09-16 Danisco Us Inc. Alpha-amylases associées à la souche bacillus megaterium dsm90, et leurs procédés d'utilisation
WO2011098531A1 (fr) 2010-02-10 2011-08-18 Novozymes A/S Variants et compositions contenant des variants à stabilité élevée en présence d'un agent chélateur
WO2013001087A2 (fr) 2011-06-30 2013-01-03 Novozymes A/S Procédé de criblage d'alpha-amylases
WO2013001078A1 (fr) 2011-06-30 2013-01-03 Novozymes A/S Variants d'alpha-amylase
WO2013004636A1 (fr) 2011-07-01 2013-01-10 Novozymes A/S Composition de subtilisine stabilisée
US20130025073A1 (en) * 2011-07-25 2013-01-31 Philip Frank Souter Detergent compositions
WO2013184577A1 (fr) 2012-06-08 2013-12-12 Danisco Us Inc. Variants d'alpha-amylase dérivés de l'alpha-amylase de cytophaga sp. amylase/ (cspamy2)
WO2014100018A1 (fr) 2012-12-19 2014-06-26 Danisco Us Inc. Nouvelle mannanase, compositions et procédés pour les utiliser
WO2014183920A1 (fr) 2013-05-17 2014-11-20 Novozymes A/S Polypeptides présentant une activité alpha-amylase
WO2014183921A1 (fr) 2013-05-17 2014-11-20 Novozymes A/S Polypeptides présentant une activité alpha-amylase
WO2014194054A1 (fr) 2013-05-29 2014-12-04 Danisco Us Inc. Métalloprotéases inédites
WO2014194032A1 (fr) 2013-05-29 2014-12-04 Danisco Us Inc. Métalloprotéases inédites
WO2014194117A2 (fr) 2013-05-29 2014-12-04 Danisco Us Inc. Métalloprotéases inédites
WO2015193488A1 (fr) 2014-06-20 2015-12-23 Novozymes A/S Métalloprotéase issue de kribbella aluminosa et compositions détergentes comprenant cette métalloprotéase
WO2016075078A2 (fr) 2014-11-10 2016-05-19 Novozymes A/S Métalloprotéases et leurs utilisations
WO2016092009A1 (fr) 2014-12-10 2016-06-16 Henkel Ag & Co. Kgaa Agent de lavage et de nettoyage solide comprenant de l'amylase
US20210009927A1 (en) * 2018-04-17 2021-01-14 Novozymes A/S Polypeptides Comprising Carbohydrate Binding Activity in Detergent Compositions And Their use in Reducing Wrinkles in Textile or Fabrics
EP3677676A1 (fr) * 2019-01-03 2020-07-08 Basf Se Composés de stabilisation d'amylases dans des liquides
WO2021115912A1 (fr) 2019-12-09 2021-06-17 Basf Se Formulations comprenant une polyéthylèneimine modifiée de manière hydrophobe et une ou plusieurs enzymes
US20220333039A1 (en) * 2019-12-23 2022-10-20 The Procter & Gamble Company Compositions comprising enzymes
WO2022063698A1 (fr) 2020-09-22 2022-03-31 Basf Se Composition liquide comprenant un aldéhyde peptidique
CN116574565A (zh) * 2023-04-24 2023-08-11 大连奥首科技有限公司 一种芯片封装用助焊剂清洗剂、其制备方法及用途

Non-Patent Citations (23)

* Cited by examiner, † Cited by third party
Title
"Detergent Cake, Dishwashing Detergents, Liquid & Paste Detergents, Enzyme Detergents, Cleaning Powder & Spray Dried Washing Powder", 2015, ENGINEERS INDIA RESEARCH INSTITUTE (EIRI
"Detergent Formulations Encyclopedia", 2016, SOLVERCHEM PUBLICATIONS
"Enzyme Nomenclature, Recommendations", 1992, article "Nomenclature Committee of the International Union of Biochemistry and Molecular Biology"
"Methods in Enzymology", vol. 160, 1988, pages: 200 - 391
"UniProt", Database accession no. 10/26/2011
BÉGUIN, P.: "Molecular Biology of Cellulose Degradation", ANNU. REV. MICROBIOL., vol. 44, 1990, pages 219248
BEGUN, P.AUBERT, J-P.: "The biological degradation of cellulose", FEMS MICROBIOLOGY REVIEWS, vol. 13, 1994, pages 25 - 58
DELMAR ET AL., ANALYTICAL BIOCHEM, vol. 99, 1979, pages 316 - 320
EL SMITH ET AL., J. BIOL CHEM, vol. 243, 1968, pages 2184 - 2191
GUPTA ET AL., APPL. MICROBIOL. BIOTECHNOL., vol. 60, 2002, pages 381 - 395
HENRISSAT, B.: "Cellulases and their interaction with cellulose", CELLULOSE, vol. 1, 1994, pages 169 - 196, XP009062749, DOI: 10.1007/BF00813506
HOFFMAN, W. S., J. BIOL. CHEM., vol. 120, 1937, pages 51
ISIDRO-LLOBET ET AL.: "Amino Acid-Protecting Groups", CHEM. REV., vol. 109, no. 6, 2009, pages 2455 - 2504, XP055559012, DOI: 10.1021/cr800323s
JA WELLS ET AL., NUCLEIC ACIDS RESEARCH, vol. 11, 1983, pages 7911 - 7925
JACOBS ET AL., NUCL. ACIDS RES, vol. 13, 1985, pages 8913 - 8926
M. H. BOYER, BIOCHEMISTRY, vol. 162, 1987, pages 311 - 316
MCCLEARY, B. V., CARBOHYDRATE RESEARCH, vol. 67, no. 1, 1978, pages 213 - 221
MILLER, G. L.: "Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugars", ANALYTICAL CHEMISTRY, vol. 31, 1959, pages 426 - 428
NEEDLEMANWUNSCH, J. MOL. BIOL., vol. 48, 1979, pages 443 - 453
PETER G. M. WUTS: "Greene's Protective Groups in Organic Synthesis", 2014, JOHN WILEY & SONS, INC
R. SIEZEN: "Subtilisin enzymes", 1996, article "Subtilases: Subtilisin-like Proteases", pages: 75 - 95
T.-M. ENVERIW.M. FOGARTY: "Microbial Enzymes and Biotechnology", 1983, APPLIED SCIENCE PUBLISHERS, article "Microbial Cellulases", pages: 183 - 224
VASANTHA ET AL., J. BACTERIOL., vol. 159, 1984, pages 811 - 819

Similar Documents

Publication Publication Date Title
US11214777B2 (en) Method for using lipase enzymes for cleaning
US11732250B2 (en) Lipase enzymes
CA3043443A1 (fr) Stabilisation d'enzymes dans des compositions
WO2021074430A1 (fr) Liquides contenant une hydrolase stable au stockage
EP4612287A1 (fr) Polypeptides présentant une activité protéasique pour utilisation dans des compositions détergentes
US20210395651A1 (en) Compounds stabilizing hydrolases in liquids
WO2024094733A1 (fr) Polypeptides présentant une activité protéasique pour utilisation dans des compositions détergentes
WO2022063699A1 (fr) Combinaison améliorée d'inhibiteur de protéase et inhibiteur de protéase ayant une enzyme secondaire
US20220112479A1 (en) Compounds stabilizing amylases in liquids
CN112840021A (zh) 在液体中稳定水解酶的化合物
WO2025093368A1 (fr) Stabilisation d'enzymes dans des compositions contenant un inhibiteur de protéase
WO2022063698A1 (fr) Composition liquide comprenant un aldéhyde peptidique
US20210115422A1 (en) Amylase enzymes
EP3677676A1 (fr) Composés de stabilisation d'amylases dans des liquides
WO2025202374A1 (fr) Polypeptides ayant une activité protéase destinés à être utilisés dans des compositions détergentes
EP4624572A1 (fr) Polypeptides ayant une activité protéase destinés à être utilisés dans des compositions détergentes
WO2025202372A1 (fr) Polypeptides ayant une activité protéase destinés à être utilisés dans des compositions détergentes
WO2025202379A1 (fr) Polypeptides ayant une activité protéase destinés à être utilisés dans des compositions détergentes
WO2025202369A1 (fr) Polypeptides ayant une activité protéase destinés à être utilisés dans des compositions détergentes
EP4389864A1 (fr) Cutinases
WO2025202370A1 (fr) Polypeptides ayant une activité protéase destinés à être utilisés dans des compositions détergentes

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 24793818

Country of ref document: EP

Kind code of ref document: A1