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EP-3725797-B1 - STABILIZED LIQUID ENZYME COMPOSITIONS

EP3725797B1EP 3725797 B1EP3725797 B1EP 3725797B1EP-3725797-B1

Inventors

  • NIELSEN, LONE, KIERSTEIN
  • MIKKELSEN, LISE MUNCH
  • FRIIS, ESBEN, PETER
  • KNÖTZEL, Jürgen, Carsten, Franz,
  • SIMONSEN, OLE
  • SOERENSEN, Lotte, Rugholm

Dates

Publication Date
20260506
Application Date
20090326

Claims (10)

  1. A liquid detergent composition comprising 1 to 95% w/w of a detersive surfactant, 0.0001 to 5% w/w of a subtilisin, and 0.00001 to 1% w/w of a peptide aldehyde of the formula B 2 -B 1 -B 0 -H wherein: H is hydrogen; B 0 is a residue of Tyr; B 1 is a single amino acid residue; and B 2 consists of one or two amino acid residues, comprising an N-terminal protection group.
  2. The liquid detergent composition of claim 1, wherein B 1 is a residue of Ala, Cys, Gly, Pro, Ser, Thr, Val, Nva or Nle.
  3. The liquid detergent composition of claim 1 or 2, wherein B 2 is a residue of Gly, Arg, or Leu comprising an N-terminal protection group.
  4. The liquid detergent composition of any of claims 1-3, wherein B 2 comprises an N-terminal protection group selected from formyl, acetyl, benzoyl, trifluoroacetyl, fluoromethoxy carbonyl, methoxysuccinyl, aromatic and aliphatic urethane protecting groups, benzyloxycarbonyl, t-butyloxycarbonyl, adamantyloxycarbonyl, p-methoxybenzyl carbonyl (MOZ), benzyl (Bn), p-methoxybenzyl (PMB) or p-methoxyphenyl (PMP).
  5. The liquid detergent composition of any of claims 1-4, wherein the peptide compound is the peptide aldehyde Z-RAY-H, Ac-GAY-H, Z-GAY-H, Z-RVY-H, Z-LVY-H, Ac-LGAY-H, Ac-FGAY-H, Ac-YGAY-H, Ac-FGVY-H or Ac-WLVY-H, wherein Z is benzyloxycarbonyl and Ac is acetyl.
  6. The liquid detergent composition of any of claims 1-5, which further comprises a second enzyme.
  7. The liquid detergent composition of any of claims 1-6, which further comprises a second enzyme selected from the group consisting of amylases, lipases, cellulases, mannanases, oxidoreductases, and lyases.
  8. The liquid detergent composition of any of claims 1-7, which further comprises a polyol.
  9. The liquid detergent composition of any of claims 1-8, which comprises 2 to 60% w/w of the detersive surfactant, 0.0005 to 1% w/w of the subtilisin, and 0.00005 to 0.2% w/w of the peptide compound.
  10. The liquid detergent composition of any of claims 1-9, which comprises 3 to 50% w/w of the detersive surfactant, 0.001 to 0.5% w/w of the subtilisin, and 0.0001 to 0.1% w/w of the peptide compound.

Description

FIELD OF THE INVENTION The present invention relates to a liquid composition comprising a subtilisin and a peptide compound as a stabilizer for the subtilisin. It also relates to a peptide compound which is useful as a stabilizer for subtilisins. BACKGROUND ART Subtilisin-type proteases are well known in liquid aqueous detergents, particularly for use on laundry washing. A generally encountered problem in such liquid detergents is the degradation by the subtilisin of other enzymes in the composition and of the subtilisin itself. Consequently, the stability of the subtilisin and other enzymes in the liquid detergent composition is reduced, resulting in a liquid detergent with a reduced wash performance. The prior art has dealt extensively with improving the storage stability of enzymes in liquid detergents, for example by adding various subtilisin inhibitors or stabilizers. Boric acid and boronic acids are known to reversibly inhibit proteolytic enzymes. The use of peptide aldehydes for stabilizing certain proteases in liquid detergents is disclosed in WO 94/04651, WO 98/13458, WO 98/13459, WO 98/13460 and WO 98/13462. More specifically, WO94/04651 discloses the use of the peptide aldehydes Phe-Gly-Ala-PheH and Phe-Gly-Ala-LeuH for stabilizing subtilisin-type proteases. WO94/04651 also discloses Leu-Leu-TyrH as a suitable peptide aldehyde for stabilizing chymotrypsin-type proteases. Furthermore, WO94/04651 proposes methyl carbamate or methyl urea as an N-terminal protecting group of the peptide aldehydes. WO98/13460 discloses the use of peptide protease inhibitors, either peptide aldehydes or trifluromethyl ketones, where the peptide chain contains 2-5 amino acids and the aldehyde/ trifluromethyl ketone is derived from the amino acids alanine, valine, isoleucine, leucine, phenylglycine, phenylalanine or homophenylalanine and where the N-terminal protection group is preferably a sulphonamide or amidophoshate. WO2007/141736, WO2007/145963 and WO2007/145964 disclose the use of a reversible peptide protease inhibitor to stabilize liquid detergent compositions. US2003/157088 describes compositions containing enzymes stabilized with inhibitors. WO 96/41638 and WO 2005/105826 disclose peptide aldehydes and ketones. SUMMARY OF THE INVENTION The present invention is defined by the claims. Any subject-matter falling outside the scope of the claims is provided for information purposes only. The inventors have surprisingly found that certain peptide aldehydes or ketone derivatives are particularly efficient for stabilizing subtilisin-type proteases in aqueous compositions such as liquid detergents, including peptide compounds with OH-substituted phenylalanine as the C-terminal residue. Accordingly, the invention provides a liquid composition comprising a detersive surfactant, a subtilisin and a peptide compound of the formula B2-B1-B0-R wherein: R is hydrogen andB1 is a single amino acid residue. B0 is a residue of Tyr and may be a phenylalanine residue with an OH substituent at the p-position and/or at a m-position; and B2 consists of one or two amino acid residues, comprising an N-terminal protection group. Alternatively, B0 may be a single amino acid residue; and B2 is a residue of Gly, Arg or Leu with an N-terminal protection group attached. The invention further provides a peptide compound of the formula B2-B1-B0-R wherein: R is hydrogen, andB1 is a single amino acid residue. B0 is a residue of Tyr; and B2 may consist of one or two amino acid residues with benzyloxycarbonyl as an N-terminal protection group, or B2 may be a residue of Gly, Arg or Leu with an N-terminal protection group attached. Alternatively, B0 may be a single amino acid residue, and B1 may be a small amino acid residue, and B2 may be a residue of Gly, Arg or Leu with an aromatic N-terminal protection group attached. DETAILED DISCLOSURE OF THE INVENTION Definitions An "amino acid residue" indicates a group with a structure like -NH-CHR-CO- written with the N-terminal at the left and the C-terminal at the right. Amino acid residues are abbreviated using standard one-letter or three-letter abbreviations, including the following abbreviations: alanine (A), phenylalanine (F), glycine (G), leucine (L), arginine (R), valine (V), tryptophan (W), tyrosine (Y). The abbreviation "Y-H" denotes tyrosinal, meaning that the C-terminal end of the tyrosine residue is converted from a carboxylic group to an aldehyde group. The tyrosinal may be prepared by known processes. Amino acids Each amino acid residue in B1 and B2 may be a natural or non-naturally occurring alpha- or beta-amino acid containing the structure -NH-(CH(R))n-C(=O)-, where n=1-2 (preferably 1) and R is selected from linear or branched and/or cyclized, substituted or unsubstituted structures from the following groups: C1-C6 alkyl; phenyl; C7-C9 alkylaryl; C4-C8 cycloalkyl. Both L- and D-forms of amino acids are included. The amino acid may be an α-amino acid such as any of the naturally occurring a