U.S. patent application number 16/749716 was filed with the patent office on 2020-05-21 for detergent composition.
This patent application is currently assigned to Novozymes A/S. The applicant listed for this patent is Novozymes A/S. Invention is credited to Esben Peter Friis, Juergen Carsten Franz Knoetzel, Lise Munch Mikkelsen, Lone Kierstein Nielsen, Ole Simonsen.
Application Number | 20200157472 16/749716 |
Document ID | / |
Family ID | 40521614 |
Filed Date | 2020-05-21 |
United States Patent
Application |
20200157472 |
Kind Code |
A1 |
Nielsen; Lone Kierstein ; et
al. |
May 21, 2020 |
Detergent Composition
Abstract
Peptide aldehydes containing a ureido group are effective as
stabilizers for protease in a detergent composition, and as
stabilizers for a second enzyme in a liquid composition with a
protease.
Inventors: |
Nielsen; Lone Kierstein;
(Lyngby, DK) ; Mikkelsen; Lise Munch; (Roedovre,
DK) ; Friis; Esben Peter; (Herlev, DK) ;
Knoetzel; Juergen Carsten Franz; (Copenhagen, DK) ;
Simonsen; Ole; (Soeborg, DK) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Novozymes A/S |
Bagsvaerd |
|
DK |
|
|
Assignee: |
Novozymes A/S
Bagsvaerd
DK
|
Family ID: |
40521614 |
Appl. No.: |
16/749716 |
Filed: |
January 22, 2020 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
13124089 |
Apr 13, 2011 |
10577568 |
|
|
PCT/EP2009/064972 |
Nov 11, 2009 |
|
|
|
16749716 |
|
|
|
|
61117989 |
Nov 26, 2008 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C11D 3/386 20130101;
C07K 5/1016 20130101; C11D 3/38663 20130101; C11D 3/33 20130101;
C11D 7/3245 20130101 |
International
Class: |
C11D 3/386 20060101
C11D003/386; C11D 7/32 20060101 C11D007/32; C11D 3/33 20060101
C11D003/33; C07K 5/107 20060101 C07K005/107 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 13, 2008 |
EP |
08169063.8 |
Claims
1-14. (canceled)
15. A liquid detergent composition comprising a surfactant, a
protease and a peptide aldehyde of the formula
P--O-A.sup.1-CO-A.sup.2-A.sup.3-A.sup.4-H wherein P is H, Me, Et or
CF.sub.3; Al is a residue of Tyr, Leu, Phe, Met, Nle or Nva;
A.sup.2 is a residue of Gly, Ala, Val, Arg, Leu or Cpd; A.sup.3 is
a residue of Val, Gly, Leu or Ile; A.sup.4 is a residue of Phe,
Tyr, Leu, Nva, Nle, Met or Arg, wherein CO is a double N-capping
group.
16. The composition of claim 15, wherein the peptide aldehyde has
the formula Phe-CO-Arg-Val-Phe-H, Phe-CO-Arg-Val-Tyr-H or
Phe-CO-Gly-Gly-Tyr-H.
17. The composition of claim 15, which further comprises a second
enzyme selected from the group consisting of lipase, amylase,
cellulase, and carbohydrase.
18. The composition of claim 15, wherein the protease is an
alkaline protease, an aspartic protease, a serine protease, a
cysteine protease, or a subtilisin.
19. A liquid enzyme detergent composition, comprising a protease at
a concentration of at least 0.1% by weight of enzyme protein and a
compound of the formula P--O-A.sup.1-CO-A.sup.2-A.sup.3-A.sup.4-H
wherein P is H, Me, Et or CF.sub.3; Al is a residue of Tyr, Leu,
Phe, Met, Nle or Nva; A.sup.2 is a residue of Gly, Ala, Val, Arg,
Leu or Cpd; A.sup.3 is a residue of Val, Gly, Leu or Ile; A.sup.4
is a residue of Phe, Tyr, Leu, Nva, Nle, Met or Arg, wherein CO is
a double N-capping group.
20. A liquid enzyme detergent composition, comprising a protease, a
second enzyme and a compound of the formula
P--O-A.sup.1-CO-A.sup.2-A.sup.3-A.sup.4-H wherein P is H, Me, Et or
CF.sub.3; Al is a residue of Tyr, Leu, Phe, Met, Nle or Nva;
A.sup.2 is a residue of Gly, Ala, Val, Arg, Leu or Cpd; A.sup.3 is
a residue of Val, Gly, Leu or Ile; A.sup.4 is a residue of Phe,
Tyr, Leu, Nva, Nle, Met or Arg, wherein CO is a double N-capping
group.
21. The composition of claim 15, wherein molar ratio of peptide
aldehyde to protease is 0.1:1.
22. The composition of claim 15, wherein molar ratio of peptide
aldehyde to protease is 100:1.
23. The composition of claim 15, wherein molar ratio of peptide
aldehyde to protease is 0.5:1 to 50:1.
24. The composition of claim 15, wherein the composition is a
laundry detergent composition.
25. The composition of claim 19, wherein the composition is a
laundry detergent composition.
26. The composition of claim 20, wherein the composition is a
laundry detergent composition.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to a liquid detergent
composition comprising a surfactant and a protease and to a liquid
enzyme composition comprising a protease and optionally a second
enzyme. More particularly, it relates to such compositions
incorporating a compound which stabilizes the protease and/or the
second enzyme. Further, the invention also relates to a stabilizing
compound for use in such compositions.
BACKGROUND OF THE INVENTION
[0002] EP 0 583 534 A, WO 94/04651 A, WO 98/13458 A and WO
2007/145963 A describe enzyme stabilization in liquid detergent
compositions comprising a reversible peptide aldehyde protease
inhibitor. WO 95/25791 A describes methods for manufacturing
protease enzymes using non-protein protease inhibitors.
[0003] .alpha.-MAPI, antipain, GE20372A and chymostatin A, B and C
are described as peptide aldehydes with activity as protease
inhibitors: R. J. Broadbridge et al., Chem. Commun. (1998), 1449.
E. Sarubbi et al., FEBS Letters, 319 (3), 253-256 (1993). I. J.
Galpin et al., Int. J. Peptide Protein Res., 23, 1984, 477-486.
SUMMARY OF THE INVENTION
[0004] The inventors have found that certain peptide aldehydes
containing a "double N-capping group" (such as ureido (N--CO--N),
oxamide (N--CO--CO--N), thioureido (N--CS--N), dithio-oxamide
(N--CS--CS--N) or thiooxamide (N--CS--CO--N) group) are effective
as stabilizers for proteases in liquid enzyme formulations and in
liquid detergent compositions, and as stabilizers for a second
enzyme in a liquid composition with a protease. They also found
that the peptide aldehydes can be released from the protease when
the enzyme formulation or detergent composition is diluted with
water. Accordingly, the invention provides a detergent composition
comprising a surfactant, a protease and a stabilizer of the
formula:
P--O-(A.sup.i-X).sub.n-A.sup.n+1-H
[0005] wherein one X is the "double N-capping group" CO, CO--CO,
CS, CS--CS or CS--CO (most preferably CO), and the other X'es are
nothing (i.e., a chemical bond),
[0006] wherein n=1-10, preferably 2-5, most preferably 3
[0007] wherein i is an integer from 1 to n
[0008] wherein each A.sup.i and A.sup.n+1 is an amino acid residue
having the structure: [0009] --NH--CR--CO--for a residue to the
right of the double N-capping group, or [0010] --CO--CR--NH-- for a
residue to the left of the double N-capping group
[0011] wherein R is H-- or an optionally substituted alkyl or
alkylaryl group which may optionally include a hetero atom and may
optionally be linked to the N atom, and
[0012] wherein P is hydrogen or any C-terminal protection
group.
[0013] The invention also provides an enzyme composition,
comprising a protease at a concentration of at least 0.1% by weight
of enzyme protein and a stabilizer of the above formula.
[0014] Furthermore, the invention provides an enzyme composition,
comprising a protease, a second enzyme and a stabilizer of the
above formula.
[0015] Finally, the invention provides a stabilizer having the
formula HO-A.sup.1-CO-A.sup.2-A.sup.3-A.sup.4-H, wherein A.sup.1 is
a Phe residue, A.sup.2 is an Arg or Gly residue, A.sup.3 is a Val
or Gly residue, and A.sup.4 is a Tyr residue.
DETAILED DESCRIPTION OF THE INVENTION
Stabilizer
[0016] The peptide aldehyde used as a protease stabilizer has the
structure P--O-(A.sup.iX).sub.n-A.sup.n+1-H with a "double
N-capping group" X=CO, CO--CO, CS, CS--CS or CS--CO. It preferably
contains four amino acid residues and preferably a ureido group
(--NH--CO--NH--) between two of the residues. Thus, the structure
is P--O-A.sup.1-CO-A.sup.2-A.sup.3-A.sup.4-H,
P--O-A.sup.1-A.sup.2-CO-A.sup.3-A.sup.4-H or
P--O-A.sup.1-A.sup.2-A.sup.3-CO-A.sup.4-H, where the amino acid
residue(s) to the right of the ureido group has/have the structure
--NH--CR--CO--, and the residue(s) to the left of the ureido group
is/are inverted to --CO--CR--NH--.
[0017] A.sup.i and A.sup.n+1, most preferably A.sup.1 and/or
A.sup.2 and/or A.sup.3 and/or A.sup.4 may be a residue of an amino
acid, e.g. an L-amino acid, particularly a naturally occurring
amino acid, L-norleucine (Nle), L-norvaline (Nva) or capreomycidine
(Cpd). In particular, A.sup.1 may be Tyr, Leu, Phe, Met, Nle or
Nva; A.sup.2 may be Gly, Ala, Val, Arg, Leu or Cpd; A.sup.3 may be
Ala, Val, Gly, Leu, or Ile; and A.sup.4 may be Phe, Tyr, Leu, Nva,
Nle, Met or Arg.
[0018] P may be H-- or the C-terminal protection group Me-, Et- or
CF.sub.3--
[0019] The stabilizer may have the formula
HO-A.sup.1-CO-A.sup.2-A.sup.3-A.sup.4-H, e.g. the following
compounds (where each amino acid is in the L-form unless
indicated):
TABLE-US-00001 Short name A.sup.1 A.sup.2 A.sup.3 A.sup.4
.alpha.-MAPI Phe Arg Val Phe .beta.-MAPI Phe Arg Val D-Phe
F-CO-RVY-H Phe Arg Val Tyr F-CO-GGY-H Phe Gly Gly Tyr F-CO-GAF-H
Phe Gly Ala Phe F-CO-GAY-H Phe Gly Ala Tyr F-CO-GAL-H Phe Gly Ala
Leu F-CO-GA-Nva-H Phe Gly Ala Nva F-CO-GA-Nle-H Phe Gly Ala Nle
Y-CO-RVY-H Tyr Arg Val Tyr Y-CO-GAY-H Tyr Gly Ala Tyr F-CS-RVF-H
Phe Arg Val Phe F-CS-RVY-H Phe Arg Val Tyr F-CS-GAY-H Phe Gly Ala
Tyr Antipain Phe Arg Val Arg GE20372A Tyr Arg Val Phe GE20372B Tyr
Arg Val D-Phe Chymostatin A Phe Cpd Leu Phe Chymostatin B Phe Cpd
Val Phe Chymostatin C Phe Cpd Ile Phe
[0020] The stabilizer may be prepared by methods known in the art,
e.g. as described by R. J. Broadbridge et al., Chem. Commun., 1998,
1449-1450; by P. Page et al., J. Org. Chem., 1999, 64, 794-799; or
by I. J. Galpin et al., Int. J. Peptide Protein Res., 23, 1984,
477-486.
[0021] The molar ratio of the inhibitor to the protease may be
0.1:1 to 100:1, e.g. 0.5:1-50:1, 1:1-25:1 or 2:1-10:1.
Optional Second Stabilizer
[0022] Optionally, the enzyme or detergent composition may comprise
one or more other protease stabilizers. Examples are boronic acid
derivatives, organic acids, protein and peptide type inhibitors and
peptide aldehydes, e.g. as described in WO 1995/029223, WO
1996/041859, WO 2006/045310, WO 2006/045310, WO 2007/113142, WO
2008/116915, WO 2009/095425 (patent application EP 08150977.0), WO
2009/118375 (patent application EP 08153299.6), WO 2007/145963, or
WO 2007/141736.
Protease
[0023] The protease may be a serine, cysteine or aspartic protease.
It may be an alkaline microbial protease, a trypsin-like protease
or a subtilisin, e.g. a subtilisin derived from Bacillus. The
protease may be subtilisin Novo, subtilisin Carlsberg, subtilisin
BPN', subtilisin 309, subtilisin 147 and subtilisin 168 (described
in WO 89/06279), trypsin (e.g. of porcine or bovine origin), the
Fusarium protease described in WO 89/06270, a protease described in
WO 1998/020115, WO 01/44452, WO 01/58275, WO 01/58276, WO
2003/006602, or WO 2004/099401, or it may have an amino acid
sequence which is at least 90%, at least 95%, at least 98% or at
least 99% identical to one of these.
[0024] Examples of commercially available proteases (peptidases)
include Kannase.TM., Everlase.TM., Esperase.TM., Alcalase.TM.,
Neutrase.TM., Durazym.TM., Savinase.TM., Ovozyme.TM.,
Liquanase.TM., Polarzyme.TM., Pyrase.TM., Pancreatic Trypsin NOVO
(PTN), Bio-Feed.TM. Pro and Clear-Lens.TM.Pro (all available from
Novozymes A/S, Bagsvaerd, Denmark). Other commercially available
proteases include Ronozyme.TM. Pro, Maxatase.TM., Maxacal.TM.,
Maxapem.TM., Opticlean.TM., Properase.TM., Purafect.TM., Purafect
Ox.TM. and Purafact Prime.TM. (available from Danisco Genencor,
Gist-Brocades, BASF, or DSM Nutritional Products).
Second Enzyme
[0025] In addition to the protease, the liquid composition may
optionally comprise one or more other enzymes, e.g. selected among
amylases, lipolytic enzymes (particularly lipases), cellulases,
mannanases and oxidoreductases.
[0026] The amylase may be an alpha-amylase of bacterial or fungal
origin, e.g. an alpha-amylase from B. licheniformis, described in
GB 1,296,839. Commercially available amylases are Duramyl.TM.,
Termamyl.TM., Stainzyme.TM., Stainzyme Plus.TM., Termamyl
Ultra.TM., Fungamyl.TM. and BAN.TM. (available from Novozymes A/S)
and Rapidase.TM., Maxamyl P.TM., Purastar and Purastar OxAm
(available from Gist-Brocades and Danisco Genencor).
[0027] The cellulase may be of bacterial or fungal origin. It may
be a fungal cellulase from Humicola insolens (U.S. Pat. No.
4,435,307) or from Trichoderma, e.g. T. reesei or T. viride.
Examples of cellulases are described in EP 0 495 257. Commercially
available cellulases include Carezyme.TM., Celluzyme.TM.,
Endolase.TM., Celluclean.TM. (available from Novozymes), Puradax,
Puradax HA, and Puradax EG (available from Danisco Genencor).
[0028] The oxidoreductase may be a peroxidase or an oxidase such as
a laccase. The peroxidase may be of plant, bacterial or fungal
origin. Examples are peroxidases derived from a strain of Coprinus,
e.g., C. cinerius or C. macrorhizus, or from a strain of Bacillus,
e.g., B. pumilus, particularly peroxidase according to WO 91/05858.
Suitable laccases herein include those of bacterial or fungal
origin. Examples are laccases from Trametes, e.g., T. villosa or T.
versicolor, or from a strain of Coprinus, e.g., C. cinereus, or
from a strain of Myceliophthora, e.g., M. thermophila.
[0029] The lipolytic enzyme may be a lipase or cutinase of
bacterial or fungal origin. Examples include a lipase from
Thermomyces lanuginosus (Humicola lanuginosa) described in EP 258
068 and EP 305 216, a Rhizomucor miehei lipase, e.g., as described
in EP 238 023, a Candida lipase, such as a C. antarctica lipase,
e.g., the C. antarctica lipase A or B described in EP 214 761, a
Fusarium oxysporum lipase (WO 98/26057), a Pseudomonas lipase such
as a P. pseudoalcaligenes and P. alcaligenes lipase, e.g., as
described in EP 218 272, a P. cepacia lipase, e.g., as described in
EP 331 376, a P. stutzeri lipase, e.g., as disclosed in BP
1,372,034, a P. fluorescens lipase, a Bacillus lipase, e.g., a B.
subtilis lipase (Dartois et al., (1993), Biochemica et Biophysica
acta 1131, 253-260), a B. stearothermophilus lipase (JP 64/744992),
B. pumilus lipase (WO 91/16422), Penicillium camenbertii lipase
(Yamaguchi et al., (1991), Gene 103, 61-67), the Geotrichum
candidum lipase (Shimada, Y. et al., (1989), J. Biochem.
[0030] 106, 383-388), and various Rhizopus lipases such as a R.
delemar lipase (Hass, M. J et al., (1991), Gene 109, 117-113), a R.
niveus lipase (Kugimiya et al., (1992), Biosci. Biotech. Biochem.
56, 716-719) and a R. oryzae lipase. Additional examples are
cutinase from Pseudomonas mendocina (WO 88/09367), cutinase from
Fusarium solani pisi (WO 90/09446) and cutinase from Humicola
insolens (WO 2001/092502). The lipolytic enzyme may be a lipase
variant, e.g. described in WO 2000/060063.
[0031] Examples of commercially available lipases include
Lipex.TM., Lipoprime.TM., Lipopan.TM., Lipopan F.TM., Lipopan
Xtra.TM., Lipolase.TM., Lipolase.TM. Ultra, Lipozyme.TM.,
Palatase.TM., Resinase.TM., Novozym.TM. 435 and Lecitase.TM. (all
available from Novozymes A/S). Other commercially available lipases
include Lumafast.TM. (Pseudomonas mendocina lipase from Danisco
Genencor); Lipomax.TM. (Ps. pseudoalcaligenes lipase from
Gist-Brocades or Danisco Genencor) and Bacillus sp. lipase from
Solvay enzymes. Further lipases are available from other suppliers
such as Lipase P "Amano" (Amano Pharmaceutical Co. Ltd.).
[0032] Examples of mannanases include Mannaway.TM. (product of
Novozymes) and MannaStar (product of Danisco Genencor).
Sequence Identity
[0033] The degree of identity between two amino acid sequences is
calculated as the number of exact matches in an alignment of the
two sequences, divided by the length of the shorter of the two
sequences. The result is expressed in percent identity. An exact
match occurs when the two sequences have identical amino acid
residues in the same positions of the overlap. The length of a
sequence is the number of amino acid residues in the sequence. The
alignment of the two amino acid sequences may be determined by
using the
[0034] Needle program from the EMBOSS package (http://emboss.org)
version 2.8.0. The Needle program implements the global alignment
algorithm described in Needleman, S. B. and Wunsch, C. D. (1970) J.
Mol. Biol. 48, 443-453. The substitution matrix used is BLOSUM62,
gap opening penalty is 10, and gap extension penalty is 0.5.
[0035] Alternatively, the alignment may be done by using the
MegAlign program (version 7) developed by DNASTAR Inc., part of the
Lasergene suite, based on Hein, J. J. (1990). "Unified approach to
alignment and phylogenies." In Methods in Enzymology, Vol. 183: pp.
626-645. Using the Jotun Hein Method and the settings GAP
PENALTY=11, GAP LENGTH PENALTY=3 for multiple alignments and
KTUPLE=2 for pairwise alignments a series of percentage identity
values can be calculated.
Detergent Composition
[0036] The detergent composition is a liquid composition. In
addition to the protease, the composition may further comprise a
second enzyme, particularly a lipase, an amylase, a cellulase or a
carbohydrase.
[0037] The invention is particularly applicable to the formulation
of liquid detergents where enzyme stability problems are
pronounced. The liquid detergent may be aqueous, typically
containing 5-95% water, e.g. 5-15% or 20-70% water and 0-20%
organic solvent (hereinafter, percentages by weight).
[0038] The detergent comprises a surfactant which may be anionic,
non-ionic, cationic, amphoteric or a mixture of these types. The
detergent will usually contain 5-30% anionic surfactant such as
linear alkyl benzene sulphonate (LAS), alpha-olefin sulphonate
(AOS), alcohol ethoxy sulphate (AES) or soap. It may also contain
3-20% anionic surfactant such as nonyl phenol ethoxylate or alcohol
ethoxylate.
[0039] The pH (measured in aqueous detergent solution) will usually
be neutral or alkaline, e.g. 7-10. The detergent may contain 1-40%
of a detergent builder such as zeolite, phosphate, phosphonate,
citrate, NTA, EDTA or DTPA, or it may be unbuilt (i.e. essentially
free of a detergent builder). It may also contain other
conventional detergent ingredients, e.g. fabric conditioners, foam
boosters, bactericides, optical brighteners and perfumes.
[0040] The detergent composition may be a fabric cleaning
compositions, hard surface cleansing compositions, light duty
cleaning compositions including dish cleansing compositions and
automatic dishwasher detergent compositions.
[0041] The liquid detergent composition may comprise from about
0.0001% to about 10%, more particularly from about 0.00015% to
about 1%, and most particularly from about 0.001% to about 0.1% of
the inhibitor
[0042] Thus, a stabilized liquid enzyme formulation typically
contains 0.5-20% by weight, particularily 1-10% by weight, of
enzyme protein (total of protease and optional second enzyme) and
0.01%-10% of the inhibitor, more particularly 0.05-5% by weight and
most particularly 0.1%-2% by weight of the inhibitor.
[0043] A liquid detergent formulation will typically contain
0.04-400 micromolar enzyme (or 1-10000 mg EP/L), more particularily
0.16-160 micromolar enzyme (4-4400 mg EP/L) and most particularly
0.8-80 (20-2200 mg EP/L) and about 1-20 times more of the
inhibitor, most particularly about 1-10 times more of the
inhibitor.
[0044] The liquid detergent composition may contain water and other
solvents as carriers. Low molecular weight primary or secondary
alcohols exemplified by methanol, ethanol, propanol, and
iso-propanol are suitable. Monohydric alcohols are preferred for
solubilizing surfactants, but polyols such as those containing from
about 2 to about 6 carbon atoms and from about 2 to about 6 hydroxy
groups (e.g., 1,3-propanediol, ethylene glycol, glycerine, and
1,2-propanediol) can also be used. The compositions may contain
from about 5% to about 90%, typically from about 10% to about 50%
of such carriers.
[0045] The detergent compositions herein will preferably be
formulated such that during use in aqueous cleaning operations, the
wash water will have a pH between about 6.8 and about 11. Finished
products are typically formulated at this range. Techniques for
controlling pH at recommended usage levels include the use of, for
example, buffers, alkalis, and acids. Such techniques are well
known to those skilled in the art.
[0046] When formulating the hard surface cleaning compositions and
fabric cleaning compositions of the present invention, the
formulator may wish to employ various builders at levels from about
5% to about 50% by weight. Typical builders include the 1-10 micron
zeolites, polycarboxylates such as citrate and oxydisuccinates,
layered silicates, phosphates, and the like. Other conventional
builders are listed in standard formularies.
EXAMPLES
Example 1
[0047] Various peptide aldehydes of the invention were produced by
a custom peptide synthesis company, all with a purity >80%. The
peptide aldehydes were dissolved in DMSO to a concentration of 10
mg/ml before use.
[0048] A model liquid detergent A was prepared for testing the
various stabilizers:
TABLE-US-00002 Component % w/w Sodium alkylethoxy sulphate (C9-15,
2EO) 6.0 Sodium dodecyl benzene sulphonate 3.0 Sodium toluene
sulphonate 3.0 Oleic acid 2.0 Primary alcohol ethoxylate (C12-15,
7EO) 3.0 Primary alcohol ethoxylate (C12-15, 3EO) 2.5 Ethanol 0.5
Monopropylene glycol 2.0 Tri-sodium citrate 2H.sub.2O 4.0
Triethanolamine 0.4 Protease (Savinase 16 LEX) 0.5 Lipase (Lipex
100L) 0.5 De-ionized water Ad 100% pH adjusted to 8.5 with NaOH
[0049] Further, the following detergents were prepared by adding
stabilizers to detergent A:
TABLE-US-00003 Stabilizer Molar ratio of (from a 10 mg/ml inhibitor
relative Detergent ID Detergent A solution) to protease B 100 g 1.5
mg F-CO-RVF-H 3 C 100 g 2.5 mg F-CO-RVF-H 5 D 100 g 5.0 mg
F-CO-RVF-H 10 E 100 g 1.5 mg F-CO-RVY-H 3 F 100 g 2.6 mg F-CO-RVY-H
5 G 100 g 5.1 mg F-CO-RVY-H 10 H 100 g 1.2 mg F-CO-GGY-H 3 J 100 g
2.0 mg F-CO-GGY-H 5 K 100 g 3.9 mg F-CO-GGY-H 10
[0050] The detergents were placed in closed glasses at 35.degree.
C. and 40.degree. C. Residual activity of lipase and protease was
measured (by comparison to a reference stored at -18.degree. C.) at
different times, using standard enzyme analytical methods available
at Novozymes (protease measured by hydrolysis of
N,N--dimethylcasein at 40.degree. C., pH 8.3 and lipase measured by
hydrolysis of p-nitrophenyl valerate at 40.degree. C., pH 7.7).
TABLE-US-00004 % residual activity Residual protease Residual
lipase activity activity 4 weeks 1 week 1 week Detergent 35.degree.
C. 40.degree. C. 35.degree. C. A (reference) 20 11 3 B (F-CO-RVF-H,
3x) 63 22 C (F-CO-RVF-H, 5x) 83 51 D (F-CO-RVF-H, 10x) 78 69 E
(F-CO-RVY-H, 3x) 62 55 17 F (F-CO-RVY-H, 5x) 81 75 53 G
(F-CO-RVY-H, 10x) 86 81 69 H (F-CO-GGY-H, 3x) 26 17 3 J
(F-CO-GGY-H, 5x) 31 20 4 K (F-CO-GGY-H, 10x) 38 28 5
[0051] From this example it is clear that the peptide aldehydes of
the invention are very efficient as protease stabilizers and as
stabilizers for a second enzyme in the presence of a protease.
* * * * *
References