CA1288368C - Enzymatic detergent composition - Google Patents
Enzymatic detergent compositionInfo
- Publication number
- CA1288368C CA1288368C CA000553755A CA553755A CA1288368C CA 1288368 C CA1288368 C CA 1288368C CA 000553755 A CA000553755 A CA 000553755A CA 553755 A CA553755 A CA 553755A CA 1288368 C CA1288368 C CA 1288368C
- Authority
- CA
- Canada
- Prior art keywords
- lipase
- detergent
- lipases
- pseudomonas
- detergent composition
- 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.)
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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
- C11D3/38627—Preparations containing enzymes, e.g. protease or amylase containing lipase
Abstract
ABSTRACT OF THE DISCLOSURE
The invention relates to a detergent composition on the basis of nonionic detergents, which includes a mixture of certain lipolytic enzymes and proteases. In this composition, the lipases remain unaffected by the proteases.
The invention relates to a detergent composition on the basis of nonionic detergents, which includes a mixture of certain lipolytic enzymes and proteases. In this composition, the lipases remain unaffected by the proteases.
Description
C 7094 ( R ) ~.2,~83~;~
ENZYMATIC DETERGENT COMPOSITION
~he present invention relates to an enzymatic detergent composition comprising as detergent active material solely a nonionic synthetic detergent, and a ~ixture of lipolytic and proteolytic enzymes.
Yroteolytic enzymes are well known in detergent compositions. Lipolytic enzymes have been mentioned in the prior art as possible enzymes for detergent compositions, but there is relatively little prior art directly concerned with lipases for inclusion in detergent compositions.
Insofar the compatibility of proteolytic and lipolytic enzymes with synthetic detergents is concerned, it is well known that nonionic synthetic detergents in general do not negatively in~luence the activity o proteolytic and lipolytic enzymes, in contrast with e.g. anionic synthetic detergents, which often negatively influence the proteases and lipases.
However, since lipases ar.e basically proteins, they would be susceptible to proteolytic action by the proteases if lipa~es were used in conjunction with proteases. It would therefore be expected that the inclu~ion of proteases in a detergent composi tion comprising lipases and a~ detergent-active material solely a nonionic detergent surfactant (which does not affect either the lipase or the protease), would cause the lipa3e activity to be destroyed by the proteolytic action of the proteases.
We have now surprisingly found that when using such compositions comprising a special class of lipases, hereinafter ~ore specifically de~ined, this expected loqs of lipolytic activity does not occur, but that, ~"
~ 88368 C 7094 (R) the activity of these lipases remains almost unaffected by the presence of the protea~es in these composition~.
The cla~s of lipa~es to be used according to the present invention embraces those lipases which show a po~itive immunological cross-reaction with the antibody of the lipase produced by the microorganism Chromobacter viscosum var. lipolyticum NRRL B 3673.
This lipase has been described in Dutch Patent Specification 154,269 of Toyo Jozo KK, and the microorganism is available to the public from the collection of the United States Department of Agriculture, Agricultural Research Service, Northern Utilization and Development Division, Peoria, Illinois, under N NRRL B 3673. This lipase will be referred to hereinafter as "Toyo Jozo" lipase. The lipases of the present invention should show a positive immunological cross-reaction with the Toyo Jozo lipase antibody, using the standard and well-known immunodiffusion procedure according to Ouchterlony (Acta. Med 5can., 133, pages 76-79 (1950)).
The preparation of the anti~erum is carried o~t as follows:
Equal volumes of 0.1 mg/ml antigen and of Freund's adjuvant (complete or incomplete) are mixed until an emulsion i5 obtained. Two female rabbits are injected with 2 ml samples of the emulsion according to the following scheme:
day O : antigen in complete Freund'~ adjuvant day 4 : antigen in complete Freund's adjuvant day 32 : antigen in incomplete Freund's adjuvant day 60 : booster of antigen in incomplete Freund's adjuvant.
The serum containing the req~ired antibody is prepared ~ 288368 C 7094 (R) . .
hy centrifugation o~ clotted blood, taken on day 67.
The titre of the anti-Toyo Jozo-lipase antiserum is determined by the inspection of precipitation of serial dilutions of antigen and antiserum according to the Ouchterlony procedure. A 25 dilution of antiserum was the dilution that still gave a visible precipitation with an antigen concentration of 0.1 mg/ml.
All lipases ~howing a positive immunological cross~
reaction with the Toyo Jozo lipase antibody as hereabove described are lipases according to the present invention. Typical examples thereof are the lipase ex Pseudomonaq fluorescens IAM 1057 (available under the trade name*Amano-P), the lipase ex Pseudomona~ fragi FERM P 1339 (available under the mano-B), lipase ex Pseudomonas nitroreducens var. lipolyticum FERM P 1338, the lipase _ ex Pseudomonas sp. available under the trade name ~Amano-CES, lipa~e ex Pseudomonas cepacia, lipases ex Chromobacter viscosum, e.g. Chromobacter viscosum var.
lipolyticum NRRL B 3673, commercially available from Toyo Jozo Co., Tagata, Japan; and further C. viscosum lipases from US Biochemical Corp., U.S.A., and Diosynth Co., The ~etherland~, and lipa9es ex P~eudomonas gladioli.
The lipase~ of the pre~ent invention are included in the detergent compoqition in such an amount that the final detergent compoqition has a lipolytic enæyme activity of from 100 to 0.005 LU/mg, preferably 25 to 0.05 LU/mg of the composition.
A Lipase Unit (LU) is that amount of lipa~e which produces l/umol o~ titratable fatty acid per minute in a pH stat. under the following conditions:
temperature 30C, pH = 9.0: ~ubstrate is an emulsion of .... ~ ... ,.. ~.. .....
~ 38368 C 709~ (R) 3.3 wt.% of olive oil and 3.3% gum arabic, in the pre~ence of 13 mmol/l Ca2~ and 20 mmol/l NaCl in 5 mmol/l Tri~-buffer.
Naturally, mixtures of the above lipases can be used.
The lipases can be used in their non-purified form or in a purified form, e.g. purified with the aid of well-known adsorption methodq, such as phenyl ~ epharose adsorption techniques.
The proteases used according to the present invention are the well-known detergent proteases and mixtures thereof. Typical examples are the subtilisin type protea~es, such as~Alcalase, ~sperase, Savinase, all available from Novo Industri A/S, Maxatase and~Maxacal, both available from Gist Brocades N.V., Kazuzase ex Showa Denko. (= API-21 = AP-l) In general, the proteases are used in such an amount that the final detergent composition has a proteolytic activity of 0.1 to 50 GU/mg, preferably 0.5 to 30 GV/mg. (GU = Glycine Unit; one GU is the amount of enzyme which under standard incubation conditions produces an amount of terminal NH2-gro~ps equivalent to 1 microgr./ml glycine.) The detergent compo~ition further comprises a~
detergent-active material solely a nonionic synthetic detergent. This type of detergent is well known in the art. Nonionics usually consist of a hydrophobic moiety derived from fatty alcohols, fatty acid amides and alkylphenols, and a hydrophilic moiety consisting of alkylene oxide units. Typical examples are the condenQation products of alkylene oxides such as ethylene oxide, propylene oxide and butylene oxide and mixtures thereo~ with C8-C18 primary or secondary, branched or straight chain alcohols, C8-Cl~ fatty e~teS ~Lra C~ r~a r~
~; , " , - , . ..
.
C 7094 (R) 336~3 acid amides, Cg-Cl8 alkylphenolq, etc. Further suitable example~ are given in M. Schick "Nonianic surfactants" (1967).
In general, the composition contains from 1-50~, usually from 2-45%, and commonly from 5-30% by weight of the nonionic detergent. Mixtures of various nonionic detergents may also be used.
The detergent composition may furthermore include the usual detergent ingredients in the usual amounts. Thus, the composition may contain from 1-60~, preferably from 5-30~ by weight of one or more organic and/or inorganic builders. Furthermore, it may contain from 1-35~ of a bleaching agent or a bleaching system comprising a bleaching agent and an activator therefor, e.g. sodium perborate with tetraacetyl ethylene diamine.
The compositions may furthermore comprise lather boosters, foam depressors, anti-corrosion agents, soil-suspending agents, sequestering agents, anti-soil redeposition agents, perfumes, dyes, solvents, stabilising agents for the enzymes and bleaching agents and so on. They may also comprise from 0.01-10~ by weight of enzymes other than lipases and proteases, such as amylases, oxidases and cellulases.
The composition~ of the present invention can be formulated in any desired form, such as powders, bars, pastes, liquids, etc.
The invention will now further be illustrated by way of Examples.
~ 8368 C 7094 (R) Example 1 Washing experiments were carried out in a Tergotometer under the following conditions:
wa3h time and temperature: 10 minutes at 40C:
three rinses with cold water water hardness: 14 and 40FH
detergent composition concentration: 2 and 6 g/l number soil/wash cycles : 4 protease concentration: 20 GU/ml lipase concentration: 1 LU/ml As lipa~es were used: Amano-P, Diosynth and a lipase ex Pseudomonas gladioli test cloth: cotton ] soiled with AS ~ /
polyester/cotton ] groundnut oil /
polyester ] milk powder The detergent composition used had the following formulation:
~ by weight C12-C14 linear primary alcohol condensed 9 with 7 moles of ethylene oxide sodium silicate 8 sodium carbonate 39 sodium sulphate 38 fluorescers, dyes, perfume~ 0.3 water q.~.
The reflectance of the test cloth~ and the residual weight percentage of fatty material on the test cloths were determined after the fourth wash cycle. The reflectance was measured in a Reflectometer at 460 nm with a UV filter in the light pathway, and the fatty matter by extracting the dried test cloths with petroleum ether, and determining the amount of ~atty matter from the weight loss of the test cloth.
~ c~en~t~s )~ac~
. .
C 7094 (R) ~3~
The following results were obtained:
Savin-Cotton PE/Cotton Polyester lipase ase~ FH R 460 % FM R 460 ~ FM R 460 ~ FM
Amano-P - 2 14 84.5 3.9 72.6 2.6 79.8 0.8 2 14 87.1 3.6 75.4 2.4 80.5 0.7 - 6 40 81.5 3.7 71.3 2.4 79.5 0.7 + 6 40 86.5 3.0 75.4 1.9 81.7 0.4 Gladioli - 2 14 31.1 4.6 68.5 4.5 74.5 4.3 + 2 14 86.5 3.6 72.3 3.8 76.2 4.0 - 6 40 76.6 4.7 68.0 3.7 75.9 3.4 + 6 40 83.8 3.6 73.3 2.8 79.1 3.0 Diosynth - 2 14 83.2 4.0 71.3 3.3 75.6 3.5 + -2 14 86.8 3.6 74.3 2.8 77.2 3.0 - 6 40 79.8 3.7 70.4 2.8 76.9 2.7 + 6 40 85.4 2.9 74.7 2.1 79.8 2.1 Cepacia - 2 14 83.8 4.1 71.2 3.4 78.3 2.8 + 2 14 86.9 3.6 75.2 2.9 79.4 2.3 - 6 40 79.6 4.1 70.0 3.2 78.8 1.9 + 6 40 85.9 3.2 74.9 2.2 81.5 1.0 _ ~ ler~o~es ~~ e f~?~r k C 7094 (R) Example 2 The procedure of Example 1 was repeated, but under the following conditions:
detergent composition concentration: 2 g/l number of soil/wash cycles : 4 wash time and temperature: 10 minutes at 40C, three rinses with cold water water hardness: 14FH
agitation: 100 rpm lipase: ex Pseudomonas gladioli 1 LU/ml protease: 20 GU/ml As proteases, Savinase, Alcalase, Maxatase, Maxacal, ~Kazuzase and Esperase were used.
The following results were obtained:
without lipase with lipase Protea~e R460 % F~ R460 ~ FM
_ . _ Savinase 69.3 5.6 87.2 2.8 Alcalase 69.3 5.4 87.2 2.8 Maxatase 70.0 5.3 87.5 2.8 Maxacal 68.9 5.7 87.1 2.8 25 Kazuzase 69.1 5.7 87.1 2.7 Esperase 68.6 5.7 87.1 2.7 ~e~o~e-S 7Lrc~G~!e rnc(rk C 7094 (R) ~.2~ 68 g Example 3 The stability of lipases according to the present invention in the presence of protea~es and varying amounts of a nonionic detergent was tested by determining the residual lipase activity with the pH-~tat. method. A composition comprising 0.7 g pentasodium triphosphate, 0.7 g sodium sulphate and 0.2 sodium silicate and varying amounts of C13-C15 alcohol, condensed with 7 and 3 moles of ethylene oxide, respectively (product A and product B) was used.
The temperature was 30C, the water hardness 14FH and the pH 9.5.
The lipase was either lipase ex Pseudomonas gladioli, or A~ano-P, at-15 LU/ml, and the protease was Savinase, at 20 GU/ml.
The following results were obtained:
_ Product A Product B _ lipa~e 9/1 i RA % RA 9/1~ RA % RA
non _ 10 min. 30_min.non 10 min. 30 min.
Pseudomonas 0 75 34 0 69 29 gladioli 0.2598 103 0.2598 100 1.0 103 103 1.0 98 105 2.0 99 102 2.0 95 90 4.0 98 98 4.0 110 108 _________,________ ________________ Amano-P 0 100 74 0.25120 116 1.0 104 101 3S 2.0 101 101 4.0 108 107 % RA = ~ residual activity o~ the lipase ~.2~33~i8 C 7094 (R) , . ~
Example 4 Example 1 was repeated, using various lipases. ~he following results were obtained:
Savin-Cotton PE/Cotton Polyester lipase ase~ FH R 460 ~ FM R 460 % FM R 460 ~ FM
No - 2 14 72.1 5.9 60.6 6.6 67.9 4.7 + 2 14 72.0 6.1 60.0 6.6 67.1 4.8 - 6 40 71.8 5.3 59.7 5.4 69.9 3.5 + 6 40 74.6 5.2 62.8 5.2 72.2 3.4 _ lipase MY - 2 14 69.2 6.0 57.8 6.4 66.1 4.8 15(ex Cand. + 2 14 72.7 5.9 60.1 6.5 67.1 5.0 cyl.) - 6 40 70.1 5.6 59.8 5.3 69.4 3.4 ~ 6 40 73.7 5.3 62.4 5.4 69.6 3.7 -lip. ~P-6 - 2 14 69.0 6.0 57.4 6.5 66.4 4.8 20(ex Aqp. + 2 14 72.1 6.0 59.5 6.6 67.3 5.0 niger) - 6 40 71.3 5.5 59.5 5.4 69.8 3.5 + 6 40 75.1 5.4 61.6 5.4 71.7 3.6 lip. SP285 - 2 14 77.8 4.5 65.8 4.1 70.2 3.6 (ex I 2 14 82.0 4.2 69.5 3.7 71.3 3.6 Fusarium - 6 40 74.8 4.8 64.1 4.4 71.8 3.2 oxysporum) + 6 40 75.0 5.0 64.8 4.4 72.0 3.4 -lip. SP225 - 2 14 74.9 4.4 64.5 4.2 76.3 2.8 30(ex Mucor + 2 14 79.3 3.5 67.4 3.8 76.6 2.5 miehei~ - 6 40 71.7 4.9 61.3 4.4 75.5 1.7 + 6 40 72.3 4.8 62.8 4.~ 76.7 1.9 _ .
Toyo Jozo - 2 14 86.2 2.7 71.3 2.4 78.6 1.7 35(lip. ex ~ 2 14 85.8 2.7 71.9 2.4 78.2 1.7 Chromobac. - 6 40 84.9 2.8 69.9 1.8 77.4 1.4 viscos~m) + 6 40 85.2 2.7 72.5 1.9 77.8 1.5 38368 c 7094 (R) Savin- Cotton PE/Cotton Polyester ~ase ase ~FHR 46096 FMR 460~6 FMR 46096 FM
Nagase - 2 1485.0 3.2 69.0 2.6 77.1 1.7 S (ex - + 21486.42.4 71.9 2.378.3 1.1 Rhiæopus) - 6 4071.7 5.1 61.7 4.5 76.0 1.5 64072.65.2 62.1 4.376.1 1.8 -C-4 (ex - 21472.15.5 60.8 5.374.4 3.4 10 Candida +2 1470.8 5.2 61.2 5.4 73.5 3.4 cylin- - 64072.15.1 61.4 4.675.4 1.7 dracea) +6 4073.2 5.3 62.4 4.6 75.4 2.0 pancreas - 2 1472.8 5.2 62.4 5.2 74.5 3.5 15 lipase ~ 21473.25.2 62.8 5.174.3 3.4 - 64072.95.3 62.3 4.675.9 1.7 + 64072.75.2 63.1 4.576.4 1.8 Esterase - 2 1472.6 5.3 61.9 5.2 75.3 3.3 20 MM (ex + 21471.95.2 62.7 4.975.4 3.2 Mucor - 64072.05.3 61~4 4.576.3 1.6 mieh~i) +6 4073.3 5.3 63.0 4.4 76.1 1.7 Amano-B - 21484.13.0 71.3 2.278.5 0.8 + 21485.83.1 71.5 2.779.2 0.9 - 64085.23.1 71.3 1.978.4 0.3 + 64086.33.0 72.8 1.979.4 0.2 Amano-CES - 2 1480.1 2.8 71.0 1.9 79.0 0.4 + 21481.42.9 73.0 1.879.7 0.4 - 64080.03.0 71.3 1.678.9 0.6 + 64082.03.0 73.3 1.779.0 0.8 Amano-CE - 2 1472.1 5.1 64.1 4.5 75.4 3.3 + 21472.75.1 66.6 4.175.8 3.3 - 64074.94.3 67.5 3.077.0 2.2 + 64077.54.0 69.0 2.977.2 2.4 -~s ~roLl~ rnQ~k .
~..2~8~68 C 7094 (R) Savin-Cotton PE/Cotton Polyester lipase ase~ FH R 460 % FM R 460 ~ FM R 460 ~ FM
lipase - 2 14 68.9 5.8 61.8 5.2 74.7 3.5 M-AP10 + 2 14 70.7 5.8 63.1 5.2 75.2 3.6 teX Mucor - 6 40 71.4 5.1 66.3 3.7 76.6 2.3 javanicus) + 6 40 74.2 4.5 66.2 4.1 75.9 2.5 lipase - 2 14 73.9 4.7 64.8 4.2 76.2 3.1 F-AP (ex + 2 14 72.1 5.2 65.5 4.7 75.7 3.4 Rhizopus - 6 40 74.7 4.6 69.4 2.4 77.1 1.9 javanicus) + 6 40 74.6 4.9 68.1 3.3 77.3 2.2 S80.000 - 2 14 72.1 5.1 66.0 4.3 76.1 3.1 (ex + 2 14 74.6 4.8 66.5 4.4 76.4 3.3 Rhizopus - 6 40 75.6 4.5 69.1 2.4 77.2 2.0 arrhizus) + 6 40 78.8 4.5 71.1 2.5 78.0 1.9 PS-54 (ex - 2 14 76.0 4.3 67.0 3.6 75.3 3.0 Thermom. + 2 14 77.1 4.4 68.9 3.7 75.6 2.9 lanu- - 6 40 78.1 4.3 69.5 2.1 76.7 2.0 ginosus) ~ 6 40 79.6 4.2 72.1 2.4 78.1 1.8 :
ex Pqeudo. - 2 14 79.6 3.2 70.3 2.0 78.4 0.4 gladioli ~ 2 14 81.1 3.0 72.1 2.1 79.9 0.4 - 6 40 79.6 3.5 69.6 1.9 77.8 0.1 + 6 40 81.0 3.4 71.7 1.8 79.7 0.2 lipase ex - 2 14 77.3 4.2 67.4 3.7 74.9 3.3 Rhizopus ~ 2 14 79.7 4.0 68.8 3.5 75.g 3.0 sp. ~ 6 40 77.4 3.7 68.1 2.4 77.8 1.7 + 6 40 80.7 4.1 71.0 2.3 78.9 1.8 -
ENZYMATIC DETERGENT COMPOSITION
~he present invention relates to an enzymatic detergent composition comprising as detergent active material solely a nonionic synthetic detergent, and a ~ixture of lipolytic and proteolytic enzymes.
Yroteolytic enzymes are well known in detergent compositions. Lipolytic enzymes have been mentioned in the prior art as possible enzymes for detergent compositions, but there is relatively little prior art directly concerned with lipases for inclusion in detergent compositions.
Insofar the compatibility of proteolytic and lipolytic enzymes with synthetic detergents is concerned, it is well known that nonionic synthetic detergents in general do not negatively in~luence the activity o proteolytic and lipolytic enzymes, in contrast with e.g. anionic synthetic detergents, which often negatively influence the proteases and lipases.
However, since lipases ar.e basically proteins, they would be susceptible to proteolytic action by the proteases if lipa~es were used in conjunction with proteases. It would therefore be expected that the inclu~ion of proteases in a detergent composi tion comprising lipases and a~ detergent-active material solely a nonionic detergent surfactant (which does not affect either the lipase or the protease), would cause the lipa3e activity to be destroyed by the proteolytic action of the proteases.
We have now surprisingly found that when using such compositions comprising a special class of lipases, hereinafter ~ore specifically de~ined, this expected loqs of lipolytic activity does not occur, but that, ~"
~ 88368 C 7094 (R) the activity of these lipases remains almost unaffected by the presence of the protea~es in these composition~.
The cla~s of lipa~es to be used according to the present invention embraces those lipases which show a po~itive immunological cross-reaction with the antibody of the lipase produced by the microorganism Chromobacter viscosum var. lipolyticum NRRL B 3673.
This lipase has been described in Dutch Patent Specification 154,269 of Toyo Jozo KK, and the microorganism is available to the public from the collection of the United States Department of Agriculture, Agricultural Research Service, Northern Utilization and Development Division, Peoria, Illinois, under N NRRL B 3673. This lipase will be referred to hereinafter as "Toyo Jozo" lipase. The lipases of the present invention should show a positive immunological cross-reaction with the Toyo Jozo lipase antibody, using the standard and well-known immunodiffusion procedure according to Ouchterlony (Acta. Med 5can., 133, pages 76-79 (1950)).
The preparation of the anti~erum is carried o~t as follows:
Equal volumes of 0.1 mg/ml antigen and of Freund's adjuvant (complete or incomplete) are mixed until an emulsion i5 obtained. Two female rabbits are injected with 2 ml samples of the emulsion according to the following scheme:
day O : antigen in complete Freund'~ adjuvant day 4 : antigen in complete Freund's adjuvant day 32 : antigen in incomplete Freund's adjuvant day 60 : booster of antigen in incomplete Freund's adjuvant.
The serum containing the req~ired antibody is prepared ~ 288368 C 7094 (R) . .
hy centrifugation o~ clotted blood, taken on day 67.
The titre of the anti-Toyo Jozo-lipase antiserum is determined by the inspection of precipitation of serial dilutions of antigen and antiserum according to the Ouchterlony procedure. A 25 dilution of antiserum was the dilution that still gave a visible precipitation with an antigen concentration of 0.1 mg/ml.
All lipases ~howing a positive immunological cross~
reaction with the Toyo Jozo lipase antibody as hereabove described are lipases according to the present invention. Typical examples thereof are the lipase ex Pseudomonaq fluorescens IAM 1057 (available under the trade name*Amano-P), the lipase ex Pseudomona~ fragi FERM P 1339 (available under the mano-B), lipase ex Pseudomonas nitroreducens var. lipolyticum FERM P 1338, the lipase _ ex Pseudomonas sp. available under the trade name ~Amano-CES, lipa~e ex Pseudomonas cepacia, lipases ex Chromobacter viscosum, e.g. Chromobacter viscosum var.
lipolyticum NRRL B 3673, commercially available from Toyo Jozo Co., Tagata, Japan; and further C. viscosum lipases from US Biochemical Corp., U.S.A., and Diosynth Co., The ~etherland~, and lipa9es ex P~eudomonas gladioli.
The lipase~ of the pre~ent invention are included in the detergent compoqition in such an amount that the final detergent compoqition has a lipolytic enæyme activity of from 100 to 0.005 LU/mg, preferably 25 to 0.05 LU/mg of the composition.
A Lipase Unit (LU) is that amount of lipa~e which produces l/umol o~ titratable fatty acid per minute in a pH stat. under the following conditions:
temperature 30C, pH = 9.0: ~ubstrate is an emulsion of .... ~ ... ,.. ~.. .....
~ 38368 C 709~ (R) 3.3 wt.% of olive oil and 3.3% gum arabic, in the pre~ence of 13 mmol/l Ca2~ and 20 mmol/l NaCl in 5 mmol/l Tri~-buffer.
Naturally, mixtures of the above lipases can be used.
The lipases can be used in their non-purified form or in a purified form, e.g. purified with the aid of well-known adsorption methodq, such as phenyl ~ epharose adsorption techniques.
The proteases used according to the present invention are the well-known detergent proteases and mixtures thereof. Typical examples are the subtilisin type protea~es, such as~Alcalase, ~sperase, Savinase, all available from Novo Industri A/S, Maxatase and~Maxacal, both available from Gist Brocades N.V., Kazuzase ex Showa Denko. (= API-21 = AP-l) In general, the proteases are used in such an amount that the final detergent composition has a proteolytic activity of 0.1 to 50 GU/mg, preferably 0.5 to 30 GV/mg. (GU = Glycine Unit; one GU is the amount of enzyme which under standard incubation conditions produces an amount of terminal NH2-gro~ps equivalent to 1 microgr./ml glycine.) The detergent compo~ition further comprises a~
detergent-active material solely a nonionic synthetic detergent. This type of detergent is well known in the art. Nonionics usually consist of a hydrophobic moiety derived from fatty alcohols, fatty acid amides and alkylphenols, and a hydrophilic moiety consisting of alkylene oxide units. Typical examples are the condenQation products of alkylene oxides such as ethylene oxide, propylene oxide and butylene oxide and mixtures thereo~ with C8-C18 primary or secondary, branched or straight chain alcohols, C8-Cl~ fatty e~teS ~Lra C~ r~a r~
~; , " , - , . ..
.
C 7094 (R) 336~3 acid amides, Cg-Cl8 alkylphenolq, etc. Further suitable example~ are given in M. Schick "Nonianic surfactants" (1967).
In general, the composition contains from 1-50~, usually from 2-45%, and commonly from 5-30% by weight of the nonionic detergent. Mixtures of various nonionic detergents may also be used.
The detergent composition may furthermore include the usual detergent ingredients in the usual amounts. Thus, the composition may contain from 1-60~, preferably from 5-30~ by weight of one or more organic and/or inorganic builders. Furthermore, it may contain from 1-35~ of a bleaching agent or a bleaching system comprising a bleaching agent and an activator therefor, e.g. sodium perborate with tetraacetyl ethylene diamine.
The compositions may furthermore comprise lather boosters, foam depressors, anti-corrosion agents, soil-suspending agents, sequestering agents, anti-soil redeposition agents, perfumes, dyes, solvents, stabilising agents for the enzymes and bleaching agents and so on. They may also comprise from 0.01-10~ by weight of enzymes other than lipases and proteases, such as amylases, oxidases and cellulases.
The composition~ of the present invention can be formulated in any desired form, such as powders, bars, pastes, liquids, etc.
The invention will now further be illustrated by way of Examples.
~ 8368 C 7094 (R) Example 1 Washing experiments were carried out in a Tergotometer under the following conditions:
wa3h time and temperature: 10 minutes at 40C:
three rinses with cold water water hardness: 14 and 40FH
detergent composition concentration: 2 and 6 g/l number soil/wash cycles : 4 protease concentration: 20 GU/ml lipase concentration: 1 LU/ml As lipa~es were used: Amano-P, Diosynth and a lipase ex Pseudomonas gladioli test cloth: cotton ] soiled with AS ~ /
polyester/cotton ] groundnut oil /
polyester ] milk powder The detergent composition used had the following formulation:
~ by weight C12-C14 linear primary alcohol condensed 9 with 7 moles of ethylene oxide sodium silicate 8 sodium carbonate 39 sodium sulphate 38 fluorescers, dyes, perfume~ 0.3 water q.~.
The reflectance of the test cloth~ and the residual weight percentage of fatty material on the test cloths were determined after the fourth wash cycle. The reflectance was measured in a Reflectometer at 460 nm with a UV filter in the light pathway, and the fatty matter by extracting the dried test cloths with petroleum ether, and determining the amount of ~atty matter from the weight loss of the test cloth.
~ c~en~t~s )~ac~
. .
C 7094 (R) ~3~
The following results were obtained:
Savin-Cotton PE/Cotton Polyester lipase ase~ FH R 460 % FM R 460 ~ FM R 460 ~ FM
Amano-P - 2 14 84.5 3.9 72.6 2.6 79.8 0.8 2 14 87.1 3.6 75.4 2.4 80.5 0.7 - 6 40 81.5 3.7 71.3 2.4 79.5 0.7 + 6 40 86.5 3.0 75.4 1.9 81.7 0.4 Gladioli - 2 14 31.1 4.6 68.5 4.5 74.5 4.3 + 2 14 86.5 3.6 72.3 3.8 76.2 4.0 - 6 40 76.6 4.7 68.0 3.7 75.9 3.4 + 6 40 83.8 3.6 73.3 2.8 79.1 3.0 Diosynth - 2 14 83.2 4.0 71.3 3.3 75.6 3.5 + -2 14 86.8 3.6 74.3 2.8 77.2 3.0 - 6 40 79.8 3.7 70.4 2.8 76.9 2.7 + 6 40 85.4 2.9 74.7 2.1 79.8 2.1 Cepacia - 2 14 83.8 4.1 71.2 3.4 78.3 2.8 + 2 14 86.9 3.6 75.2 2.9 79.4 2.3 - 6 40 79.6 4.1 70.0 3.2 78.8 1.9 + 6 40 85.9 3.2 74.9 2.2 81.5 1.0 _ ~ ler~o~es ~~ e f~?~r k C 7094 (R) Example 2 The procedure of Example 1 was repeated, but under the following conditions:
detergent composition concentration: 2 g/l number of soil/wash cycles : 4 wash time and temperature: 10 minutes at 40C, three rinses with cold water water hardness: 14FH
agitation: 100 rpm lipase: ex Pseudomonas gladioli 1 LU/ml protease: 20 GU/ml As proteases, Savinase, Alcalase, Maxatase, Maxacal, ~Kazuzase and Esperase were used.
The following results were obtained:
without lipase with lipase Protea~e R460 % F~ R460 ~ FM
_ . _ Savinase 69.3 5.6 87.2 2.8 Alcalase 69.3 5.4 87.2 2.8 Maxatase 70.0 5.3 87.5 2.8 Maxacal 68.9 5.7 87.1 2.8 25 Kazuzase 69.1 5.7 87.1 2.7 Esperase 68.6 5.7 87.1 2.7 ~e~o~e-S 7Lrc~G~!e rnc(rk C 7094 (R) ~.2~ 68 g Example 3 The stability of lipases according to the present invention in the presence of protea~es and varying amounts of a nonionic detergent was tested by determining the residual lipase activity with the pH-~tat. method. A composition comprising 0.7 g pentasodium triphosphate, 0.7 g sodium sulphate and 0.2 sodium silicate and varying amounts of C13-C15 alcohol, condensed with 7 and 3 moles of ethylene oxide, respectively (product A and product B) was used.
The temperature was 30C, the water hardness 14FH and the pH 9.5.
The lipase was either lipase ex Pseudomonas gladioli, or A~ano-P, at-15 LU/ml, and the protease was Savinase, at 20 GU/ml.
The following results were obtained:
_ Product A Product B _ lipa~e 9/1 i RA % RA 9/1~ RA % RA
non _ 10 min. 30_min.non 10 min. 30 min.
Pseudomonas 0 75 34 0 69 29 gladioli 0.2598 103 0.2598 100 1.0 103 103 1.0 98 105 2.0 99 102 2.0 95 90 4.0 98 98 4.0 110 108 _________,________ ________________ Amano-P 0 100 74 0.25120 116 1.0 104 101 3S 2.0 101 101 4.0 108 107 % RA = ~ residual activity o~ the lipase ~.2~33~i8 C 7094 (R) , . ~
Example 4 Example 1 was repeated, using various lipases. ~he following results were obtained:
Savin-Cotton PE/Cotton Polyester lipase ase~ FH R 460 ~ FM R 460 % FM R 460 ~ FM
No - 2 14 72.1 5.9 60.6 6.6 67.9 4.7 + 2 14 72.0 6.1 60.0 6.6 67.1 4.8 - 6 40 71.8 5.3 59.7 5.4 69.9 3.5 + 6 40 74.6 5.2 62.8 5.2 72.2 3.4 _ lipase MY - 2 14 69.2 6.0 57.8 6.4 66.1 4.8 15(ex Cand. + 2 14 72.7 5.9 60.1 6.5 67.1 5.0 cyl.) - 6 40 70.1 5.6 59.8 5.3 69.4 3.4 ~ 6 40 73.7 5.3 62.4 5.4 69.6 3.7 -lip. ~P-6 - 2 14 69.0 6.0 57.4 6.5 66.4 4.8 20(ex Aqp. + 2 14 72.1 6.0 59.5 6.6 67.3 5.0 niger) - 6 40 71.3 5.5 59.5 5.4 69.8 3.5 + 6 40 75.1 5.4 61.6 5.4 71.7 3.6 lip. SP285 - 2 14 77.8 4.5 65.8 4.1 70.2 3.6 (ex I 2 14 82.0 4.2 69.5 3.7 71.3 3.6 Fusarium - 6 40 74.8 4.8 64.1 4.4 71.8 3.2 oxysporum) + 6 40 75.0 5.0 64.8 4.4 72.0 3.4 -lip. SP225 - 2 14 74.9 4.4 64.5 4.2 76.3 2.8 30(ex Mucor + 2 14 79.3 3.5 67.4 3.8 76.6 2.5 miehei~ - 6 40 71.7 4.9 61.3 4.4 75.5 1.7 + 6 40 72.3 4.8 62.8 4.~ 76.7 1.9 _ .
Toyo Jozo - 2 14 86.2 2.7 71.3 2.4 78.6 1.7 35(lip. ex ~ 2 14 85.8 2.7 71.9 2.4 78.2 1.7 Chromobac. - 6 40 84.9 2.8 69.9 1.8 77.4 1.4 viscos~m) + 6 40 85.2 2.7 72.5 1.9 77.8 1.5 38368 c 7094 (R) Savin- Cotton PE/Cotton Polyester ~ase ase ~FHR 46096 FMR 460~6 FMR 46096 FM
Nagase - 2 1485.0 3.2 69.0 2.6 77.1 1.7 S (ex - + 21486.42.4 71.9 2.378.3 1.1 Rhiæopus) - 6 4071.7 5.1 61.7 4.5 76.0 1.5 64072.65.2 62.1 4.376.1 1.8 -C-4 (ex - 21472.15.5 60.8 5.374.4 3.4 10 Candida +2 1470.8 5.2 61.2 5.4 73.5 3.4 cylin- - 64072.15.1 61.4 4.675.4 1.7 dracea) +6 4073.2 5.3 62.4 4.6 75.4 2.0 pancreas - 2 1472.8 5.2 62.4 5.2 74.5 3.5 15 lipase ~ 21473.25.2 62.8 5.174.3 3.4 - 64072.95.3 62.3 4.675.9 1.7 + 64072.75.2 63.1 4.576.4 1.8 Esterase - 2 1472.6 5.3 61.9 5.2 75.3 3.3 20 MM (ex + 21471.95.2 62.7 4.975.4 3.2 Mucor - 64072.05.3 61~4 4.576.3 1.6 mieh~i) +6 4073.3 5.3 63.0 4.4 76.1 1.7 Amano-B - 21484.13.0 71.3 2.278.5 0.8 + 21485.83.1 71.5 2.779.2 0.9 - 64085.23.1 71.3 1.978.4 0.3 + 64086.33.0 72.8 1.979.4 0.2 Amano-CES - 2 1480.1 2.8 71.0 1.9 79.0 0.4 + 21481.42.9 73.0 1.879.7 0.4 - 64080.03.0 71.3 1.678.9 0.6 + 64082.03.0 73.3 1.779.0 0.8 Amano-CE - 2 1472.1 5.1 64.1 4.5 75.4 3.3 + 21472.75.1 66.6 4.175.8 3.3 - 64074.94.3 67.5 3.077.0 2.2 + 64077.54.0 69.0 2.977.2 2.4 -~s ~roLl~ rnQ~k .
~..2~8~68 C 7094 (R) Savin-Cotton PE/Cotton Polyester lipase ase~ FH R 460 % FM R 460 ~ FM R 460 ~ FM
lipase - 2 14 68.9 5.8 61.8 5.2 74.7 3.5 M-AP10 + 2 14 70.7 5.8 63.1 5.2 75.2 3.6 teX Mucor - 6 40 71.4 5.1 66.3 3.7 76.6 2.3 javanicus) + 6 40 74.2 4.5 66.2 4.1 75.9 2.5 lipase - 2 14 73.9 4.7 64.8 4.2 76.2 3.1 F-AP (ex + 2 14 72.1 5.2 65.5 4.7 75.7 3.4 Rhizopus - 6 40 74.7 4.6 69.4 2.4 77.1 1.9 javanicus) + 6 40 74.6 4.9 68.1 3.3 77.3 2.2 S80.000 - 2 14 72.1 5.1 66.0 4.3 76.1 3.1 (ex + 2 14 74.6 4.8 66.5 4.4 76.4 3.3 Rhizopus - 6 40 75.6 4.5 69.1 2.4 77.2 2.0 arrhizus) + 6 40 78.8 4.5 71.1 2.5 78.0 1.9 PS-54 (ex - 2 14 76.0 4.3 67.0 3.6 75.3 3.0 Thermom. + 2 14 77.1 4.4 68.9 3.7 75.6 2.9 lanu- - 6 40 78.1 4.3 69.5 2.1 76.7 2.0 ginosus) ~ 6 40 79.6 4.2 72.1 2.4 78.1 1.8 :
ex Pqeudo. - 2 14 79.6 3.2 70.3 2.0 78.4 0.4 gladioli ~ 2 14 81.1 3.0 72.1 2.1 79.9 0.4 - 6 40 79.6 3.5 69.6 1.9 77.8 0.1 + 6 40 81.0 3.4 71.7 1.8 79.7 0.2 lipase ex - 2 14 77.3 4.2 67.4 3.7 74.9 3.3 Rhizopus ~ 2 14 79.7 4.0 68.8 3.5 75.g 3.0 sp. ~ 6 40 77.4 3.7 68.1 2.4 77.8 1.7 + 6 40 80.7 4.1 71.0 2.3 78.9 1.8 -
Claims (2)
1. A detergent composition comprising from 1-50%
by weight of one or more nonionic detergent surfactants as the sole detergent surfactant, proteases in an amount such that the detergent composition has a proteolytic activity of 0.1-50 Glycine Units per milligram, and lipases in an amount such that the detergent composition has a lipolytic activity of 0.005-100 Lipase Units per milligram, the lipases being those which show a positive immunological cross-reaction with the antibody of the lipase produced by Chromobacter viscosum var. lipolyticum NRRL-B 3673.
by weight of one or more nonionic detergent surfactants as the sole detergent surfactant, proteases in an amount such that the detergent composition has a proteolytic activity of 0.1-50 Glycine Units per milligram, and lipases in an amount such that the detergent composition has a lipolytic activity of 0.005-100 Lipase Units per milligram, the lipases being those which show a positive immunological cross-reaction with the antibody of the lipase produced by Chromobacter viscosum var. lipolyticum NRRL-B 3673.
2. A composition according to claim 1, wherein the lipase is obtained from Pseudomonas fluorescens, Pseudomonas fragi, Pseudomonas nitroreducens var.
lipolyticum, Pseudomonas cepacia, Pseudomonas gladioli and Chromobacter viscosum.
lipolyticum, Pseudomonas cepacia, Pseudomonas gladioli and Chromobacter viscosum.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB868629535A GB8629535D0 (en) | 1986-12-10 | 1986-12-10 | Enzymatic detergent composition |
| GB8629535 | 1986-12-10 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1288368C true CA1288368C (en) | 1991-09-03 |
Family
ID=10608785
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000553755A Expired - Fee Related CA1288368C (en) | 1986-12-10 | 1987-12-08 | Enzymatic detergent composition |
Country Status (9)
| Country | Link |
|---|---|
| EP (1) | EP0271153B1 (en) |
| JP (1) | JPH0696717B2 (en) |
| AU (1) | AU605806B2 (en) |
| BR (1) | BR8706682A (en) |
| CA (1) | CA1288368C (en) |
| DE (1) | DE3761910D1 (en) |
| ES (1) | ES2014465B3 (en) |
| GB (1) | GB8629535D0 (en) |
| ZA (1) | ZA879296B (en) |
Families Citing this family (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4933287A (en) * | 1985-08-09 | 1990-06-12 | Gist-Brocades N.V. | Novel lipolytic enzymes and their use in detergent compositions |
| GB8629536D0 (en) * | 1986-12-10 | 1987-01-21 | Unilever Plc | Enzymatic detergent composition |
| DK571587D0 (en) * | 1987-11-02 | 1987-11-02 | Novo Industri As | ENZYMATIC DETERGENT COMPOSITION |
| BE1001436A3 (en) * | 1988-02-22 | 1989-10-31 | Synfina Sa | New lipase and detergent compositions containing. |
| US5733763A (en) * | 1988-08-19 | 1998-03-31 | Novo Nordisk A/S | Enzyme granulate formed of an enzyme-containing core and an enzyme-containing shell |
| GB8826110D0 (en) * | 1988-11-08 | 1988-12-14 | Unilever Plc | Enzyme-containing detergent compositions |
| DK78189D0 (en) * | 1989-02-20 | 1989-02-20 | Novo Industri As | ENZYMOUS GRANULATE AND PROCEDURE FOR PREPARING THEREOF |
| US4950417A (en) * | 1989-05-01 | 1990-08-21 | Miles Inc. | Detergent formulations containing alkaline lipase derived from Pseudomonas plantarii |
| GB8915658D0 (en) * | 1989-07-07 | 1989-08-23 | Unilever Plc | Enzymes,their production and use |
| US5658871A (en) * | 1989-07-07 | 1997-08-19 | Lever Brothers Company, Division Of Conopco, Inc. | Microbial lipase muteins and detergent compositions comprising same |
| JPH0489897A (en) * | 1990-07-30 | 1992-03-24 | Solvay Enzyme Prod Inc | Detergent compound containing alkaline lipase |
| CA2145176C (en) * | 1992-09-25 | 1999-07-13 | Beatrijs Lutgarde Aloysia De Smet | Detergent composition comprising lime soap dispersant and lipase enzymes |
| ES2169726T3 (en) * | 1992-09-25 | 2002-07-16 | Procter & Gamble | USE OF A LIME SOAP DISPERSANT IN A DETERGENT COMPOSITION THAT INCLUDES LIPASE ENZYMES. |
| US5772786A (en) * | 1993-08-13 | 1998-06-30 | The Procter & Gamble Company | Detergent composition comprising lime soap dispersant and lipase enzymes |
| DE19954181A1 (en) * | 1999-11-10 | 2001-05-31 | Eppendorf Geraetebau Netheler | Use of non-ionic surfactants to carry out enzymatic reactions |
| AU2009357432B2 (en) | 2009-12-28 | 2013-10-31 | Toyota Jidosha Kabushiki Kaisha | Hood structure of vehicle |
| CN110804506B (en) * | 2019-11-13 | 2021-06-01 | 领先生物农业股份有限公司 | Microbial wool detergent and using method thereof |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE1619087A1 (en) * | 1967-08-14 | 1969-10-02 | Henkel & Cie Gmbh | Surfactant combinations which can be used as laundry detergents and detergents or auxiliary washing agents containing them |
| GB1273545A (en) * | 1968-06-24 | 1972-05-10 | Albright & Wilson | Multi-enzyme cleaning compositions |
| DK289083A (en) * | 1983-06-23 | 1984-12-24 | Novo Industri As | LIPASE, PROCEDURE FOR PREPARING THEREOF AND ITS APPLICATION |
| GB8514707D0 (en) * | 1985-06-11 | 1985-07-10 | Unilever Plc | Enzymatic detergent composition |
| GB8514708D0 (en) * | 1985-06-11 | 1985-07-10 | Unilever Plc | Enzymatic detergent composition |
| DK154572C (en) * | 1985-08-07 | 1989-04-24 | Novo Industri As | ENZYMATIC DETERGENT ADDITIVE, DETERGENT AND METHOD FOR WASHING TEXTILES |
| GB8629536D0 (en) * | 1986-12-10 | 1987-01-21 | Unilever Plc | Enzymatic detergent composition |
-
1986
- 1986-12-10 GB GB868629535A patent/GB8629535D0/en active Pending
-
1987
- 1987-12-02 EP EP87202385A patent/EP0271153B1/en not_active Expired - Lifetime
- 1987-12-02 DE DE8787202385T patent/DE3761910D1/en not_active Expired - Fee Related
- 1987-12-02 ES ES87202385T patent/ES2014465B3/en not_active Expired - Lifetime
- 1987-12-08 AU AU82226/87A patent/AU605806B2/en not_active Ceased
- 1987-12-08 CA CA000553755A patent/CA1288368C/en not_active Expired - Fee Related
- 1987-12-09 JP JP62311792A patent/JPH0696717B2/en not_active Expired - Lifetime
- 1987-12-09 BR BR8706682A patent/BR8706682A/en not_active IP Right Cessation
- 1987-12-10 ZA ZA879296A patent/ZA879296B/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| DE3761910D1 (en) | 1990-04-19 |
| JPH0696717B2 (en) | 1994-11-30 |
| BR8706682A (en) | 1988-07-19 |
| AU8222687A (en) | 1988-06-16 |
| GB8629535D0 (en) | 1987-01-21 |
| ZA879296B (en) | 1989-08-30 |
| EP0271153A2 (en) | 1988-06-15 |
| EP0271153A3 (en) | 1988-08-10 |
| ES2014465B3 (en) | 1990-07-16 |
| JPS63161084A (en) | 1988-07-04 |
| AU605806B2 (en) | 1991-01-24 |
| EP0271153B1 (en) | 1990-03-14 |
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