EP0233721A2 - Zusammensetzungen zur Entfernung von Proteinverunreinigungen - Google Patents

Zusammensetzungen zur Entfernung von Proteinverunreinigungen Download PDF

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Publication number
EP0233721A2
EP0233721A2 EP87300845A EP87300845A EP0233721A2 EP 0233721 A2 EP0233721 A2 EP 0233721A2 EP 87300845 A EP87300845 A EP 87300845A EP 87300845 A EP87300845 A EP 87300845A EP 0233721 A2 EP0233721 A2 EP 0233721A2
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EP
European Patent Office
Prior art keywords
dcr
protease
reagent
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.)
Ceased
Application number
EP87300845A
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English (en)
French (fr)
Other versions
EP0233721A3 (de
Inventor
Pushkaraj J. Lad
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.)
Genencor Inc
Original Assignee
Genencor Inc
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 Genencor Inc filed Critical Genencor Inc
Publication of EP0233721A2 publication Critical patent/EP0233721A2/de
Publication of EP0233721A3 publication Critical patent/EP0233721A3/de
Ceased legal-status Critical Current

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    • 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

Definitions

  • the invention relates to the removal of stains using detergent compositions. More specifically, compositions which are capable of removing protein-containing stains, such as blood and food, are disclosed.
  • proteases used will function at the relatively high pH values (7.5-10.5) that are conducive to the performance of commonly used detergents.
  • Typical commercial detergents rely for their cleaning power on highly basic substances such as trisodium phosphate, detergent substances such as the long-chain alkyl or aryl sulfonates, and the protease additives such as subtilisin or other proteolytic enzymes.
  • compositions of the present invention provide a more effective release of protein-containing stains by causing disulfide bond cleavage in addition to proteolysis in the context of a detergent preparation.
  • the combination of proteolytic enzymes with disulfide cleavage reagents has been used previously in an effort to destroy proteins in research preparations, such as in the preparation of nucleic acids, or to fragment the target proteins, as in the performance of Cleveland gels for mapping peptides.
  • Use of a combination of a disulfide cleaving reducing agent, such as calcium thioglycolate with a protease to dissolve hair in clogged drains was disclosed in US 4,540,506. Up to 10% detergent could be added to the compositions used for this purpose.
  • patent 4,294,087 describes a method to dissociate and recover hair from animal hides using successive cleavage of disulfide bridges followed by protease treatment.
  • British patent application GB 2,139,260A published 7 November 1984, discloses the simultaneous use of peroxidant bleach and a protease in detergent compositions, wherein the presence of manganese ion permitted this composition to remain effective at lower temperature range than those previously used.
  • decomposition of the peroxide and action of the bleach requires temperatures above 70°C; even in the presence of these ions (other than manganese), temperatures above 60°C are required.
  • the manganese ion-containing compositions disclosed in the British patent may contain up to 50% detergent.
  • the invention provides an effective method and detergent composition for the removal of blood, food, and other protein-containing stains from fabrics, regardless of whether washing is prompt or delayed over a period of days or weeks.
  • the composition utilizes ordinary detergent preparations, but with the addition of both a protease and effective amount of a substance capable of cleaving disulfide bonds.
  • the resulting preparations are operable at temperatures of less than 70°C and even at temperatures below 55°C.
  • the invention is directed to a detergent composition which comprises an effective amount of a protease and of a substance capable of cleaving disulfide bonds (a disulfide cleavage reagent or DCR) in admixture with a suitable detergent formulation.
  • a suitable detergent formulation which comprises an effective amount of a protease and of a substance capable of cleaving disulfide bonds (a disulfide cleavage reagent or DCR) in admixture with a suitable detergent formulation.
  • the invention relates to methods for preparing the detergent compositions.
  • Figure 1 shows a comparison of a stained cloth swatch washed using the method of the invention with similar swatches washed with detergent alone, with detergent plus protease alone, or with detergent plus cleavage reagent alone.
  • the essential components of the composition are a protease, a substance or mixture of substances capable of cleaving disulfide bonds, and a detergent formulation carrier.
  • the protease is one of, or a mixture of general specificity proteolytic enzymes which are available commercially or otherwise are known in the art. While in theory any protease can be used, more favorable embodiments are those which have pH optima suitable for use in the presence of the mildly basic environments associated with detergent compositions. Thus, while pepsin and papain, for example, are marginally workable, enzymes with higher pH optima such as trypsin, chymotrypsin, subtilisin, and the like, are preferred. The preferred pH range is 7-12, more preferably 10-12. A number of commercially available enzyme preparations useful in detergents are also appropriate. Such enzymes include those trademarked Alcalase, Maxatase, Maxacal, and Esperase.
  • subtilisin prepared from Bacillus amyloliquefaciens , disclosed in European patent application publication no. 130,756, published 9 January 1985.
  • the compositions of the invention may include any protease or mixtures thereof, and the most effective choice will depend on the conditions of use, including pH, temperature, length of wash time, and the presence or absence of particular inhibitors with respect to the subject enzyme.
  • oxidizing agents include hydrogen peroxide, performic acid, sodium perborate, and oxidizing bleaches.
  • Effective reducing agents include dithiothreitol (DTT), ⁇ -mercaptoethanol (BME), sodium borohydride, and the like.
  • Alternate reagents which are not easily classified include mercuric chloride, nitroprusside, tributylphosphine, and phosphothiolate.
  • a particularly useful cleavage reagent is sodium sulfite, while results in sulfitolysis of the disulfide according to the reaction: R-S-S-R + SO ⁇ R-S-SO + ⁇ SR.
  • the equi­librium of this reaction may be shifted by removal of the thiol anion using heavy metal ions or oxidizing agents.
  • the oxidizing power may be provided by aeration if the wash solution is agitated, as is typically the case in conventional washing machines, or an oxidizing agent, such as CuSO4 or sodium perborate, may be added.
  • the remainder of the composition is that of a conventional detergent formulation, typically containing a base such as trisodium phosphate, detergents such as linear alkyl benzene sulfonates, alkyl ethoxylated sulfate, sulfated linear alcohol or ethoxylated linear alcohol, or soap, and other components such as builders and whiteners.
  • a base such as trisodium phosphate
  • detergents such as linear alkyl benzene sulfonates, alkyl ethoxylated sulfate, sulfated linear alcohol or ethoxylated linear alcohol, or soap
  • other components such as builders and whiteners.
  • detergent compositions may be in solid, granular, or liquid form. Of course, mixtures of bases and detergents can be used, as well.
  • Typical detergent compositions are disclosed in, for example, U.S.
  • the protease forms 0.01- 3% wt/wt of the detergent compositions of the invention, and the disulfide-cleaving substance, 10-40% wt/wt thereof.
  • the amounts present depend, of course, on the nature of the protease and of the disulfide cleavage reagent, the dilution of the detergent in the wash solution, and the conditions of wash. However, the ranges given are generally typical.
  • fabrics containing proteinaceous stains are treated with the combination (simultaneous or sequential) of a protease and disulfide cleaving reagent at suitable pH, temperature, and time of wash.
  • a protease and disulfide cleaving reagent at suitable pH, temperature, and time of wash.
  • the preferred times and temperatures are those generally utilized in household washing machines, neighborhood laundromats, and professional laundry services, since in order to be commercially practical, the process needs to be conducted under conditions ordinarily available to the user. It is to be noted that despite the temperature range here employed (less than 70°C, and even 55°C and below), no heavy metal ion catalyst, such as Mn(II), is required or desirable.
  • conventional washing procedures using commercial detergents are used and the protease and disulfide-cleaving substance are provided, either separately or together, as an additive, much in the manner of the methods in which bleach is used.
  • these may be added along with the detergent at the beginning of the wash cycle or at some intermediate point, for example, after approximately half of the wash cycle has been completed. If handled in this way, assuming an approximately 1:500 dilution of a solid detergent composition (approximately 2 mg/ml of the solid), arbitrary amounts of the protease and DCR may be added without the upper limit imposed by the dilution.
  • the protease is added to a final concentration of approximately 1-50 ⁇ g/ml of wash solution. In the case of the DCR, however, larger amounts than would be permitted by the dilution of the detergent may be desirable. For example, cleavage of disulfide bonds using sodium borohydride may conveniently be carried out with concentrations as high as 0.2 M reagent in the present of similar quantities of buffer (Lundbald, R.L., et al, Chemical Reagents for Protein Modification , supra).
  • the protease and disulfide-cleaving substance are added to the original detergent composition, and the process is conducted as a standard wash procedure using these modified detergents.
  • the detergent composition will correspond to that described above, but the amount of the composition can also be varied over the range of approximately 0.5 mg/ml-10 mg/ml or greater of the wash solution, depending, again, on the conditions of the wash solution and procedure, and on the solubilities of the detergent components.
  • the inclusion of the DCR and protease in the detergent limits the concentrations of these components in accordance with the dilution of the detergent.
  • a maximum concentration of 5 mg/ml DCR in the resulting wash solution is an upper limit.
  • concentration of DCR in the detergent will be less than 50%, mandating even lower concentration of the DCR.
  • the detergent compositions of the invention contain mostly detergent, relatively smaller amounts of DCR, and quite small amounts of protease, which is especially desirable in view of the cost of enzymic components.
  • the preparation will contain 60-90% “detergent” (surface active substances but including the additives such as builders and whiteners), 0.01-3% protease, and approximately 10-40% disulfide cleavage reagent.
  • disulfide cleavage reagent may be added to a prewash, followed by a detergent containing the protease, or addition of the detergent containing disulfide cleavage reagent may be followed or preceded by treatment with protease.
  • the following protocols are exemplary:
  • Increase of absorbance is a measure of protein extracted from test scratches by the test reagent containing disulfide-cleaving ⁇ -mercaptoethanol (BME) or dithiothreitol (DIT) using methods known in the art. The results are shown in Table 1 below.
  • Example 9 The conditions of treatment (water wash, incubation, rinse, protein determination) were as in Example 9, but these samples had a high initial protein content. The results are shown in Table 3.

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  • 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)
  • Detergent Compositions (AREA)
EP87300845A 1986-02-12 1987-01-30 Zusammensetzungen zur Entfernung von Proteinverunreinigungen Ceased EP0233721A3 (de)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US82877386A 1986-02-12 1986-02-12
US828773 1986-02-12
US88522486A 1986-07-14 1986-07-14
US885224 1986-07-14

Publications (2)

Publication Number Publication Date
EP0233721A2 true EP0233721A2 (de) 1987-08-26
EP0233721A3 EP0233721A3 (de) 1988-08-31

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Family Applications (1)

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EP87300845A Ceased EP0233721A3 (de) 1986-02-12 1987-01-30 Zusammensetzungen zur Entfernung von Proteinverunreinigungen

Country Status (3)

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EP (1) EP0233721A3 (de)
AU (1) AU6870187A (de)
DK (1) DK71087A (de)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0425018A2 (de) * 1989-10-27 1991-05-02 The Procter & Gamble Company Verfahren und Formulierung unter Verwendung von Endoglycosidase vom Typ II
US5258304A (en) * 1989-10-27 1993-11-02 Genencor International, Inc. Method of removing microorganisms from surfaces with Type II endoglycosidase
US5356803A (en) * 1989-10-27 1994-10-18 Genencor International, Inc. Antimicrobial composition containing Type II endoglycosidase and antimicrobial agent
EP2219687A2 (de) * 2007-10-17 2010-08-25 American Sterilizer Company Prionen-deaktivierende zusammensetzung und anwendungsverfahren dafür

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE1948653A1 (de) * 1969-09-26 1971-04-01 Henkel & Cie Gmbh Eiweissloesende Wasch-,Waschhilfs- und Reinigungsmittel
US3741901A (en) * 1970-08-07 1973-06-26 Pabst Brewing Co Washing compositions and process
US4421664A (en) * 1982-06-18 1983-12-20 Economics Laboratory, Inc. Compatible enzyme and oxidant bleaches containing cleaning composition

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE1948653A1 (de) * 1969-09-26 1971-04-01 Henkel & Cie Gmbh Eiweissloesende Wasch-,Waschhilfs- und Reinigungsmittel
US3741901A (en) * 1970-08-07 1973-06-26 Pabst Brewing Co Washing compositions and process
US4421664A (en) * 1982-06-18 1983-12-20 Economics Laboratory, Inc. Compatible enzyme and oxidant bleaches containing cleaning composition

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0425018A2 (de) * 1989-10-27 1991-05-02 The Procter & Gamble Company Verfahren und Formulierung unter Verwendung von Endoglycosidase vom Typ II
EP0425018A3 (en) * 1989-10-27 1991-10-02 The Procter & Gamble Company Method and formulation employing type ii endoglycosidase
US5238843A (en) * 1989-10-27 1993-08-24 Genencor International, Inc. Method for cleaning a surface on which is bound a glycoside-containing substance
US5258304A (en) * 1989-10-27 1993-11-02 Genencor International, Inc. Method of removing microorganisms from surfaces with Type II endoglycosidase
US5356803A (en) * 1989-10-27 1994-10-18 Genencor International, Inc. Antimicrobial composition containing Type II endoglycosidase and antimicrobial agent
EP2219687A2 (de) * 2007-10-17 2010-08-25 American Sterilizer Company Prionen-deaktivierende zusammensetzung und anwendungsverfahren dafür
EP2219687B1 (de) * 2007-10-17 2017-03-08 American Sterilizer Company Prionen-deaktivierende zusammensetzung und anwendungsverfahren dafür

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Publication number Publication date
DK71087A (da) 1987-08-13
DK71087D0 (da) 1987-02-12
AU6870187A (en) 1987-08-13
EP0233721A3 (de) 1988-08-31

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