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
  • C11D3/38645—Preparations containing enzymes, e.g. protease or amylase containing cellulase
• • 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 present invention relates to a detergent additive and a detergent composition comprising a cellulase and a protease in the presence of which the cellulase exhibits improved stability properties.
• cellulases are sensitive to the action of proteases, i.e. that in the presence of proteases commonly employed in detergents, cellulases are degraded to lower molecular weight polypeptides resulting in inactivation of the cellulase enzymes.
• proteases When cellulases are employed in liquid detergent compositions, it has typically been necessary to protect them from proteolytic degradation.
• the present invention relates to a detergent additive comprising a cellulase and a protease, the protease being one which has a higher degree of specificity than Bacillus lentus serine protease.
• the term "higher degree of specificity” is defined as a protease which degrades human insulin to fewer components than does the B. lentus serine protease under the following conditions: 0.5 ml of a 1 mg/ml solution of human insulin in B and R buffer, pH 9.5, is incubated with 75 ⁇ l enzyme solution of 0.6 CPU (cf. Novo Nordisk Analysis Method No. AF 228/1) per litre for 120 min. at 37°C, and the reaction is quenched with 50 ⁇ l 1N HCl.
• the present invention relates to a detergent composition
• a detergent composition comprising a cellulase and a protease as defined above.
• subtilisin Novo or a variant thereof (e.g. a variant described in US 4,914,031), a protease derivable from Nocardiopsis dassonvillei NRRL 18133 (described in WO 88/03947), a serine protease specific for glutamic and aspartic acid, producible by Bacillus licheniformis (this protease is described in detail in co-pending International patent application No. PCT/DK91/00067), or a trypsin-like protease producible by Fusarium sp., DSM 2672 (this protease is described in WO 89/06270).
• the cellulase included in the detergent additive or detergent composition of the invention may be any enzyme exhibiting activity and may, for instance be derived from a strain of Humicola , Fusarium , Myceliophthora , Thermonospora , Bacillus or Streptomyces .
• the cellulase is an endoglucanase, i.e. an enzyme which shows affinity for cellulose and which attacks amorphous regions of low crystallinity in cellulose fibres.
• a particular endoglucanase which it has been found favourable to combine with a protease as defined above is one which is immunoreactive with an antibody raised against a highly purified ⁇ 43 kD endoglucanase derived from Humicola insolens, DSM 1800.
• One such enzyme which is decribed in detail in co-pending Danish patent application No. DK 1159/90 has the amino acid sequence shown in the appended Sequence Listing ID# 1. For the present purpose, this enzyme is referred to in the following as " ⁇ 43 kD endoglucanase".
• the detergent additive according to the invention may suitably be in the form of a non-dusting granulate, stabilized liquid or protected enzyme.
• Non-dusting granulates may be produced e.g. according to US 4,106,991 and 4,661,452 (both to Novo Industri A/S) and may optionally be coated by methods known in the art.
• Liquid enzyme preparations may, for instance, be stabilized by adding a polyol such as propylene glycol, a sugar or sugar alcohol, lactic acid or boric acid according to established methods. Other enzyme stabilizers are well known in the art.
• Protected enzymes may be prepared according to the method disclosed in EP 238,216.
• the detergent additive may suitably contain 0.001-1.0 mg of cellulase and 0.001-1.0 mg of protease per gram of the additive. It will be understood that the detergent additive may further include one or more other enzymes, such as a lipase, peroxidase or amylase, conventionally included in detergent additives.
• the detergent composition of the invention additionally comprises a surfactant which may be of the anionic, non-ionic, cationic, amphoteric, or zwitterionic type as well as mixtures of these surfactant classes.
• a surfactant which may be of the anionic, non-ionic, cationic, amphoteric, or zwitterionic type as well as mixtures of these surfactant classes.
• anionic surfactants are linear alkyl benzene sulfonates (LAS), alpha olefin sulfonates (AOS), alcohol ethoxy sulfates (AES) and alkali metal salts of natural fatty acids.
• Detergent compositions of the invention may contain other detergent ingredients known in the art as e.g. builders, bleaching agents, bleach activators, anti-corrosion agents, sequestering agents, anti soil-redeposition agents, perfumes, enzyme stabilizers, etc.
• the detergent composition of the invention may be formulated in any convenient form, e.g. as a powder or liquid.
• the enzyme may be stabilized in a liquid detergent by inclusion of enzyme stabilizers as indicated above.
• the pH of a solution of the detergent composition of the invention will be 7-12 and in some instances 7.0-10.5.
• Other detergent enzymes such as proteases, lipases or amylases may be included in the detergent compositions of the invention, either separately or in a combined additive as described above.
• the cellulase is typically present in a concentration corresponding to 0.0005-20 mg of enzyme protein per litre of washing liquor, and the protease is typically present in a concentration corresponding to 0.0005-20 mg per litre of washing liquor.
• the detergent composition of the invention is usually employed in concentrations of 0.5 - 20 g/l in the washing solution. In general, it is most convenient to add the detergent additive in amounts of 0.1 - 5% w/w or, preferably, in amounts of 0.2 - 2% of the detergent composition.
• the detergent additive or detergent composition of the invention may be used to reduce the rate at which cellulose-containing fabrics become harsh or to reduce the harshness of cellulose-containing fabrics, or to provide colour clarification of coloured cellulose-containing fabrics in a manner known per se (cf. for instance WO 89/00069).
• Endoglucanase 12 CMCU/g of detergent
• Proteases 0.2 mg/g of detergent
• the residual activity of the endoglucanase after 7 days of incubation with the respective proteases was determined in terms of its CMCase activity (CMCU).
• the CMCase activity was determined as follows: A substrate solution of 30 g/l CMC (Hercules 7 LFD) in deionized water was prepared. The enzyme sample to be determined was dissolved in 0.01 M phosphate buffer, pH 7.5. 1.0 ml of the enzyme solution and 2.0 ml of a 0.1 M phosphate buffer, pH 7.5, were mixed in a test tube, and an enzyme reaction was initiated by adding 1.0 ml of the substrate solution to the test tube. The mixture was incubated at 40°C for 20 minutes, after which the reaction was stopped by adding 2.0 ml of 0.125 M trisodium phosphate.12H 2 O. A blind sample was prepared without incubation.
• a ferricyanide solution (1.60 g of potassium ferricyanide and 14.0 g of trisodium phosphate.12H 2 O in 1 1 of deionized water) was added to a test sample as well as to a blind immediately followed by immersion in boiling water and incubation for 10 minutes. After incubation, the samples were cooled with tap water. The absorbance at 420 nm was measured, and a standard curve was prepared with glucose solution.
• CMCase unit is defined as the amount of enzyme which, under the conditions specified above, forms an amount of reducing carbohydrates corresponding to 1 ⁇ mol of glucose per minute.
• the storage stability of the endoglucanase of the invention was determined as its residual activity (in CMCU%) under the conditions indicated above. Protease Residual Activity (%) + boric acid - boric acid Glu/Asp specific 105 93 Trypsin-like 77 63 B. lentus serine 57 24 Subtilisin Novo 63 55

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• 1991
  • 1991-04-22 DK DK91737A patent/DK73791D0/da not_active Application Discontinuation
• 1992
  • 1992-04-10 BR BR9205921A patent/BR9205921A/pt not_active Application Discontinuation
  • 1992-04-10 EP EP92909626A patent/EP0581839B1/en not_active Revoked
  • 1992-04-10 JP JP4508964A patent/JPH06506712A/ja active Pending
  • 1992-04-10 WO PCT/DK1992/000116 patent/WO1992018599A1/en not_active Application Discontinuation
  • 1992-04-10 ES ES92909626T patent/ES2090636T3/es not_active Expired - Lifetime
  • 1992-04-10 DE DE69212150T patent/DE69212150T2/de not_active Revoked
  • 1992-04-10 DK DK92909626.1T patent/DK0581839T3/da active
  • 1992-04-10 AT AT92909626T patent/ATE140262T1/de not_active IP Right Cessation
  • 1992-04-21 MX MX9201826A patent/MX9201826A/es unknown
• 1993
  • 1993-10-21 FI FI934656A patent/FI934656A/fi unknown

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