Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1994-10-26
1997-01-28
Naff, David M.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435219, 435220, 435222, 435264, 510530, C12N 950, C12N 952, C12N 954, C12N 956, D06M 1600
Patent
active
055977200
DESCRIPTION:
BRIEF SUMMARY
CROSS-REFERENCE TO RELATION APPLICATIONS
This application is a continuation of PCT/DK93/00183 filed May 26, 1993, which is incorporated herein by reference.
TECHNICAL FIELD
This invention relates to proteases derived from strains of Bacillus sp. More specifically, the invention is directed towards a novel alkaline protease derived from a strain of Bacillus sp. PD498, as well as isolated biologically pure cultures of Bacillus sp. PD498.
Moreover, the invention is directed towards a process for the preparation of the protease, detergent additives and detergent compositions comprising the protease of the invention, and the use of this protease in washing processes.
BACKGROUND ART
Detergent enzymes have been marketed for more than 20 years and are now well established as normal detergent ingredients in both powder and liquid detergents all over the world. With the trend towards lower washing temperature, s detergent enzyme consumption has increased during late years. Enzymes used in washing formulations comprise proteases, lipases, amylases, cellulases, as well as other enzymes, or mixtures hereof. Commercially most important are proteases.
Detergent proteases have been developed by isolation of proteases found in nature followed by testing in detergent formulations. Most detergent of proteases are obtained from members of the genus Bacillus. Currently new types of proteases enter the market, offering the possibility of giving a better cost/performance ratio at various specified conditions.
Examples of commercial protease products are ALCALASE.TM., ESPERASE.TM. and SAVINASE.TM., all supplied by Novo Nordisk A/S, Denmark. These and similar enzyme products from other commercial sources are active in detergent solutions, i.e. at pH values in the range of from 8 to 11 and in the presence of sequestering agents, surfactants and bleaching agents such as sodium borate. The ALCALASE.TM. protease is produced by strains of the species Bacillus licheniformis. The ESPERASE.TM. and SAVINASE.TM. proteases are obtained by cultivation of strains of alkalophilic Bacilli.
SUMMARY OF THE INVENTION
According to the present invention novel detergent proteases are provided.
In its first aspect, the invention provides a protease having an apparent molecular weight of 34 kD as determined by SDS-PAGE, a pI of approximately 9.3, a pH optimum in the range of pH 9-11 determined at 25.degree. C. (with casein as substrate), a temperature optimum in the range of 40.degree.-55.degree. C. determined at pH 9.5 (with casein as substrate), and immunochemical properties identical or partially identical to those of a protease derived from Bacillus sp. PD498, NCIMB No. 40484.
In another aspect, the invention provides a protease having an apparent molecular weight of 34 kD as determined by SDS-PAGE, a pI of approximately 9.3, a pH optimum in the range of pH 9-11 determined at 25.degree. C. (with casein as substrate), a temperature optimum in the range of 40.degree.-55.degree. C. determined at pH 9.5 (with casein as substrate), and being obtainable from a strain of Bacillus sp. PD498, or from another host organism carrying the gene encoding a protease having immunochemical properties identical or partially identical to those of the protease derived from Bacillus sp. PD498.
In a third aspect, the invention provides an isolated biologically pure culture of a strain of Bacillus sp. PD498. In a more specific aspect, a strain of Bacillus sp. PD498, NCIMB No. 40484, or a mutant or a variant thereof, is provided.
In a fourth aspect, the invention provides a process for the preparation of the protease, which process comprises cultivation of a protease producing strain of Bacillus sp. PD498 in a suitable nutrient medium, containing carbon and nitrogen sources and inorganic salts, followed by recovery of the desired enzyme. In a more specific aspect, Bacillus sp. PD498, NCIMB No. 40484, or a mutant or a variant thereof, or another host organism carrying the gene encoding a protease having immunochemical properties identical or partially identical to
REFERENCES:
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patent: 4480037 (1984-10-01), Ichishima et al.
patent: 4764470 (1988-08-01), Durham et al.
patent: 4771003 (1988-09-01), Stellwag et al.
patent: 5362414 (1994-11-01), Outtrup et al.
Aaslyng Dorrit A.
Dambmann Claus
Outtrup Helle
Agris, Esq. Cheryl H.
Naff David M.
Novo Nordisk A S
Weber Jon P.
Zelson Esq. Steve T.
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