Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1993-09-28
1995-12-05
Naff, David M.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
25217412, 252DIG12, 435188, 4352541, 435911, C12N 958, C12N 114, C12N 996, C11D 1700
Patent
active
054728657
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention is in the field of detergent proteases. More specifically, the invention is directed towards novel alkaline proteases derived from strains of imperfect fungi of the genus/genera Dendryphiella and/or Scolecobasidium. Moreover, the invention is directed towards a process for preparation of the proteases, the use of the proteases as detergent enzyme, and detergent compositions comprising the proteases of the invention.
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 temperature washing, 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 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 AS, 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 perborate. 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 obtainable from a strain of the genus Dendryphiella or Scolecobasidium.
In a more specific aspect, the invention provides a protease having a pl of approximately 9.1, a molecular weight of approximately 18-20 kD, being active towards haemoglobin, casein, skimmed milk and Suc-Ala-Ala-Pro-Phe-pNA, and having immunochemical properties identical or partially identical to those of a protease derived from the strain D. arenaria, DSM No. 6260, or the strain D. salina, DSM No. 6331.
In another specific aspect, the invention provides a protease having a pl>9.5, a molecular weight of approximately 28 kD, and immunochemical properties identical or partially identical to those of a protease derived from Dendryphiella arenaria, DSM No. 6260.
In its second aspect, the invention provides a process for the preparation of the proteases comprising cultivation of a protease producing strain of the genus Dendryphiella or Scolecobasidiurn in a suitable nutrient medium, containing carbon and nitrogen sources and inorganic salts, followed by recovery of the desired enzyme.
In its third aspect, the invention provides a process for the preparation of the protease which process comprises isolating a DNA fragment encoding the protease, combining the DNA fragment with an appropriate expression signal in an appropriate plasmid vector, introducing the plasmid vector into an appropriate host either as an autonomously replicating plasmid or integrated into the chromosome, cultivating the host organism under conditions leading to expression of the protease, and recovering of the protease from the culture medium.
In its fourth aspect, the invention claims the use of the proteases as detergent enzymes. In a more specific embodiment of this aspect, the invention provides detergent compositions comprising the proteases. In another specific embodiment of this aspect, the detergent compositions are provided in the form of a detergent additive, preferably a non-dusting granulate or a stabilized liquid.
DETAILED DISCLOSURE OF THE INVENTION
The new deter
REFERENCES:
MacDonald et al., Can. J. Bot. , vol. 60, 1982, pp. 838-844.
Michaelis et al., Mycolgia, 79(4), 1987, pp. 514-518.
Christiansen Margrethe
Lindegaard Poul
Pedersen Kim B.
Lowney Karen A.
Meller Mike
Naff David M.
Novo Nordisk A S
Zelson Steve T.
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