Alkaline protease, process for the production thereof, use there

Details Alkaline protease, process for the production thereof, use there Alkaline protease, process for the production thereof, use there

1998-03-25

1999-11-09

Achutamurthy, Ponnathapura

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

4352525, 43525231, C12N 954, C12N 120

Patent

active

059812559

DESCRIPTION:

BRIEF SUMMARY
TECHNICAL FIELD

This invention relates to a novel alkaline protease. More particularly, it relates to an alkaline protease which is stable in surfactants and heat resistant, its manufacturing method, its use in detergents, and microorganisms producing alkaline protease.


BACKGROUND ART

In recent years, environmental pollution has become a social issue. With regulation of the use of phosphate as an ingredient in detergents, an enzyme has been mixed with a detergent to improve detergency. Detergents with an enzyme such as protease, amylase, cellulase and lipase are currently marketed. Particularly, protease can degrade organic stains of clothes, 10 to 40% of which are caused by proteins and which cannot be cleaned with conventional detergents. Protease has become an essential ingredient improving detergency.
As protease for detergents, there are many enzymes originated from microorganisms which are active in the alkaline pH range, and proteases such as Kazusase (Showa Denko K. K.), Savinase (Novo), Maxacal (Gist), Alcalase (Novo) and Biosam (Showa Denko K. K.) have been used. With spreading of automatic dish washers, an enzyme usable for automatic dish washers is being required. Protease is effective on protein stains on dishes from egg yolk, dairy products, etc. However, generally inactivation of enzymes is accelerated in an aqueous solution at high temperature. Thus, an enzyme having an advantage in stability is required. Although Esperase (Novo), etc. have been currently used for this purpose, these enzymes do not have entirely satisfactory activity and stability. The protease which is known to be the most stable in surfactants at present is a protease produced by Bacillus strain SD 521 (patent application number 191781 of 1991). However, an even more stable protease is desired.
Therefore, the object of the present invention is to provide an alkaline protease more stable in the presence of a surfactant, manufacturing method of the alkaline protease, use of the protease for detergent, etc., and a microorganism producing the protease.


STATEMENT OF THE INVENTION

As a result of our devoted study, we found that protease API-26 produced by Bacillus sp. SD 114, a strain of Bacillus, has excellent stability and is suitable for detergent, and we completed the present invention.
Accordingly, the present invention provides a novel alkaline protease, its manufacturing method, its use, and a microorganism producing the protease. following (a) to (c): a surfactant solution (50 mM Atkins-Pantin borate buffer, pH 10, 0.1 mM EDTA, and 500 ppm of linear alkylbenzensulfonate LAS) a buffer solution (50 mM Atkins-Pantin borate buffer, pH 10, and 0.1 mM EDTA) a surfactant solution (50 mM Atkins-Pantin borate buffer, pH 10, 0.1 mM EDTA, and 500 ppm of LAS) properties: in as a substrate at 30.degree. C. for 10 minutes. 30.degree. C. for 24 hours. reacted with casein as a substrate at pH 10 for 10 minutes. electrophoresis. gel electrophoresis. by microorganisms belonging to Bacillus. microorganism belonging to Bacillus, which is Bacillus sp. SD 114 (FERM BP-5736). specified in the above item 4), and satisfying at least one of the conditions specified in the following (a) to (c): a surfactant solution (50 mM Atkins-Pantin borate buffer, pH 10, 0.1 mM EDTA, and 500 ppm of LAS) a buffer solution (50 mM Atkins-Pantin borate buffer, pH 10, and 0.1 mM EDTA) a surfactant solution (50 mM Atkins-Pantin borate buffer, pH 10, 0.1 mM EDTA, and 500 ppm of LAS) said protease from a culture where a microorganism belonging to Bacillus or its variant producing a protease specified in the above items 1) to 5) produced by a microorganism belonging to Bacillus, which is Bacillus sp. SD 114 (FERM BP-5736) above items 1) to 5) is contained is contained in the above items 1) to 5) is contained reacting a protease specified in any of the above items 1) to 5) with protein or peptides


BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 Graph presenting the optimal pH range for the enzyme of the invention.
FIG. 2 Graph presenting the stable pH range for the enzy

REFERENCES:
patent: 3905869 (1975-09-01), Hidaka et al.
patent: 5387518 (1995-02-01), Sawayanagi et al.

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