Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1988-08-30
1994-05-17
Robinson, Douglas W.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435 711, 435187, 435188, 4352521, 4352532, 25217412, C12P 2104, C12N 998, C12N 952, C12N 120
Patent
active
053127480
DESCRIPTION:
BRIEF SUMMARY
This invention relates to a protease-producing strain of Nocardiopsis, to a process for preparing alkaline protease, to an alkaline protease preparation derived from Nocardiopsis, to a detergent additive and to an enzymatic detergent additive.
BACKGROUND OF THE INVENTION
Proteolytic enzymes produced by cultivation of microorganism strains from the genus Bacillus in suitable nutrient media are widely used in detergent compositions. Examples of such commercially available proteinase products are ALCALASE.RTM., ESPERASE.RTM., SAVINASE.RTM., all supplied by NOVO INDUSTRI A/S, Denmark. These and the similar Bacillus derived enzyme products from other suppliers are enzymatically active in detergent solutions, at pH values in the range of from 8 to 11 and in the presence of the sequestering agents, surfactants and bleaching agents normally present in detergent solutions.
The protease in ALCALASE.RTM. is produced by cultivating strains of species Bacillus licheniformis. The proteases in ESPERASE.RTM. and SAVINASE.RTM. are obtainable by cultivation of alkalophilic Bacillus species, such as the strains NCIB 10147 and NCIB 10309, respectively. The temperature optima of the commercially available alkaline proteases is about 60.degree. C. However, these commercial enzymes exhibit a relatively lower activity at room temperature.
BRIEF STATEMENT OF THE INVENTION
This invention relates to the production, isolation, characterization and use of novel alkaline proteases obtained from actinomycete microorganisms. The proteases of this invention are isolated from the actinomycete Nocardiopsis sp. strain 10R and Nocardiopsis dassonvillei strain M58-1 which have not been previously known to produce alkaline proteases. The proteases described in this invention are useful as detergent additives for laundering. The strain ZIMET 43647 of the species Nocardiopsis dassonvillei described by GDR patent No. DD 200,432 8 is known to produce protease for fibrinolytic and biological-sludge-clarifying applications. However, the strain was not available to the public as of the date hereof, the type strain does not produce protease, and no other strain of this species is known to produce protease. Further the aforementioned patented protease has a temperature profile and pH optimum much different from that of proteases in the invention.
In FIG. 3, the pH profiles for the Nocardiopsis sp. proteases of this invention were compared to that reported for the protease from patent strain ZIMET 43647. Under the same experimental conditions, strains 10R and M58-1 both have pH optima of 8, while that of the ZIMET 43647 protease is 9-10. In addition, the proteases of this invention have broader pH optima, showing at least 60% of the maximum activity between pH 7-11. The ZIMET 43647 protease possesses only 50% of maximum activity at pH 7 and 0% of maximum activity at pH 11. The expression of high proteolytic activity at pH 11 is especially important for detergent application.
The temperature profiles for the proteases of this invention were compared to that reported for the protease from ZIMET 43647 in FIG. 4. Between 40.degree. and 50.degree. C., the ZIMET 43647 protease exhibits 70-100% of its optimum activity, which is greater than that shown by the proteases of this invention, i.e. 25-65%.
In addition to the aforementioned differences, protease production in the strains of this invention responds differently to the addition of mineral salts. The strains of this invention produce the same level of protease activity whether or not mineral salts are included in the growth medium. The ZIMET 43647 strain is reported to show an improved response upon addition of mineral salts to the growth medium, possibly indicating the presence of a metalloprotease. As shown clearly in FIG. 2b, the protease of this invention has no detectable metalloprotease activity.
The Nocardiopsis sp. strain 10R and Nocardiopsis dassonvillei strain M58-1 proteases of this invention are especially effective as detergent additives at low laundering temperature, e.g. in cool water,
REFERENCES:
patent: 3652399 (1972-03-01), Isono et al.
patent: 4511490 (1985-04-01), Stanislowski et al.
Miyoshita, K; Mikami, Y.; Arai, T.; International Journal of Systematic Bacteriology, "Alkalophilic Actinomycete", Nocardiopsisdassonville subsp. prosing. Nov., Isolated from Soil, 34(4), pp. 405-409, 1984.
Beck Carol M.
Liu Chi-Li
Overholt Janet M.
Strobel, Jr. Robert J.
Lambiris Elias J.
Novo Nordisk A S
Robinson Douglas W.
Ware Deborah K.
Zelson Steve T.
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