Protease with low thermostability derived from Mucor pusillus

Details Protease with low thermostability derived from Mucor pusillus Protease with low thermostability derived from Mucor pusillus

1992-10-08

1994-07-26

Wax, Robert A.

Chemistry: molecular biology and microbiology

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

Hydrolase

435219, 435931, C12N 950, C12N 958

Patent

active

053326687

ABSTRACT:
A mutant fungus strain which produces a protease with low thermostability and low productivity was made from protease producing fungi and the gene coding for mutant enzyme is isolated from the mutant strain. A promoter which can function in yeast is ligated to the gene and inserted into a plasmid replicable in yeast, and the resulting plasmid is introduced into yeast. The yeast is cultured, thereby producing the mutant enzyme. Furthermore, a gene expressing an enzyme with a far lower thermostability is prepared by site-directed mutagenesis, which is introduced in yeast, thereby producing an enzyme with more distinctively reduced thermostability.

REFERENCES:
Hiramatsu, R. et al.; J. Biol. Chem. 264: 16862-16866 (1989).
Branner, S. et al.; Ann. N.Y. Acad. Sciences, vol. 434, pp. 340-342.
Darnell, J. et al.; Molecular Cell Biology, Scientific American Books, N.Y. (1984), pp. 53-55.
Brock, T. D. et al.; Biology of Microorganisms, Fourth Ed., Prentice-Hall Inc., N.J. (1984), pp. 302-312.
Beppu, T., Database WPIL, Week 8801 (1987).
Beppu, et al., Modification of milk-clotting aspartic proteases, chymosin and Mucor rennin Chemical Abstracts, vol. 113, No. 23, 207369z Columbus, Ohio.
Aikawa, et al., Advances in Experimental Medicine and Biology, vol. 306, 1991, New York US, pp. 233-242.
Branner-Jorgensen, et al.; Neth. Milk Dairy J. 35 (1981) 361-364; "Reduced thermostability of modified Mucor miehei rennet".
Suzuki, et al.; Portein Engineering vol. 2, No. 7, pp. 563-569, 1989; "Alteration of catalytic properties of chymosin by site-directed mutagenesis".

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