PRODUCTION AND CHARACTERIZATION OF A THERMOSTABLE PROTEASE PRODUCED BY AN ASPOROGENOUS MUTANT OF BACILLUS-STEAROTHERMOPHILUS

被引：16

作者：

ZAMOST, BL

BRANTLEY, QI

ELM, DD

BECK, CM

机构：

[1] New Products Research and Development, NOVO Laboratories, Inc., Danbury, CT

来源：

JOURNAL OF INDUSTRIAL MICROBIOLOGY | 1990年 / 5卷 / 05期

关键词：

Enzyme characterization; Ethyl methanesulfonate; Metalloprotease; Mutagenesis; Thermolysin;

D O I：

10.1007/BF01578205

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

An asporogenous mutant of Bacillus stearothermophilus (TPM-8) which produces 4-fold higher levels of a thermostable neutral protease than does wild-type strain 308-1 was obtained by mutagenesis with ethyl methanesulfonate. The protease produced by both the mutant and wild-type strain is a metalloprotease requiring Zn2+ and Ca2+ for activity and thermostability, respectively. It has a temperature optimum of 80°C at pH 7.0 and is highly thermostable, retaining 60% of its activity after 60 min at 85°C. The properties of the enzyme are similar to those of thermolysin. © 1990 Society for Industrial Microbiology.

引用

页码：303 / 312

页数：10

共 25 条

[1]

Chopra A.K., Mathur D.K., Isolation, screening and characterization of thermophilic Bacillus species isolated from dairy products, J. Appl. Bacteriology, 57, pp. 262-271, (1984)

[2]

Cook W.R., Ramaley R.F., Properties of a naturally occurring asporogenous variant of Bacillus caldolyticus, capable of growth in minimal glucose medium, Spores VII, pp. 171-175, (1978)

[3]

Emmanuiloua E.I., Toda K., Alpha-amylase production in batch and continuous cultures by Bacillus caldolyticus, Appl. Microbiol. Biotechnol., 19, pp. 301-305, (1984)

[4]

Endo S., Studies of protease produced by thermophilic bacteria, Hakka Kogaku Zasshi, 40, pp. 346-353, (1962)

[5]

Feder J., A spectrophotometric assay for neutral protease, Biochem. Biophys. Res. Commun., 32, pp. 326-332, (1968)

[6]

Feder J., Kuo M.J., Wildi B.S., Thermolysin, a thermostable neutral protease, Dev. Ind. Microbiol., 18, pp. 267-272, (1977)

[7]

Feder J., Schuck J.M., Studies on the Bacillus subtilus neutral protease and Bacillus thermoproteolyticus thermolysin-catalyzed hydrolysis of dipeptide substrates, Biochemistry, 9, pp. 2784-2791, (1970)

[8]

Fujii M., Takagi M., Imanaka T., Aiba S., Molecular cloning of a thermostable neutral protease gene from Bacillus stearothermophilus into a vector plasmid and its expression in Bacillus stearothermophilus and Bacillus subtilis, J. Bacteriol., 154, pp. 831-837, (1983)

[9]

Imanaka T., Fujii M., Aramori I., Aiba S., Transformation of Bacillus stearothermophilus with plasmid DNA and characterization of shuttle vector plasmids between Bacillus stearothermophilus and Bacillus subtilis, J. Bacteriol., 149, pp. 824-830, (1982)

[10]

Imanaka T., Shibazaki M., Takagi M., A new way of enhancing the thermostability of proteases, Nature, 324, pp. 695-697, (1986)

← 1 2 3 →