Enhanced thermal stability of an alkaline protease, AprP, isolated from a Pseudomonas sp by mutation at an autoproteolysis site, Ser-331

被引：26

作者：

Jang, JW

Ko, JH

Kim, EK

Jang, WH

Kang, JH

Yoo, OJ

机构：

[1] Korea Adv Inst Sci & Technol, Biomed Res Ctr, Yusong Gu, Taejon 305701, South Korea

[2] Inje Univ, Paik Inje Mem Inst Biomed Sci, Pusanjin Gu, Pusan 614735, South Korea

来源：

BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY | 2001年 / 34卷 / 02期

关键词：

autoproteolytic degradation; catalytic efficiency; thermostability;

D O I：

10.1042/BA20010027

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 [生物化学与分子生物学]; 081704 [应用化学];

摘要：

The thermal stability of the alkaline protease extracellular subtilisin-type serine protease (AprP) from Pseudomonas sp. KFCC 10818 was improved by altering an amino acid residue at an autoproteolytic cleavage site. N-terminal sequence analysis of the autoproteolytic products of the protein revealed the presence of two cleavage sites, Ser-307 and Ser-331. To increase the thermal stability of the enzyme, serine residues of these sites were replaced with aspartate. The S331D mutant enzyme was successfully purified and characterized whereas the S307D mutant was not. The half-lives of the S331D mutant at 55 degreesC and 60 degreesC were 1.5 and 2.4 times longer than that of the wild-type enzyme, respectively. In addition, the catalytic efficiency was also enhanced.

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页码：81 / 84

页数：4

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