Thermostabilization of porcine kidney D-amino acid oxidase by a single amino acid substitution

被引:12
|
作者
Bakke, M
Setoyama, C
Miura, R
Kajiyama, N
机构
[1] Kikkoman Foods Inc, Div Res & Dev, Noda, Chiba 2780037, Japan
[2] Kumamoto Univ, Grad Sch Med Sci, Dept Mol Enzymol, Kumamoto 8608556, Japan
关键词
porcine kidney D-amino acid oxidase; increased thermal stability; decreased K-m value; random mutagenesis;
D O I
10.1002/bit.20754
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
(D)-amino acid oxidase (DAO) is of considerable practical importance, such as bioconversion and enzymatic assay. In this study, we succeeded in obtaining a thermostable mutant DAO from porcine kidney by a single amino acid substitution. This mutant enzyme, F42C, was stable at 55 degrees C, while the wild-type enzyme was stable only up to 45 degrees C. The K, values of F42C for (D)-amino acids was about half of those of the wild-type enzyme. This mutant DAO with improved stability and affinity for its substrates is advantageous for the determination of (D)-amino acids. (c) 2005 Wiley Periodicals, Inc.
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页码:1023 / 1027
页数:5
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