Stabilization of penicillin G acylase from Escherichia coli:: Site-directed mutagenesis of the protein surface to increase multipoint covalent attachment

被引:90
|
作者
Abian, O
Grazú, V
Hermoso, J
González, R
García, JL
Fernández-Lafuente, R
Guisán, JM
机构
[1] Univ Autonoma Madrid, CSIC, Inst Catalisis & Petroleoquim, Dept Biocatalisis, E-28049 Madrid, Spain
[2] CSIC, Inst Quim Fis Rocasolano, E-28006 Madrid, Spain
[3] CSIC, Inst Fermentac Ind, E-28006 Madrid, Spain
[4] CSIC, Ctr Invest Biol, Madrid 28008, Spain
关键词
D O I
10.1128/AEM.70.2.1249-1251.2004
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
Three mutations on the penicillin acylase surface (increasing the number of Lys in a defined area) were performed. They did not alter the enzyme's stability and kinetic properties; however, after immobilization on glyoxyl-agarose, the mutant enzyme showed improved stability under all tested conditions (e.g., pH 2.5 at 4degreesC, pH 5 at 60degreesC, pH 7 at 55degreesC, or 60% dimethylformamide), with stabilization factors ranging from 4 to 11 compared with the native enzyme immobilized on glyoxyl-agarose.
引用
收藏
页码:1249 / 1251
页数:3
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