大肠杆菌青霉素G酰化酶的稳定性：对蛋白质表面进行定点诱变以增加多点共价连接
Stabilization of penicillin G acylase from Escherichia coli: site-directed mutagenesis of the protein surface to increase multipoint covalent attachment.
作者信息
Abian Olga, Grazú Valeria, Hermoso Juan, González Ramón, García José Luis, Fernández-Lafuente Roberto, Guisán José Manuel
机构信息
Departamento de Biocatalisis Instituto de Catálisis y Petroleoquímica, CSIC, Universidad Autónoma de Madrid, 28049 Madrid, Spain.
出版信息
Appl Environ Microbiol. 2004 Feb;70(2):1249-51. doi: 10.1128/AEM.70.2.1249-1251.2004.
Abstract
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 4 degrees C, pH 5 at 60 degrees C, pH 7 at 55 degrees C, or 60% dimethylformamide), with stabilization factors ranging from 4 to 11 compared with the native enzyme immobilized on glyoxyl-agarose.
摘要
对青霉素酰化酶表面进行了三个突变（增加特定区域赖氨酸的数量）。这些突变并未改变该酶的稳定性和动力学性质；然而，在固定于乙醛酸琼脂糖上后，突变酶在所有测试条件下（例如，4℃下pH 2.5、60℃下pH 5、55℃下pH 7或60%二甲基甲酰胺）均表现出更高的稳定性，与固定于乙醛酸琼脂糖上的天然酶相比，稳定因子在4至11之间。
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