Stabilization of Penicillin G Acylase from Escherichia coli : Site-Directed Mutagenesis of the Protein Surface To Increase Multipoint Covalent Attachment
Open Access
- 1 February 2004
- journal article
- research article
- Published by American Society for Microbiology in Applied and Environmental Microbiology
- Vol. 70 (2) , 1249-1251
- https://doi.org/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°C, pH 5 at 60°C, pH 7 at 55°C, or 60% dimethylformamide), with stabilization factors ranging from 4 to 11 compared with the native enzyme immobilized on glyoxyl-agarose.Keywords
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