Stabilization of Penicillin G Acylase from Escherichia coli : Site-Directed Mutagenesis of the Protein Surface To Increase Multipoint Covalent Attachment-Reference-Cited by-同舟云学术

Stabilization of Penicillin G Acylase from Escherichia coli : Site-Directed Mutagenesis of the Protein Surface To Increase Multipoint Covalent Attachment

Published:2004-02 Issue:2 Volume:70 Page:1249-1251
ISSN:0099-2240
Container-title:Applied and Environmental Microbiology
language:en
Short-container-title:Appl Environ Microbiol

Author:

Abian Olga¹,Grazú Valeria¹,Hermoso Juan²,González Ramón³,García José Luis⁴,Fernández-Lafuente Roberto¹,Guisán José Manuel¹

Affiliation:

1. Departamento de Biocatalisis Instituto de Catálisis y Petroleoquímica, CSIC, Universidad Autónoma de Madrid, 28049 Madrid

2. Instituto de Química-Física Rocasolano

3. Instituto de Fermentaciones Industriales, CSIC, 28006 Madrid

4. Centro de Investigaciones Biológicas, CSIC, 28008 Madrid, Spain

Abstract

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.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Link

https://journals.asm.org/doi/pdf/10.1128/AEM.70.2.1249-1251.2004

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