Generic placeholder image

Protein & Peptide Letters

Editor-in-Chief

ISSN (Print): 0929-8665
ISSN (Online): 1875-5305

Biochemical and Thermostability Features of Acetyl Esterase Aes from Escherichia coli

Author(s): T. Farias, L. Mandrich, M. Rossi and G. Manco

Volume 14, Issue 2, 2007

Page: [165 - 169] Pages: 5

DOI: 10.2174/092986607779816113

Price: $65

Abstract

Previously we characterized an acetyl-esterase from Escherichia coli, formally Aes, from a thermodynamic point of view in comparative studies with thermophilic homologs. Since the enzyme appeared unusually resistant to the thermal denaturation we analysed the kinetic behaviour with respect to the temperature. The enzyme displays a surprising optimal temperature at 65 °C, showing a specific activity of 250 U/mg using pNP-butanoate as substrate, but a low kinetic stability at the same temperature (t1/2 of inactivation=5 min). By a random mutagenesis approach we searched for mutated versions of Aes with increased thermostability. We found the mutant T74A, which shows the same specific activity of wild type but a t1/2 of inactivation of 30 min at 65 °C.

Keywords: Aes, acetyl-esterase from Escherichia coli, HSL family, Hormone Sensitive lipase family of the esterase/lipase superfamily, SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis


Rights & Permissions Print Export Cite as
© 2023 Bentham Science Publishers | Privacy Policy