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Enhanced fructose oxidase activity in a galactose oxidase variant

Lookup NU author(s): Dr Susan Firbank

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Abstract

Galactose oxidase (GO; EC 1.1.3.9) catalyses the oxidation of a wide range of primary alcohols including mono-, oligo- and polysaccharides. High-resolution structures have been determined for GO, but no structural information is available for the enzyme with bound substrate or inhibitor. Previously, computer-aided docking experiments have been used to develop a plausible model for interactions between GO and the D-galactose substrate. Residues implicated in such interactions include Arg330, Gln406, Phe464, Phe194 and Trp290. In the present study we describe an improved expression system for recombinant GO in the methylotrophic yeast Pichia pastoris. We use this system to express variant proteins mutated at Arg330 and Phe464 to explore the substrate binding model. We also demonstrate that the Arg330 variants display greater fructose oxidase activity than does wild-type GO.

Publication metadata

Author(s): Deacon SE, Mahmoud K, Spooner RK, Firbank SJ, Knowles PF, Phillips SE, McPherson MJ

Publication type: Article

Publication status: Published

Journal: ChemBioChem

Year: 2004

Volume: 5

Issue: 7

Pages: 972-979

ISSN (print): 1439-4227

ISSN (electronic): 1439-7633

Publisher: Wiley - VCH Verlag GmbH & Co. KGaA

URL: http://dx.doi.org/10.1002/cbic.200300810

DOI: 10.1002/cbic.200300810

Notes: Comparative Study

Journal Article

Research Support, Non-U.S. Gov't

Germany

a European journal of chemical biology

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