Rational Engineering of a Flavoprotein Oxidase for Improved Direct Oxidation of Alcohols to Carboxylic Acids

Molecules

Silvia M. Glueck¹  Christoph K. Winkler¹  Kurt Faber²  Mathias Pickl²  Marco W. Fraaije³ 

[1] Austrian Centre of Industrial Biotechnology, ACIB GmbH c/o Department of Chemistry, University of Graz, Heinrichstrasse 28, A-8010 Graz, Austria;Department of Chemistry, University of Graz, Heinrichstrasse 28, A-8010 Graz, Austria;Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands;

关键词: biocatalysis;    alcohol oxidation;    aldehyde oxidation;    flavoprotein oxidase;    protein design;   

DOI  :  10.3390/molecules22122205

来源: DOAJ

【 摘 要 】

The oxidation of alcohols to the corresponding carbonyl or carboxyl compounds represents a convenient strategy for the selective introduction of electrophilic carbon centres into carbohydrate-based starting materials. The O2-dependent oxidation of prim-alcohols by flavin-containing alcohol oxidases often yields mixtures of aldehyde and carboxylic acid, which is due to “over-oxidation” of the aldehyde hydrate intermediate. In order to directly convert alcohols into carboxylic acids, rational engineering of 5-(hydroxymethyl)furfural oxidase was performed. In an attempt to improve the binding of the aldehyde hydrate in the active site to boost aldehyde-oxidase activity, two active-site residues were exchanged for hydrogen-bond-donating and -accepting amino acids. Enhanced over-oxidation was demonstrated and Michaelis–Menten kinetics were performed to corroborate these findings.

【 授权许可】

Unknown