Rational Engineering of a Flavoprotein Oxidase for Improved Direct Oxidation of Alcohols to Carboxylic Acids.
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.
['Pickl|Mathias|M|', 'Winkler|Christoph K|CK|0000-0003-3068-9817', 'Glueck|Silvia M|SM|', 'Fraaije|Marco W|MW|', 'Faber|Kurt|K|']
D000429:Alcohol Oxidoreductases D000438:Alcohols D000447:Aldehydes D002264:Carboxylic Acids D002384:Catalysis D020134:Catalytic Domain D004926:Escherichia coli D005415:Flavins D005420:Flavoproteins D005662:Furaldehyde D006860:Hydrogen Bonding D007700:Kinetics D010084:Oxidation-Reduction D011487:Protein Conformation
flavoprotein oxidase alcohol oxidation aldehyde oxidation biocatalysis protein design