title: Biocatalytic Properties and Structural Analysis of Eugenol Oxidase from Rhodococcus jostii RHA1: A Versatile Oxidative Biocatalyst.
abstract: Eugenol oxidase (EUGO) from Rhodococcus jostii RHA1 had previously been shown to convert only a limited set of phenolic compounds. In this study, we have explored the biocatalytic potential of this flavoprotein oxidase, resulting in a broadened substrate scope and a deeper insight into its structural properties. In addition to the oxidation of vanillyl alcohol and the hydroxylation of eugenol, EUGO can efficiently catalyze the dehydrogenation of various phenolic ketones and the selective oxidation of a racemic secondary alcohol-4-(1-hydroxyethyl)-2-methoxyphenol. EUGO was also found to perform the kinetic resolution of a racemic secondary alcohol. Crystal structures of the enzyme in complexes with isoeugenol, coniferyl alcohol, vanillin, and benzoate have been determined. The catalytic center is a remarkable solvent-inaccessible cavity on the si side of the flavin cofactor. Structural comparison with vanillyl alcohol oxidase from Penicillium simplicissimum highlights a few localized changes that correlate with the selectivity of EUGO for phenolic substrates bearing relatively small p-substituents while tolerating o-methoxy substituents.
authors.affiliation: Molecular Enzymology, Groningen Biomolecular Sciences and, Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG, Groningen, NL.|Department of Biology and Biotechnology, University of Pavia, Via Ferrata 1, 27100, Pavia, Italy.
authors.forename: Quoc-Thai
authors.initials: QT
authors.lastname: Nguyen
authors.affiliation: Departmento de Química Orgánica, Universidad de Sevilla, c/Profesor García González 1, 41012, Sevilla, Spain.
authors.forename: Gonzalo
authors.initials: G
authors.lastname: de Gonzalo
authors.affiliation: Department of Biology and Biotechnology, University of Pavia, Via Ferrata 1, 27100, Pavia, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Binda
authors.affiliation: Stratingh Institute for Chemistry, University of Groningen, Nijenborgh 4, 9747 AG, Groningen, NL.
authors.forename: Ana
authors.initials: A
authors.lastname: Rioz-Martínez
authors.affiliation: Department of Biology and Biotechnology, University of Pavia, Via Ferrata 1, 27100, Pavia, Italy. andrea.mattevi@unipv.it.
authors.forename: Andrea
authors.initials: A
authors.lastname: Mattevi
authors.affiliation: Molecular Enzymology, Groningen Biomolecular Sciences and, Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG, Groningen, NL. m.w.fraaije@rug.nl.
authors.forename: Marco W
authors.initials: MW
authors.lastname: Fraaije
mesh_terms: D055162:Biocatalysis; D020134:Catalytic Domain; D006899:Mixed Function Oxygenases / Q000737:chemistry* / Q000378:metabolism; D015394:Molecular Structure; D010084:Oxidation-Reduction; D010636:Phenols / Q000378:metabolism; D012240:Rhodococcus / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D010636:Phenols; D006899:Mixed Function Oxygenases; C520320:eugenol oxidase, Rhodococcus
keywords: biocatalysis; dehydrogenation; enzyme structures; kinetic resolution; oxidases