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Updated: Sep 17, 2025

Monitoring the Reductive and Oxidative Half-Reactions of a Flavin-Dependent Monooxygenase using Stopped-Flow Spectrophotometry
Published on: March 18, 2012
Discovery of a monomeric flavin-containing opine dehydrogenase from bacteria
Seiya Watanabe1,2,3, Kentaroh Yoshiwara1, Mizue Date2
1Department of Bioscience, Graduate School of Agriculture, Ehime University, 3-5-7 Tarumi, Matsuyama, Ehime, Japan.
Abstract:
Opines, such as nopaline and octopine, are specifically produced in the crown gall by Agrobacterium tumefaciens, and specifically catabolized by opine dehydrogenase (OpnDH) from them. Known OpnDH consists of α-, β-, and γ-subunits, which contain FAD, FMN, and 2 iron-sulfur clusters as cofactors. We herein identified a novel type of FAD-containing OpnDH from Aureimonas altamirensis (AaOdhB3), consisting of a single polypeptide and only ∼22% sequence identity with the β-subunit of the heteromeric enzyme. AaOdhB3, a monomeric structure, exhibited strict specificity toward nopaline. A crystallographic analysis revealed that the overall structure and binding mode of FAD were similar to those of D-amino acid oxidase superfamily members. A site-directed mutagenic analysis and structural comparisons with other members of this protein family indicated the significance of Arg312 (and Arg247) for substrate recognition, and fructosyl-amino acid oxidase, related to santhopine metabolism from A. tumefaciens, was phylogenetically and physiologically close to OdhB3.
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