Structure-Based Redesign of a Methanol Oxidase into an “Aryl Alcohol Oxidase” for Enzymatic Synthesis of Aromatic Flavor Compounds

Journal article (2023)

Authors

Bin Wu South China University of Technology
S. Wang University of Chinese Academy of Sciences, Chinese Academy of Sciences
Yunjian Ma South China University of Technology
Shuguang Yuan Chinese Academy of Sciences
F. Hollmann BT/Biocatalysis - AS
Yonghua Wang South China University of Technology
Research Group
BT/Biocatalysis (AS) (TU Delft)
Copyright: © 2023 Bin Wu, S. Wang, Yunjian Ma, Shuguang Yuan, F. Hollmann, Yonghua Wang
DOI
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https://doi.org/10.1021/acs.jafc.3c01069
Alcohol oxidase Flavor compounds Molecular modification Substrate selectivity
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Published Date

2023

Language

English

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Faculty

Applied Sciences

Department

BT/Biotechnologie

Research Group

BT/Biocatalysis

Abstract

Alcohol oxidases (AOxs) catalyze the aerobic oxidation of alcohols to the corresponding carbonyl products (aldehydes or ketones), producing only H2O2 as the byproduct. The majority of known AOxs, however, have a strong preference for small, primary alcohols, limiting their broad applicability, e.g., in the food industry. To broaden the product scope of AOxs, we performed structure-guided enzyme engineering of a methanol oxidase from Phanerochaete chrysosporium (PcAOx). The substrate preference was extended from methanol to a broad range of benzylic alcohols by modifying the substrate binding pocket. A mutant (PcAOx-EFMH) with four substitutions exhibited improved catalytic activity toward benzyl alcohols with increased conversion and kcat toward the benzyl alcohol from 11.3 to 88.9% and from 0.5 to 2.6 s-1, respectively. The molecular basis for the change of substrate selectivity was analyzed by molecular simulation.