The Biocatalytic Synthesis of Syringaresinol from 2,6-Dimethoxy-4-allylphenol in One-Pot Using a Tailored Oxidase/Peroxidase System

Abstract

Syringaresinol was synthesized in a one-pot conversion containing eugenol oxidase (EUGO) and horseradish peroxidase (HRP) using the relatively cheap 2,6-dimethoxy-4-allylphenol as a substrate. This conversion is fully coupled as the hydrogen peroxide generated from the reaction of EUGO with the substrate is utilized by the HRP to convert the formed sinapyl alcohol into syringaresinol. To improve the performance of EUGO on 2,6-dimethoxy-4-allylphenol, structure-inspired enzyme engineering was performed. This yielded the I427A EUGO mutant that is significantly more efficient with 2,6-dimethoxy-4-allylphenol. The I427A EUGO mutant together with HRP were capable of efficiently producing syringaresinol as a major product. After optimization and upscaling the conversion to a semipreparative scale (1 gr), syringaresinol was obtained in 81% yield.