Screening methods for enzyme-mediated alcohol oxidation

Martina L Contente,Kurt Puentener,Irene Marzuoli,Hans Iding,Steven P Hanlon,Francesca Paradisi,Dennis Wetzl

doi:10.1038/s41598-022-07008-7

Martina L Contente, Kurt Puentener + Show 5 more

Open Access

https://doi.org/10.1038/s41598-022-07008-7

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Abstract

Alcohol oxidation for the generation of carbonyl groups, is an essential reaction for the preparation of fine chemicals. Although a number of chemical procedures have been reported, biocatalysis is a promising alternative for more sustainable and selective processes. To speed up the discovery of novel (bio)catalysts for industrial applications, efficient screening approaches need to be established. Here, we report on an enzyme-mediated alcohol oxidation screening platform to rapidly detect the activities and selectivities of three classes of biocatalysts; ketoreductases (KREDs), alcohol oxidases (AlcOXs) and laccase-mediator systems (LMSs) with diverse substrates.

Highlights

Alcohol oxidation for the generation of carbonyl groups, is an essential reaction for the preparation of fine chemicals
We report on an enzyme-mediated alcohol oxidation screening platform to detect the activities of three classes of biocatalysts (KREDs, alcohol oxidases (AlcOXs) and laccase-mediator systems (LMSs))
All the samples were used as cell-free extracts (CFEs); no enzyme purification was carried out

Summary

Introduction

Alcohol oxidation for the generation of carbonyl groups, is an essential reaction for the preparation of fine chemicals. We report on an enzyme-mediated alcohol oxidation screening platform to rapidly detect the activities and selectivities of three classes of biocatalysts; ketoreductases (KREDs), alcohol oxidases (AlcOXs) and laccase-mediator systems (LMSs) with diverse substrates. Among the enzymes available for alcohol oxidations, the superfamily of ketoreductases (KREDs), alcohol oxidases (AlcOxs) and laccase-mediator systems (LMSs) are the most commonly employed. AlcOXs represent another class of enzymes useful for alcohol oxidation Unlike reductases, they do not rely on nicotinamide cofactors but the reducing equivalents formed during the oxidation reaction, are transferred to molecular oxygen generating H 2O2 in stoichiometric a mounts[13,14]. We report on an enzyme-mediated alcohol oxidation screening platform (colorimetric and HPLC assays) to detect the activities of three classes of biocatalysts (KREDs, AlcOXs and LMSs)

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