Discovery, Characterization and Synthetic Potential of Two Novel Bacterial Alcohol Oxidases

The search for novel synthetic tools to prepare industrial chemicals in a safer and greener manner is a continuing challenge in synthetic chemistry. In this manuscript, we report the discovery, characterization, and synthetic potential of two novel aryl-alcohol oxidases from bacteria which are able to oxidize a variety of aliphatic and aromatic alcohols in high efficiencies (up to 4970 min-1mM-1). Crystal structures revealed unusually wide-open entrance to the active-site pockets compared to that previously described for traditional fungal aryl-alcohol oxidases, which could correlate with differences in substrate scope, catalytic efficiency, and other functional properties. Preparative-scale reactions and ability to operate at high substrate loadings also demonstrate the potential of these enzymes in synthetic chemistry with turnover numbers > 30000. Moreover, their availability as soluble and active recombinant proteins enabled their use as cell-free extracts which further highlights their potential for the large-scale production of carbonyl compounds.