Expanding the Application of Alcohol Dehydrogenases in Pharmaceutical Chemistry: A Focus on Piperidone Synthesis

Abstract

AbstractAlcohol dehydrogenases (ADHs) have garnered recognition for their potential in the synthesis of pivotal pharmaceutical compounds. However, their utilization in the context of piperidone synthesis remains an area ripe for exploration. In this study, we examine the performance of an alcohol dehydrogenase derived from Corynebacterium glutamicum (CgADH) using a substrate analogue functional screening (SAFS) method to elucidate its substrate specificity. To improve the catalytic activity of CgADH, a phenylalanine/alanine‐scanning and iterative saturation mutation (PAS‐ISM) method was used. The most active variant, I151F/I195A, exhibited a remarkable 10.6‐fold increase in catalytic activity compared to the wild‐type. Structural analysis revealed that the introduction of residues 151F and 195A led to a remodeling of the substrate‐binding pocket, enabling additional p‐π hydrophobic interactions with the substrate, ultimately promoting a more favorable substrate binding pose. This study introduces the SAFS screening method, which enables the identification of enzymes with no sequence homology to known enzymes. Furthermore, the application of PAS‐ISM presents an efficient approach for the engineering of alcohol dehydrogenases. These findings open up promising avenues to expand the utility of ADHs in the synthesis of piperidone, thereby advancing the field of pharmaceutical chemistry.