High-Throughput F NMR Chiral Analysis for Screening and Directed Evolution of Imine Reductases.

Biocatalysis is an essential tool for asymmetric synthesis, significantly enhancing the production of chiral molecules. As advancements in protein screening and engineering rapidly evolve, the demand for efficient, rapid chiral analysis methods has intensified. Addressing this need, we introduce a high-throughput F NMR-based assay that provides comprehensive insights into the enantioselectivity, stereopreference, and yields of biocatalytic reactions. This assay has been successfully applied to screen imine reductases, showcasing its efficacy in the directed evolution for synthesizing an intermediate of the anti-Parkinson drug, rotigotine. Our method offers substantial promise for propelling forward the fields of biocatalysis and synthetic biology by accelerating the assessment of stereochemical outcomes in biocatalytic processes.