High-Throughput 19F NMR Chiral Analysis for Screening and Directed Evolution of Imine Reductases

Shucheng Song; Chenyang Wang; Wenjing Bao; Zhenchuang Xu; Jian Wu; Liang Lin; Yanchuan Zhao

doi:10.1021/acscentsci.5c00498

High-Throughput ¹⁹F NMR Chiral Analysis for Screening and Directed Evolution of Imine Reductases

ACS Cent Sci. 2025 Jun 3;11(7):1094-1102. doi: 10.1021/acscentsci.5c00498. eCollection 2025 Jul 23.

Authors

Shucheng Song¹, Chenyang Wang², Wenjing Bao², Zhenchuang Xu², Jian Wu³, Liang Lin¹, Yanchuan Zhao^{2

3}

Affiliations

¹ State Key Laboratory of Chemical Biology, Shanghai Institute of Organic Chemistry, University of Chinese Academy of Sciences, 345 Ling-Ling Road, Shanghai 200032, China.
² State Key Laboratory of Fluorine and Nitrogen Chemistry and Advanced Materials and Shanghai Hongkong Joint Laboratory in Chemical Synthesis, Shanghai Institute of Organic Chemistry, University of Chinese Academy of Sciences, Chinese Academy of Sciences, 345 Ling-Ling Road, Shanghai 200032, China.
³ Instrumental Analysis Center, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China.

Abstract

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 ¹⁹F 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.