Synergistic CRISPR–Cas9 Host Engineering and Enzyme Evolution for Enantioselective Synthesis of a Vibegron Pyrrolidine Intermediate

Key Points

• This research aims to improve the synthesis of chiral pyrrolidine intermediates for vibegron production using enzyme engineering and CRISPR technology.
• Utilized directed evolution to create enzyme variants
• Employed imine reductase for asymmetric reduction of imines
• Integrated whole cell catalysis for cleaner synthesis
• Evaluated conversion rates and diastereomeric excess
• Achieved 94% conversion of the imine substrate
• Obtained over 99% diastereomeric excess in products
• Established a scalable and robust biocatalytic platform

Abstract