Engineering of Reductive Aminases for Asymmetric Synthesis of Enantiopure Rasagiline

Kai Zhang; Yuanzhi He; Jiawei Zhu; Qi Zhang; Luyao Tang; Li Cui; Yan Feng

doi:10.3389/fbioe.2021.798147

Engineering of Reductive Aminases for Asymmetric Synthesis of Enantiopure Rasagiline

Front Bioeng Biotechnol. 2021 Dec 22:9:798147. doi: 10.3389/fbioe.2021.798147. eCollection 2021.

Authors

Kai Zhang¹, Yuanzhi He¹, Jiawei Zhu¹, Qi Zhang¹, Luyao Tang¹, Li Cui¹, Yan Feng¹

Affiliation

¹ State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.

Abstract

Reductive aminases (RedAms) for the stereoselective amination of ketones represent an environmentally benign and economically viable alternative to transition metal-catalyzed asymmetric chemical synthesis. Here, we report two RedAms from Aspergillus calidoustus (AcRedAm) and bacteria (BaRedAm) with NADPH-dependent features. The enzymes can synthesize a set of secondary amines using a broad range of ketone and amine substrates with up to 97% conversion. To synthesize the pharmaceutical ingredient (R)-rasagiline, we engineered AcRedAm through rational design to obtain highly stereoselective mutants. The best mutant Q237A from AcRedAm could synthesize (R)-rasagiline with >99% enantiomeric excess with moderate conversion. The features of AcRedAm and BaRedAm highlight their potential for further study and expand the biocatalytic toolbox for industrial applications.

Keywords: chiral amine; rasagiline; rational design; reductive aminase; site saturation mutagenesis.