Controlling Stereopreferences of Carbonyl Reductases for Enantioselective Synthesis of Atorvastatin Precursor

• DOI: 10.1021/acscatal.0c05607
• Publication Date: 2021-02-13T01:49:17Z
• Authors: Feng Cheng; Yi Chen; Shuai Qiu; Qiu-Yao Zhai; Hua-Tao Liu; Shu-Fang Li; Chun-Yue Weng; Ya-Jun Wang; Yu-Guo Zheng
• Keywords: Kluyveromyces marxianus; Sequence (biology); Protein Engineering

Carbonyl reductase (CR)-catalyzed asymmetric reduction offers an approach for producing t-butyl 6-cyano-(3R,5R)-dihydroxyhexanoate ((3R,5R)-1b), which serves as a key building block in atorvastatin (Liptor). However, controlling the stereopreference of CR with desired selectivity remains challenging because natural CRs usually exhibit Prelog preference. Moreover, transferring knowledge from engineered anti-Prelog to other is difficult. Herein, residues that regulate Kluyveromyces marxianus (KmCR) were identified by semirational engineering toward 6-cyano-(5R)-hydroxy-3-oxohexanoate ((5R)-1a). A structural switch consists was discovered, and related features summarized predict analyzing information multiple-sequence alignment structure-available carefully. According obtained knowledge, simultaneous mutation four enabled conversion Prelog-selectivity KmCR into complete (dep > 99% (R) (3R,5R)-1b). stereopreferences 11 share 20–40% sequence identities predicted successfully experimentally. The gained this protein study on has universal significance (5R)-1a.