Rationally Engineering the Cofactor Specificity of LfSDR1 for Biocatalytic Synthesis of the Key Intermediate of Telotristat Ethyl. Issue 21 (10th October 2022)

• Record Type: Journal Article
• Authors: Li, Hengyu; Zhang, Wenhe; Che, Changli; Wang, Huibin; Jia, Yutian; Gao, Xiao; Jia, Xian; Qin, Bin; You, Song
• Is Part Of: ChemCatChem. Volume 14:Issue 21(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-10-10
• Subjects: short-chain dehydrogenase/reductase -- cofactor specificity -- cofactor regeneration -- telotristat ethyl; Catalysis -- Periodicals; 541.39505
• DOI: 10.1002/cctc.202201035
• Languages: English
• ISSNs: 1867-3880

Abstract

Abstract: Switching cofactor preference of oxidoreductases from NADPH to NADH by rational engineering, replacing the expensive cofactor NADP + with the cheap cofactor NAD +, is a focus of attention in the industrial application of oxidoreductases. This study focuses on the reversal of cofactor preference for short‐chain dehydrogenases/reductases (SDRs). Combined with bioinformatics analyses and in silico analyses, a small and smart mutant library (Mu1‐Mu3) of Lf SDR1 was rationally designed and constructed. Thus, the excellent NADH‐dependent recombinant Lf SDR1‐V186A/G92V/E141L/G38D/T15A variant (Mu2) was obtained. Meanwhile, novel enzymatic processes for synthesis of the key intermediates [( R )‐2 and ( S )‐4 ] of telotristat ethyl and crizotinib were successfully created, which mainly relied on Mu2 coupled with an FDH‐catalyzed cofactor regeneration system. A co‐expressed E. coli whole‐cell biocatalyst containing the genes of Mu2 and Pp FDH was developed to reduce ketones 1 and 3 . Finally, ketone 1 was almost completely converted into the product ( R )‐2 with a space‐time yield of 115.7 g⋅L −1 ⋅d −1 and a 98.8 % ee value. Abstract : Biocatalytic Synthesis : This study focuses on the reversal of cofactor preference for short‐chain dehydrogenases/reductases and the excellent NADH‐dependent recombinant Lf SDR1‐V186A/G92V/E141L/G38D/T15A variant (Mu2) was obtained. Meanwhile, a co‐expressed E. coli whole‐cell biocatalyst containing the genes of Mu2 and Pp FDH was developed to reduce ketone 1 . Finally, ketone 1 was almost completely converted into the product ( R )‐2 with a space‐time yield of 115.7 g⋅L −1 ⋅d −1 and a 98.8 % ee value.