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An Ammonium-Formate-Driven Trienzymatic Cascade for ω-Transaminase-Catalyzed (R)-Selective Amination

J Org Chem.. 2019; 
Chen FF1, Zhang YH1, Zhang ZJ1, Liu L1, Wu JP2, Xu JH1, Zheng GW1.
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Codon Optimization The genes of formate dehydrogenase (FDH) from Candida boidinii (FDH, a variant with mutation C23S),20 At-ω-TA from Aspergillus terreus, 21 and BbAmDH (named BbAmDH here, developed by Bommarius and coworkers using a phenylalanine dehydrogenase as scaffold)11b were respectively synthesised after codon optimization using OptimumGeneTM algorithm, inserted into pET-28a(+) plasmid in GenScript (Nanjing, China), and subsequently expressed in E. coli BL21 (DE3) cells. ATA117-Mut11 gene15 (Gene ID: JA717225.1) was cloned into plasmid pETDuet-1 and overexpressed in E. coli BL21 (DE3) cells. 1H NMR and 13C NMR analysis were conducted on a Bruker Avance 400 MHz spectrometer. Get A Quote

摘要

(R)-Amination mediated by (R)-specific ω-transaminases generally requires costly d-alanine in excess to obtain the desired chiral amines in high yield. Herein, a one-pot, trienzymatic cascade comprising an (R)-specific ω-transaminase, an amine dehydrogenase, and a formate dehydrogenase was developed for the economical and eco-friendly synthesis of (R)-chiral amines. Using inexpensive ammonium formate as the sole sacrificial agent, the established cascade system enabled efficient ω-transaminase-mediated (R)-amination of various ketones, with high conversions and excellent ee (>99%); water and CO2 were the only waste products.

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