The Journal of Organic Chemistry, Journal Year: 2025, Volume and Issue: unknown
Published: Jan. 21, 2025
Aldolases, especially 2-deoxyribose-5-phosphate aldolase (DERA) enzymes, have been widely employed to access key chiral precursors for various active pharmaceutical ingredients (APIs). This has enabled by expanding their substrate scope toward non-natural acceptors and donors via protein engineering. In this study, we endeavored broaden the acceptor of DERA from Geobacillus sp. (DERAGeo) heteroaryl aldehydes through a rational engineering approach. We successfully performed iterative saturation mutagenesis DERAGeo, resulting in two enantiocomplementary variants, viz., (R)-selective DERAGeo-S185G (S)-selective DERAGeo-T12I/S185A, with enhanced catalytic efficiencies enantioselectivities. Remarkably, natural enantioselectivity DERAGeo was reversed single mutation (S185G). The synthetic applicability mutants demonstrated conducting aldol reactions on semipreparative scale, which both (R)- (S)-enantiomers aldols were isolated high yields (up 99%) excellent enantiopurities 99:1 e.r.).
Language: Английский