Kinetic Resolution of Nearly Symmetric 3-Cyclohexene-1-carboxylate Esters Using a Bacterial Carboxylesterase Identified by Genome Mining

Abstract

A new bacterial carboxylesterase (CarEst3) was identified by genome mining and found to efficiently hydrolyze racemic methyl 3-cyclohexene-1-carboxylate (rac-CHCM) with a nearly symmetric structure for the synthesis of (S)-CHCM. CarEst3 displayed a high substrate tolerance and a stable catalytic performance. The enantioselective hydrolysis of 4.0 M (560 g•L −1 ) rac-CHCM was accomplished, yielding (S)-CHCM with a >99% ee, a substrate to catalyst ratio of 1400 g•g −1 , and a space-time yield of 538 g•L −1 •d −1 .

Enhanced stereodivergent evolution of carboxylesterase for efficient kinetic resolution of near-symmetric esters through machine learning

“…The intricate stereochemical structures of enzymes often manifest in distinctive spatial, hydrophobic, and electrostatic properties, particularly within the active center, forming the basis for high stereoselectivity [5][6][7] . However, enzymes face challenges maintaining high stereoselectivity, especially when confronted with substrates boasting nearly symmetric structures [8][9][10] . These substrates are commonly deemed "hard-to-bediscriminated," not only by chemical catalysts but also by biocatalysts 11,12 .…”

Asymmetric Synthesis of Optically Active 3-Cyclohexene-1-carboxylic Acid Utilizing Lactic Ester as a Chiral Auxiliary in the Diastereoselective Diels–Alder Reaction

“…6 It has been reported that more than 2 or 3 recrystallization processes are required to afford the salt with high diastereoselectivity. Meanwhile, Tanyeli and Turkut, 7 and Xu, Ni, and co-workers 8 individually reported biotransformations from the racemic 3-cyclohexene-1-carboxylic acid methyl ester 2. In this chiral resolution strategy, generally, discarding at least half of the starting racemic mixture is inevitable.…”