Carboxylesterases catalyze the enantioselective synthesis of chiral carboxylic acids and esters but exhibit limited enantioselectivity, particularly toward ethyl 3-cyclohexene-1- carboxylate (CHCE). Here, the authors establish a high-throughput method to generate a high-quality dataset for carboxylesterase AcEst1 from Acinetobacter sp. JNU9335 using CHCE as the substrate, use it to train a machine learning predictor and design two stereocomplementary mutants to synthesize both enantiomers of chiral CHCE.
- Zhe Dou
- Xuanzao Chen
- Guochao Xu
