RESUMEN

A gene encoding an esterase from Bacillus aryabhattai (BaCE) was identified, synthesized and efficiently expressed in the Escherichia coli system. A semi-rational protein engineering was applied to further improve the enzyme's enantioselectivity. Under the guidance of the molecular docking result, a single mutant BaCE-L86Q and a double mutant BaCE-L86Q/G284E were obtained, with its Emax value 6.4 times and 13.9 times of the wild-type BaCE, respectively. The recombinant BaCEs were purified and characterized. The overwhelming E value demonstrated that BaCE-L86Q/G284E was a promising biocatalyst for the biological resolution to prepare (S)-indoline-2-carboxylic acid.

Asunto(s)

Ácidos Carboxílicos , Esterasas , Bacillus , Escherichia coli/genética , Escherichia coli/metabolismo , Esterasas/metabolismo , Indoles , Simulación del Acoplamiento Molecular , Ingeniería de Proteínas