1. Directed evolution of an enantioselective lipase with broad substrate scope for hydrolysis of alpha-substituted esters.
- Author
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Engström K, Nyhlén J, Sandström AG, and Bäckvall JE
- Subjects
- Candida enzymology, Catalytic Domain, Hydrolysis, Kinetics, Lipase chemistry, Models, Molecular, Phenylpropionates chemistry, Phenylpropionates metabolism, Stereoisomerism, Substrate Specificity, Directed Molecular Evolution methods, Esters chemistry, Esters metabolism, Lipase genetics, Lipase metabolism
- Abstract
A variant of Candida antarctica lipase A (CalA) was developed for the hydrolysis of alpha-substituted p-nitrophenyl esters by directed evolution. The E values of this variant for 7 different esters was 45-276, which is a large improvement compared to 2-20 for the wild type. The broad substrate scope of this enzyme variant is of synthetic use, and hydrolysis of the tested substrates proceeded with an enantiomeric excess between 95-99%. A 30-fold increase in activity was also observed for most substrates. The developed enzyme variant shows (R)-selectivity, which is reversed compared to the wild type that is (S)-selective for most substrates.
- Published
- 2010
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