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Towards Quantitative Computer‐Aided Studies of Enzymatic Enantioselectivity: The Case of Candida antarctica Lipase A
Author(s) -
Frushicheva Maria P.,
Warshel Arieh
Publication year - 2012
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201100600
Subject(s) - candida antarctica , enantioselective synthesis , lipase , enantiomer , sampling (signal processing) , hydrolysis , biocatalysis , enzyme , chemistry , stereoselectivity , stereochemistry , computational biology , combinatorial chemistry , computer science , biochemistry , biology , catalysis , ionic liquid , filter (signal processing) , computer vision
Abstract The prospect for consistent computer‐aided refinement of stereoselective enzymes is explored by simulating the hydrolysis of enantiomers of an α‐substituted ester by wild‐type and mutant Candida antarctica lipase A, using several strategies. In particular, we focused on the use of the empirical valence bond (EVB) method in a quantitative screening for enantioselectivity, and evaluate both k cat and k cat / K M of the R and S stereoisomers. We found that an extensive sampling is essential for obtaining converging results. This requirement points towards possible problems with approaches that use a limited conformational sampling. However, performing the proper sampling appears to give encouraging results and to offer a powerful tool for the computer‐aided design of enantioselective enzymes. We also explore faster strategies for identifying mutations that will help in augmenting directed‐evolution experiments, but these approaches require further refinement.

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