title: Towards quantitative computer-aided studies of enzymatic enantioselectivity: the case of Candida antarctica lipase A.
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.
authors.affiliation: Department of Chemistry, University of Southern California, Los Angeles, CA 90089-1062, USA.
authors.forename: Maria P
authors.initials: MP
authors.lastname: Frushicheva
authors.forename: Arieh
authors.initials: A
authors.lastname: Warshel
mesh_terms: D000215:Acylation; D002175:Candida / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D017076:Computer-Aided Design; D008049:Lipase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D009154:Mutation; D013237:Stereoisomerism; D013816:Thermodynamics
chemical_list: D008049:Lipase