Towards Quantitative Computer Aided Studies of Enzymatic Enantioselectivity: The case of Candida antarctica lipase A

• Published in: ChemBioChem
• Main Authors: Frushicheva, Maria P., Warshel, Arieh
• Format: Text
• Language: English
• Published: 2011
• Subjects: Article; Antarc*; Antarctica
• Online Access: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3414264; http://www.ncbi.nlm.nih.gov/pubmed/22190449; https://doi.org/10.1002/cbic.201100600

The prospect for consistent computer aided refinement of stereoselective enzymes is explored by simulating of the hydrolysis of enantiomers of an α-substituted ester by the wild type and mutants of CALA, using several strategies. In particular we focus on the use of the empirical valence bond (EVB) method in a quantitative screening for enantioselectivity, evaluating both kcat and kcat/KM in the R and S stereoisomers. It is found that an extensive sampling is essential for obtaining converging results. This requirement points out 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 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 requires further refinement.