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Enantioselective Enzymes by Computational Design and In Silico Screening
Author(s) -
Wijma Hein J.,
Floor Robert J.,
Bjelic Sinisa,
Marrink Siewert J.,
Baker David,
Janssen Dick B.
Publication year - 2015
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201411415
Subject(s) - enantioselective synthesis , epoxide hydrolase , in silico , chemistry , substrate (aquarium) , steric effects , stereoselectivity , directed evolution , combinatorial chemistry , enzyme , stereochemistry , mutant , biochemistry , catalysis , biology , ecology , gene , microsome
Computational enzyme design holds great promise for providing new biocatalysts for synthetic chemistry. A strategy to design small mutant libraries of complementary enantioselective epoxide hydrolase variants for the production of highly enantioenriched ( S , S )‐diols and ( R , R )‐diols is developed. Key features of this strategy (CASCO, catalytic selectivity by computational design) are the design of mutations that favor binding of the substrate in a predefined orientation, the introduction of steric hindrance to prevent unwanted substrate binding modes, and ranking of designs by high‐throughput molecular dynamics simulations. Using this strategy we obtained highly stereoselective mutants of limonene epoxide hydrolase after experimental screening of only 37 variants. The results indicate that computational methods can replace a substantial amount of laboratory work when developing enantioselective enzymes.