Enantioselective Enzymes by Computational Design and In Silico Screening

• Published in: Angewandte Chemie Vol. 127; no. 12; pp. 3797 - 3801
• Main Authors: Wijma, Hein J., Floor, Robert J., Bjelic, Sinisa, Marrink, Siewert J., Baker, David, Janssen, Dick B.
• Format: Journal Article
• Language: English; German
• Published: Weinheim WILEY-VCH Verlag 16-03-2015; WILEY‐VCH Verlag; Wiley Subscription Services, Inc
• Subjects: Binding; Biokatalyse; Chemistry; Computation; Computer simulation; Computerchemie; Enzymes; Epoxid-Hydrolase; Molecular dynamics; Mutations; Screening; Screening-Methoden; Stereoselektivität; Strategy
• ISSN: 0044-8249; 1521-3757
• DOI: 10.1002/ange.201411415

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.
Enzyme vom Reißbrett könnten vielversprechende Biokatalysatoren werden. Eine computerchemische Strategie liefert, nach dem experimentellen Screening von nur 37 Varianten, enantiodivergente und hoch enantioselektive Mutanten der Limonenepoxid‐Hydrolase. Folglich können computerchemische Methoden bei der Entwicklung enantioselektiver Enzyme eine große Menge an Laborarbeit einsparen.