Laboratory evolution of an epoxide hydrolase - Towards an enantioconvergent biocatalyst

0366238 - MBÚ 2012 RIV NL eng J - Článek v odborném periodiku
Kotík, Michael - Archelas, A. - Faměrová, Veronika - Oubrechtová, Pavla - Křen, Vladimír
Laboratory evolution of an epoxide hydrolase - Towards an enantioconvergent biocatalyst.
Journal of Biotechnology. Roč. 156, č. 1 (2011), s. 1-10. ISSN 0168-1656. E-ISSN 1873-4863
Grant CEP: GA ČR GAP207/10/0135
Výzkumný záměr: CEZ:AV0Z50200510
Klíčová slova: Directed evolution * Regioselectivity * Enantioconvergence
Kód oboru RIV: EE - Mikrobiologie, virologie
Impakt faktor: 3.045, rok: 2011

We performed a laboratory evolution study with the epoxide hydrolase from Aspergillus niger M200. This enzyme exhibits no enantioconvergence with the substrates styrene oxide or para-chlorostyrene oxide, i.e. racemic vicinal diols are produced from the racemic substrates. After saturation mutagenesis, screening by chiral gas chromatography revealed enzyme variants with improved enantioconvergence as manifested by an increased enantiomeric excess of the diol product. Nine amino acid exchanges accumulated in the active site and the substrate access tunnel over the course of 5 productive rounds of iterative saturation mutagenesis, resulting in an enantioconvergent epoxide hydrolase variant
Trvalý link: http://hdl.handle.net/11104/0201293