Directed evolution of metagenome-derived epoxide hydrolase for improved enantioselectivity and enantioconvergence

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F13%3A00422915" target="_blank" >RIV/61388971:_____/13:00422915 - isvavai.cz</a>
Result on the web
<a href="http://dx.doi.org/10.1016/j.molcatb.2013.02.006" target="_blank" >http://dx.doi.org/10.1016/j.molcatb.2013.02.006</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.molcatb.2013.02.006" target="_blank" >10.1016/j.molcatb.2013.02.006</a>

Result language
angličtina
Original language name
Directed evolution of metagenome-derived epoxide hydrolase for improved enantioselectivity and enantioconvergence
Original language description
We performed a directed evolution study with a metagenome-derived epoxide hydrolase (EH), termed Kau2. Homology models of Kau2 were built; we selected one of them and used it as a guide for saturation mutagenesis experiments targeted at specific residueswithin the large substrate binding pocket. During the molecular evolution process, we found several enzyme variants with higher enantioselectivity or enhanced enantioconvergence toward para-Chlorostyrene oxide. Improved enantioselectivities by up to a factor of 5, reaching an E-value of up to 130 with the R-enantiomer as the residual epoxide, were achieved by replacing amino acid pairs at the positions 110 and 113, or 290 and 291, which are positions located in the vicinity of two presumed binding sites for the epoxide enantiomers. The (R)-para-Chlorophenylethane-1,2-diol product exhibited a high enantiomeric excess (ee) of 97% at 50% conversion of the racemic epoxide for the most enantioselective variant. Further, five amino acid subs
Czech name
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Czech description
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Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
CE - Biochemistry
OECD FORD branch
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Project
<a href="/en/project/GAP207%2F10%2F0135" target="_blank" >GAP207/10/0135: Investigations into regioselectivity of evolved epoxide hydrolases</a><br>
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year
2013
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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