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

Kód výsledku v 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>

Výsledek na webu

<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>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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

Název v anglickém jazyce

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

Popis výsledku anglicky

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

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CE - Biochemie

OECD FORD obor

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Projekt

<a href="/cs/project/GAP207%2F10%2F0135" target="_blank" >GAP207/10/0135: Probádání regioselektivit u evolučně získaných epoxidhydrolas</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2013

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Molecular Catalysis B-Enzymatic

ISSN

1381-1177

e-ISSN

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Svazek periodika

91

Číslo periodika v rámci svazku

JUL 2013

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

8

Strana od-do

44-51

Kód UT WoS článku

000318194300007

EID výsledku v databázi Scopus

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