Novosphingobium aromaticivorans로 부터 신규 광학특이적 Epoxide Hydrolase 분석

Title
Novosphingobium aromaticivorans로 부터 신규 광학특이적 Epoxide Hydrolase 분석
Alternative Title
Cloning and Characterization of a Novel Enantioselective Epoxide Hydrolase from Novosphingobium aromaticivorans
Author(s)
우정희; 강지현; 강성균; 황영옥; 김상진
KIOST Author(s)
Kang, Sung Gyun(강성균)
Publication Year
2008-04-01
Abstract
A gene encoding a putative epoxide hydrolase (EHase) was identified by analyzing an open reading frame (ORF) of genome sequence of Novosphingobium aromaticivorans is available in public. N. aromaticivorans of revealed the presence of a putataive microsomal EHase (mEHase) containing the characteristic catalytic triad composed of Asp177, Glu328, and His355 as well as the oxyanion hole common to all mEHase. The enantioselective EHase gene (neh) was cloned, and the recombinant EHase (rNEH) could be purified to apparent homogeneity by one step of metal affinity chromatography, and further characterized. The purified NEH showed enantioselective hydrolysis toward styrene oxide, glycidyl phenyl ether, epoxybutane, and epichlorohydrin. The optimal pH and temperature for the EHase activity toward styrene oxide were pH 6.5 and 45 ℃, respectively. The purified NEH could preferentially hydrolyze (R)-styrene oxide with enantiomeric excess (ee) of more than 99% and 11.7% yield after 20 min incubation. The determination of 1-phenyl-1,2-ethanediol configuration derived from styrene oxide confirmed the enantioselective hydrolyzing activity of the purified NEH. No substrate inhibition was observed at initial substrate concentrations as high as 200 mM styrene oxide. Acknowledgement: This work was supported by KORDI in-house program (PE98230) and the Marine and Extreme Genome Research Center Program, Ministry of Marine Affairs and Fisheries.
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/30086
Bibliographic Citation
2008한국생물공학회 춘계국제학술대회, pp.160, 2008
Publisher
한국생물공학회
Type
Conference
Language
English
Publisher
한국생물공학회
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