Applied Chemistry for Engineering (공업화학)

The Korean Society of Industrial and Engineering Chemistry (한국공업화학회)
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Enantioselective Kinetic Resolution of Racemic Styrene Oxide using Recombinant Marine Fish Epoxide Hydrolase of Mugil cephalus

해양 어류 Mugil cephalus 유래의 에폭사이드 가수분해효소를 이용한 라세믹 styrene oxide의 입체선택적 분할 반응

  • Choi, Sung Hee (Dept. of Food Science and Biotechnology, Kyungsung University) ;
  • Kim, Hee Sook (Dept. of Food Science and Biotechnology, Kyungsung University) ;
  • Lee, Eun Yeol (Dept. Of Chem. Eng., Kyung Hee University)
  • 최성희 (경성대학교 공과대학 식품생명공학과) ;
  • 김희숙 (경성대학교 공과대학 식품생명공학과) ;
  • 이은열 (경희대학교 환경.응용화학대학 화학공학과)
  • Received : 2008.05.08
  • Accepted : 2008.09.16
  • Published : 2008.10.10
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Abstract

The microsomal epoxide hydrolase gene (referred to as mMCEH) of Mugil cephalus was cloned by PCR, and then inserted to pColdI and pET-21b(+) vector, respectively. The recombinant E. coli possessing the recombinant plasmids exhibited the enantioperference toward (R)-styrene oxide. When enantioselective kinetic resolutions were conducted with 20 mM racemic styrene oxide, enantiopure (S)-styrene oxide was obtained with high enantiopurity more than 99% enantiomeric excess (ee) and 24.50% yield by using the recombinant E. coli harboring pET-21b(+)/mMCEH.

해양 어류인 Mugil cephalus로부터 에폭사이드 가수분해효소(epoxide hydrolase, EH) 유전자를 PCR을 이용하여 클로닝하고, pColdI 및 pET-21b(+) 발현벡터에 삽입시켜 재조합 Escherichia coli 생촉매를 개발하여 각각에 대하여 가수분해 활성을 비교하였다. 재조합 E. coli 생촉매 $10mg\;dcw\;mL^{-1}$을 사용하여 20 mM 라세믹 styrene oxide를 입체선택적 가수분해를 한 결과, (R)-styrene oxide 기질에 대해서 입체선택성을 보였다. pET-21b(+)를 발현벡터로 사용하여 M. cephalus의 EH 유전자를 저온 발현시킨 재조합 E. coli 생촉매를 사용하여, 약 40 min 반응을 통해 광학순도 99% ee (enantiomeric excess) 이상인 (S)-styrene oxide를 최종 수율 24.5% (이론수율 50%)로 제조할 수 있었다.

Keywords

Acknowledgement

Supported by : 국토해양부

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