KSBB Journal

  • 제23권5호
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  • Pages.369-374
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  • 2008
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  • 1225-7117(pISSN)
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  • 2288-8268(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Rhodotorula glutinis 유래의 고효율 재조합 Epoxide Hydrolase를 이용한 라세믹 Styrene Oxide의 비대칭 광학분할

Asymmetric resolution of racemic styrene oxide using recombinant Escherichia coli harboring epoxide hydrolase of Rhodotorula glutinis

  • 박규덕 (경성대학교 공과대학 식품생명공학과) ;
  • 최성희 (경성대학교 공과대학 식품생명공학과) ;
  • 김희숙 (경성대학교 공과대학 식품생명공학과) ;
  • 이은열 (경희대학교 환경.응용화학대학 화학공학과)
  • Park, Kyu-Deok (Dept. of Food Science and Biotechnology, Kyungsung Univ.) ;
  • Choi, Sung-Hee (Dept. of Food Science and Biotechnology, Kyungsung Univ.) ;
  • Kim, Hee-Sook (Dept. of Food Science and Biotechnology, Kyungsung Univ.) ;
  • Lee, Eun-Yeol (Dept. of Chemical Engineering, Kyung Hee Univ.)
  • 발행 : 2008.10.31
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초록

Rhdotorula glutinis epoxide hydrolase 유전자를 pColdI 벡터 와 pET-21b(+) 벡터에 재조합하여 제작한 E. coli를 생촉매로 사용하여 라세믹 styrene oxide에 대하여 회분식 가수분해 반응을 실시하였다. pET-21b(+)/RgEH 재조합 플라스미드 DNA를 가진 E. coli를 $15^{\circ}C$에서 저온 배양할 때 수용성 단백질 형태로 가장 많이 발현되었고, 입체선택적 가수분해 활성과 촉매 안정성이 가장 좋았다. 라세믹 styrene oxide 20 mM에 대하여 반응온도 $30^{\circ}C$에서는 반응시간 20분 동안에 수율 24.0%로 (S)-styrene oxide를 얻은 반면에, 반응온도를 $10^{\circ}C$로 낮추고 0.5% (w/v) Tween 20을 첨가하고 반응시키면 광학순도 99.0% ee 이상의 (S)-styrene oxide을 46.0%의 수율로 얻을 수 있었다. 최적조건에서 E 값은 6.68이었으며, 100 mM의 라세믹 styrene oxide에 대해서는 반응시간 50분에 이론 수율 50% 대비 40%의 높은 수율로 (S)-styrene oxide를 얻을 수 있었다.

The effects of reaction temperature and the addition of various detergents on the enantioselective hyrolysis activity of the recombinant Escherichia coli containing the epoxide hydrolase (EH) gene of Rhodotorula glutinis were investigated for the production of enantiopure styrene oxide. The recombinant E. coli harboring the EH gene from R. glutinis exhibited the enantiopreference toward (R)-styrene oxide with the maximum hydrolytic activity of $165.04{\mu}mol/min/mg$ of dry cell weight (dcw). The addition of 0.5% (w/v) Tween 20 at $10^{\circ}C$ increased the initial hydrolysis rate and enantioselectivity by 1.45-fold and 2.0-fold, respectively. Enantiopure (S)-styrene oxide was prepared with 99% ee enantiopurity and 46.0% yield (theoretical yield=50%) from 20 mM racemic styrene oxide.

키워드

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