KSBB Journal

  • 제20권4호
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  • Pages.311-316
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  • 2005
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  • 1225-7117(pISSN)
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  • 2288-8268(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Bioinformatics를 활용한 토양미생물인 Gordonia westfalica Epoxide Hydrolase 생촉매 개발 및 Chiral Epoxides 제조 특성 분석

Bioinformatics based Identification and Characterization of Epoxide Hydrolase of Gordonia westfalica for the Production of Chiral Epoxides

  • 이수정 (경성대학교 공과대학 식품공학과) ;
  • 이은정 (경성대학교 공과대학 식품공학과) ;
  • 김희숙 (경성대학교 공과대학 식품공학과) ;
  • 이은열 (경성대학교 공과대학 식품공학과)
  • Lee Soo Jung (Department of Food Science and Technology, Kyungsung University) ;
  • Lee Eun Jung (Department of Food Science and Technology, Kyungsung University) ;
  • Kim Hee Sook (Department of Food Science and Technology, Kyungsung University) ;
  • Lee Eun Yeol (Department of Food Science and Technology, Kyungsung University)
  • 발행 : 2005.08.01
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초록

EH의 catalytic nucleophile residue, His-Asp로 구성된 charge relay system, oxyanion hole 등의 EH 관련 conserved domain의 아미노산 공통 서열을 참고로 하여 G. westfalica megaplasmid로부터 putative EH를 선별할 수 있었다. Bioinformatics를 기반으로 스크리닝한 G. westfalica에 의한 라세믹 styrene oxide 기질에 대한 입체선택성 가수분해 반응에 있어 중요 반응 parameter들인, pH 및 온도 등이 초기 가수분해반응속도에 미치는 영향을 분석하고, 최적 회분식 반응조건을 결정하였다. 최적 반응조건인 pH 7, 반응 온도 $30^{\circ}C$, 생촉매량 40 mg의 조건에서 약 5시간 20분간 반응을 통해 20 mM 라세믹 기질로부터 광학순도 $100\%$ ee인 (S)-styrene oxide를 $36.5\%$의 수율로 얻을 수 있었다.

Epoxide hydrolases (EHs) are versatile biocatalysts for the preparation of chiral epoxides by enantioselective hydrolysis from racemic epoxides. Various microorganisms were identified to possess a EH activity by multiple sequence alignment and analysis of conserved domain sequence from genomic and megaplasmid sequence data. We successfully isolated Gordonia westfalica possessing EH activity from various microbial strains from culture type collections. G. westfalica exhibited (R)-styrene oxide preferred enantioselective hydrolysis activity. Chiral (S)-styrene oxide with high optical purity $(>\;99\%)\;ee)$ and yield of $36.5\%$ was obtained from its racemate using whole-cell of G. westfalica.

키워드

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