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광학활성 Phenyl Oxirane 제조용 유전자 재조합 생촉매 개발

Development of Recombinant Saccharomyces cerevisiae Expressing Epoxide Hydrolase for the Preparation of Chiral Phenyl Oxirane

  • 이수정 (경성대학교 공과대학 식품공학과) ;
  • 이은정 (경성대학교 공과대학 식품공학과) ;
  • 김초희 (경성대학교 공과대학 식품공학과) ;
  • 이지원 (경성대학교 공과대학 식품공학과) ;
  • 김희숙 (경성대학교 공과대학 식품공학과) ;
  • 이은열 (경성대학교 공과대학 식품공학과)
  • 발행 : 2003.02.01

초록

Aspergillus niger #33로부터 RT-PCR 및 PCR방법을 이용하여 epoxide hydrolase (EH)유전자를 클로닝 하고 염기서 열을 분석한 결과 A. niger LCP521 유래의 EB와 85%수준의 유사성을 가지고 있었다. 클로닝된 EH 유전자를 Sacrharomyces cerevisiae에 형질전환 시킨 후 galactose를 inducer로 사용하여 발현시켰다. 유전자 재조합 S. cerevisiae는 라세믹 phenyl oxirane 기질에 대하여 입체선택적 가수분해능이 있음을 확인할 수 있었으며, 이러한 유전자 재조합 EH는 광학활성 에폭사이드 제조를 위한 생촉매로 응용될 수 있을 것이다.

Epoxide hydrolase (EH) gene from Aspergillus niger #33 was cloned from cDNA library generated by reverse transcriptase-polymerase chain reaction (RT-PCR). The nucleotide sequence analysis revealed that the deduced amino acid sequence was almost similiar to that of A. niger LCP521 previously reported. The cloned EH gene was transformed into Saccharomyces cerevisiae and expressed by addition of galactose. The recombinant S. cerevisiae showed hydrolytic activity toward racemic phenyl oxirane substrate based on chiral GC analysis, and can be used as a potential biocatalyst for the preparation of chiral phenyl oxirane.

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참고문헌

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피인용 문헌

  1. Biocatalytic preparation of chiral epichlorohydrins using recombinantPichia pastoris expressing epoxide hydrolase ofRhodotorula glutinis vol.9, pp.1, 2004, https://doi.org/10.1007/BF02949324