Korean Chemical Engineering Research

The Korean Institute of Chemical Engineers (한국화학공학회)
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Stabilization of HRP Using Hsp90 in Water-miscible Organic Solvent

Hsp90을 이용한 유기용매에서의 과산화효소 안정화 연구

  • Chung, Ja Hee (Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University) ;
  • Choi, Yoo Seong (School of Chemical and Biological Engineering, Seoul National University) ;
  • Song, Seung Hoon (Bio-MAX institute, Seoul National University) ;
  • Yoo, Young Je (School of Chemical and Biological Engineering, Seoul National University)
  • 정자희 (서울대학교 협동과정 생물화학공학전공) ;
  • 최유성 (서울대학교 화학생물공학부) ;
  • 송승훈 (서울대학교 생명공학공동연구원) ;
  • 유영제 (서울대학교 화학생물공학부)
  • Received : 2006.01.03
  • Accepted : 2006.02.03
  • Published : 2006.02.28
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Abstract

Enzymes in organic media afford many advantages such as chiral synthesis and resolution, modification of fats and oils and production of biodegradable polymers. However, the nature of solvents influences the activity and stability of enzymes, and the presence of organic solvents always constitute a risk of enzyme inactivation. Heat-shock protein Hsp90, one of the molecular chaperone, was applied for understanding of enzyme inactivation and for increasing of enzyme stability in water-miscible organic solvent. Hsp90 showed stabilization effect on HRP in the 30％ of DMSO, in the 30％ and 50％ of dioxane. Hsp90 also showed reactivation effect on the inactivated HRP by water-miscible organic solvent such as dioxane and DMSO. In addition, structural analysis using fluorescence spectrophotometry and circular dichroism showed that exposure of HRP in water-miscible organic solvent caused appreciable conformational changes and enzyme inactivation, and the unfolded HRP by water-miscible organic solvent was refolded by Hsp90.

유기용매에서 효소를 이용하면 다양한 선택적 반응을 쉽게 수행할 수 있어 산업적 적용 가능성이 매우 높지만, 효소의 안정성 저하는 아직까지 큰 문제 중의 하나로 남아있다. 유기용매에서 효소 반응시 효소의 실활 원인과 효소의 안정화 방법 연구를 위하여 단백질의 folding에 관여하는 molecular chaperone의 일종인 heat-shock protein Hsp90을 이용하여, 대표적인 유기용매 반응시스템에서의 과산화효소 HRP 안정성 향상 연구를 수행하였다. 그 결과 Hsp90은 30％ DMSO, 30％ 및 50％ dioxane 완충용액에서 HRP의 실활 방지 효과를 보였고, 실활된 효소의 재생에도 탁월한 효과를 보였다. 그리고 형광분석과 CD(circular dichroism)에 의한 구조분석을 수행하여 Hsp90이 유기용매에 의해 unfolding되어 있는 효소를 다시 refolding하는 데 기여함을 알았다.

Keywords

References

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