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Language: korean

Conflict of Interest: In relation to this article, we declare that there is no conflict of interest.

Publication history: Received January 3, 2006
Accepted February 3, 2006

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/bync/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Hsp90을 이용한 유기용매에서의 과산화효소 안정화 연구

Stabilization of HRP Using Hsp90 in Water-miscible Organic Solvent

정자희 최유성¹ 송승훈² 유영제^1†

Ja Hee Chung Yoo Seong Choi¹ Seung Hoon Song² Young Je Yoo^1†

서울대학교 협동과정 생물화학공학전공, 151-742 서울시 관악구 신림동 산 56-1 ¹서울대학교 화학생물공학부, 151-742 서울시 관악구 신림동 산 56-1 ²서울대학교 생명공학공동연구원, 151-742 서울시 관악구 신림동 산 56-1

Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University, San 56-1, Shilim-dong, Gwanak-gu, Seoul 151-742, Korea ¹School of Chemical and Biological Engineering, Seoul National University, San 56-1, Shilim-dong, Gwanak-gu, Seoul 151-742, Korea ²Bio-MAX institute, Seoul National University, San 56-1, Shilim-dong, Gwanak-gu, Seoul 151-742, Korea

Korean Chemical Engineering Research, February 2006, 44(1), 92-96(5), NONE Epub 10 March 2006

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Abstract

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.

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.

Keywords

Hsp90 HRP Enzyme Activity Enzyme Stability Reactivation Water-miscible Organic Solvent

References

Laane C, Boeren S, Vos K, Veeger C, Biotechnol. Bioeng., 30(1), 81 (1987)
Lee SM, Yeo JS, Park K, Yoo YJ, Kor. J. Biotechnol. Bioeng., 15(1), 22 (2000)
Ryu K, Dordick JS, Biochemistry, 31(9), 2588 (1992)
Park BJ, Lee CH, Koo YM, Korean J. Chem. Eng., 22(3), 425 (2005)
Bell G, Halling PJ, Moor BD, Rees DG, Trends Biotechnol., 13(11), 468 (1995)
Schmitke JL, Stern LJ, Klibanov AM, Proc. Natl. Acad. Sci. USA, 95(22), 12918 (1998)
Jaenicke R, Buchner J, Chemtract: Biochem. Mol. Biol., 4, 1 (1993)
Wiech H, Buchner J, Zimmermann R, Jakob U, Nature, 358(6382), 169 (1992)
Freeman B, Morimoto R, Embo J., 15(12), 2969 (1996)
Kim JR, Choi YS, Yoo YJ, Biotechnol. Lett., 22(6), 486 (2000)
Xu Y, Lindquist S, Proc. Natl. Acad. Sci. USA, 90, 7074 (1993)
Jakob U, Buchner J, Trends Biochem. Sci., 19(5), 205 (1994)
Buchner J, Faseb J., 10(1), 10 (1996)