Earticle

초록

영어

Proteins have wide applications in the medical, industrial and agricultural fields. They are usually produced in huge quantities in host cells such as Escherichia coli. However, high-level expression of recombinant proteins in E. coli often results in accumulation of insoluble aggregates known as inclusion bodies (IBs), thus requiring further solubilization, refolding and purification procedures to achieve functionally active products. Molecular chaperones have been applied successfully to refold various proteins both in vivo and in vitro, opening a new era in protein refolding. However, the exact function of individual molecular chaperones and the interaction between target protein and molecular chaperones are still unclear. We demonstrated that refolding cocktail comprising ClpB/DnaKJE with ATP regeneration system and a refolding additive, PEG, could significantly enhance the refolding efficiency of heat-denatured malate dehyrogenase (MDH). To further clarify the individual or synergistic roles of each chaperone, various molecular chaperones including His-ClpB, His-DnaK, His-DnaJ, His-GrpE, His-Gro EL, His-Gro ES, and His-Trigger Factor were recently cloned, expressed and purified, respectively. By using different temperature for heat-denaturation of MDH, protein aggregates of varying size were produced. Following the characterization of these protein aggregates (e.g. susceptibility to proteolysis and kinetics of solubilization), chaperoning efficiencies of the His-tagged molecular chaperones on the renaturation of the protein aggregates are being studied systematically. It is expected that our study will provide a better understanding of chaperone-assisted refolding process, thereby facilitating engineering implementation of a novel refolding strategy, folding-like-refolding approach based on refolding cocktail.

키워드

저자

• Lin ZHAI [ School of Chemical Engineering, Sungkyunkwan University, Suwon, 440-746, Republic of Korea. ]
• Jiryang JANG [ School of Chemical Engineering, Sungkyunkwan University, Suwon, 440-746, Republic of Korea. ]
• Sung-Eun KIM [ School of Chemical Engineering, Sungkyunkwan University, Suwon, 440-746, Republic of Korea. ]
• Woo Seok CHOE [ School of Chemical Engineering, Sungkyunkwan University, Suwon, 440-746, Republic of Korea. ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576