Earticle

초록

영어

Gloshedobin is one of promising therapeutic thrombin like enzymes, potentially useful for the treatment of blood clotting disorders through their anti-coagulant function. However, it is difficult to obtain a sufficient amount of the enzymes from the nature, so we use Escherichia coli to offer a route for the rapid and economical production of recombinant proteins. However, over-expression of recombinant gloshedobin in E.coli led to intracellular accumulation as solid aggregates known as inclusion bodies (IBs). We would like to develop novel refolding cocktail based refolding strategies by harnessing in vitro realization of a molecular chaperone network for renaturation of solubilized but denatured gloshedobin in a manner to mimic cellular folding
machinery.[1] In order to redesign the traditional batch dilution refolding schemes with the use of refolding cocktail containing recyclable molecular chaperones,[2] two types of beads functionalized with various molecular chaperones (e.g. DnaK, DnaJ, GrpE, ClpB, TF, GroEL, GroES) are prepared : 1) metal chelating beads loaded with in-house cloned His-tagged molecular chaperones, and 2) CNBr-activated Sepharose 4B covalently conjugated with chaperones. To address any plausible alteration on the chaperoning activity of each chaperone following the bead coupling, each chaperone is cloned to harbor the 6xHis-tag at either N- or C-terminus. Following exploration of chaperoning efficacy of individual bead attached chaperones on the refolding of gloshedobin, a recyclable refolding cocktail comprising essential chaperones at an optimal stoichiometry will be prepared to compare the efficiency of cocktail assisted
refolding strategy with that of the conventional batch dilution refolding strategy.

키워드

저자

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

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576