Protein Refolding System Using Holo-Chaperonin from the Thermophilic Bacterium Thermus thermophilus

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概要

• 論文の詳細を見る

• Thermus thermophilus holo-chaperonin(holo-cpn), a cpn60 14-mer-cpn10 7-mer complex, which was overproduced in Escherichia coli, was easily purified from a crude cell extract by heat treatment. The refolding of guanidine hydrochloride(GdnHCI)unfolded enzymes in the absence and presence of T. thermophilus holo-cpn was studied using Leuconostoc mesenteroides glucose-6-phoshate dehydrogenase(G6PD)and bovine deoxyribonuclease I(DNaseI)as model enzymes. T. thermophilus holo-cpn significantly enhanced the recovery of the enzyme activities at around 30-40℃, which is the same temperature range within which the enzymes spopntaneously recovered their highest activity. Since the complex formed by T. thermophilus cpn60 and cpn10 was stable, it could be separated from G6PD and DNase I in the refolding mixture by ultrafiltration using membranes with molecular weight cut-offs of 300kDa and 100kDa, respectively, and successfully reused. To increase the final concentration of renatured enzyme, step-wise addition of unfolded enzyme into the refolding mixture containing T. thermophilus holo-cpn was found to be effective. A combination of step-wise addition of unfolded enzyme and recycling of T. thermophilus holo-cpn using the ultrafiltration system significantly increased both the recovery of enzyme activity and the final concentration of renatured enzyme.

• 社団法人日本生物工学会の論文
• 1998-06-25

著者

• FUKUDA HIDEKI

  Division of Molecular Science, Graduate School of Science and Technology, Kobe University

• Fukuda Hideki

  Organization Of Advanced Science And Technology Kobe University

• Fukuda H

  Kobe Univ. Kobe

• Fukuda Hideki

  Department Of Molecular Science And Material Engineering Graduate School Of Science And Technology K

• Yohda M

  Tokyo Univ. Agriculture And Technol. Tokyo

• Yohda M

  Inst. Physical And Chemical Res. (riken) Saitama Jpn

• Fukuda H

  Kobe Univ. Kobe Jpn

• Fukuda Hideki

  Division Of Molecular Science Graduate School Of Science And Technology Kobe University

• KOHDA JIRO

  Division of Molecular Science, Graduate School of Science and Technology, Kobe University

• TESHIMA TADANARU

  Division of Bioscience, Kobe University

• KONDO AKIHIKO

  The Graduate School of Science and Technology, and Department of Chemical Science and Engineering, F

• TAGUCHI HIDEKI

  Research Laboratory of Resources Utilization, Tokyo Institute of Technology

• YOHDA MASAFUMI

  Biochemical Systems Laboratory, The Institute of Physical and Chemical Research(RIKEN)

• ENDO ISAO

  Biochemical Systems Laboratory, The Institute of Physical and Chemical Research(RIKEN)

• Miyashita Yukiko

  Department Of Biotechnology And Life Science Tokyo University Of Agriculture And Technology

• Endo I

  Inst. Physical And Chemical Res.(riken) Saitama

• Endo I

  Inst. Physical And Chemical Res. (riken) Saitama Jpn

• Endo Isao

  Biochemical Systems Laboratory The Institute Of Physical And Chemical Research (riken)

• Kohda J

  Hiroshima City University

• Kohda Jiro

  Division Of Molecular Science Graduate School Of Science And Technology Kobe University:(present Add

• Taguchi Hideki

  Research Laboratory Of Resources Utilization Tokyo Institute Of Technology

• Taguchi Hideki

  Research Laboratory For Surface Science Faculty Of Science Okayama University

• Yohda Masafumi

  Biochemical Systems Laboratory The Institute Of Physical And Chemical Research(riken)

• Teshima Tadanaru

  Division Of Bioscience Kobe University

• Fukuda Hideo

  Dept. Appl. Microbial Technology Kumamoto Institute Of Technology

• Yoshida Mamiko

  Precision Systems Science Co. Ltd.

• Fukuda Hideki

  Division Of Bioscience The Graduate School Of Science And Technology Kobe University

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