Refolding of Denatured and Reduced Lysozyme with Cysteine/Cystine Red/Ox Solution in Diafiltration
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概要
- 論文の詳細を見る
Refolding of reduced and denatured protein in vitro has been an important issue for both basic research and applied biotechnology. Refolding at low protein concentration requires large volumes of refolding buffer. Diafiltration method is useful to control the denaturant and red/ox reagents in the refolding solution. We constructed a refolding procedure for high concentrations of reduced and denatured lysozyme of about 10 mg/ml (700 μM) on linear reduction of urea concentration in diafiltration, 0.8 mM cystine and 8 mM cysteine under nitrogen at 2 atm. This method can obtain about 90% refolding yield at 700 μM and almost 100% in 350 μM lysozyme. The refolding yields during the diafiltration can be simulated using the competitive reaction between the refolding and aggregation. In the red/ox control with cysteine and cystine, a rate order of aggregation reaction of near 2 is obtained.
- 社団法人 化学工学会の論文
著者
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吉井 英文
鳥取大学大学院工学研究科 化学・生物応用工学専攻
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Linko P
Department Of Chemical Technology Helsinki University Of Technology
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ITO Daisuke
Department of Surgery and Surgical Basic Science, Kyoto University
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Furuta Takeshi
Tottori University
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YOSHII Hidefumi
Department of Biotechnology, Tottori University
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FURUTA Takeshi
Department of Biotechnology, Tottori University
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LINKO Pekka
Department of Chemical Technology, Helsinki University of Technology
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YONEHARA Takahiro
Department of Biotechnology, Tottori University
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Yoshii H
Tottori University
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Yoshii H
Tottori Univ. Tottori Jpn
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Yoshii H
Toyama National Coll. Technol. Toyama Jpn
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Yoshii Hidefumi
Department Of Biochemical Engineering Toyama National College Of Technology
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Furuta T
Tottori University
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Furuta Takeshi
Department Of Biotechnology Faculty Of Engineering Tottori University
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Ito D
Graduate School Of Engineering Osaka Prefecture University Department Of Applied Materials Science
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Linko Pekka
Department Of Chemical Technology Helsinki University Of Technology
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Yoshii Hidefumi
Department Of Applied Biological Science Kagawa University
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Yonehara Takehiro
Department Of Biotechnology Tottori University
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Ito Daisuke
Department Of Mechanical Science And Engineering Nagoya University
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Ito Daisuke
Department Of Applied Materials Science. Graduate School Of Eneineerine. Osaka Prefecture University
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Ito Daisuke
Department Of Advanced Bioscience Faculty Of Agriculture Kinki University
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ITO Daisuke
Department of Biotechnology, Tottori University
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