Effective renaturation of reduced lysozyme by gentle removal of urea

Yoshitake Maeda, Hiroshi Koga, Hidenori Yamada, Tadashi Ueda, Taiji Imoto

研究成果: Contribution to journalArticle査読

74 被引用数 (Scopus)

抄録

To increase the folding yield of concentrated reduced lysozyme, we developed a renaturation method by means of dialysis from concentrated urea with redox agents. After lysozyme was incubated in the reducing buffer (8 M urea solution) with oxidized glutathione, renaturation of reduced lysozyme was started by dialysis against the dialyzing buffer containing 8 M urea with redox agents. The urea concentration of the dialyzing bottle was gradually diluted with dialyzing buffer without urea at a flow rate of 0.1 ml/min by high pressure pump. Using this systematic dialysis, a concentration as high as 5 mg/ml of reduced lysozyme could be renaturated in 80% yield, while the folding yield was <5% even at a concentration of 1 mg/ml using a conventional rapid dilution method [Goldberg et al. (1991) Biochemistry, 30, 2790-2797]. Therefore, it was concluded that gentle removal of urea from denatured proteins, dissolved in concentrated urea solution, by means of dialysis should be useful to renature denatured proteins effectively.

本文言語英語
ページ(範囲)201-205
ページ数5
ジャーナルProtein Engineering, Design and Selection
8
2
DOI
出版ステータス出版済み - 2 1995

All Science Journal Classification (ASJC) codes

  • バイオテクノロジー
  • バイオエンジニアリング
  • 生化学
  • 分子生物学

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