Protein refolding by reversed micelles utilizing solid-liquid extraction technique

Yukihisa Hashimoto, Tsutomu Ono, Masahiro Goto, T. Alan Hatton

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

This article reports that a reversed micellar solution is useful for refolding proteins directly from a solid source. The solubilization of denatured RNase A, which had been prepared by reprecipitation from the denaturant protein solution, into reversed micelles formulated with sodium di-2-ethylhexyl sulfosuccinate (AOT) has been investigated by s solid-liquid extraction system. This method is an alternative to the ordinary protein extraction in reversed micelles based on the liquid-liquid extraction. The solid-liquid extraction method was found to facilitate the solubilization of denatured proteins more efficiently in the reversed micellar media than the ordinary phase transfer method of liquid extraction. The refolding of denatured RNase A entrapped in reversed micelles was attained by adding a redox reagent (reduced and oxidized glutathion). Enzymatic activity of RNase A was gradually recovered with time in the reversed micelles. The denatured RNase A was completely refolded within 30 h. In addition, the efficiency of protein refolding was enhanced when reversed micelles were applied to denatured RNase A containing a higher protein concentration that, in the case of aqueous media, would lead to protein aggregation. The solid-liquid extraction technique using reversed micelles affords better scale-up advantages in the direct refolding process of insoluble protein aggregates.

Original languageEnglish
Pages (from-to)620-623
Number of pages4
JournalBiotechnology and Bioengineering
Volume57
Issue number5
DOIs
Publication statusPublished - Mar 5 1998
Externally publishedYes

Keywords

  • DNA
  • Protein refolding
  • RNase A
  • Reversed micelles
  • Solid-liquid extraction

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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