Initial protein concentration and residual denaturant concentration strongly affect the batch refolding of hen egg white lysozyme

Andrew D Guise, Julian B Chaudhuri

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The effects of several variables on the refolding of hen egg white lysozyme have been studied. Lysozyme was denatured in both urea and guanidine hydrochloride (GuHCl), and batch refolded by diln. (100-1000-fold) into 0.1 M Tris-HCl, pH 8.2, 1 mM EDTA, 3 mM reduced glutathione, and 0.3 mM oxidized glutathione. Refolding was found to be sensitive to temp., with the highest refolding yield obtained at 50 Deg. The apparent activation energy for lysozyme refolding was found to be 56 kJ/mol. Refolding by diln. results in low concns. of both denaturant and reducing agent species. It was found that the residual concns. obtained during diln. (100-fold diln.: [GuHCl] = 0.06 mM, [DTT] = 0.15 mM) were significant and could inhibit lysozyme refolding. This study has also shown that the initial protein concn. (1-10 mg/mL) that is refolded is an important parameter. In the presence of residual GuHCl and DTT, higher refolding yields were obtained when starting from higher initial lysozyme concns. This trend was reversed when residual denaturant components were removed from the refolding buffer. [on SciFinder (R)]
Original languageEnglish
Pages (from-to)410-418
Number of pages9
JournalBiotechnology and Bioprocess Engineering
Volume6
Issue number6
Publication statusPublished - 2001

Keywords

  • Protein folding
  • initial protein concn. and residual denaturant concn. strongly affect the batch refolding of hen egg white lysozyme)
  • PROC (Process) (recombinant
  • protein refolding denaturant concn
  • Temperature effects (initial protein concn. and residual denaturant concn. strongly affect the batch refolding of hen egg white lysozyme)
  • lysozyme refolding guanidine concn
  • Proteins Role
  • PEP (Physical
  • CPS (Chemical process)
  • engineering or chemical process)

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