TITLE:
Efficient Protein Refolding Using Surfactants at High Final Protein Concentration
AUTHORS:
Hidetaka Noritomi, Yoshiyuki Kato, Satoru Kato
KEYWORDS:
Protein Refolding; Dilution Method; Surfactant; Hen Egg-White Lysozyme
JOURNAL NAME:
Journal of Surface Engineered Materials and Advanced Technology,
Vol.4 No.1,
January
16,
2014
ABSTRACT: The refolding
of denatured hen egg white lysozyme (HEWL) was examined by surfactants at a
high final refolded HEWL
concentration (1 mg/mL). Hexadecyltrimethylammonium bromide (CTAB) and sucrose
fatty acid monoester (DK-SS) were used to dissolve denatured HEWL without
denaturants such as guanidine hydrochloride (GuHCl) and urea. When denatured HEWL was perfectly
dissolved in buffer solutions containing surfactants and dithiothreitol (DTT), the concentration of
CTAB was about one-twentieth times less than that of DK-SS. The concentration
of CTAB strongly affected the refolding yield, and the maximum refolding yield
was obtained at 0.88 mM CTAB, which is around the critical micelle
concentration of CTAB. The refolding yield was influenced by the molar ratio of
oxidized glutathione (GSSG) to DTT, and the maximum refolding yield was obtained when [GSSG]/[DTT] was 1.5. The refolding
yield was markedly dependent upon the solution pH of HEWL, and exhibited 80% at
pH 5.2.