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Efficient Protein Refolding Using Surfactants at High Final Protein Concentration

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DOI: 10.4236/jsemat.2014.41002    4,310 Downloads   6,009 Views   Citations

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.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

H. Noritomi, Y. Kato and S. Kato, "Efficient Protein Refolding Using Surfactants at High Final Protein Concentration," Journal of Surface Engineered Materials and Advanced Technology, Vol. 4 No. 1, 2014, pp. 9-13. doi: 10.4236/jsemat.2014.41002.

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