Efficient Protein Refolding Using Surfactants at High Final Protein Concentration

Hidetaka Noritomi; Yoshiyuki Kato; Satoru Kato

doi:10.4236/jsemat.2014.41002

Home > Journals > Chemistry & Materials Science | Engineering > JSEMAT

JSEMAT> Vol.4 No.1, January 2014

Share This Article:

Open Access

Efficient Protein Refolding Using Surfactants at High Final Protein Concentration

Abstract Full-Text HTML

Download as PDF (Size:245KB) PP. 9-13

DOI: 10.4236/jsemat.2014.41002 4,419 Downloads 6,198 Views Citations

Author(s) Leave a comment

Hidetaka Noritomi, Yoshiyuki Kato, Satoru Kato

Affiliation(s)

Department of Applied Chemistry, Tokyo Metropolitan University, Tokyo, Japan.

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.

KEYWORDS

Protein Refolding; Dilution Method; Surfactant; Hen Egg-White Lysozyme

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.

References

[1]	M. J. Rosen, “Surfactants and Interfacial Phenomena,” 3rd Edition, John Wiley & Sons, Hoboken, 2004. http://dx.doi.org/10.1002/0471670561

[2]	F. Franks, “Protein Biotechnology,” Humana Press Inc., New York, 1993. http://dx.doi.org/10.1007/978-1-59259-438-2

[3]	E. D. B. Clark, “Refolding of Recombinant Proteins,” Current Opinion in Biotechnology, Vol. 9, No. 2, 1998, pp. 157-163. http://dx.doi.org/10.1016/S0958-1669(98)80109-2

[4]	T. E. Creighton, “Proteins Structures and Molecular Properties,” 2nd Edition, W. H. Freeman and Company, New York, 1993.

[5]	H. Noritomi, T. Takasugi and S. Kato, “Refolding of Denatured Lysozyme by Water-In-Oil Microemulsions of Sucrose Fatty Acid Esters,” Biotechnology Letters, Vol. 30, No. 4, 2008, pp. 689-693. http://dx.doi.org/10.1007/s10529-007-9587-z

[6]	P. Jollès, “Lysozymes: Model Enzymes in Biochemistry and Biology,” 1st Edition, Birkhäuser Verlag, Basel, 1996.

[7]	V. P. Saxena and D. B. Wetlaufer, “Formation of Three-Dimensional Structure in Proteins. Rapid Nonenzymatic Reactivation of Reduced Lysozyme,” Biochemistry, Vol. 9, No. 25, 1970, pp. 5015-5022. http://dx.doi.org/10.1021/bi00827a028

[8]	Y.-C. Chen, W.-T. Lin, J. W. Wu and H.-S. Liu, “Efficient Lysozyme Refolding at a High Final Concentration and a Low Dilution Factor,” Process Biochemistry, Vol. 47, No. 12, 2012, pp. 1883-1888. http://dx.doi.org/10.1016/j.procbio.2012.06.026

[9]	E. D. Goddard and K. P. Ananthapadmanabhan, “Interactions of Surfactants with Polymers and Proteins,” CRC Press, New York, 1993.