关键词: 蛋白质复性, 重组人溶菌酶, 重组β-甘露聚糖酶, CTAB, β-CD

Abstract: The feasibility of surfactant assisted refolding, which was performed by diluting denatured protein sample into a refolding buffer containing a surfactant of appropriate concentration, was examined by using recombinant human lysozyme (rhLys) and recombinant β-mannanase (rMan) as protein samples.The effectiveness of CTAB, an artificial chaperone consisting of CTAB and β-CD, as well as other kinds of surfactants was investigated respectively.The effects of these surfactants on refolding yield and product distribution were determined by using biological activity assay, non-reductive SDS-PAGE and fluorescence emission spectrometry. A significant increase in refolding yield occurred for rhLys when using CTAB and the artificial chaperone, compared to that obtained by using direct dilution. Moreover, the refolding of rhLys presented similar process characteristics to that of native lysozyme (Lys).In both cases, the yield and product distribution depended on the molar ratio of surfactant to protein. The artificial chaperone worked also for rMan, particularly at a high concentration of the denatured rMan. It was shown that the charge property and concentration of surfactant, and the concentration of protein being refolded are major parameters determining the success of refolding process. The results described above are helpful to both the understanding of process mechanism and the application of this new refolding method.

Key words: 蛋白质复性, 重组人溶菌酶, 重组β-甘露聚糖酶, CTAB, β-CD