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以重组人溶菌酶(rhLys)与重组β甘露聚糖酶(rMan)为体系,综合采用活性测定、非还原性SDSPAGE以及荧光发射光谱分析等方法研究了十六烷基三甲基溴化铵(CTAB)、CTAB与β环糊精(βCD)组成的人工分子伴侣以及其他种类复性助剂对重组蛋白质复性过程的影响.结果表明CTAB及人工分子伴侣均可有效地辅助rhLys复性,且rhLys与鸡卵清溶菌酶(Lys)呈现出类似的复性过程特性;人工分子伴侣可显著地提高rMan在高浓度下的复性率;表面活性剂带电性质、表面活性剂与蛋白质的摩尔比以及变性蛋白质的浓度等是影响复性率及复性产品分布的主要因素.这些结果对于此类复性技术应用于重组蛋白体系时的工艺选择和优化提供了重要的依据和参考.
The recombinant human lysozyme (rhLys) and recombinant β-mannanase (rMan) as the system, the comprehensive determination of cetyltrimethylammonium bromide by using activity determination, non-reducing SDSPAGE and fluorescence emission spectrometry, CTAB), CTAB and β-cyclodextrin (βCD) and other kinds of refolding aids on the refolding process of recombinant protein.The results show that CTAB and artificial chaperones can effectively assist the refolding of rhLys, and rhLys and Lysozyme (Lys) showed a similar process of refolding. The artificial chaperone significantly increased the renaturation rate of rMan at high concentration. The charged nature of surfactant, the molar ratio of surfactant to protein As well as the concentration of denatured protein are the main factors affecting the refolding rate and the distribution of refolding products.These results provide an important basis and reference for the process selection and optimization of such refolding technology when applied to recombinant protein system.