荧光相图法研究变性蛋白溶菌酶在脲和盐酸胍溶液中的复性过程

Studies on the Refolding Procedure of Protein Lysozyme Denatured by Urea and Guanidine Hydrochloride by "Phase Diagram" Method of Fluorescence

[目的]研究了变性体系无还原剂和有还原剂存在时,变性蛋白溶菌酶在脲和盐酸胍溶液中的复性过程。[方法]利用荧光相图法。[结果]当变性体系中无还原剂存在时,变性蛋白溶菌酶在脲和盐酸胍溶液中的复性过程均符合"四态模型";而当变性体系中有还原剂存在时,变性蛋白溶菌酶在脲和盐酸胍溶液中的复性过程均符合"二态模型"。当变性体系中无还原剂存在时,蛋白溶菌酶的复性过程较复杂;比较变性和复性过程推测,非还原脲和盐酸胍诱导的蛋白溶菌酶变性及复性过程并不是2个完全可逆的过程,而还原脲和盐酸胍诱导的蛋白溶菌酶变性及复性过程有可能是可逆的。[结论]该研究可为开发合成杀菌谱广、成本低和安全性高的溶菌酶提供帮助。

[Objective] The refolding procedure of non-reducing and reduced protein lysozyme in urea and guanidine hydrochloride solution were studied.[Method] The ＂phase diagram＂ method of fluorescence was used.[Result] In the absence of reducing agent,the refolding procedure followed a four-state model,while in the presence of the reducing agent,the refolding procedure followed a two-state model.The refolding procedure of non-reducing protein lysozyme in urea and guanidine hydrochloride solution was more complicated.Through the comparison between the unfolding and refolding procedures of protein lysozyme in urea and guanidine hydrochloride solution,it was found that the refolding procedure of non-reducing protein lysozyme could be considered to be irreversible to their unfolding procedure,while the refolding procedure of reduced protein lysozyme could be considered to be reversible to their unfolding procedure.[Conclusion] The study can provide help for developing and synthetizing the lysozyme with a broad spectrum of disease control,low cost and high safety.