人工伴侣促进溶菌酶复性动力学  被引量:5

Kinetics of Lysozyme Refolding Facilitated by Artificial Chaperones

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作  者:董晓燕[1] 王颖[1] 孙彦[1] 

机构地区:[1]天津大学化工学院生物化工系,天津300072

出  处:《高校化学工程学报》2002年第3期306-310,共5页Journal of Chemical Engineering of Chinese Universities

基  金:教育部优秀年轻教师基金资助项目。

摘  要:利用复性和聚集竞争反应动力学模型描述自发复性和人工伴侣系统(十六烷基三甲基溴化铵与β-环糊精)促进溶菌酶复性动力学。在酶浓度为0.5-2.0mg·mL^-1、盐酸胍浓度为0.52.2mol·L^-1范围内,系统分析了盐酸胍浓度和人工伴侣浓度对复性动力学的影响。与自发复性相似,人工伴侣促进溶菌酶复性亦符合3级聚集反应动力学。人工伴侣系统的主要作用是抑制聚集体的生成速率,人而达到提高复性收率的效果。在盐酸胍浓度小于1.2mol·L^-1时。人工伴侣系统和盐酸胍对促进变性溶菌酶复性具有协同作用,两者共同作用对促进蛋白质复性具有显著效果。A kinetic model based on the competition between protein folding and aggregation was employed to express the refolding process of denatured/reduced lysozyme by direct dilution and that facilitated by the artificial chaperone system (ACS), i.e. cetyltrimethylammonium bromide and b -cyclodextrin. In the lysozyme concentration of 0.5~2.0 mgmL-1 and guanidinium chloride (GdmCl) concentration of 0.5~2.2 molL-1, the effect of GdmCl and ACS concentrations on the refolding kinetics was investigated. It was found that the aggregation reaction of the ACSassisted refolding can ben described as a third order reaction, similar to that of the unassisted refolding. The primary role of the ACS was to suppress the aggregation reaction rate, which leads to the increase of refolding yield. Since at GdmCl concentrations less than 1.2mol稬-1, the ACS and GdmCl work cooperatively in enhancing protein refolding, the ACS cooperation with relatively low denaturant concentration can be used to achieve a higher refolding yield for the protein refolding process.

关 键 词:体外复性 人工伴侣 动力学 溶菌酶 十六烷基三甲基溴化铵 蛋白质 Β-环糊精 

分 类 号:Q51[生物学—生物化学] Q556.2

 

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