L-精氨酸对提高蛋白质复性与纯化率的作用及其分析

Effect and Analysis of the Addition of L-arginine in the Renaturation and Purification Process of Denatured Protein

分别以还原变性的溶菌酶(Lys)和大肠杆菌(Escherichia coli)中表达的重组人酪氨酸激酶配体(Recombined human Flt3ligand,rhFL)为不同来源的变性蛋白,利用荧光光谱技术分析L-精氨酸(L-arginine,L-Arg)作为小分子添加剂时,蛋白质折叠液相色谱(PFLC)法对变性蛋白质的复性效果,以及变性蛋白复性过程中的复性效率、荧光强度与蛋白构象变化的规律与关系,为解决包涵体蛋白质难于复性和复性效率低的问题提供可靠的数据.

The denatured lysozyme（Lys）and the recombined human Flt3 ligand（rhFL）which express in Escherichia coli are different sources for denatured protein.The L-arginine as a small molecular additive in liquid chromatography for protein folding（PFLC）is used to analyze the result of refolding denatured protein by fluorescence spectroscopy technology.The analysis includes the relationship of the renaturation efficiency,the fluorescence intensity and the change of protein conformation in the denatured protein folding process.The results provide reliable data for solving the difficulties in refolding proteins from inclusion bodies and their low refolding efficiency.