对大肠杆菌异源表达乳酸片球菌素包涵体复性方法的研究

Research of the refolding of a heterologous pediocin expressed in Escherichia coli

本研究对E.coli BL21(DE3)基因工程菌高效表达的乳酸片球菌素融合蛋白包涵体进行了提取和变性溶解,并采用稀释复性,反稀释复性,透析复性,CTAB辅助复性和CTAB和β-CD结合辅助复性五种方法对乳酸片球菌素融合蛋白包涵体的变性溶液进行了复性的研究。结果表明CTAB和β-CD结合辅助复性的复性方法优于其它的复性方法,可以实现以较高浓度的包涵体变性蛋白溶液,较小体积的复性液来获得较高浓度的复性产物和达到相对稳定的复性率。

The inclusion body of the recombinant pediocin fusion protein was produced by the excess expression of gene engineering E. coil BL21 （ DE3 ）.The inclusion body was separated, denatured, and then refolded using direct dilution, reversed dilution, dialysis, CTAB assisted, and CTAB and ^-CD assisted refolding.The results showed that CTAB and #-CD assisted refolding have advantage over other refolding methods.Through this method,a higher concentration of denatured inclusion protein and a smaller volume renatured solution were used. And a higher concentration of refolding product and a relatively stable refolding rate were achieved.