离子交换层析纯化蔗糖酶实验方法改进研究

Studies on Method Development of Invertase Ion Exchange Chromatogram Purification

通过对不同离子交换层析方法的比较 ,进行酵母蔗糖酶纯化实验改进 ,采用 Q Sepharose离子交换介质 ;流动相经 5 0 min由 0 .0 5 MTris- HCLp H7.3缓冲液到 1MNa CL的 0 .0 5 MTris- HCL p H7.3缓冲液 ,进行梯度洗脱分离 ,分离效果最好 ,蔗糖酶与杂蛋白得到了有效的分离 ,回收率为 93.2 %,是现有实验的 3.17倍 ,浓缩倍数为 2 .91,是现有实验的 1.4 3倍 ,提高了实验效果。

A comparison was made in different ion exchange chromatogram to develop test methods carried experiments of the yeast's invertase purification. It uses Q Sepharose ion exchange media; start buffer A: 0.05 Mtris-HCL, pH7.3; elution buffer B:0.05MTris-HCL+1MNaCL, pH7.3; Gradient: 0~100% elution buffer B in 10 column volumes. The result shows that the recovery rate reachs 93.2%(3.17 times that of the present labs' data) and the concentration fold is 2.91(1.43 times that of the present data). Investase and other impurity protein are separated efficiently.