刺桐胰蛋白酶抑制剂的色谱复性研究

Study on chromatographic renaturation of Erythrina trypsin inhibitor

目的 研究在大肠杆菌中表达的刺桐胰蛋白酶抑制剂 (Erythrinatrypsininhibitor,ETI)的色谱复性。方法 使用透析、阴离子交换和凝胶色谱法对ETI复性进行研究 ,比较 3种复性方法的相对复性率及蛋白质回收率。结果 3种复性方法的复性率分别为 2 0 . 1% ,32 . 1%和 5 0 . 8% ,蛋白质回收率分别为 16 . 6 % ,15 . 1%和 5 5 . 2 %。结论 凝胶色谱复性优于其他两种方法。色谱复性使蛋白质的复性与纯化同步进行 ,简化了操作程序 ,提高了产品的回收率。

OBJECTIVE: To study the renaturation of Erythrina trypsin inhibitor(ETI) expressed in Escherichia coli by means of chromatography. METHODS: Renaturation of ETI was performed by dialysis, anion exchange chromatography and gel filtration. The relative renaturation ratios and relative recoveries of protein of the three methods were compared. RESULTS: Dialysis, anion exchange chromatography and gel filtration renatured ETI with relative renaturation ratios of 20.1%, 32.1% and 50.8% and relative recoveries of protein 16.6%, 15.1% and 55.2%, respectively. CONCLUSION: The renaturation by gel filtration appeared to be high efficient than the other two methods. The means of chromatographic renaturation can renature and purify the protein at the same time, thus simplify the procedure of operation and promote the recovery ratio of the product.