核糖核酸酶Onconase的稀释复性条件优化研究

Study on Optimization of Dilution Renaturation Conditions of RNase Onconase

采用稀释复性方法从包涵体中复性得到有活力的Onconase蛋白,并与传统的醋酸沉淀复性方法进行了比较,考察了复性温度、尿素浓度、氧化型谷胱甘肽(GSSG)加入时间对复性效果的影响。结果表明,在最佳复性温度为4℃、采用最佳复性体系(先加GSSG的含2mol·L-1尿素的稀释复性缓冲溶液)时,复性纯化后Onconase含量和回收率明显高于醋酸沉淀复性法。

The dilution renaturation method was appliedto obtain active Onconase from inclusion and com- pared with the traditional acetic acid sinking method. The effects of renaturation temperature, urea concentra- tion and addition time of GSSG on renaturation efficiency were explored. The results showed that,the Onconase content and recovery were obviously higher than that of acetic acid sinking method after renaturation and purifi- cation under the optimum renaturation temperature of 4 ℃ with the optimum renaturation system composed of dilution optimum renaturation buffer solution containing 2 mol ＇ L-1 urea which was added after GSSG.