利用电泳滴定曲线选择多核苷酸磷酸化酶快速离子交换层析的最佳条件

Use of electrophoretic titration curve for predicting optimal chromatographic conditions for purification of polynucleotide phosphorylase by fast ion-exchange chromatography

目的 :选择多核苷酸磷酸化酶快速离子交换层析的最佳条件。方法 :制备大肠杆菌提取物的电泳滴定曲线 ,根据该曲线选择快速离子交换层析MonoSI柱的洗脱缓冲液的pH。结果 :根据该滴定曲线选择pH 8.0作为快速离子交换层析MonoSI柱的洗脱缓冲液的pH ,在该条件下只一步层析 ,得到的多核苷酸磷酸化酶的比活性提高2 50 0倍。结论

OBJECTIVE:Selection of optimal chromatographic conditions for purification of polynucleotide phosphorylase in fast ion-exchange chromatography.METHODS:Preparation of electrophoretic titration curve of E. coli extract and prediction of optimal conditions for fast ion-exchange chromatography.RESULTS:According to the electrophoretic titration curve of E. coli extract,the buffer with pH 8.0 was chosen as eluent for the fast ion-exchange chromatography on Mono SI column.Polynucleotide phosphorylase with a specific activity of 2500-fold higher than that of the original extract was obtained in one step.CONCLUSION:Electrophoretic titration curve could be used to predict optimal chromatographic conditions for ion-exchange chromatography.