基于大肠杆菌蛋白质质谱鉴定的阳离子交换色谱优化研究  

Optimization of conditions for the separation with strong cation exchange chromatography based on Escherichia coli protein for mass spectrometry identification

作　　者：朱平川[1] 岑卫健[1] 付强[1] 胡炜[1] 卢洁[2] 

机构地区：[1]亚热带农业生物资源保护与利用国家重点实验室,广西大学,广西南宁530004 [2]广西大学生命科学与技术学院,广西南宁530004

出　　处：《湖北大学学报（自然科学版）》2014年第5期435-438,共4页Journal of Hubei University：Natural Science

基　　金：国家自然科学基金项目(31360459);广西大学科研基金项目(XJZ120280)资助

摘　　要：以大肠杆菌蛋白质的胰蛋白酶切产物为研究对象,对蛋白质的提取效率进行初步探讨,重点是对多维色谱分离中常用的强阳离子交换色谱的分离条件,包括缓冲液的种类、pH值,洗脱盐的种类、有机溶剂的比例以及上样量进行了考察和比较.结果表明:功率为400W,超声时间为15min,蛋白质的提取效率达到最高.在强阳离子交换色谱分离线性上样量范围内,10mmol/L磷酸二氢钾(pH=3.0)作为缓冲体系,在氯化钾溶液作为洗脱盐、流动相中乙腈的体积分数为30%的条件下进行梯度洗脱时,可获得最佳的分离结果.As a research of tryptic digestion products from Escherichia coli protein , on the extraction efficiency of protein were studied in this paper ,The focus is on the separation conditions of strong cation used multidimensional chromatography exchange chromatography . The conditions included the buffer species ,pH ,the type of the salt in mobile phase ,the proportion of organic solvent added in the elution solution ,and the loading amount of the sample .The results showed that ,in the power of 400 W ,ultrasonic time was 15 minutes ,achieved the highest extraction efficiency of protein . In the strong cation exchange chromatography separation of linear sample volume range ,10 mmol/L potassium dihydrogen phosphate （pH = 3 .0 ） of mobile phases and 30% （V/V ） organic solvent in mobile phases as buffer system ,as washing desalination in potassium chloride solution ,can obtain the best separation results .

关 键 词：大肠杆菌蛋白质 蛋白提取 强阳离子交换色谱 多肽分离 

分 类 号：Q814[生物学—生物工程]