一种快速分离纯化金葡萄球菌肠毒素C_2的方法  

A rapid method for the separation and purification of staphylococcal enterotoxin C_2

作　　者：李洪义[1,2] 陈彦竹[2] 杨宏丽[2] 陈晓辉[1] 毕开顺[1] 

机构地区：[1]沈阳药科大学药学院,辽宁沈阳110016 [2]沈阳协合生物制药股份有限公司,辽宁沈阳110179

出　　处：《沈阳药科大学学报》2009年第7期574-578,共5页Journal of Shenyang Pharmaceutical University

摘　　要：目的建立一种简单快捷的方法,用来从金黄色葡萄球菌的发酵液(以下简称金葡液)中纯化分离肠毒素C2(staphylococcal enterotoxin C2,SEC2)。方法金葡液首先经超滤浓缩,之后再分别经阳离子交换层析和阴离子交换层析进行选择性分离SEC2,用SDS-PAGE电泳、HPLC法检测纯度及相对分子质量,飞行质谱测定精确分子质量,对其进行N-端氨基酸序列分析。结果分离得到的SEC2电泳纯及色谱分析纯度均达到质量分数98%以上,经各项理化指标检测均与文献一致,在等电点上表现出微不均匀性,等电点pI为7.49和6.74,该蛋白的分子质量为27.58 ku,N-端氨基酸序列分析与文献报道的SEC2序列一致(ESQPDPTPDELHKSS),最大吸收波长为277.5 nm。结论用该方法得到的SEC2纯度高,回收率质量分数高达50%,适宜于大批量制备SEC2。Objective To develop a simple and convenient method to purify staphylococcal enterotoxin C2 （ SEC2 ） from the supematant of staphylococcus aureus culture. Methods SEC2 was purified by ultrafiltration, anion ion-exchange chromatography, and cation ion-exchange chromatography sequentially. The molecular weight of SEC2 was detected by the SDS-PAGE and flight time mass-spectrum. The purity of SEC2 was detected by the SDS-PAGE and high performance liquid chromatography. The purified SEC2 was verified by amino acids sequence analysis. Results The purity of purified SEC2 was greater than 98% （w） which was consistent with literature, including microheterogeneity of isoelectric point, pI 7. 49 and 6. 74, the molecular weight was 27.58 KD. The amino acids sequence of purified SEC2 was consistent with what was reported in literatures（ESQPDPTPDELHKSS）. The maximum absorption wavelength of purified SEC2 was 277.5 nm. Conclusions The method is rapid, simple and convenient with high SEC2 purity, the recovery is more than 50% （w）. This method is suitable to large scale preparation of SEC2.

关 键 词：金葡菌肠毒素C2(SEC2) 阴离子交换层析 阳离子交换层析 微不均匀性 

分 类 号：R917[医药卫生—药物分析学]