Walton C M, Wu C H, Wu G Y
Department of Medicine, Division of Gastroenterology-Hepatology, Farmington, Connecticut 06030, USA.
Protein Expr Purif. 1999 Mar;15(2):243-5. doi: 10.1006/prep.1998.1022.
A simple and convenient method for the purification of the hemolytic toxin listeriolysin O (LLO) from Listeria monocytogenes is described. Supernatants from bacteria cultures were purified by application to a CH2 spiral cartridge concentrator (Amicon) and ion exchange chromatography. A critical step is removal of contaminating RNA. The purified proteins had characteristics described for bacterial thiol-activated hemolysins: activation by a reducing agent (DTT) and inactivation by cholesterol. In addition, the molecular weight of 58, 000 and pH-dependent hemolytic activity of this purified protein are consistent with the previously published characteristics of LLO.
本文描述了一种从单核细胞增生李斯特菌中纯化溶血毒素李斯特菌溶血素O（LLO）的简单便捷方法。细菌培养物的上清液通过应用CH2螺旋柱浓缩器（密理博公司）和离子交换色谱法进行纯化。一个关键步骤是去除污染的RNA。纯化后的蛋白质具有细菌硫醇激活溶血素的特征：被还原剂（二硫苏糖醇）激活，被胆固醇灭活。此外，这种纯化蛋白质的分子量为58,000以及pH依赖性溶血活性与LLO先前发表的特征一致。
