霍乱弧菌非O1型溶血素变体的两步纯化及部分特性分析

Two-step purification and partial characterization of a variant of the Vibrio cholerae non-O1 hemolysin.

作者信息

McCardell B A, Kothary M H, Madden J M

机构信息

Division of Virulence Assessment, Center for Food Safety and Applied Nutrition, FDA, Washington, DC 20204, USA.

出版信息

FEMS Microbiol Lett. 1999 Nov 15;180(2):177-82. doi: 10.1111/j.1574-6968.1999.tb08793.x.

DOI:10.1111/j.1574-6968.1999.tb08793.x

PMID:10556709

Abstract

A two-step purification method using ammonium sulfate precipitation and gel filtration was developed for the purification of a variant of the El Tor hemolysin/cytolysin from supernatant fluids of a Vibrio cholerae non-O1 human isolate (strain 2194c). The toxin displayed delayed elution from a Sephacryl gel filtration column, eluting at between two and three column volumes. The molecular mass and isoelectric point of the purified 2194c toxin were 60 kDa and 5. 3, respectively. The N-terminal amino acid sequence was ASPAPANSETNTLPHVAFYI. Purified toxin was cytolytic for Chinese hamster ovary cells and erythrocytes from several animal species.

摘要

开发了一种两步纯化方法，该方法使用硫酸铵沉淀和凝胶过滤，用于从霍乱弧菌非O1型人体分离株（菌株2194c）的上清液中纯化埃尔托溶血素/细胞毒素的变体。该毒素从Sephacryl凝胶过滤柱上洗脱延迟，在两到三个柱体积之间洗脱。纯化的2194c毒素的分子量和等电点分别为60 kDa和5.3。N端氨基酸序列为ASPAPANSETNTLPHVAFYI。纯化的毒素对中国仓鼠卵巢细胞和几种动物物种的红细胞具有细胞溶解作用。