Purification and Properties of Clostridium botulinum Type F Toxin
Purification and Properties of Clostridium botulinum Type F Toxin
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F型肉毒梭菌毒素的纯化及性质
DOI：
10.1128/am.29.5.598-603.1975
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发表时间：
1975
期刊：
影响因子：
--
通讯作者：
H. Sugiyama
中科院分区：
文献类型：
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作者：
K. Yang;H. Sugiyama
纯化蛋白水解朗厄兰菌株的F型肉毒梭菌毒素。用 (NH4)2SO4 沉淀整个培养物中的毒素。沉淀物提取物依次在pH 6.0的二乙氨基乙基纤维素、pH 4.9的O-(羧甲基)纤维素、pH 8.1的Sephadex G-200、pH 4.9的季氨基乙基-Sephadex、最后pH 8.1的二乙氨基乙基纤维素上进行色谱分离。该程序回收了起始培养物中检测到的毒素的 14%。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、双凝胶扩散血清学和等电聚焦检测，该毒素是均质的。纯化的毒素通过凝胶过滤得到的分子量为150,000，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳得到的分子量为155,000。比毒性为每吸光度 (278 nm) 单位 9.6 × 106 平均致死剂量。当纯化的毒素用二硫化物还原剂处理并在十二烷基硫酸钠-聚丙烯酰胺凝胶上电泳时，发现分子量为 105,000 和 56,000 的亚单位。当纯化的F型和E型毒素与用纯化毒素制备的免疫类毒素获得的抗毒素反应时，证明了相互交叉中和。
Clostridium botulinum type F toxin of proteolytic Langeland strain was purified. Toxin in whole cultures was precipitated with (NH4)2SO4. Extract of the precipitate was successively chromatographed on diethylaminoethyl-cellulose at pH 6.0, O-(carboxymethyl) cellulose at pH 4.9, Sephadex G-200 at pH 8.1, quaternary aminoethyl-Sephadex at pH 4.9, and finally diethylaminoethyl-cellulose at pH 8.1. The procedure recovered 14% of the toxin assayed in the starting culture. The toxin was homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, double gel diffusion serology, and isoelectric focusing. Purified toxin had a molecular weight of 150,000 by gel filtration and 155,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Specific toxicity was 9.6 × 106 mean lethal doses per absorbancy (278 nm) unit. Sub-units of 105,000 and 56,000 molecular weight are found when purified toxin is treated with a disulfide reducing agent and electrophoresed on sodium dodecyl sulfate-polyacrylamide gels. Reciprocal cross neutralizations were demonstrated when purified type F and E toxins were reacted with antitoxins which were obtained with immunizing toxoids prepared with purified toxins.