Yang K H, Sugiyama H
Appl Microbiol. 1975 May;29(5):598-603. doi: 10.1128/am.29.5.598-603.1975.
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, and finally diethylaminoethyl-cellulose at pH 8.1. The procedure recovered 14 percent 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 X 10-6 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.
对蛋白水解性朗格兰德菌株的F型肉毒梭菌毒素进行了纯化。用硫酸铵沉淀全培养物中的毒素。沉淀提取物先后在pH 6.0的二乙氨基乙基纤维素、pH 4.9的羧甲基纤维素上进行层析，最后在pH 8.1的二乙氨基乙基纤维素上进行层析。该方法回收了起始培养物中所测毒素的14%。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、双向凝胶扩散血清学和等电聚焦法检测，该毒素是均一的。通过凝胶过滤法测得纯化毒素的分子量为150,000，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法测得为155,000。比毒性为每吸光度（278nm）单位9.6×10⁻⁶平均致死剂量。当用二硫键还原剂处理纯化毒素并在十二烷基硫酸钠-聚丙烯酰胺凝胶上进行电泳时，可发现分子量为105,000和56,000的亚基。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，可证明存在相互交叉中和作用。
