Cherniak R, Frederick H M
Infect Immun. 1977 Mar;15(3):765-71. doi: 10.1128/iai.15.3.765-771.1977.
Several closely related capsular polysaccharides were isolated from a strain of Clostridium perfringens Hobbs 9 type A by extraction of encapsulated cells with cold 0.85% NaCl. The soluble polymers were precipitated with alcohol and purified by (NH4)2SO4 fractionation, enzymatic digestion with papain and ribonuclease, and chromatography on diethylaminoethyl-Sephadex A25. The polysaccharides were composed mainly of glucose, galactose, and galactosamine. The major fraction contained these constituents (representing 77% of the dry weight) in a molar ratio of 1:1.6:1.1. All of the fractions contained phosphate and peptide material that was not removed during purification. The polysaccharides were closely related but not identical as indicated by double-diffusion-in-gel experiments. Immunoelectrophoresis in agarose demonstrated that the polysaccharides had identical mobilities and that no resolution into additional fractions occurred. The immunological activity of all the purified polysaccharides was destroyed by periodate oxidation but was unaffected by protease.
通过用冷的0.85%氯化钠提取荚膜化细胞,从一株A型产气荚膜梭菌霍布斯9中分离出几种密切相关的荚膜多糖。将可溶性聚合物用酒精沉淀,并用硫酸铵分级分离、木瓜蛋白酶和核糖核酸酶酶解以及在二乙氨基乙基-葡聚糖A25上进行色谱法纯化。这些多糖主要由葡萄糖、半乳糖和氨基半乳糖组成。主要部分含有这些成分(占干重的77%),摩尔比为1:1.6:1.1。所有部分都含有在纯化过程中未去除的磷酸盐和肽物质。如凝胶双向扩散实验所示,这些多糖密切相关但不相同。琼脂糖免疫电泳表明,这些多糖具有相同的迁移率,并且没有再分离出其他部分。所有纯化多糖的免疫活性都被高碘酸盐氧化破坏,但不受蛋白酶影响。