Sakaguchi G, Uemura T, Riemann H P
Appl Microbiol. 1973 Nov;26(5):762-7. doi: 10.1128/am.26.5.762-767.1973.
Purification of Clostridium perfringens type A enterotoxin from sporulated cells was simplified. The method consisted of precipitation of the enterotoxin from the extract of sonically treated cells at 40% saturation of ammonium sulfate at pH 7, differential solubilization in 0.02 M phosphate buffer, pH 6.7, and repeated gel filtration on Sephadex G-200. The purified enterotoxin was at least 98% pure in ultracentrifugation, polyacrylamide gel electrophoresis, and agar gel double diffusion. Recovery was over 74% from the sporulated cell extract. The toxin had biological activities of at least 4,700 mouse intravenous minimal lethal doses/mg of N, 3,900 capillary permeability-increasing U/mg of N in the guinea pig skin, and 210 rabbit intestinal loop distension U/mg of N. The toxin, containing no hexose, lipid, or nucleic acid, appeared to be identical in sedimentation constant, isoelectric point, and ultraviolet absorption spectrum to the toxin purified previously by different procedures.
A型产气荚膜梭菌肠毒素从芽孢形成细胞中的纯化过程得到了简化。该方法包括在pH 7的条件下，用40%饱和度的硫酸铵从经超声处理的细胞提取物中沉淀肠毒素，在pH 6.7的0.02 M磷酸盐缓冲液中进行分级溶解，并在Sephadex G - 200上反复进行凝胶过滤。纯化后的肠毒素在超速离心、聚丙烯酰胺凝胶电泳和琼脂凝胶双向扩散中纯度至少为98%。从芽孢形成细胞提取物中的回收率超过74%。该毒素具有至少4700小鼠静脉注射最小致死剂量/毫克氮的生物活性、3900豚鼠皮肤毛细血管通透性增加单位/毫克氮的生物活性以及210兔肠袢扩张单位/毫克氮的生物活性。该毒素不含己糖、脂质或核酸，其沉降常数、等电点和紫外吸收光谱似乎与先前通过不同方法纯化的毒素相同。
