Levdikova G A, Klimacheva L V, Ispolatovskaia M V, Bulatova T I, Anisimova L I
Biokhimiia. 1979 Aug;44(8):1392-400.
A new method of isolation of highly purified Cl. botulinum toxin of E type from the cultural fluid of strain 188 centrifugates was developed. The method allows to isolate the toxin both in a precursor and in activated forms with a yield of 10--15%. The method includes fractionation by ammonium sulfate, ultrafiltration and subsequent column chromatography on DEAE-cellulose, Sephadex G-200 and DEAE-Sephadex A-50. The preparations were found homogeneous during polyacrylamide gel electrophoresis and immunoprecipitation in agar with antitoxic horse serum. The potential specific toxicity of the preparations is 1--1,2.10(7) DLM/mg of protein. The molecular weight of the toxin is about 160 000; the molar extinction coefficient is equal to 278 nm. The isoelectric point lies around pH 6.0. The highly purified Cl. botulinum toxin of E type was found stable upon storage.
已开发出一种从188株离心培养液中分离高纯度E型肉毒杆菌毒素的新方法。该方法能够以10%-15%的产率分离出前体形式和活化形式的毒素。该方法包括硫酸铵分级分离、超滤以及随后在DEAE-纤维素、葡聚糖G-200和DEAE-葡聚糖A-50上进行柱色谱分离。在聚丙烯酰胺凝胶电泳和用抗毒素马血清进行的琼脂免疫沉淀过程中,发现这些制剂是均一的。制剂的潜在比毒性为1-1.2×10⁷DLM/毫克蛋白质。毒素的分子量约为160000;摩尔消光系数等于278纳米。等电点在pH 6.0左右。发现高纯度的E型肉毒杆菌毒素在储存时是稳定的。