Booth B A, Boesman-Finkelstein M, Finkelstein R A
Infect Immun. 1984 Sep;45(3):558-60. doi: 10.1128/iai.45.3.558-560.1984.
Unnicked cholera enterotoxin was isolated from culture supernatants of Vibrio cholerae 569B by either rapid processing of flask-grown cultures or by growing and processing fermentor cultures in the presence of ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetra acetic acid, an inhibitor of the previously described V. cholerae hemagglutinin/protease. When unnicked cholera enterotoxin was incubated with purified hemagglutinin/protease, the unnicked A subunit was converted to a molecular weight consistent with that of the A1 subunit as demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and its specific activity for Y1 adrenal cells increased.
通过对摇瓶培养物进行快速处理,或者在乙二醇 - 双(β - 氨基乙基醚)-N,N,N',N'-四乙酸(一种先前描述的霍乱弧菌血凝素/蛋白酶的抑制剂)存在的情况下培养和处理发酵罐培养物,从霍乱弧菌569B的培养上清液中分离出未切割的霍乱肠毒素。当未切割的霍乱肠毒素与纯化的血凝素/蛋白酶一起孵育时,如通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳所证明的,未切割的A亚基转化为与A1亚基一致的分子量,并且其对Y1肾上腺细胞的比活性增加。