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.
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