The use of the sac culture method for production of relatively large amounts of enterotoxins A (SEA), D (SED), and E (SEE) for use in purification of the enterotoxins was investigated. One-hundred milliliters of medium per sac, made from Union Carbide sausage casing, with a shaking speed of 150 rpm was chosen as the optimum condition for the enterotoxin production. Brain heart infusion (BHI) broth was the medium of choice for production of SEA by strain FRI-722, with over 100 μg/ml being produced. Strain 1151m produced 20 μg/ml of SED in BHI medium which was 10 times that produced in shake flasks. The production of SEE was best in the Biosate + yeast extract medium, with adequate amounts being produced for purification.

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