PRODUCTION AND PURIFICATION OF CLOSTRIDIUM-PERFRINGENS ALPHA-TOXIN USING A PROTEIN-HYPERPRODUCING STRAIN, BACILLUS-BREVIS-47

Clostridium perfringens alpha-toxin was produced in a protein-hyperpro ducing strain, Bacillus brevis 47, by cloning the gene into the constr ucted expression-secretion vector which has the multiple promoters and the signal peptide coding region of an outer cell wall protein gene. The amount of alpha-toxin produced by the B. brevis 47 transformant ca rrying the gene was approximately 10 times greater than that produced by a B. subtilis transformant carrying the toxin gene. Biological acti vities and the N-terminal amino acid sequence of the toxin secreted by the B. brevis 47 transformant were identical to those of wildtype alp ha-toxin.