Proteolysis of Clostridium perfringens type A enterotoxin during purification.
The small satellite bands of enterotoxin frequently seen in polyacrylamide gels following purification of Clostridium perfringens enterotoxin were found to be due to endogenous protease activity and were not present if phenylmethylsulfonyl fluoride (PMSF; 1 mM) and EDTA (10 mM) were used in the purification protocol. The use of PMSF was avoided by passing gel filtration-purified enterotoxin material through DEAE-Sephacel. This modified protocol resulted in an 11.4-fold purification of enterotoxin and a 26.8% yield. Contrary to previous reports (B. R. Dasgupta and M. W. Pariza, Infect. Immun. 38: 592-597, 1982), if PMSF and EDTA were included during purification, we were unable to detect the novel enterotoxin ET-1 produced by strain NCTC 10240. C. perfringens proteases cleaved homogeneous enterotoxin into two additional fragments, suggesting that ET-1 was a product of endogenous protease action during purification.
Park
K B
KB
Department of Food Science, University of Massachusetts, Amherst 01003.
Labbé
R G
RG
D001427:Bacterial Toxins / Q000302:isolation & purification* / Q000378:metabolism; D002850:Chromatography, Gel; D003016:Clostridium perfringens / Q000201:enzymology / Q000378:metabolism*; D004492:Edetic Acid / Q000494:pharmacology; D004768:Enterotoxins / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology
D001427:Bacterial Toxins; D004768:Enterotoxins; D011480:Protease Inhibitors; D004492:Edetic Acid; D010447:Peptide Hydrolases