Purification and characterization of Aeromonas hydrophila beta-hemolysin.
Aeromonas hydrophila hemolysin was excreted in our culture conditions during the stationary growth phase. The toxin was purified to homogeneity by a three-step method: ultrafiltration, acid precipitation in the presence of RNA and anion exchange chromatography with FPLC apparatus. Beta-hemolysin is a protein not associated with lipids, carbohydrates or nucleic acids whose subunit mol. wt is 51,000. The mol. wt determined by polyacrylamide gel electrophoresis suggests that the molecule is in a trimeric form. The toxin is thermolabile and inactivated by proteolytic enzymes such as trypsin, chymotrypsin, pronase, subtilisin and proteinase K. Antibodies raised against the beta-hemolysin neutralize both hemolytic and cytotoxic activities. When injected at high dose, this purified hemolytic protein causes a positive rabbit ileal loop test, thus indicating that beta-hemolysin could be the main virulence factor involved in intestinal symptoms.
Bloch
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Laboratoire de Toxinologie Bacterienne, Institut de Bactériologie, Faculté de Médecine, Strasbourg, France.
Monteil
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D000333:Aeromonas / Q000032:analysis*; D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification*; D002470:Cell Survival / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D004768:Enterotoxins / Q000302:isolation & purification*; D004912:Erythrocytes / Q000187:drug effects; D006460:Hemolysin Proteins / Q000302:isolation & purification* / Q000494:pharmacology; D006358:Hot Temperature; D006801:Humans; D007120:Immunochemistry; D005779:Immunodiffusion; D007525:Isoelectric Focusing; D007931:Leucyl Aminopeptidase; D009113:Muramidase; D010447:Peptide Hydrolases; D000516:alpha-Amylases
D001426:Bacterial Proteins; D004768:Enterotoxins; D006460:Hemolysin Proteins; D000516:alpha-Amylases; D009113:Muramidase; D010447:Peptide Hydrolases; D007931:Leucyl Aminopeptidase