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Proteases of Clostridium Botulinum
Clostridium botulinum proteaser. VI. Virkning av proteaser og proteaseinhibitorer på utvikling av toksisitet i voksende kulturer av Clostridium botulinum
VI. The Role of Trypsin, Clostridium Botulinum Proteases and Protease Inhibitors in the Formation and Activation of Toxin in Growing Cultures of Clostridium Botulinum
Acta Veterinaria Scandinavica volume 15, pages 487–506 (1974)
Abstract
In this study the influence of bovine serum protease inhibitors, trypsin and proteases produced by different types of Clostridium botulinum has been investigated. Trypsin and botulinum proteases had the capability of increasing the toxicity in growing cultures in Clostridium botulinum types A, B and E. Trypsin increased the toxin level to a greater extent than proteases from Clostridium botulinum types A, B, C and F. Protease inhibitors did not influence the toxin formation to any extent compared with the controls. The combined effects of proteases and protease inhibitors on the development of toxin in Clostridium botulinum type B were also investigated by adding proteases and protease inhibitors to the same culture at different time intervals. Protease inhibitors did not reduce the toxicity of the cultures as compared to the controls. Altogether a complex relationship seems to exist between protoxin, toxin, proteases and inhibitors in the culture, and the order and time sequence of addition seem to be of importance. The results obtained in this investigation indicate that proteases of Clostridium botulinum play a part in the formation and/or activation of toxin in growing cultures of proteolytic strains such as Clostridium botulinum types A and B. As to the activation of protoxin and progenitor toxin produced by non-proteolytic Clostridium botulinum types B and E, botulinum proteases showed a marked capability of increasing the toxicity in these cultures. Trypsinization may be valuable for the detection of Clostridium botulinum types A and B in foods, as well as for type E, where it is commonly used.
Sammendrag
Det er utført undersøkelser over virkning av proteaser (trypsin og Clostridium botulinum proteaser) og protease inhibitorer på toksindannelsen hos voksende kulturer av Clostridium botulinum A, B, C og E. Trypsin økte toksisiteten i kulturer av Clostridium botulinum type A, B og E mer enn hva Clostridium botulinum proteasene gjorde. Protease inhibitorer hindret ikke toksindannelsen hos Clostridium botulinum sammenlignet med kontroll. Den proteolytiske type C stammen som ble undersøkt dannet ikke toksin hverken med eller uten naervær av protease eller protease inhibitorer. Videre undersøkte man den kombinerte effekt av proteaser og protease inhibitorer ved å tilsette disse til den samme kultur, men ved forskjellige tidspunkter. Proteaseinhibitorene hindret ikke toksinutvikling sammenlignet med kontroll, og proteaser tilsatt kulturer hvor inhibitorer var til stede økte toksisiteten i løpet av 2–3 timer, med et derpå følgende fall. Dersom man tilsatte proteaser ved starten av forsøket, økte dette toksisiteten i forhold til kontroll, og protease inhibitorer tilsatt denne kultur etter 24 og 48 timer hemmet ikke den videre toksinutvikling selv om proteolytisk aktivitet i kulturen ble hemmet fuUstendig. Pä den annen side synes det som om inhibitorerne hemmet degradering av toksinet. I det hele tatt synes undersøkelsene å vise et komplekst forhold mellom protoksin og toksin og hvordan proteolytiske enzymer og inhibitorer influerer disse. Resultatene indikerer at Clostridium botulinum proteaser og trypsin influerer danneise og/eller aktivering av toksin hos Clostridium botulinum type A og proteolytisk type B. Når det gjelder aktivering av progenitor toksin hos Clostridium botulinum type E og ikke proteolytisk type B, så økte toksisiteten etter tilsetting av proteaser fra Clostridium botulinum. Trypsin hadde imidlertid en bedre aktiverende effekt. Ut fra de foreliggende data er det grunn til å påpeke at trypsin-tilsetting, som er vanlig à benytte til næringsmidler eller kulturer som mistenkes å inneholde type E, også bør forsøkes til kulturer eller næringsmidler som mistenkes å inneholde Clostridium botulinum type A og B. Tilsetting av protease inhibitor hemmet ikke toksindannelsen hos Clostridium botulinum type A og B.
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Tjaberg, T.B. Proteases of Clostridium Botulinum. Acta Vet Scand 15, 487–506 (1974). https://doi.org/10.1186/BF03547221
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DOI: https://doi.org/10.1186/BF03547221