Optimization of alpha toxin production from clostridium perfringens type a under bioreactor conditions

Clostridium Perfringens type A, is a C. perfringens species that produces the highest amount of alpha toxin. The substance it secretes (known as alpha toxin) is phospholipase C (PLC), an enzyme in protein structure. C. perfringens type A is a species that has the ability to infect humans and animals with this alpha toxin it secretes. It causes some diseases in humans and animals. It is an important pathogen for animal health. It causes significant economic losses in the livestock sector. Since these diseases progress with toxication, they cannot be treated and they progress very quickly and cause high rates of death. Vaccination is the most effective and only method to prevent clostridial diseases. In this study, it is aimed to optimize alpha toxin production from C. perfringens type A, in bioreactor conditions. For this purpose, two different strains, a reference strain and a strain isolated in our country, were used. In the study, the effects of different pH and temperature conditions, different media components and different nutrient media, nitrogen feeding of the nutrient media, and different inoculation rates on alpha toxin production were investigated. The incubation period was determined as 24 hours at 37 oC for the production culture. Bioreactor conditions were determined as 100 rpm, 37 oC, and pH value 6,5-7. It was determined by toxin titration tests that the amount of alpha toxin was maximum between 6-8. th hours of incubation. It has been observed that alpha toxin production is higher between 6.5-7 pH value of the medium. It was concluded that nitrogen feeding should be done at certain hours to the nutrient medium. According to toxin titration tests, at the specified parameter conditions, a result of over 100 mld has been reached. vii With this produced alpha toxin, it is aimed to obtain an inactivated vaccine (by inactivating with formaldehyde) in order to protect against the diseases caused by this bacterium. For this purpose, more alpha toxin will be secreted and this alpha toxin will be used as vaccine active substance (antigen).