Combined analysis of tertiary structure and function(s) of Clostridium botulinum type C and D phospholipase C

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Clostridium botulinum cultures are classified into seven types, types A to G, based on the antigenicity of the neurotoxins produced. Of these seven types, only types C and D produce phospholipase C (lecithinase, PLC) in addition to the neurotoxin. Phospholipase of C. perfringens is known as α-toxin, which is the best characterized of all clostridial phospholipases and is an important agent of gas gangrene. The toxin, which exhibits phospholipase C and sphingomyelinase (SMase) activities, causes hemolysis, necrosis, and death. Here we determined the PLC genes of C. botulinum type C and D (Cbotα and Dbotα) by using inverse PCR and primer walking, and expressed as a GST-fusion protein. We compare the enzymatic and biological activities (egg-yolk activity, hemolysis, and mouse lethality) of the purified protein of C. perfringens α-toxin, Cbotα, and Dbotα.