Botulinum neurotoxin production

Eklund, M.W., Poysky, F.T. and Habig, W.H., Bacteriophages and plasmids in Clostridium botulinum and Clostridium tetani and their relationship to production of toxins, in Simpson, L.L., ed., Botulinum Neurotoxin and Tetanus Toxin, Academic Press, New York, pp. 25-51, 1989. 

Segelke B, Knapp M, Kadkhodayan S, Balhorn R, Rupp B (2004) Crystal structure of Clostridium botulinum neurotoxin protease in a product-bound state evidence for noncanonical zinc protease activity. Proc Natl Acad Sci U S A 101 6888-93... 

Daniels-Holgate, P., Dolly, J. (1996). Productive and nonproductive binding of botulinum neurotoxin A to motor nerve endings are distinguished by its heavy chain. J. Neurosci. Res. 44 263-71. 

CeU-based assay for stability and potency of botulinum neurotoxin type A products In vitro... 

Since botulinum is a neurotoxin, large-scale production should meet biosafety level 3 containment and strict handling to avoid the possible human intoxication. Fermentation is carried out in complex medium containing casein hydrolysate, yeast extract, and glucose for 4 days. Then, botulinum toxin is precipitated with 3 N sulfuric acid followed by purification with DEAE-Sephadex column. Botulinum toxin was crystallized then and purified by additional steps using various affinity and size exclusion chromatography. Purity and quality were checked by animal testing. Production and purification steps vary with Clostridium strain used and botulinum type to be produced. ... 

Infant botulism has been observed in babies of 3-35 weeks where ingestion of C. botulinum spores leads to the organisms growth and production of the neurotoxin in the intestine (Miduara and Amon, 1976 Pickett et al., 1976). Recently, non-botulinum clostridial species, such as Clostridium butyricum and C. barati have been found to be responsible for human botulism cases (McCrosckey et al., 1988 Amon, 1992). In addition, botulism has been held responsible for several sudden infant death syndrome (SIDS) cases (Amon, 1992). 

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