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Purification and Characterization of Neurotoxin Complex from a Dual Toxin Gene Containing Clostridium Botulinum Strain PS-5 |
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| Authors: | Ajay K Singh Amita Sachdeva Jeffrey A DeGrasse Timothy R Croley Larry H Stanker David Hodge Shashi K Sharma |
| Institution: | 1. Divisions of Microbiology, Center for Food Safety and Applied Nutrition, Food and Drug Administration, College Park, MD, 20740, USA 2. Analytical Chemistry, Center for Food Safety and Applied Nutrition, Food and Drug Administration, College Park, MD, 20740, USA 3. Western Regional Research Center, Agricultural Research Service, United States Department of Agriculture, Albany, CA, 94170, USA 4. Department of Homeland Security, Washington, DC, 20528, USA |
| Abstract: | Botulinum neurotoxins are produced as a toxin complex (TC) which consists of neurotoxin (NT) and neurotoxin associated proteins. The characterization of NT in its native state is an essential step for developing diagnostics and therapeutic countermeasures against botulism. The presence of NT genes was validated by PCR amplification of toxin specific fragments from genomic DNA of Clostridium botulinum strain PS-5 which indicated the presence of both serotype A and B genes on PS-5 genome. Further, TC was purified and characterized by Western blotting, Digoxin-enzyme linked immunosorbent assay, endopeptidase activity assay, and Liquid chromatography–Mass spectrometry. The data showed the presence of serotype A specific neurotoxin. Based on the analysis of neurotoxin genes and characterization of TC, PS-5 strain appears as a serotype A (B) strain of C. botulinum which produces only serotype A specific TC in the cell culture medium. |
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