Skip to main content
NCBI home page
Applied Microbiology logoLink to Applied Microbiology
. 1975 May;29(5):598–603. doi: 10.1128/am.29.5.598-603.1975

Purification and Properties of Clostridium botulinum Type F Toxin

K H Yang 1, H Sugiyama 1
PMCID: PMC187043  PMID: 807160

Abstract

Clostridium botulinum type F toxin of proteolytic Langeland strain was purified. Toxin in whole cultures was precipitated with (NH4)2SO4. Extract of the precipitate was successively chromatographed on diethylaminoethyl-cellulose at pH 6.0, O-(carboxymethyl) cellulose at pH 4.9, Sephadex G-200 at pH 8.1, quaternary aminoethyl-Sephadex at pH 4.9, and finally diethylaminoethyl-cellulose at pH 8.1. The procedure recovered 14% of the toxin assayed in the starting culture. The toxin was homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, double gel diffusion serology, and isoelectric focusing. Purified toxin had a molecular weight of 150,000 by gel filtration and 155,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Specific toxicity was 9.6 × 106 mean lethal doses per absorbancy (278 nm) unit. Sub-units of 105,000 and 56,000 molecular weight are found when purified toxin is treated with a disulfide reducing agent and electrophoresed on sodium dodecyl sulfate-polyacrylamide gels. Reciprocal cross neutralizations were demonstrated when purified type F and E toxins were reacted with antitoxins which were obtained with immunizing toxoids prepared with purified toxins.

Full text

PDF
598

Images in this article

603Image
on p.603

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andrews P. The gel-filtration behaviour of proteins related to their molecular weights over a wide range. Biochem J. 1965 Sep;96(3):595–606. doi: 10.1042/bj0960595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beers W. H., Reich E. Isolation and characterization of Clostridium botulinum type B toxin. J Biol Chem. 1969 Aug 25;244(16):4473–4479. [PubMed] [Google Scholar]
  3. Boroff D. A., Fleck U. Statistical analysis of a rapid in vivo method for the titration of the toxin of Clostridium botulinum. J Bacteriol. 1966 Nov;92(5):1580–1581. doi: 10.1128/jb.92.5.1580-1581.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DasGupta B. R., Boroff D. A., Cheong K. Isolation of chromatographically pure toxin of Clostridium botulinum type B. Biochem Biophys Res Commun. 1968 Sep 30;32(6):1057–1063. doi: 10.1016/0006-291x(68)90137-x. [DOI] [PubMed] [Google Scholar]
  5. DasGupta B. R., Sugiyama H. A common subunit structure in Clostridium botulinum type A, B and E toxins. Biochem Biophys Res Commun. 1972 Jul 11;48(1):108–112. doi: 10.1016/0006-291x(72)90350-6. [DOI] [PubMed] [Google Scholar]
  6. Dasgupta B. R., Berry L. J., Boroff D. A. Purification of Clostridium botulinum type A toxin. Biochim Biophys Acta. 1970 Aug 21;214(2):343–349. doi: 10.1016/0005-2795(70)90011-5. [DOI] [PubMed] [Google Scholar]
  7. Eklund M. W., Poysky F. T., Wieler D. I. Characteristics of Clostridium botulinum type F isolated from the Pacific Coast of the United States. Appl Microbiol. 1967 Nov;15(6):1316–1323. doi: 10.1128/am.15.6.1316-1323.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kitamura M., Sakaguchi S., Sakaguchi G. Significance of 12S toxin of Clostridium botulinum type E. J Bacteriol. 1969 Jun;98(3):1173–1178. doi: 10.1128/jb.98.3.1173-1178.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Knox J., Brown W. P., Spero L. The role of sulfhydryl groups in the activity of type A botulinum toxin. Biochim Biophys Acta. 1970 Aug 21;214(2):350–354. doi: 10.1016/0005-2795(70)90012-7. [DOI] [PubMed] [Google Scholar]
  10. MOLLER V., SCHEIBEL I. Preliminary report on the isolation of an apparently new type of CI. botulinum. Acta Pathol Microbiol Scand. 1960;48:80–80. doi: 10.1111/j.1699-0463.1960.tb04741.x. [DOI] [PubMed] [Google Scholar]
  11. Midura T. F., Nygaard G. S., Wood R. M., Bodily H. L. Clostridium botulinum type F: isolation from venison jerky. Appl Microbiol. 1972 Aug;24(2):165–167. doi: 10.1128/am.24.2.165-167.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sugiyama H., Das Gupta R., Yang K. H. Disulfide-toxicity relationship of botulinal toxin types A, E, and F. Proc Soc Exp Biol Med. 1973 Jul;143(3):589–591. doi: 10.3181/00379727-143-37372. [DOI] [PubMed] [Google Scholar]
  13. Sugiyama H., DasGupta B. R., Yang K. H. Toxicity of purified botulinal toxin fed to mice. Proc Soc Exp Biol Med. 1974 Nov;147(2):589–591. doi: 10.3181/00379727-147-38394. [DOI] [PubMed] [Google Scholar]
  14. Sugiyama H., Oishi I., Dasgupta B. R. Evaluation of type A botulinal toxin assays that use antitoxin to crystalline toxin. Appl Microbiol. 1974 Feb;27(2):333–336. doi: 10.1128/am.27.2.333-336.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  16. Williams-Walls N. J. Clostridium botulinum Type F: Isolation from Crabs. Science. 1968 Oct 18;162(3851):375–376. doi: 10.1126/science.162.3851.375. [DOI] [PubMed] [Google Scholar]

Articles from Applied Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES