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Infect Immun. Oct 1974; 10(4): 750–756.
PMCID: PMC423017

Purification and Some Properties of Progenitor Toxins of Clostridium botulinum Type B


Purification of progenitor toxin of Clostridium botulinum type B strain Okra was undertaken by sequential steps of acid precipitation, extraction, ammonium sulfate precipitation, ribonuclease digestion, acid precipitation, protamine treatment, sulphopropyl-Sephadex chromatography, and Sephadex G-200 gel filtration. Two different molecular-sized toxins, named large (L) and medium (M) toxins, were obtained. L toxin was centrifugally homogeneous but electrophoretically heterogeneous. It contained 2.5 × 108 to 3.0 × 108 mean lethal doses per mg of nitrogen, and its sedimentation constant was 16S. M toxin was centrifugally and electrophoretically homogeneous. It contained 5.5 × 108 to 6.0 × 108 mean lethal doses per mg of nitrogen, and its sedimentation constant was 12S. The presence of both L and M toxins in spent culture was demonstrated. It seems justified, therefore, to call both progenitor toxins. Both consisted of toxic and nontoxic components. The toxic components of L and M toxins appeared to be identical with each other. The nontoxic component of L toxin was 12S and possessed a hemagglutinin activity of about 0.5% that of type A crystalline toxin; that of M toxin was 7S and possessed no hemagglutinin activity. They were antigenically related but not identical.

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Selected References

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  • Beers WH, Reich E. Isolation and characterization of Clostridium botulinum type B toxin. J Biol Chem. 1969 Aug 25;244(16):4473–4479. [PubMed]
  • Boroff DA, 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. [PMC free article] [PubMed]
  • DasGupta BR, Boroff DA. Separation of toxin and hemagglutinin from crystalline toxin of Clostridium botulinum type A by anion exchange chromatography and determination of their dimensions by gel filtration. J Biol Chem. 1968 Mar 10;243(5):1065–1072. [PubMed]
  • DasGupta BR, Boroff DA, Cheong K. Isolation of chromatographically pure toxin of Clostridium botulinum type B. Biochem Biophys Res Commun. 1968 Sep 30;32(6):1057–1063. [PubMed]
  • DUFF JT, KLERER J, BIBLER RH, MOORE DE, GOTTFRIED C, WRIGHT GG. Studies on immunity to toxins of Clostridium botulinum. II. Production and purification of type B toxin for toxoid. J Bacteriol. 1957 May;73(5):597–601. [PMC free article] [PubMed]
  • Gerwing J, Dolman CE, Kason DV, Tremaine JH. Purification and characterization of Clostridium botulinum type B oxin. J Bacteriol. 1966 Feb;91(2):484–487. [PMC free article] [PubMed]
  • Kitamura M, Sakaguchi S, Sakaguchi G. Purification and some properties of Clostridium botulinum type-E toxin. Biochim Biophys Acta. 1968 Oct 21;168(2):207–217. [PubMed]
  • Kitamura M, Sakaguchi S, Sakaguchi G. Significance of 12S toxin of Clostridium botulinum type E. J Bacteriol. 1969 Jun;98(3):1173–1178. [PMC free article] [PubMed]
  • Lamanna C, Glassman HN. The Isolation of Type B Botulinum Toxin. J Bacteriol. 1947 Nov;54(5):575–584. [PMC free article] [PubMed]
  • LAMANNA C, LOWENTHAL JP. The lack of identity between hemagglutinin and the toxin of type A botulinal organism. J Bacteriol. 1951 Jun;61(6):751–752. [PMC free article] [PubMed]
  • Lamanna C, Sakaguchi G. Botulinal toxins and the problem of nomenclature of simple toxins. Bacteriol Rev. 1971 Sep;35(3):242–249. [PMC free article] [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • REISFELD RA, LEWIS UJ, WILLIAMS DE. Disk electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature. 1962 Jul 21;195:281–283. [PubMed]
  • Sakaguchi G, Sakaguchi S, Kondo H. Rapid bioassay for Clostridium botulinum type-E toxins by intravenous injection into mice. Jpn J Med Sci Biol. 1968 Dec;21(6):369–378. [PubMed]
  • Sakaguchi G, Sakaguchi S, Kozaki S, Sugii S, Oishi I. Cross reaction in reversed passive hemagglutination between Clostridium botulinum type A and B toxins and its avoidance by the sue of anti-toxic component immunoglobulin isolated by affinity chromatography. Jpn J Med Sci Biol. 1974 Jun;27(3):161–172. [PubMed]
  • WAGMAN J, BATEMAN JB. The behavior of the bolulinus toxins in the ultracentrifuge. Arch Biochem Biophys. 1951 May;31(3):424–430. [PubMed]

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