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Volume 31, Issue 1

The Fractionation of -Antigen on Cellulose Ion Exchangers Free

Abstract

SUMMARY

A low-protein medium is described for the production of reasonable yields of ) -prototoxin. The -antigen in young cultures on this medium was virtually non-toxic, 99.8% being in the form of prototoxin. Its purity, estimated in terms of the protein content of crystalline -prototoxin, was 50%. Fractionation of concentrates of the antigen on columns of diethylaminoethyl- and carboxy-methyl-celluloses suggested that a number of proteins intermediate between -prototoxin and -toxin were present which differed from the latter in their isoelectric points in their activation ratios. The concentration and number of these intermediates increased as the incubation of cultures was prolonged. Since they could be fully converted to -toxin by trypsin it is probable that they were formed from -prototoxin by proteolytic or spontaneous degradation. In toxicity tests, death caused by unactivated antigen may be delayed. This evidence suggested that the intermediates were intrinsically non-toxic but could become activated in the animal body. The activation ratios might then be regarded as a measure of the ease with which the intermediates could be converted to toxin Physical data and the amino acid composition of crystalline -prototoxin are presented.

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© Society for General Microbiology, 1962
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1963-04-01
2024-04-20
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References

  1. Bennetts H. W. 1932; Enterotoxaemia. Abstract fro. Bulletin 57. Aust. Counc. Sci. Industr. Res
  2. Bosworth T. J., Glover R. E. 1934-35; The effects on C. welchii toxins of a substance present in the normal intestine. Univ. of Cambridge Inst. Anim. Path. 4th Report79
  3. Bullen J. J., Scarisbrick R. 1957; Enterotoxaemia of sheep. Experimental reproduction of the disease. J. Path. Bact 73:495
     [Google Scholar]
  4. Gladstone G. P., Fildes P. 1940; A simple culture medium for general use without meat extract or peptone. Brit. J. exp. Path 21:161
     [Google Scholar]
  5. Hirs C. H. W. 1956; The oxidation of ribonuclease with performic acid. J. biol. Chem 219:611
     [Google Scholar]
  6. Holiday E. R. 1936; Spectrophotometry of proteins. 1. Absorption spectra of tyrosine and tryptophan and their mixtures. 2. Estimation of tyrosine and tryptophan in proteins. Biochem. J 30:1795
     [Google Scholar]
  7. Kohn J. 1959; A simple method for the concentration of fluids containing protein. Nature, Lond 183:1055
     [Google Scholar]
  8. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. J. biol. Chem 193:265
     [Google Scholar]
  9. Markham R. 1942; A steam distillation apparatus suitable for micro-Kjeldahl analysis. Biochem. J 36:790
     [Google Scholar]
  10. Meisel H., Albricht H., Rymkiewicz D. 1960; Toxin, prototoxin and phospholipase C in culture liquids, vegetative cells and spores o. CI. perfringens Type D. Bull. acad. Polon. sci. Sér. sci. biol 8:497
     [Google Scholar]
  11. Oakley C. L., Warrack G. H., van Heyningen W. E. 1946; The collagenase (k toxin) of C. welchii Type A. J. Path. Bact 58:229
     [Google Scholar]
  12. Oncley J. L. 1941; Evidence from physical chemistry regarding the size and shape of protein molecules from ultracentrifugal, diffusion, viscosity, dielectric dispersion and double refraction of flow. Ann. N.Y. Acad. Sci 41:121
     [Google Scholar]
  13. Orlans E. S., Richards C. B., Jones V. E. 1960; C. welchii epsilon toxin and antitoxin. Immunology 3:28
     [Google Scholar]
  14. Pope C. G., Stevens M. F. 1939; The determination of amino nitrogen using a copper method. Biochem. J 33:1070
     [Google Scholar]
  15. Schachmann H. K. 1957; Ultracentrifugation, diffusion and viscometry. Meth. Enzymol 4:32
     [Google Scholar]
  16. Spackman D. H., Stein W. H., Moore S. 1958; Automatic recording apparatus for use in the chromatography of amino acids. Anal. Chem 30:1190
     [Google Scholar]
  17. Thomson R. O. 1962; Crystallin. ε-prototoxin from C. welchii. Nature, Lond 193:69
     [Google Scholar]
  18. Trautmann R., Crampton C. F. 1959; Application of the Archibald principle for the ultracentrifugal determination of the molecular weight in urea solutions of histone fractions from calf thymus. J. Amer. chem. Soc 81:4036
     [Google Scholar]
  19. Turner A. W., Rodwell A. W. 1943; The epsilon toxin o. C. welchii Type D. 1. Proteolytic conversion of ε-prototoxin into ε-toxin by trypsin and other proteases. 2. Mechanism of its development in cultures through the action of extracellular proteinases upon ε-prototoxin. Aust. J. exp. Biol. med. Sci 21:1727
     [Google Scholar]
  20. Verwoerd D. W. 1960; Isolasie van die protoksien va. C. welchii Type D. J. S. Afr. Vet. Med. Ass 31:195
     [Google Scholar]
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The Fractionation of Clostridium welchii e-Antigen on Cellulose Ion Exchangers
Microbiology 31, 79 (1963); https://doi.org/10.1099/00221287-31-1-79
/content/journal/micro/10.1099/00221287-31-1-79
/content/journal/micro/10.1099/00221287-31-1-79
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