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. 1980 May;28(2):469–474. doi: 10.1128/iai.28.2.469-474.1980

Escherichia coli heat-stable enterotoxin: rapid method of purification and some characteristics of the toxin.

R Lallier, S Lariviere, S St-Pierre
PMCID: PMC550959  PMID: 6995310

Abstract

In this study we used Escherichia coli strain F11(P155) of porcine origin. The heat-stable enterotoxin (ST) was produced with a batch fermentor under agitation (500 rpm) and forced aeration (5 liters/min) in Casamino Acids yeast extract medium containing 0.2% glucose. The pH varied from 7.2 to 7.8. The maximum amount of ST was obtained after 7 h of growth. ST was purified by ammonium sulfate precipitation, ultrafiltration on Amicon membranes, and chromatography in Bio-Gel P-4. The enterotoxin, which was purified approximately 1,000 times, was active at nanogram levels. On 20% polyacrylamide gel electrophoresis, ST exhibited an Rf of 0.6 ST was recovered from the gel slices by eluting in buffer and testing the activity in suckling mice, since no band appeared on the gel after staining with Coomasie brilliant blue R, Schiff reagent, or red oil. ST was resistant at pH 2 to 10 and at 100 degrees C for 15 min, but it was inactivated at 121 degrees C; it did not lose biological activity after treatment with pronase, lipase, or amylase. In suckling mice antiserum obtained from rabbits or goats immunized with ST neutralized the enterotoxin activity of a cell-free supernatant of purified ST.

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Selected References

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

  1. Alderete J. F., Robertson D. C. Purification and chemical characterization of the heat-stable enterotoxin produced by porcine strains of enterotoxigenic Escherichia coli. Infect Immun. 1978 Mar;19(3):1021–1030. doi: 10.1128/iai.19.3.1021-1030.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alderete J. F., Robertson D. C. Repression of heat-stable enterotoxin synthesis in enterotoxigenic Escherichia coli. Infect Immun. 1977 Sep;17(3):629–633. doi: 10.1128/iai.17.3.629-633.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burgess M. N., Bywater R. J., Cowley C. M., Mullan N. A., Newsome P. M. Biological evaluation of a methanol-soluble, heat-stable Escherichia coli enterotoxin in infant mice, pigs, rabbits, and calves. Infect Immun. 1978 Aug;21(2):526–531. doi: 10.1128/iai.21.2.526-531.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Callahan L. T., 3rd, Richardson S. H. Biochemistry of Vibrio cholerae virulence. 3. Nutritional requirements for toxin production and the effects of pH on toxin elaboration in chemically defined media. Infect Immun. 1973 Apr;7(4):567–572. doi: 10.1128/iai.7.4.567-572.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dean A. G., Ching Y. C., Williams R. G., Harden L. B. Test for Escherichia coli enterotoxin using infant mice: application in a study of diarrhea in children in Honolulu. J Infect Dis. 1972 Apr;125(4):407–411. doi: 10.1093/infdis/125.4.407. [DOI] [PubMed] [Google Scholar]
  6. Dorner F. Escherichia coli enterotoxin. Purification and partial characterization. J Biol Chem. 1975 Nov 25;250(22):8712–8719. [PubMed] [Google Scholar]
  7. Evans D. G., Evans D. J., Jr, Pierce N. F. Differences in the response of rabbit small intestine to heat-labile and heat-stable enterotoxins of Escherichia coli. Infect Immun. 1973 Jun;7(6):873–880. doi: 10.1128/iai.7.6.873-880.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Finkelstein R. A., Atthasampunna P., Chulasamaya M., Charunmethee P. Pathogenesis of experimental cholera: biologic ativities of purified procholeragen A. J Immunol. 1966 Mar;96(3):440–449. [PubMed] [Google Scholar]
  9. Giannella R. A. Suckling mouse model for detection of heat-stable Escherichia coli enterotoxin: characteristics of the model. Infect Immun. 1976 Jul;14(1):95–99. doi: 10.1128/iai.14.1.95-99.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gyles C. L. Relationships among heat-labile enterotoxins of Escherichia coli and Vibrio cholerae. J Infect Dis. 1974 Mar;129(3):277–283. doi: 10.1093/infdis/129.3.277. [DOI] [PubMed] [Google Scholar]
  11. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  12. Larivière S., Gyles C. L., Barnum D. A. A comparative study of the rabbit and pig gut loop systems for the assay of Escherichia coli enterotoxin. Can J Comp Med. 1972 Oct;36(4):319–328. [PMC free article] [PubMed] [Google Scholar]
  13. Moon H. W. Pathogenesis of enteric diseases caused by Escherichia coli. Adv Vet Sci Comp Med. 1974;18(0):179–211. [PubMed] [Google Scholar]
  14. Mullan N. A., Burgess M. N., Newsome P. M. Characterization of a partially purified methanol-soluble heat-stable Escherichia coli enterotoxin in infant mice. Infect Immun. 1978 Mar;19(3):779–784. doi: 10.1128/iai.19.3.779-784.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sack R. B., Gorbach S. L., Banwell J. G., Jacobs B., Chatterjee B. D., Mitra R. C. Enterotoxigenic Escherichia coli isolated from patients with severe cholera-like disease. J Infect Dis. 1971 Apr;123(4):378–385. doi: 10.1093/infdis/123.4.378. [DOI] [PubMed] [Google Scholar]
  16. Sack R. B. Human diarrheal disease caused by enterotoxigenic Escherichia coli. Annu Rev Microbiol. 1975;29:333–353. doi: 10.1146/annurev.mi.29.100175.002001. [DOI] [PubMed] [Google Scholar]
  17. Schenkein I., Green R. F., Santos D. S., Maas W. K. Partial purification and characterization of a heat-labile enterotoxin of Escherichia coli. Infect Immun. 1976 Jun;13(6):1710–1720. doi: 10.1128/iai.13.6.1710-1720.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Smith H. W., Gyles C. L. The effect of cell-free fluids prepared from cultures of human and animal enteropathogenic strains of Escherichia coli on ligated intestinal segments of rabbits and pigs. J Med Microbiol. 1970 Aug;3(3):403–409. doi: 10.1099/00222615-3-3-403. [DOI] [PubMed] [Google Scholar]
  19. Smith H. W., Gyles C. L. The relationship between two apparently different enterotoxins produced by enteropathogenic strains of Escherichia coli of porcine origin. J Med Microbiol. 1970 Aug;3(3):387–401. doi: 10.1099/00222615-3-3-387. [DOI] [PubMed] [Google Scholar]
  20. Smith H. W., Halls S. Studies on Escherichia coli enterotoxin. J Pathol Bacteriol. 1967 Apr;93(2):531–543. doi: 10.1002/path.1700930212. [DOI] [PubMed] [Google Scholar]
  21. 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]

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