Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1986 Apr;52(1):45–49. doi: 10.1128/iai.52.1.45-49.1986

Purification and characterization of Vibrio cholerae non-O1 heat-stable enterotoxin.

M Arita, T Takeda, T Honda, T Miwatani
PMCID: PMC262195  PMID: 3957432

Abstract

A toxin which causes rapid fluid accumulation in a suckling mouse assay and which was produced by Vibrio cholerae non-O1 was investigated. The toxin was purified from the culture supernatant of V. cholerae non-O1 (strain A-5) by ammonium sulfate fractionation, hydroxyapatite treatment, ethanol extraction, column chromatographies on SP-Sephadex C-50 and DEAE-Sephadex A-25, and high-pressure liquid chromatography on a Lichrosorb RP-8 column. About 1.4 X 10(5)-fold purification was achieved, with a recovery of about 12%. Although the crude preparation was heat labile, the purified toxin was heat stable. The minimum effective dose of purified toxin was about 5 ng in the suckling mouse assay. The amino acid composition of the purified toxin was determined to be Asp(3), Glu(1), Ala(1), half-Cys(6), Ile(2), Leu(1), Phe(1), and Pro(1). These data show the production of a new type of heat-stable enterotoxin (NAG-ST) by V. cholerae non-O1.

Full text

PDF
45

Selected References

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

  1. Aimoto S., Takao T., Shimonishi Y., Hara S., Takeda T., Takeda Y., Miwatani T. Amino-acid sequence of a heat-stable enterotoxin produced by human enterotoxigenic Escherichia coli. Eur J Biochem. 1982 Dec 15;129(2):257–263. doi: 10.1111/j.1432-1033.1982.tb07047.x. [DOI] [PubMed] [Google Scholar]
  2. Aldová E., Láznicková K., Stepánková E., Lietava J. Isolation of nonagglutinable vibrios from an enteritis outbreak in Czechoslovakia. J Infect Dis. 1968 Feb;118(1):25–31. doi: 10.1093/infdis/118.1.25. [DOI] [PubMed] [Google Scholar]
  3. Blake P. A., Weaver R. E., Hollis D. G. Diseases of humans (other than cholera) caused by vibrios. Annu Rev Microbiol. 1980;34:341–367. doi: 10.1146/annurev.mi.34.100180.002013. [DOI] [PubMed] [Google Scholar]
  4. Craig J. P., Yamamoto K., Takeda Y., Miwatani T. Production of cholera-like enterotoxin by a Vibrio cholerae non-O1 strain isolated from the environment. Infect Immun. 1981 Oct;34(1):90–97. doi: 10.1128/iai.34.1.90-97.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Johnson W. M., Lior H. Cytotoxicity and suckling mouse reactivity of Aeromonas hydrophila isolated from human sources. Can J Microbiol. 1981 Oct;27(10):1019–1027. doi: 10.1139/m81-159. [DOI] [PubMed] [Google Scholar]
  7. MCINTYRE O. R., FEELEY J. C., GREENOUGH W. B., 3rd, BENENSON A. S., HASSAN S. I., SAAD A. DIARRHEA CAUSED BY NON-CHOLERA VIBRIOS. Am J Trop Med Hyg. 1965 May;14:412–418. doi: 10.4269/ajtmh.1965.14.412. [DOI] [PubMed] [Google Scholar]
  8. Madden J. M., Nematollahi W. P., Hill W. E., McCardell B. A., Twedt R. M. Virulence of three clinical isolates of Vibrio cholerae non-O-1 serogroup in experimental enteric infections in rabbits. Infect Immun. 1981 Aug;33(2):616–619. doi: 10.1128/iai.33.2.616-619.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nishibuchi M., Seidler R. J. Medium-dependent production of extracellular enterotoxins by non-O-1 Vibrio cholerae, Vibrio mimicus, and Vibrio fluvialis. Appl Environ Microbiol. 1983 Jan;45(1):228–231. doi: 10.1128/aem.45.1.228-231.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nishibuchi M., Seidler R. J., Rollins D. M., Joseph S. W. Vibrio factors cause rapid fluid accumulation in suckling mice. Infect Immun. 1983 Jun;40(3):1083–1091. doi: 10.1128/iai.40.3.1083-1091.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Pai C. H., Mors V. Production of enterotoxin by Yersinia enterocolitica. Infect Immun. 1978 Mar;19(3):908–911. doi: 10.1128/iai.19.3.908-911.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Takao T., Hitouji T., Aimoto S., Shimonishi Y., Hara S., Takeda T., Takeda Y., Miwatani T. Amino acid sequence of a heat-stable enterotoxin isolated from enterotoxigenic Escherichia coli strain 18D. FEBS Lett. 1983 Feb 7;152(1):1–5. doi: 10.1016/0014-5793(83)80469-4. [DOI] [PubMed] [Google Scholar]
  14. Takao T., Tominaga N., Shimonishi Y., Hara S., Inoue T., Miyama A. Primary structure of heat-stable enterotoxin produced by Yersinia enterocolitica. Biochem Biophys Res Commun. 1984 Dec 28;125(3):845–851. doi: 10.1016/0006-291x(84)91360-3. [DOI] [PubMed] [Google Scholar]
  15. Takeda Y., Takeda T., Yano T., Yamamoto K., Miwatani T. Purification and partial characterization of heat-stable enterotoxin of enterotoxigenic Escherichia coli. Infect Immun. 1979 Sep;25(3):978–985. doi: 10.1128/iai.25.3.978-985.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Yamamoto K., Takeda Y., Miwatani T., Craig J. P. Evidence that a non-O1 Vibrio cholerae produces enterotoxin that is similar but not identical to cholera enterotoxin. Infect Immun. 1983 Sep;41(3):896–901. doi: 10.1128/iai.41.3.896-901.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES