Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1986 Jul;167(1):57–65. doi: 10.1128/jb.167.1.57-65.1986

Characterization of anguibactin, a novel siderophore from Vibrio anguillarum 775(pJM1).

L A Actis, W Fish, J H Crosa, K Kellerman, S R Ellenberger, F M Hauser, J Sanders-Loehr
PMCID: PMC212840  PMID: 3013839

Abstract

Anguibactin, a siderophore produced by cells of Vibrio anguillarum 775 harboring the pJM1 plasmid, has now been isolated from the supernatants of iron-deficient cultures. This iron-reactive material was purified by adsorption onto an XAD-7 resin and subsequent gel filtration on a Sephadex LH-20 column. The resulting neutral compound produced an ion at m/z 348 in mass spectrometry and contained one sulfur, four oxygen, and four nitrogen atoms as determined by elemental analysis. Its strong UV absorbance and blue fluorescence were suggestive of a phenolic moiety. In colorimetric reactions anguibactin behaved like a catechol. The catechol assignment was supported by the appearance of a new absorption band at 510 nm in the ferric complex and by the appearance of peaks at 1,367, 1,447, 1,469, and 1,538 cm-1 in the resonance Raman spectrum. In addition, the infrared spectrum gave evidence of a secondary amide function, but no free carboxylic acid or hydroxamic acid groups were observed. A third iron-ligating group was suggested by the liberation of three protons during iron binding; mass spectrometry of the resulting material yielded a molecular ion characteristic of a 1:1 complex of ferric anguibactin. The purified anguibactin exhibited specific growth-promoting activity under iron-limiting conditions for a siderophore-deficient mutant of V. anguillarum 775(pJM1). A novel structure for anguibactin was indicated by the failure of a large number of known siderophores and synthetic chelators to yield a similar type of specific cross-feeding in the V. anguillarum bioassay.

Full text

PDF
57

Selected References

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

  1. Actis L. A., Potter S. A., Crosa J. H. Iron-regulated outer membrane protein OM2 of Vibrio anguillarum is encoded by virulence plasmid pJM1. J Bacteriol. 1985 Feb;161(2):736–742. doi: 10.1128/jb.161.2.736-742.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barnum D. W. Spectrophotometric determination of catechol, epinephrine, dopa, dopamine and other aromatic vic-diols. Anal Chim Acta. 1977 Mar;89(1):157–166. doi: 10.1016/S0003-2670(01)83081-6. [DOI] [PubMed] [Google Scholar]
  3. Bullen J. J., Rogers H. J., Griffiths E. Role of iron in bacterial infection. Curr Top Microbiol Immunol. 1978;80:1–35. doi: 10.1007/978-3-642-66956-9_1. [DOI] [PubMed] [Google Scholar]
  4. Cisar J. O., Fryer J. L. An epizootic of vibriosis in chinook salmon. Wildl Dis. 1969 Apr;5(2):73–76. doi: 10.7589/0090-3558-5.2.73. [DOI] [PubMed] [Google Scholar]
  5. Cox C. D., Graham R. Isolation of an iron-binding compound from Pseudomonas aeruginosa. J Bacteriol. 1979 Jan;137(1):357–364. doi: 10.1128/jb.137.1.357-364.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cox C. D., Rinehart K. L., Jr, Moore M. L., Cook J. C., Jr Pyochelin: novel structure of an iron-chelating growth promoter for Pseudomonas aeruginosa. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4256–4260. doi: 10.1073/pnas.78.7.4256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Crosa J. H. A plasmid associated with virulence in the marine fish pathogen Vibrio anguillarum specifies an iron-sequestering system. Nature. 1980 Apr 10;284(5756):566–568. doi: 10.1038/284566a0. [DOI] [PubMed] [Google Scholar]
  8. Crosa J. H., Actis L. A., Mitoma Y., Perez-Casal J., Tolmasky M. E., Valvano M. A. Plasmid-mediated iron sequestering systems in pathogenic strains of Vibrio anguillarum and Escherichia coli. Basic Life Sci. 1985;30:759–774. doi: 10.1007/978-1-4613-2447-8_52. [DOI] [PubMed] [Google Scholar]
  9. Crosa J. H., Hodges L. L. Outer membrane proteins induced under conditions of iron limitation in the marine fish pathogen Vibrio anguillarum 775. Infect Immun. 1981 Jan;31(1):223–227. doi: 10.1128/iai.31.1.223-227.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Crosa J. H., Hodges L. L., Schiewe M. H. Curing of a plasmid is correlated with an attenuation of virulence in the marine fish pathogen Vibrio anguillarum. Infect Immun. 1980 Mar;27(3):897–902. doi: 10.1128/iai.27.3.897-902.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Crosa J. H., Schiewe M. H., Falkow S. Evidence for plasmid contribution to the virulence of fish pathogen Vibrio anguillarum. Infect Immun. 1977 Nov;18(2):509–513. doi: 10.1128/iai.18.2.509-513.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Crosa J. H. The relationship of plasmid-mediated iron transport and bacterial virulence. Annu Rev Microbiol. 1984;38:69–89. doi: 10.1146/annurev.mi.38.100184.000441. [DOI] [PubMed] [Google Scholar]
  13. Emery T. A convenient assay for siderochrome hydrolytic enzymes. Anal Biochem. 1976 Mar;71(1):294–299. doi: 10.1016/0003-2697(76)90039-7. [DOI] [PubMed] [Google Scholar]
  14. Gaber B. P., Miskowski V., Spiro T. G. Resonance Raman scattering from iron(3)- and copper(II)-transferrin and an iron(3) model compound. A spectroscopic interpretation of the transferrin binding site. J Am Chem Soc. 1974 Oct 30;96(22):6868–6873. doi: 10.1021/ja00829a010. [DOI] [PubMed] [Google Scholar]
  15. Neilands J. B. Microbial iron compounds. Annu Rev Biochem. 1981;50:715–731. doi: 10.1146/annurev.bi.50.070181.003435. [DOI] [PubMed] [Google Scholar]
  16. Neilands J. B. Siderophores. Adv Inorg Biochem. 1983;5:137–166. [PubMed] [Google Scholar]
  17. Rioux C., Jordan D. C., Rattray J. B. Colorimetric determination of catechol siderophores in microbial cultures. Anal Biochem. 1983 Aug;133(1):163–169. doi: 10.1016/0003-2697(83)90238-5. [DOI] [PubMed] [Google Scholar]
  18. Salama S., Stong J. D., Neilands J. B., Spiro T. G. Electronic and resonance Raman spectra of iron(III) complexes of enterobactin, catechol, and N-methyl-2,3-dihydroxybenzamide. Biochemistry. 1978 Sep 5;17(18):3781–3785. doi: 10.1021/bi00611a017. [DOI] [PubMed] [Google Scholar]
  19. Sjöberg B. M., Loehr T. M., Sanders-Loehr J. Raman spectral evidence for a mu-oxo bridge in the binuclear iron center of ribonucleotide reductase. Biochemistry. 1982 Jan 5;21(1):96–102. doi: 10.1021/bi00530a017. [DOI] [PubMed] [Google Scholar]
  20. Walter M. A., Potter S. A., Crosa J. H. Iron uptake system medicated by Vibrio anguillarum plasmid pJM1. J Bacteriol. 1983 Nov;156(2):880–887. doi: 10.1128/jb.156.2.880-887.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Weinberg E. D. Iron withholding: a defense against infection and neoplasia. Physiol Rev. 1984 Jan;64(1):65–102. doi: 10.1152/physrev.1984.64.1.65. [DOI] [PubMed] [Google Scholar]

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

RESOURCES