1887

Abstract

The proposal that nitrite exerts its inhibitory effect on anaerobic bacteria by direct interaction with the iron-sulphur proteins of the phosphoroclastic system was investigated. The effects of nitrate, nitrite with or without ascorbate, and nitric oxide on the growth of in liquid cultures at pH 7·4, on the rates of hydrogen production, and on the activities of the enzymes pyruvate-ferredoxin oxidoreductase and hydrogenase, and of ferredoxin were investigated. In agreement with previous studies, nitrate was the least effective inhibitor of cell growth, and nitric oxide the most effective. Nitrite reductase activity was very low in , indicating that the presence of external reducing agents would be necessary for the reduction of nitrite to nitric oxide. Inhibition by nitrite was enhanced by ascorbate; 0·5 m-nitrite with 10 m-ascorbate stopped growth completely. In partially-purified preparations 4·1 m-NaNO and equimolar ascorbate caused complete inactivation of hydrogenase activity but only partial (up to 78 %) inactivation of pyruvate-ferredoxin oxidoreductase. This agreed with the loss of hydrogen production observed with nitrite Inhibition occurred within 5 min, and was irreversible in each case. Electron paramagnetic resonance (EPR) spectroscopy showed that paramagnetic [Fe(NO)(SR)] species were formed during growth in the presence of nitrite, and were associated with cells. However, the intensity of these EPR signals did not correlate with the inhibition of cell growth. The [4Fe-4S] clusters in ferredoxin were shown by EPR spectroscopy to be resistant to treatment with 3·6 m-NaNO and 3·6 m-ascorbate. It is concluded that the effects of nitrite on pre-formed ironsulphur proteins are not convincing as a basis for the lethal effects on bacterial cells.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-136-10-2067
1990-10-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/136/10/mic-136-10-2067.html?itemId=/content/journal/micro/10.1099/00221287-136-10-2067&mimeType=html&fmt=ahah

References

  1. Adams M.W.W., Eccleston E., Howard J.B. 1989; Iron-sulphur clusters of hydrogenase I and hydrogenase II of Clostridium pasteurianum. Proceedings of the National Academy of Sciences of the United States of America 86:4932–4936
    [Google Scholar]
  2. Angermaier L., Simon H. 1983; On the reduction of aliphatic and aromatic nitro compounds by Clostridia; the role of ferredoxin in its stabilisation. Hoppe-Seyler’s Zeitschrift für Physiologische Chemie 364S:961–975
    [Google Scholar]
  3. Asan T., Solberg M. 1976; Inhibition of Clostridium perfringens by heated combinations of nitrite, sulfur and ferrous or ferric ions. Applied and Environmental Microbiology 31:49–52
    [Google Scholar]
  4. Baird-Parker A.C., Baillie M.A.H. 1973; The inhibition of Clostridium botulinum by nitrite and sodium chloride. Proceedings of the International Symposium on Nitrite Meat Products II:77–91
    [Google Scholar]
  5. Berlier Y., Fauque G.D., Le Gall J., Choi E.S., Peck H.D., Lespinat P.A. 1987; Inhibition studies of three classes of Desulfovibrio hydrogenase - application to the further characterisation of the multiple hydrogenases found in D. vulgaris Hilden- borough. Biochemical and Biophysical Research Communications 146:147–154
    [Google Scholar]
  6. Bonner W.D.JR Blum H., Rich P.R., Salerno J.C. 1978; Further observation on a mitochrondrial metallocompound and on a ferrous-EDTA-NO model compound. In Frontiers of Biological Energetics pp. 997–1001 London: Academic Press.;
    [Google Scholar]
  7. Butler A.R., Glidewell C., Li M.-H. 1988; Nitrosyl complexes of iron-sulphur clusters. In Advances in Inorganic Chemistry pp. 335–393 Sykes A. G. London: Academic Press.;
    [Google Scholar]
  8. Carpenter C.E., Reddy D.S., Cornforth D.S. 1987; Inactivation of clostridial ferredoxin and PFR by sodium nitrite. Applied and Environmental Microbiology 53:549–552
    [Google Scholar]
  9. James E.W., Kell D.B., Lovitt R.W., Morris J.G. 1988; Electrosynthesis and electroanalysis using Clostridium sporogenes. Bioelectrochemistry and Bioenergetics 20:21–32
    [Google Scholar]
  10. Jones R.W., Gray T.A., Garland P.B. 1976; A study on the permeability of the cytoplasmic membrane of E.coli to reduced and oxidised benzyl viologen and methyl viologen cations; complications in the use of viologens as redox mediators for membrane bound enzymes. Biochemical Society Transactions 4:671–673
    [Google Scholar]
  11. Knight T.M., Forman D., Al-Dabbagh S.A., Doll R. 1987; Estimation of dietary intake of nitrate and nitrite in Great Britain. Food and Chemical Toxicology 25:277–285
    [Google Scholar]
  12. Larkworthy L.F., Turnbull M.H., Yavari A. 1977; Possible role of iron II amino acid complexes in nitrite depletion and inhibitor formation in cured meats. Chemistry and IndustryMay21401–402
    [Google Scholar]
  13. Mcdonald C.C., Phillips W.D., Mower H.F. 1965; An electron spin resonance study of some complexes of iron, nitric oxide and anionic ligands. Journal of the American Chemical Society 87:3319–3326
    [Google Scholar]
  14. Meyer C.L., Roos J.W., Papoutsakis L. 1986; CO gassing leads to alcohol production and butyrate uptake without acetone formation in continuous cultures of Clostridium acetobutylicum. Applied Microbiology and Biotechnology 24:159–167
    [Google Scholar]
  15. O’Leary V., Solberg M. 1976; Effect of sodium nitrite inhibition on intracellular thiol groups and on activity of certain glycolytic enzymes in Clostridium perfringens. Applied and Environmental Microbiology 31:208–212
    [Google Scholar]
  16. Payne M.J., Cammack R. 1986; Inhibition by nitrite of the phosphoroclastic system of Clostridium sporogenes. Biochemical Society Transactions 14:1283–1284
    [Google Scholar]
  17. Payne M.J., Glidewell C., Cammack R. 1990; Interaction of iron-thiol-nitrosyl compounds with the phosphoroclastic system of Clostridium sporogenes. Journal of General Microbiology 136:2077–2087
    [Google Scholar]
  18. Perigo J.A., Roberts T.A. 1968; Inhibition of clostridia by nitrite. Journal of Food Technology 3:91–94
    [Google Scholar]
  19. Perigo J.A., Whiting E., Bashford T.E. 1967; Observations on the inhibition of vegetative cells of Clostridium sporogenes by nitrite which has been autoclaved in a laboratory medium, discussed in the context of sub-lethally processed cured meats. Journal of Food Technology 2:377–397
    [Google Scholar]
  20. Pierson M.D., Smoot L.A. 1981; Nitrite, nitrite alternatives and the control of Clostridium botulinum in cured meats. CRC Critical Reviews in Food Science and Nutrition 17:141–187
    [Google Scholar]
  21. Reddy D., Lancaster J.R.JR Cornforth D.P. 1983; Nitrite inhibition of Clostridium botulinum ; Electron spin resonance detection of iron-nitric oxide complexes. Science 221:769–770
    [Google Scholar]
  22. Rich P.R., Salerno J.C., Leigh J.S., Bonner W.D.JR. 1978; A spin 3/2 ferrous-NO derivative of an iron containing moiety associated with Neurospora crassa and higher plant mitochondria. FEBS Letters 93:323–326
    [Google Scholar]
  23. Roberts T.A., Ingram M. 1966; The effect of sodium chloride, potassium nitrate and sodium nitrite on the recovery of heated bacterial spores. Journal of Food Technology 1:147–163
    [Google Scholar]
  24. Tompkin R.B., Christiansen L.N., Shaparis A.B. 1978a; Antibotulinal role of isoascorbate in cured meats. Journal of Food Science 43:1368–1370
    [Google Scholar]
  25. Tompkin R.B., Christiansen L.N., Shaparis A.B. 1978b; Enhancing nitrite inhibition of Clostridium botulinum with iso-ascorbate in perishable canned cured meat. Applied and Environmental Microbiology 35:59–61
    [Google Scholar]
  26. Tompkin R.B., Christiansen L.N., Shaparis A.B. 1978C; The effect of iron on botulinal inhibition in perishable canned cured meat. Journal of Food Technology 13:521–527
    [Google Scholar]
  27. Uyeda K., Rabinowitz J.C. 1971; Pyruvate: ferredoxin oxidoreductase.IV.Studies on the reaction mechanism. Journal of Biological Chemistry 246:3120–3125
    [Google Scholar]
  28. Vanin A.F., Varich V.Ya. 1981; Nitrosyl non-heme iron complexes in animal tissues. Studia Biophysica 86:177–185
    [Google Scholar]
  29. Woods L.F.J., Wood J.M. 1982; A note on the effect of nitrite inhibition on the metabolism of Clostridium botulinum. Journal of Applied Bacteriology 52:109–110
    [Google Scholar]
  30. Woods L.F.J., Wood J.M., Gibbs P.A. 1981; The involvement of nitric oxide in the inhibition of the phosphoroclastic system in Clostridium sporogenes by sodium nitrite. Journal of General Microbiology 125:399–406
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-136-10-2067
Loading
/content/journal/micro/10.1099/00221287-136-10-2067
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error