, Volume 79, Issue 4, pp 439–444 | Cite as

Activation of the antioxidant complex in Pseudomonas aurantiaca—Producer of phenazine antibiotics

  • E. G. VeremeenkoEmail author
  • N. P. Maksimova
Experimental Articles


Two Pseudomonas aurantiaca mutant strains overproducing phenazine antibiotics (synthesis levels of 210 and 410 mg/l, respectively) along with wild-type bacteria (production level of 71–75 mg/l) and a phz mutant not producing phenazines were used to study the changes in the activity of the antioxidant complex components, that is, catalase, superoxide dismutase (SOD), glutathione reductase, and NADH oxidase; glutathione concentration (in both reduced and oxidized forms); and activity of acyl-CoA synthetase, the key enzyme of cell metabolism.

Bacterial producers were found to respond to an increase in intra- and extracellular phenazines by induction of catalase, SOD, glutathione reductase, and glutathione synthesis. However, while in the case of catalase and glutathione reductase this trend was observed in all the strains under study, the activity of SOD at a high level of phenazine synthesis (in particular, 410 mg/l) decreased somewhat, probably due to high its sensitivity to high concentrations of H2O2 generated by phenazines. Decrease in SOD activity was compensated by increase in the synthesis rates of glutathione and glutathione reductase. NADH oxidase was shown to be practically uninvolved in formation of P. aurantiaca response toward phenazine accumulation, and acyl-CoA synthetase activity was found to decrease.

Key words

phenazine antibiotics catalase superoxide dismutase glutathione reductase NADH oxidase acyl-CoA synthetase 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Laursen, J.B. and Nielsen, J., Phenazine Natural Products: Biosynthesis, Synthetic Analogues, and Biological Activity, Chem. Rev., 2004, vol. 104, pp. 1663–1686.CrossRefPubMedGoogle Scholar
  2. 2.
    Kerr, J.R., Phenazine Pigments: Antibiotics and Virulence Factors, Infect. Dis. Rev., 2000, vol. 2, pp. 184–194.Google Scholar
  3. 3.
    Price-Whelan, A., Dietrich, L.P., and Newman, D.K., Rethinking “Secondary” Metabolism: Physiological Roles for Phenazine Antibiotics, Nat. Chem. Biol., 2006, vol. 2, no. 2, pp. 71–78.CrossRefPubMedGoogle Scholar
  4. 4.
    Look, D.C., Stoll, L.L., Romig, S.A., Humlicek, A., Britigan, B.E., and Denning, G.M., Pyocyanin and Its Precursor Phenazine-1-Carboxylic Acid Increase IL-8 and Intercellular Adhesion Molecule-1 Expression in Human Airway Epithelial Cells by Oxidant-Dependent Mechanisms, J. Immunol., 2005, vol. 175, pp. 4017–4023.PubMedGoogle Scholar
  5. 5.
    Hassan, H.M. and Fridovich, I., Mechanism of Antibiotic Action of Pyocyanine, J. Bacteriol., 1980, vol. 141, no. 1, pp. 156–163.PubMedGoogle Scholar
  6. 6.
    Kim, K.-J. and Kim, T.-S., Isolation and Characterization of Soil Microorganisms Producing Acyl CoA Synthetase Inhibitor, Yakhak Hoeji, 1996, vol. 40, pp. 713–719.Google Scholar
  7. 7.
    Black, P.N., Faergeman, J.N., and DiRusso, C.C., Long-Chain Acyl-CoA-Dependent Regulation of Gene Expression in Bacteria, Yeast and Mammals, J. Nutr., 2000, vol. 130, pp. 305–309.Google Scholar
  8. 8.
    Rao, Y.M. and Sureshkumar, G.K., Oxidative-Stress-Induced Production of Pyocyanin by Xanthomonas campestris and Its Effect on the Indicator Target Organism, Escherichia coli, J. Ind. Microbiol. Biotechnol., 2000, vol. 25, pp. 266–272.CrossRefGoogle Scholar
  9. 9.
    Hassett, D.J., Charniga, L., Bean, K., Ohman, D.E., and Cohen, M.S., Response of Pseudomonas aeruginosa to Pyocyanin: Mechanisms of Resistance, Antioxidant Defenses, and Demonstration of a Manganese-Cofactored Superoxide Dismutase, Infect. Immun., 1992, vol. 60, no. 2, pp. 328–336.PubMedGoogle Scholar
  10. 10.
    Miyoshi, A., Rochat, T., Gratadoux, J., Loir, Y., Oliveira, S., Langella, P., and Azavedo, V., Oxidative Stress in Lactococcus lactis, Gen. Mol. Res., 2003, vol. 2, no. 4, pp. 348–359.Google Scholar
  11. 11.
    Kyoung-Ja, K., Phenazine 1-Carboxylic Acid Resistance in 1-Carboxylic Acid Producing Bacillus sp. B-6, J. Biochem Mol. Biol., 2000, vol. 33, no. 4, pp. 332–336.Google Scholar
  12. 12.
    Veremeenko, E.G., Production of Pseudomonas aurantiaca B-162 Regulatory Mutants Resistant to Toxic Analogues of Aromatic Amino Acids, Mat. Mezhd. Konf. “Ot klassicheskikh metodov genetiki i selektsii k DNK-tekhnologiyam”, (Proc. Int. Conf. “From Classical Methods of Genetics and Selection to DNA Technologies”), Gomel’, 2007, p. 112.Google Scholar
  13. 13.
    Levitch, M.E. and Stadtman, E.R., A Study of the Biosythesis of Phenazine-1-Carboxylic Acid, Arch. Biochem. Biophys., 1964, vol. 106, pp. 194–199.CrossRefPubMedGoogle Scholar
  14. 14.
    Lorenzo, V., Herrero, M., and Timmis, K., Transposon Vectors Containing Non-Antibiotic Resistance Selection Markers for Cloning and Stable Chromosomal Insertion of Foreign Genes in Gram-Negative Bacteria, J. Bacteriol., 1990, vol. 172, no. 11, pp. 6557–6567.PubMedGoogle Scholar
  15. 15.
    Kostyuk, V.A., Potapovich, A.I., and Kovaleva, Zh.V., A Rapid and Sensitive Method for Determination of Superoxide Dismutase Activity Based on Quercetin Oxidatrion, Vopr. Med. Khim., 1990, vol. 36, no. 2, pp. 88–91.Google Scholar
  16. 16.
    Aebi, H., Catalase in vitro, Methods Enzymol., 1984, vol. 105, pp. 121–126.CrossRefPubMedGoogle Scholar
  17. 17.
    Lopez, F., Kleerebezem, M., and Hugenholtz, J., Cofactor Engineering: a Novel Approach to Metabolic Engineering in Lactococcus lactis by Control of NADH Oxidase, J. Bacteriol., 1998, vol. 180, no. 15, pp. 3804–3808.Google Scholar
  18. 18.
    Senft, A., Dalton, T., and Shertzer, H., Determining Glutathione and Glutathione Disulfide Using the Fluorescence Probe o-Phthalaldehyde, Anal. Biochem., 2000, vol. 280, pp. 80–86.CrossRefPubMedGoogle Scholar
  19. 19.
    Li, Y., Hugenholtz, J., Abee, T., and Molenaar, D., Glutathione Protects Lactococcus lactis against Oxidative Stress, Appl. Eviorn. Microbiol., 2003, vol. 69, no. 10, pp. 5739–5745.CrossRefGoogle Scholar
  20. 20.
    Bradford, J.K., A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding, Anal. Biochem., 1976, vol. 72, pp. 248–254.CrossRefPubMedGoogle Scholar
  21. 21.
    Hiroshi, T., Kazuaki, I., and Satoshi, O., Inhibition of Acyl-CoA Synthetase by Triacsins, Biochim. Biophys. Acta, 1987, vol. 921, pp. 595–598.Google Scholar
  22. 22.
    Feklistova, I.N., Synthesis of Aromatic Antibiotics in Pseudomonas aurantiaca B-162 Extended Abstract of Cand. Sci. (Biol.) Dissertation, Minsk: Belarus State Univ., 2006.Google Scholar
  23. 23.
    Levitch, M.E., Regulation of Aromatic Amino Acid Biosynthesis in Phenazine-Producing Strains, J. Bacteriol., 1970, vol. 103, no. 1, pp. 16–19.PubMedGoogle Scholar
  24. 24.
    Hertel, C., Schmidt, G., Fischer, M., Oellers, K., and Hammes, W., Oxygen-Dependent Regulation of the Expression of the Catalase Gene katA of Lactobacillus sakei LTH677, Appl. Environ. Microbiol., 1998, vol. 64, no. 4, pp. 1359–1365.PubMedGoogle Scholar
  25. 25.
    Vance, C. and Miller, K.A., Novel Insights Into the Basis for Escherichia coli Superoxide Dismutase’s Metal Ion Specificity from Mn-Substituted FeSOD and Its Very High Em, Biochemistry, 2001, no. 40, pp. 13079–13087.Google Scholar
  26. 26.
    Miller, A., Handbook of Metalloproteins: Fe Superoxide Dismutase, Chichester: John Wiley & Sons, Ltd, 2001, pp. 668–682.Google Scholar
  27. 27.
    Chesney, J.A., Eaton, J.W., and Manorey, J.R., Bacterial Glutathione: a Sacrificial Defense against Chlorine Compounds, J. Bacteriol., 1995, vol. 178, no. 7, pp. 2131–2135.Google Scholar
  28. 28.
    Vergauwen, B., Pauwels, F., and Van Beeumen, J., Glutatione and Catalase Provide Defenses for Protection Against Respiration-Generated Hydrogen Peroxide in Haemophilus infuenzae, J. Bacteriol., 2003, vol. 185, no. 18, pp. 5555–5562.CrossRefPubMedGoogle Scholar
  29. 29.
    Riccillo, P.M., Muglia, C.I., Bruijn, F.J., Roe, A.J., Booth, I.R., and Agular, O.M., Glutatione Is Involved in Environmental Stress Responses in Rhizobium tropici, Including Acid Tolerence, J. Bacteriol., 2000, vol. 182, no. 6, pp. 1748–1753.CrossRefPubMedGoogle Scholar
  30. 30.
    Ochi, T., Hydrogen Peroxide Increases the Activity of γ-Glutamylcysteine Synthetase in Cultured Chinese Hamster V79 Cells, Ach. Toxicol., 1995, vol. 70, no. 2, pp. 96–103.CrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2010

Authors and Affiliations

  1. 1.Belarusian State UniversityMinskBelarus

Personalised recommendations