Journal of Chemical Ecology

, Volume 29, Issue 10, pp 2369–2378 | Cite as

Comparative Benzene-Induced Fatty Acid Changes in a Rhodococcus Species and Its Benzene-Sensitive Mutant: Possible Role of Myristic and Oleic Acids in Tolerance

  • Tony Gutiérrez
  • Robert P. Learmonth
  • Peter D. Nichols
  • Iain Couperwhite

Abstract

A Gram positive bacterium of the genus Rhodococcus was isolated from a contaminated site in Sydney, Australia, for its ability to tolerate and degrade high concentrations of benzene. To identify fatty acids that may impart this Rhodococcus sp. with tolerance to toxic solvents, a benzene-sensitive strain, labeled M2b, was isolated using EMS mutagenesis. A comparative analysis of fatty acid profiles showed that strain M2b was unable to increase its saturated:unsaturated ratio of fatty acids to the level achieved by the w-t strain when both strains were challenged with benzene. This was due to M2b's increased abundance of myristic acid, and decreased abundance of oleic acid. In addition, by measuring the generalized polarization of the fluorescent membrane probe laurdan using fluorescence spectroscopy, we have shown for the first time the effects of an aromatic hydrocarbon on the membrane fluidity of a Rhodococcus sp. The fluidity of the membranes increased after only 0.5 hr of exposure to benzene, thus suggesting the partitioning of benzene within the lipid bilayer. The response of this Rhodococcus sp. to benzene may suggest a mechanism for how other microorganisms survive when toxic solvents are released within the vicinity of their environment.

Benzene membrane fluidity bioremediation fatty acids myristic acid oleic acid Rhodococcus cell membrane solvent tolerance 

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References

  1. Boden, N., Jones, S. A., and Sixl, F. 1991. On the use of deuterium nuclear magnetic resonance as a probe of chain packing in lipid bilayers. Biochemistry 30:2146–2155.Google Scholar
  2. Claus, D. and Walker, N. 1964. The decomposition of toluene by soil bacteria. J. Gen. Microbiol. 36:107–122.Google Scholar
  3. Cullis, P. R. and Hope, M. J. 1985. Physical properties and functional roles of lipid membranes, pp. 25–72, in J. E. Vance and D. E. Vance, (Eds.) Biochemistry of Lipids and Membranes. Benjamin/Cummings, California.Google Scholar
  4. Dunkelblum, E., Tan, S. H., and Silk, R. J. 1985. Double-bond location in monounsaturated fatty acids by dimethyl disulfide derivatization and mass spectrometry. J. Chem. Ecol. 11:265–277.Google Scholar
  5. Eisenstadt, E., Carlton, B. C., and Brown, B. J. 1994. Gene manipulation, pp. 297–316, in, P. Gerhardt (Ed.)Methods for General and Molecular Bacteriology. American Society for Microbiology, Washington, DC.Google Scholar
  6. Gutiérrez, J. A., Nichols, P., and Couperwhite, I. 1999. Changes in whole cell-derived fatty acids induced by benzene and occurrence of the unusual 16:1ω6c in Rhodococcus sp. 33. FEMS Microbiol. Lett. 176:213–218.Google Scholar
  7. Hamilton, J. T. G., Mcroberts, W. C., Larkin, M. J., and Harper, D. B. 1995. Long-chain haloalkanes are incorporated into fatty acids by Rhodococcus rhodochrous NCIMB 13064. Microbiology 141:2611–2617.Google Scholar
  8. Heipieper, H. J., Weber, F. J., Sikkema, J., Keweloh, H., and de Bont, J. A. M. 1994. Mechanisms of resistance of whole cells to toxic organic solvents. Trends Biotechnol. 12:409–415.Google Scholar
  9. Learmonth, R. P. and Gratton, E. 2002. Assessment of membrane fluidity in individual yeast cells by laurdan generalized polarization and multi-photon scanning flourescence microscopy, pp. 241–252, in R. Kraayenhof, A. J.-W. G. Visser, and H.-G. Gerritsen (Eds). Flourescence Spectroscopy, Imaging and Probes—New Tools in chemical, Physical and Life Sciences, Springer, Heidelberg.Google Scholar
  10. Middlehoven, W., Koorevaar, M., and Schuur, G. 1992. Degradation of benzene compounds by yeasts in acidic soils. Plant Soil 145:37–43.Google Scholar
  11. Nichols, P. D., Guckert, J. B., and White, D. C. 1986. Determination of monounsaturated fatty acid double-bond position and geometry for microbial monocultures and complex consortia by capillary GC–MS of their dimethyl disulfide adducts. J. Microbiol. Methods 5:49–55.Google Scholar
  12. Paje, M. L., Neilan, B., and Couperwhite, I. 1997. A Rhodococcus species that thrives on medium saturated with liquid benzene. Microbiology 143:2975–2981.Google Scholar
  13. Parasassi, T., de Stasio, G., D'ubaldo, A., and Gratton, E. 1990. Phase fluctuation in phospholipid membranes revealed by Laurdan fluorescence. Biophys. J. 57:1179–1186.Google Scholar
  14. Siegrist, R. L. 1992. Volatile organic compounds in contaminated soils: The nature and validity of the measurement process. J. Hazard. Mater. 29:3–15.Google Scholar
  15. Sikkema, J., de Bont, J. A. M., and Poolman, B. 1994. Interactions of cyclic hydrocarbons with biological membranes. J. Biol. Chem. 269:8022–8028.Google Scholar
  16. Sikkema, J., de Bont, J. A. M., and Poolman, B. 1995. Mechanisms of membrane toxicity of hydrocarbons. Microbiol. Rev. 59:201–222.Google Scholar
  17. Simon, S. A., Mcdaniel, R. V., and Mcintosh, T. J. 1982. Interaction of benzene with micelles and bilayers. J. Phys. Chem. 86:1449–1456.Google Scholar
  18. Tsitko, I. V., Zaitsev, G. M., Lobanok, A. G., and Salkinoja-Salonen, M. S. 1999. Effect of aromatic compounds on cellular fatty acid composition of Rhodococcus opacus. Appl. Environ. Microbiol. 65:853–855.Google Scholar
  19. Ward, A. J. L., Rananavare, S. B., and Friberg, S. E. 1986. Solvation changes induced in alyotropic liquid crystal containing solubilized benzene. Langmuir 2:373–375.Google Scholar
  20. Warhurst, A. M. and Fewson, C. A. 1994. Biotransformations catalyzed by the genus Rhodococcys. Crit. Rev. Botechnol. 14:29–73.Google Scholar
  21. Weber, F. J. and de Bont, J. A. M. 1996. Adaptation mechanisms of microorganisms to the toxic effects of organic solvents on membranes. Biochem. Acta 1286:225–245.Google Scholar

Copyright information

© Plenum Publishing Corporation 2003

Authors and Affiliations

  • Tony Gutiérrez
    • 1
  • Robert P. Learmonth
    • 2
  • Peter D. Nichols
    • 3
  • Iain Couperwhite
    • 1
  1. 1.School of Microbiology and ImmunologyThe University of New South WalesSydneyAustralia
  2. 2.Department of Biological and Physical Sciences, and Center for Rural and Environmental BiotechnologyUniversity of Southern QueenslandToowoombaAustralia
  3. 3.CSIRO Marine ResearchTasmaniaAustralia

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