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.
Similar content being viewed by others
References
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.
Claus, D. and Walker, N. 1964. The decomposition of toluene by soil bacteria. J. Gen. Microbiol. 36:107–122.
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.
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.
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.
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.
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.
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.
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.
Middlehoven, W., Koorevaar, M., and Schuur, G. 1992. Degradation of benzene compounds by yeasts in acidic soils. Plant Soil 145:37–43.
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.
Paje, M. L., Neilan, B., and Couperwhite, I. 1997. A Rhodococcus species that thrives on medium saturated with liquid benzene. Microbiology 143:2975–2981.
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.
Siegrist, R. L. 1992. Volatile organic compounds in contaminated soils: The nature and validity of the measurement process. J. Hazard. Mater. 29:3–15.
Sikkema, J., de Bont, J. A. M., and Poolman, B. 1994. Interactions of cyclic hydrocarbons with biological membranes. J. Biol. Chem. 269:8022–8028.
Sikkema, J., de Bont, J. A. M., and Poolman, B. 1995. Mechanisms of membrane toxicity of hydrocarbons. Microbiol. Rev. 59:201–222.
Simon, S. A., Mcdaniel, R. V., and Mcintosh, T. J. 1982. Interaction of benzene with micelles and bilayers. J. Phys. Chem. 86:1449–1456.
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.
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.
Warhurst, A. M. and Fewson, C. A. 1994. Biotransformations catalyzed by the genus Rhodococcys. Crit. Rev. Botechnol. 14:29–73.
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.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Gutiérrez, T., Learmonth, R.P., Nichols, P.D. et al. Comparative Benzene-Induced Fatty Acid Changes in a Rhodococcus Species and Its Benzene-Sensitive Mutant: Possible Role of Myristic and Oleic Acids in Tolerance. J Chem Ecol 29, 2369–2378 (2003). https://doi.org/10.1023/A:1026286700855
Issue Date:
DOI: https://doi.org/10.1023/A:1026286700855