Mikrobiol. Z. 2021; 83(4):15-23.
doi: https://doi.org/10.15407/microbiolj83.04.015

Fatty Acid Composition of Rhodococcus aetherivorans Cells During Phenol Assimilation

T.M. Nogina, L.A. Khomenko, V.S. Pidgorskyi, M.A. Kharkhota

Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine

The ability of microorganisms to survive in unfavorable conditions and maintain their biodegradation activity is mainly associated with changes in the composition of their cellular lipids. One of the factors of negative impact on cells can be their interaction with petroleum hydrocarbons, especially monoaromatic compounds, which are toxic to microorganisms even in small amounts. Aim. To research the changes in the cell fatty acids composition of Rhodococcus aetherivorans UCM Ac-602 strain during phenol degradation. Methods. The cultivation of the strain was carried out in a liquid mineral medium with initial phenol concentration 0.75, 1.0 and 1.5 g/L as a sole carbon and energy source. Cells grown in medium with n-hexadecane (7.5 g/L) and glucose (5.0 g/L) were used as controls. Fatty acid methyl esters were obtained by hydrolysis of cells in a 5% solution of acetyl chloride in methanol, followed by extraction with a mixture of ether-hexane. Identification of methyl esters was performed using a gas chromatography-mass spectrometry system Agilent 6800N/5973 inert (Agilent Technologies, US). The fatty acid content was determined using AgilentChemStation software. Results. Among the cellular fatty acids of R. aetherivorans UCM Ac-602 the straight-chain saturated hexadecanoic (C16:0), unsaturated hexadecenoic (C16:1 cis-9) and octadecenoic (C18:1 cis-9) acids as well as branched 10-methyl octadecanoic (tuberculostearic) (10-Me-C18:0) acid were dominated during growth on phenol and glucose. While in n-hexadecane grown cells main components of fatty acids pool were saturated tetradecanoic (C14:0) and hexadecanoic (C16:0) and unsaturated hexadecenoic (C16:1 cis-9) acids. The quantitative ratio of individual fatty acids of R. aetherivorans UCM Ac-602 cells differed depending on the substrate and incubation time. Under the influence of high phenol concentrations (1.5 g/L) there was a threefold increase in the ratio of straight-chain saturated to unsaturated fatty acids in comparison to cells grown on glucose and double increase compared to those grown on n-hexadecane. The amounts of 10-Мe-C18:0 fatty acid in cells grown on phenol were 1.8–3.2-fold higher in comparison to cells grown on glucose and 38.3–60.3-fold higher compared to those grown on n-hexadecane. In addition, the content of this acid in cells increased with increasing the time of incubation on phenol. Conclusions. A significant increase in the ratio of straightchain saturated to unsaturated fatty acids in the cells of R. aetherivorans UCM Ac-602 strain during growth on phenol and n-hexadecane in comparison to cells grown on glucose, as well as significant increase of methyl-branched (10-Me-C18:0) acid amount in phenol grown cells indicates the possible involvement of these fatty acids in the adaptation of the strain to the assimilation of toxic substances.

Keywords: Rhodococcus aetherivorans, fatty acids, phenol, n-hexadecane, glucose.

Full text (PDF, in English)

  1. Konings WN, Albers S-V, Koning S, Driessen AJM. The cell membrane plays a crucial role in survival of bacteria and archaea in extreme environments. Antonie Van Leeuwenhoek. 2002; 81:61–72. https://doi.org/10.1023/A:1020573408652
  2. De Carvalho CCCR. Adaptation of Rhodococcus to Organic Solvents. In: Alvarez HM, editor. Biology of Rhodococcus. Microbiology Monographs. 2nd ed. Springer Nature Switzerland AG; 2019. p. 103–135. https://doi.org/10.1007/978-3-030-11461-9_5
  3. Heipieper HJ, Weber FJ, Sikkema J, Keweloh H, de Bont JAM. Mechanisms of resistance of whole cells to toxic organic solvents. Trends in Biotechnology. 1994; 12(10):409–415. https://doi.org/10.1016/0167-7799(94)90029-9
  4. De Carvalho CCCR, Caramujo MJ. The various roles of fatty acids. Molecules. 2018; 23(10):2583. https://doi.org/10.3390/molecules23102583
  5. Ivshina IB. [Bacteria of the genus Rhodococcus (immunodiagnostics, detection, biodiversity)]. A dissertation in the form of a scientific report for the degree of doctor of biological sciences. Perm; 1997. Russian.
  6. Yoon JH, Cho YG, Kang SS, Kim SB, Lee ST, Park YH. Rhodococcus koreensis sp. nov., a 2,4-dinitrophenol-degrading bacterium. Int J Syst Evol Microbiol. 2000; 50(3):1193–1201. https://doi.org/10.1099/00207713-50-3-1193
  7. Nogina T, Fomina M, Dumanskaya T, et al. A new Rhodococcus aetherivorans strain isolated from lubricant-contaminated soil as a prospective phenol-biodegrading agent. Appl Microbiol Biotechnol. 2020; 104:3611–3625. https://doi.org/10.1007/s00253-020-10385-6
  8. Keweloh H, Weyrauch G, Rehm HJ. Phenolinduced membrane changes in free and immobilized Escherichia coli. Appl Microbiol Biotechnol. 1990; 33:66–71. https://doi.org/10.1007/BF00170572
  9. Heipieper HJ, Keweloh H, Rehm HJ. Influence of phenols on growth and membrane permeability of free and immobilized Escherichia coli. Appl Environ Microbiol. 1991; 57(4):1213–1217. https://doi.org/10.1128/aem.57.4.1213-1217.1991
  10. Keweloh H, Diefenbach R, Rehm HJ. Increase of phenol tolerance of Escherichia coli by alterations of the fatty acid composition of the membrane lipids. Arch Microbiol. 1991; 157(1):49–53. https://doi.org/10.1007/BF00245334
  11. Heipieper HJ, Diefenbach R, Keweloh H. Conversion of cis unsaturated fatty acids to trans, a possible mechanism for the protection of phenol-degrading Pseudomonas putida P8 from substrate toxicity. Appl Environ Microbiol. 1992; 58(6):1847–1852. https://doi.org/10.1128/aem.58.6.1847-1852.1992
  12. Whyte LG, Slagman SJ, Pietrantonio F, Bourbonnière L, Koval SF, Lawrence JR, Inniss WE, Greer CW. Physiological adaptations involved in alkane assimilation at low temperatures by Rhodococcus sp. strain Q15. Appl Environ Microbiol. 1999; 65(7):2961–2968. https://doi.org/10.1128/AEM.65.7.2961-2968.1999
  13. Mrozik A, Łabużek S, Piotrowska-Seget Z. Changes in fatty acid composition in Pseudomonas putida and Pseudomonas stutzeri during naphthalene degradation. Microbiological Research. 2005; 160:149–157. https://doi.org/10.1016/j.micres.2004.11.001
  14. Patent UA N124128 for a utility model. Pidgorsky VS, Nogina TM, Dumanskaya TU, Khomenko LA. [The strain of actinobacteria Rhodococcus aetherivorans is a phenol destructor]. Publ. 03.26.2018 Bull. N6. Ukrainian.
  15. Shumkova ES, Solyanikova IP, Plotnikova EG, Golovleva LA. Phenol degradation by Rhodococcus opacus strain 1G. Appl Biochem Microbiol. 2009; 45(1):43–49. https://doi.org/10.1134/S0003683809010086
  16. Larkin MJ, Kulakov LA, Allen CC. Biodegradation and Rhodococcus: masters of catabolic versatility. Curr Opin Biotechnol. 2005; 16:282–290. https://doi.org/10.1016/j.copbio.2005.04.007
  17. Martínková L, Uhnáková B, Pátek M, Nesvera J, Kren V. Biodegradation potential of the genus Rhodococcus. Environ Int. 2009; 35(1):162–177. https://doi.org/10.1016/j.envint.2008.07.018
  18. Goodfellow M, Jones AL, Maldonado LA, Salanitro J. Rhodococcus aetherivorans sp. nov., a new species that contains methyl t-butyl etherdegrading actinomycetes. Syst Appl Microbiol. 2004; 27(1):61–65. https://doi.org/10.1078/0723-2020-00254
  19. Tajima T, Hayashida N, Matsumura R, Omura A, Nakashimada Y, Kato J. Isolation and characterization of tetrahydrofuran-degrading Rhodococcus aetherivorans strain M8. Process Biochem. 2012; 47(11):1665–1669. https://doi.org/10.1016/j.procbio.2011.08.009
  20. Inoue D, Tsunoda T, Yamamoto N, Ike M, Sei K. 1,4-Dioxane degradation characteristics of Rhodococcus aetherivorans JCM 14343. Biodegradation. 2018; 29(3):301–310. https://doi.org/10.1007/s10532-018-9832-2
  21. Auffret M, Labbé D, Thouand G, Greer CW, Fayolle-Guichard F. Degradation of a mixture of hydrocarbons, gasoline, and diesel oil additives by Rhodococcus aetherivorans and Rhodococcus wratislaviensis. Appl Environ Microbiol. 2009; 75(24):7774–7782. https://doi.org/10.1128/AEM.01117-09
  22. Frascari D, Pinelli D, Nocentini M, Fedi S, Pii Y, Zannoni D. Chloroform degradation by butane-grown cells of Rhodococcus aetherivorans BCP1. Appl Microbiol Biotechnol. 2006; 73:421–428. https://doi.org/10.1007/s00253-006-0433-3
  23. Garba L, Ali MSM, Oslan SN, Abd Rahman RNZR. Review on fatty acid desaturases and their roles in temperature acclimatisation. J Appl Sci. 2017; 17(6):282–295. https://doi.org/10.3923/jas.2017.282.295
  24. Tsitko IV, Zaitsev GM, Lobanok AG, Salkinoja-Salonen MS. Effect of aromatic compounds on cellular fatty acid composition of Rhodococcus opacus. Appl Environ Microbiol. 1999; 65(2):853–855. https://doi.org/10.1128/AEM.65.2.853-855.1999
  25. Lindström F, Thurnhofer S, Vetter W, Gröbner G. Impact on lipid membrane organization by free branched-chain fatty acids. Phys Chem Chem Phys. 2006; 8:4792–4797. https://doi.org/10.1039/B607460J
  26. Ascenzi JM, Vestal JR. Regulation of fatty acid biosynthesis by hydrocarbon substrates in Mycobacterium convolutum. J Bacteriol. 1979; 137(1):384–390. https://doi.org/10.1128/jb.137.1.384-390.1979
  27. King DH, Perry JJ. The origin of fatty acids inthe hydrocarbon-utilizing microorganism Mycobacterium vaccae. Can J Microbiol. 1975; 21(1):85–89. https://doi.org/10.1139/m75-012
  28. Atakishieva YAYU, Ismayilova LM. [Lipogenesis of a hydrocarbon-degrading bacterium Rhodococcus erythropolis GH]. In: Materials of the international scientific-practical conference biotechnology of microorganisms; 2019 Nov 27–29; Minsk, Belarus. Minsk: Ecoperspectiva, 2019. p. 24–7. Russian.
  29. Murínová S, Dercová K. Response mechanisms of bacterial degraders to environmental contaminants on the level of cell walls and cytoplasmic membrane. Int J Microbiol. 2014; 1–16. https://doi.org/10.1155/2014/873081