Simultaneous Biosurfactant Production and Hydrocarbon Biodegradation by the Resident Aerobic Bacterial Flora of Oil Production Skimmer Pit at Elevated Temperature and Saline Conditions

Chuma C. Okoro, Akhil Agrawal, Cameron Callbeck


Six aerobic thermo- and halotolerant bacterial isolates from an oil production skimmer pit were evaluated for their ability to produce biosurfactants and degrade petroleum hydrocarbons simultaneously under elevated temperature and saline conditions. Phylogenetic analysis using 16S rRNA sequencing revealed that the six bacterial isolates used in the study (SKP-1, SKP-2, SKP-3, SKP-4, SKP-5 and SKP-6) were most homologous to the gamma-proteobacteria Pseudomonas sp. VS-1, Pseudomonas aeruginosa strain S2QPS8, Serratia marcescens strain A4, Pseudomonas stutzeri, Pseudomonas stutzeri strain RA10 and Pseudomonas stutzeri strain BOD-3 respectively. Using previously sterilized skimmer pit sample as the sole nutrient, carbon and energy sources and at an elevated temperature of 450C and salinity (chloride) level of 6012 mg L-1, all the bacterial isolates in a mixed culture were able to grow, produce biosurfactants and degrade petroleum hydrocarbons simultaneously by removing about 92% of residual TPH in the skimmer pit within 2 weeks of exposure. This study suggests that in-situ bioremediation procedure using the resident aerobic bacterial flora of the skimmer pit that are thermotolerant and halotolerant can be developed to degrade the petroleum hydrocarbon contaminants in-situ. This bioremediation procedure can be a more attractive and cost effective option than the costly thermal treatment option that is currently in operation in the industry.



Bioremediation; Biosurfactant; Skimmer Pit; Thermotolerant Bacteria; Halotolerant Bacteria, Petroleum Hydrocarbon


Agrawal, A.; Park, H.S.; Nathoo, S.; Gieg, L.M.; Jack, T.R.; Miner, K.; Ertmoed, R.; Benko, A. and Voordouw, G. 2012. Toluene depletion in produced oil contributes to souring control in a field subjected to nitrate injection. Environmental Science and Technology 46: 1285-1292.

Anyanwu, C.U.; Obi, S.K.C. and Okolo, B.N. 2011. Lipopeptide bio-surfactant production by Serratia marcescens NSK-1 Strain isolated from petroleum-contaminated soil. Journal of Applied Sciences and Research 7(1): 79-87.

Atlas, R.M. 1981. Microbial degradation of petroleum hydrocarbons; an environmental perspective. Microbiological Reviews 45 (1): 180-209.

Bovdoloi, N.K. and Konwar, B.K. 2009. Bacterial biosurfactants in enhancing solubility and metabolism of petroleum hydro-carbons. Journal of Hazardous Materials 170 (1): 495-505.

Bradford, M.M. 1976. A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of dye binding. Analytical Biochemistry 72: 248-254

Cameotra, S.S. and Makkar, R. 1998. Synthesis of biosurfactants in extreme conditions. Applied Microbiology and Biotechnology 50: 520-529.

Desai, J.D. and Banat, I.M. 1997. Microbial production of surfactants and their commercial potential. Microbiology and Molecular Biology Reviews 61(1): 47-64.

Eaton, A.D.; Clesceri, L.S. and Greenberg, A.E. 1995. Standard Methods for Examination of Water and Wastewater. 19th edition. United Books Press, Batimore, MD. 1126 pages.

Guerra-Santos, L.H.; Kappedi, O. and Fiechter, A. 1984. Pseudo-monas aeruginosa biosurfactant production in a continuous culture with glucose as a carbon source. Applied and Environmental Microbiology 48: 301-305.

Huang, Yi-Chen; Wei, Yu-Hong; Jo, Shu-Chang and Chin,Chi-Lai. 2009. Biosurfactant enhanced removal of total petroleum hydrocarbon from contaminated soil. Journal of Hazardous Materials 167(1-3): 609-614.

Ijah, U. 1998. Studies on relative capabilities of bacterial and yeast isolates from tropical soil in degrading crude oil. Waste Management 18: 293-299.

Inakolu, S.; Hung, H. and Shieve, G.S. 2004. Biosurfactant enhancement of microbial degradation of various structural classes of hydrocarbon in mixed waste systems. Environmental Engineering and Science 24(4): 463-469.

Javis, F.G. and Johnson, M.J. 1949. A glycolipid produced by Pseudomonas aeruginosa. Journal of American Chemical Society 71: 4124-4126.

Kates, M. 1972. Techniques in Lipidology. North-Holland Publishing, New York. 610 pages.

Kumar, M.; Leon, V.L.; Materano, A. D.; Ilzins, O.A., and Luis, L. 2008. Biosurfactant production and hydrocarbon degradation by halotolerant and thermotolerant Pseudomonas sp. World Journal of Microbiology and Biotechnology 24(7):1047-1057.

Lugowski, A. J.; Palamter, G.A.;, Boose, T.R. and Merriman, J.E. 1997. Biodegradation process for detoxifying liquid streams. Patent US 565 6169. August 12, 1997.

Margesin, R. and Schinner, F. 2001. Biodegradation and Bioreme-diation of hydrocarbons in extreme environments. Applied Microbiology and Biotechnology 56: 650-663.

Mills, A.L.; Breuil, C. and Colwell, R.R. 1978. Enumeration of petroleum degrading marine and estuarine microorganisms by most probable number method. Canadian Journal of Microbiology 24: 552-557.

Mnif, S.; Chankha, M.; Labat, M. and Sayadi, S. 2011. Simultaneous hydrocarbon biodegradation and biosurfactant production by oil field selected bacteria. Journal of Applied Microbiology 111: 523-536.

Muller, R.; Antranikian, G.; Malony, S. and Shamp, R. 1998. Thermophilic degradation of environmental pollutants. Pages 155-169, In: Antranikian, G. (Editor) Biotechnology of Extre-mophiles. Advances in Biochemical Engineering and Bio-technology, Volume 61. Springler, Berlin.

Niehaus, F.; Bertoldo, C.; Kahler, M. and Antranikian, G. 1999. Extremophiles as a source of novel enzymes for industrial application. Applied Microbiology and Biotechnology 51: 711-729.

Okoro, C.C. 1999. Microbial Degradation of Hydrocarbons in Produced Water From Crude Oil Production Operations in Escravos Tank Farm. Ph.D. Thesis. University of Lagos, Nigeria. 269 pages.

Okoro, C.C. 2009. Biosurfactant-enhanced remediation of hydro-carbon contaminated mangrove swamp. International Journal of Biological and Chemical Sciences 3(1): 63-74.

Okoro, C.C. 2010. Enhanced bioremediation of hydrocarbon contaminated mangrove swamp in the Nigerian oil-rich Niger-Delta using sea water microbial inocula amended with crude biosurfactants and micronutrients. Journal of Nature and Science 8 (8): 195-206.

Roldan-Carrillo, T.; Martinez-Garcia, X.; Zapta-Penaso, I.; Castrorena-Cortes, G.; Reyes-Avila, J.; Mayol-Castino, M. and Olguin-Lora, P. 2011. Evaluation of the effect of nutrient ratios on biosurfactant production by Serratia marcescens using a boxbehuken design. Colloids and Surfaces B: Biointerfaces 86: 284-389.

Rosenberg, E.; Zuckerberg, A.; Rubinovitz, C. and Gutnick, D.L. 1979. Emulsifier Arthrobacter, RAG-1. Isolation and emul-sifying properties. Applied Environmental Microbiology 37: 402-408.

Rosenberg, E.; Rubinovitz, C.; Gottlieb, A.; Rosenhak, S. and Ron, E.Z. 1988. Production of biodispersan by Acinetobacter calcoaceticus A2. Applied Environmental Microbiology 54: 317-322.

Song, R.; Hua, Z.; Li, H. and Chen, J. 2006. Biodegradation of petroleum hydrocarbons by two Pseudomonas aeruginosa strains with different uptake modes. Journal of Environmental Science and Health 41(4): 733-748

Spiro, R.G. 1966. Analysis of sugars found in glycoproteins. Pages 7-9, In: Colowick, S.P. and Kaplan, N. (Editors) Enzymology, Volume 8. Academic Press, New York.

Umeji, A.A.; Onwura, I.N. and Anyanwu, C.U. 2010. Isolation and characterization of biosurfactants produced by diculture of Pseudomonas sp. and Azotobacter vinelandii. Nigerian Journal of Biochemistry and Molecular Biology 25(2): 78-85.

Whyte, L.G.; Bourbonnieve, C.; Bellerose, C. and Green, C.W. 1999. Bioremediation assessment of hydrocarbon-contaminated soils from the high arctic. Bioremediation 5(3): 69-79.

Full Text: PDF


  • There are currently no refbacks.

Creative Commons License
This work is licensed under a Creative Commons Attribution 3.0 License.

COPYRIGHT of this Journal vests fully with the National Instional Institute of Ecology. Any commercial use of the content on this site in any form is legally prohibited.