A Pseudomonas aeruginosa Strain Ikw1 Produces an Unusual Polymeric Surface-Active Compound in Waste Frying Oil-Minimal Medium

A Pseudomonas aeruginosa Strain Ikw1 Produces an Unusual Polymeric Surface-Active Compound in Waste Frying Oil-Minimal Medium

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Author(s): Maurice George Ekpenyong, Sylvester Peter Antai, Atim David Asitok

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DOI: 10.18483/ijSci.1064 283 780 108-123 Volume 5 - Jun 2016


A Gram-negative bacterium isolated from the sub-surface water of Ikang River, Niger Delta region, Nigeria produced an unusual biosurfactant in waste frying oil-minimal medium. Cultural and biochemical characterizations as well as 16S rRNA sequencing identified the bacterium as a strain of Pseudomonas aeruginosa with 100% sequence homology with Pseudomonas aeruginosa strain HNYM41. Biochemical characterizations, thin layer chromatography (TLC), high performance liquid chromatography (HPLC) and Fourier transform-infrared (FT-IR) spectrometry identified the active compound as a glycolipopeptide (peptidoglycolipid) composed of 40.36% carbohydrates, 20.16% proteins and 34.56% lipids. The biosurfactant reduced surface tension of water from 72.00 to 24.62 dynes/cm at a critical micelle concentration (CMC) of 20.80 mg/L indicating excellent effectiveness and efficiency properties. Commendable oil-washing property (79.92% oil recovery) with an elution rate of 0.68 mL/min at 70°C, foaming and foam stability, excellent emulsification activity in kerosene, crude oil and palm oil and a significant (P = 0.000; R2 = 0.9901) oil solubilization property indicate excellent oil recovery, detergency and remediation potentials of the biosurfactant. Oil displacement, emulsifying and antimicrobial activities of the compound were relatively stable at relevant temperatures, pH and NaCl levels suggesting suitability for applications in hydrophobic compound remediation, emulsion stabilization and preservation of formulations.


Pseudomonas aeruginosa strain IKW1, glycolipopeptide, enhanced oil recovery, emulsification, stability


  1. Al-Araji, L., Rahman, R.N.Z.R.A., Basri, M. and Salleh, A.B. (2007) Microbial surfactants, Asian Pacific Journal of Molecular Biology and Biotechnology, 15, 99-105.
  2. Banat, I.M., Franzetti, A., Gandolfi, I., Bestetti, G., Martinotti, M.G., Fracchia, L., Smyth, T.J. and Marchant, R. (2010) Microbial biosurfactants production, applications and future potential, Applied Microbiology and Biotechnology, 87, 427- 444. http://dx.doi.org/10.1007/s00253-010-2589-0
  3. Pacwa-Plociniczak, M., Plaza, G.A., Piotrowska-Seget, Z. and Cameotra S.S. (2011) Environmental applications of biosurfactants: recent advances. International Journal of Molecular Sciences, 12, 633-654. http://dx.doi.org/10.3390/ijms12010633
  4. Sachdev, D.P. and Cameotra, S.S. (2013) Biosurfactants in agriculture. Applied Microbiology and Biotechnology, 97, 1005-1016. http://dx.doi.org/10.1007/s00253-012-4641-8
  5. Lawniczak, L., Marecik, R. and Chrzanowski, L. (2013) Contribution of biosurfactants to natural or induced bioremediation. Applied Microbiology and Biotechnology, 97, 2327-2339. http://dx.doi.org/10.1007/s00253-013-4740-1
  6. Morikawa, M., Daido, H., Takao, T., Murata, S., Shimonishi, Y. and Imanaka, T. (1993) A new lipopeptide biosurfactant produced by Arthrobacter sp. Strain MIS38. Journal of Bacteriology, 175, 6459-6466.
  7. Desai, J.D. and Banat, I.M. (1997). Microbial production of surfactants and their commercial potential, Microbiology and Molecular Biology Reviews, 61, 47-64.
  8. Thavasi, R.S., Jayalakshmi, L., Balasubramaian, T. Banat I M. (2007) Biosurfactants production by Corynebacterium kutscheri from waste motor lubricant oil and peanut oil cake. Letters in Applied Microbiology, 45, 686-691. http://dx.doi.org/10.1111/j.1472-765X.2007.02256.x
  9. Banat, I.M., Makkar, R.S. and Cameotra, S.S. (2000) Potential commercial applications of microbial surfactants. Applied Microbiology and Biotechnology, 53, 495-508. http://dx.doi.org/10.1007/s002530051648
  10. Mulligan, C.N. (2005) Environmental applications for biosurfactants. Environmental Pollution, 133, 183-198. http://dx.doi.org/10.1016/j.envpol.2004.06.009
  11. Franzetti, A., Tamburini, E. and Banat, I.M. (2010) Applications of biological surface active compounds in remediation technologies. Advances in Experimental Medicine and Biology, 672, 121-34. http://dx.doi.org/10.1007/978-1-4419-5979-9_9
  12. Lima, T.M., Procópio, L.C., Brandão, F.D, Leão, B.A., Tótola, M.R. and A.C. Borges, A.C. (2011) Evaluation of bacterial surfactant toxicity towards petroleum-degrading microorganisms. Bioresource Technology, 102, 2957–2964. http://dx.doi.org/10.1016/j.biortech.2010.09.109
  13. Bodour, A.A., Drecs, K.P. and Maier, R.M. (2003) Distribution of biosurfactant-producing bacteria in undisturbed and contaminated arid southwestern soils. Applied and Environmental Microbiology, 69, 3280-3287. http://dx.doi.org/10.1128/AEM.69.6.3280-3287.2003
  14. Satpute, S.K., Bhawsar, B.D., Dhakephalkar, P.K. and Chopade, B.A. (2008) Assessment of different screening methods for selecting biosurfactant producing marine bacteria. Indian Journal of Marine Sciences, 37, 243-250.
  15. Daverey, A., Pakshirajan, K. and Sangeetha, P. (2009) Sophorolipids production by Candida bombicola using synthetic dairy wastewater, International Journal of Environmental Science and Engineering, 1, 173-175.
  16. Savergave, L.S., Gadre, R.V., Vaidya, B.K. and Narayanan, K. (2011) Strain improvement and statistical media optimization for enhanced erythritol production with minimal by-products from Candida magnoliae mutant R23. Biochemical Engineering Journal, 55, 92-100. http://dx.doi.org/10.1016/j.bej.2011.03.009
  17. Kiran, G.S., Selvin, J., Manilal, A. and Sujith, S. (2011) Biosurfactants as green stabilizers for the biological synthesis of nanoparticles. Critical Reviews in Biotechnology, 31, 354-364. http://dx.doi.org/10.3109/07388551.2010.539971
  18. Mabrouk, M.E.M., Youssif, E.M. and Sabry, S.A. (2014) Biosurfcatant production by a newly isolated soft coral-associated marine Bacillus sp.E34: Statistical optimization and characterization, Life Science Journal, 11, 756-768.
  19. Nitschke, M., Ferraz, C. and Pastore, G.M. (2004) Selection of microorganisms for biosurfactant production using agro-industrial wastes. Brazilian Journal of Microbiology, 35, 81-85. http://dx.doi.org/10.1590/S1517-83822004000100013
  20. Rodrigues, L.R., Teixeira, J.A. and Oliveira, R. (2006) Low-cost fermentative medium for biosurfactant production by probiotic bacteria. Biochemical Engineering Journal, 32, 135-142. http://dx.doi.org/10.1016/j.bej.2006.09.012
  21. Thavasi, R.S., Subramanyam Nambaru, V.R.M., Jayalakshmi, S., Balasubramanian, T. and Banat, I.M. (2011) Biosurfactant production by Pseudomonas aeruginosa from renewable resources. Indian Journal of Microbiology, 51, 30-36. http://dx.doi.org/10.1007/s12088-011-0076-7
  22. Asitok, A.D. and Antai, S.P. (2006) Petroleum hydrocarbon utilization and biosurfactant production by Pseudomonas and Bacillus species. Nigerian Journal of Microbiology, 20, 824-831.
  23. Lindahl, M., Faris, A., Wadstrom, T. and Hjerten, S. (1981) A new test based on salting out to measure relative surface hydrophobicity of bacterial cells. Biochemistry and Biophysics Acta, 677, 471- 476. http://dx.doi.org/10.1016/0304-4165(81)90261-0
  24. Queiroga, C.L., Nascimento, L.R. and Serra, G.E. (2003) Evaluation of paraffins biodegradation and biosurfactant production by Bacillus subtilis in the presence of crude oil. Brazilian Journal Microbiology, 34, 321-324. http://dx.doi.org/10.1590/S1517-83822003000400006
  25. Dubois, M., Gilies, K.A., Hamilton, J.K., Rebers, P.A. and Smith, F. (1956) Colorimetric method for determination of sugar and related substances, Analytical Chemistry, 28, 350-356. http://dx.doi.org/10.1021/ac60111a017
  26. Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Analytical Biochemistry, 72, 248-254. http://dx.doi.org/10.1016/0003-2697(76)90527-3
  27. Folch, J.M., Lees, M. and Stanly, H.S. (1956) A simple method for the isolation and quantification of total lipids from animal tissues. Journal of Biological Chemistry, 226, 497–509
  28. Fernandes P A V, Renata de Arruda I, Botelho dos Santos A F A, Albertina de Araujo A, Maior A M S and Ximenes, E.A. (2007) Antimicrobial activity of surfactants produced by Bacillus subtilis R14 against multidrug-resistant bacteria. Brazilian Journal of Microbiology, 38, 704-709. http://dx.doi.org/10.1590/S1517-83822007000400022
  29. Berry, C., Dilantha Fernando, W.G., Loewen, P.C. and de Kievit, T.R. (2010) Lipopeptides are essential for Pseudomonas sp. DF41 biocontrol of Sclerotinia sclerotiorum. Biological Control, 55, 211-218. http://dx.doi.org/10.1016/j.biocontrol.2010.09.011
  30. Kiran, G.S., Thomas, T.A., Selvin, J., Sabarathnam, B. and Lipton, A.P. (2010) Optimization and characterization of a new lipopeptide biosurfactant produced by marine Brevibacterium aureum MSA13 in solid state culture. Bioresource Technology, 101, 2389-2396. http://dx.doi.org/10.1016/j.biortech.2009.11.023
  31. Whang, L.M., Liu, P.W.G., Ma, C.C. and Cheng, S.S. (2008) Application of biosurfactants; rhamnolipid and surfactin, for enhanced biodegradation of diesel-contaminated water and soil, Journal of Hazardous Materials, 151 155-163. http://dx.doi.org/10.1016/j.jhazmat.2007.05.063
  32. Chen, Y., Yang, C., Chang, M., Ciou, Y. and Huang, Y. (2010) Foam properties and detergent abilities of the saponins Camellia oleifera, International Journal of Molecular Sciences, 11, 4417-4425. http://dx.doi.org/10.3390/ijms11114417
  33. Abouseoud, M., Maachi, R., Amrane, A., Boudergua, S. and Nabi, A. (2008) Evaluation of different carbon and nitrogen sources in production of biosurfactant by Pseudomonas fluorescens, Desalination, 223, 143-151. http://dx.doi.org/10.1016/j.desal.2007.01.198
  34. Makkar, R.S., Cameotra, S.S. and Banat, I.M. (2011) Advances in utilization of renewable substrates for biosurfactant production, AMB Express, 1, 5. http://dx.doi.org/10.1186/2191-0855-1-5
  35. Karanth, N.G.K., Deo, P.G. and Veenanadig, N.K. (1999) Microbial biosurfactants and their importance, Current Science, 77, 116-126.
  36. Onbasli, D. and Aslim, B. (2009) Biosurfactant production in sugar beet molasses by some Pseudomonas spp. Journal of Environmental Biology, 30, 161-163
  37. Koronelli, T.V., Komarova, T.I and Denisov, luV. (1983) Chemical composition and role of Pseudomonas aeruginosa peptidoglycolipid in hydrocarbon assimilation, Mikrobiologiia, 52, 767-770
  38. Ilori, M.O. and Amund, D.I. (2001) Production of a peptidoglycolipid bioemulsifier from Pseudomonas aeruginosa grown on hydrocarbon. Z Naturforsch C, 56, 547-552. http://dx.doi.org/10.1515/znc-2001-7-812
  39. Hisatsuka, K., Nakahara, T., Minoda, Y. and Yamada, K. (1977). Formation of protein-like activator for n-alkane oxidation and its properties, Agricultural and Biological Chemistry, 41, 445-450. http://dx.doi.org/10.1271/bbb1961.41.445
  40. http://dx.doi.org/10.1080/00021369.1977.10862518
  41. Cohen, Y. (2002) Bioremediation of oil by marine microbial mats. International Microbiology, 5, 189-193. http://dx.doi.org/10.1007/s10123-002-0089-5
  42. Sekhon, K.K.., Khanna, S. and Cameotra, S.S. (2012) Biosurfactant production and potential correlation with esterase activity. Journal of Petroleum and Environmental Biotechnology, 3, 333-343.
  43. Gupta, R., Gupta, N. and Rathi, P. (2004) Bacterial lipases: an overview of production, purification and biochemical properties. Applied Microbiology and Biotechnology, 64, 763-781. http://dx.doi.org/10.1007/s00253-004-1568-8

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