Bioremediation of crude oil by Bacillus licheniformis in the presence of different concentration nanoparticles and produced biosurfactant

  • H. S. El-SheshtawyEmail author
  • W. Ahmed
Original Paper


This research work focuses on testing the bacterial strain Bacillus licheniformis for the bioremediation capacity of the crude oil. A biosurfactant and two different nanoparticles with different concentrations (0.05, 0.1, 0.2 g/100 ml) were applied separately to enhance the biodegradation process. The optimum biodegradation of crude oil was demonstrated at 60% of microcosms containing biosurfactant and nanoparticles after 7 days. The bacterial strain is highly potential to consume the total paraffins (iso- and n-paraffins) in crude oil samples. Accordingly, the best biodegradation of total paraffins was observed in microcosms containing (0.2 g) of Fe2O3, Zn5(OH)8Cl2 (nps) and biosurfactant separately. Additionally, the consumption of specific member rings of polyaromatics depends on the type and the concentration of nanoparticles. Thus, this bacterial strain was considered as a good candidate to be applied in the bioremediation process of petroleum-contaminated sites using biosurfactant and specific concentration of (Fe2O3 and Zn5OH8Cl2) nanoparticles.


Bacillus licheniformis Bioremediation Biosurfactants Nanoparticles Crude oil 



This work was sponsored by the Egyptian Petroleum Research Institute (EPRI), Cairo, Egypt. The author would like to extend thanks to all members of the Application of Biotechnology in the Field of Petroleum Industry Lab, Process, Design and Development Department (EPRI).


  1. American Society for Testing and Materials (1999) Standard test method for separation of representative aromatics and nonaromatics fractions of high boiling oils by elution chromatography. D-2549:246–251Google Scholar
  2. American Society for Testing and Materials (2001a) Standard practice for density, relative density (specific gravity), or API gravity of crude petroleum and liquid petroleum products by hydrometer method, Annual Book of ASTM Standards (petroleum products and lubricants). D-1298:484–488Google Scholar
  3. American Society for Testing and Materials (2001b) Standard test method for pour point of petroleum oil, Annual Book of ASTM Standards (petroleum products and lubricants). D-97:57–67Google Scholar
  4. American Society for Testing and Materials (2001c) Standard test method for flash point by Pensky-Martens closed tester, Annual Book of ASTM Standards (petroleum products and lubricants). D-93:28–40Google Scholar
  5. American Society for Testing and Materials (2001d) Standard test method for kinematic viscosity of transparent and opaque liquids and the calculation of dynamic viscosity, Annual Book of ASTM Standards (petroleum products and lubricants). D-445:168–173Google Scholar
  6. American Society for Testing and Materials (2001e) Standard test method for sulfur in petroleum and petroleum products by energy-dispersive X-ray fluorescence spectrometry, Annual Book of ASTM Standards (petroleum products and lubricants). D-4294:348–350Google Scholar
  7. Atlas RM, Atlas MC (1991) Biodegradation of oil and bioremediation of oil spills. Curr Opin Biotechnol 2:440–443CrossRefGoogle Scholar
  8. Azim A, Davood Z, Ali F, Mohammad RM, Dariush N, Shahram T, Majid M, Nasim B (2009) Synthesis and characterization of gold nanoparticles by tryptophane. Am J Appl Sci 6:691–695CrossRefGoogle Scholar
  9. Banat IM, Makkar RS, Cameotra SS (2000) Microbial production of surfactants and their commercial potential. Appl Microbiol Biotechnol 53:495–508CrossRefGoogle Scholar
  10. Chang FW, Roselin LS, Ou TC (2008) Hydrogen production by partial oxidation of methanol over bimetallic Au-Ru/Fe2O3 catalysts. Appl Cat A Gen 334:147–155CrossRefGoogle Scholar
  11. Cooper DG, Goldenberg BG (1987) Surface active agents from two Bacillus species. Appl Environ Microbiol 53(2):224–229Google Scholar
  12. Desai JD, Banat IM (1997) Microbial production of surfactants and their commercial potential. Microbiol Mol Rev 61:47–64Google Scholar
  13. Deziel E, Paquette G, Villemur R, Lepine F, Bisaillon J (1996) Biosurfactant production by a soil Pseudomonas strain growing on polycyclic aromatic hydrocarbons. Appl Environ Microbiol 62:1908–1912Google Scholar
  14. El-Bastawissy AM, Moustafa YM, Zakaria AI, Sidky NM, El-Sheshtawy HS (2004) Growth kinetics and biodegradation potentials of five pure petroleum samples utilizing bacteria on different hydrocarbon substrates. Egy J Appl Sci 19(3B):583–593Google Scholar
  15. El-Sheshtawy HS, Doheim MM (2014) Selection of Pseudomonas aeruginosa for biosurfactant production and studies of its antimicrobial activity. Egy J Petr 23:1–6CrossRefGoogle Scholar
  16. El-Sheshtawy HS, Khalil NM, Ahmed W, Abdallah RI (2014) Monitoring of oil pollution at Gemsa Bay and bioremediation capacity of bacterial isolates with biosurfactants and nanoparticles. Marine Poll Bull 87:191–200CrossRefGoogle Scholar
  17. Guha S, Jaffe P (1996) Biodegradation kinetics of phenanthrene partitioned into the micellar phase of nonionic surfactants. Environ Sci Technol 30:605–611CrossRefGoogle Scholar
  18. Haghighat S, Akhavan A, Assadi MM, Pasdar SH (2008) Ability of indigenous Bacillus licheniformis and Bacillus subtilis in microbial EOR. Int J Environ Sci Tech 5(3):385–390CrossRefGoogle Scholar
  19. Institute of Petroleum, IP (1995) Standard methods for analysis and testing of petroleum and related products. Characterization of pollutants-High resolution gas chromatography method 318:1–4Google Scholar
  20. Lai B, Khanna S (1996) Degradation of crude oil by Acinetobacter calcoaceticus and Alcaligenes odorans. J Appl Bacteriol 81:355–362Google Scholar
  21. Leahy JG, Colwell RR (1990) Microbial degradation of hydrocarbons in the environment. Microbiol Rev 54:305–309Google Scholar
  22. Litt G, Almquist G (2009) An investigation of CuO/Fe2O3 catalysts for the gas-phase oxidation of ethanol. Appl Cat B Environ 90:10–17CrossRefGoogle Scholar
  23. Maneerat S (2005) Biosurfactants from marine microorganisms. J Sci Technol 27(6):1263–1272Google Scholar
  24. Mostafa YM, Abdalh RI, Abd ElNaby IM (1997) Levels of hydrocarbons and source identification of pollutants contaminating Ismailya Canal. J Edu 22:705–716Google Scholar
  25. Nezahat B, Nebahat D, Dilhan MK (2009) Conversion of biomass to fuel: transesterification of vegetable oil to biodiesel using KF loaded nano-g-Al2O3 as catalyst. Appl Catal B 89:590–596CrossRefGoogle Scholar
  26. Nitschke M, Pastore G (2006) Production and properties of a surfactant obtained from Bacillus subtilis grown on cassava wastewater. Biores Technol 97(2):336–344CrossRefGoogle Scholar
  27. Paudyn K, Rutter A, Rowe RK, Poland JS (2008) Remediation of hydrocarbon contaminated soils in the Canadian arctic by landfarming. Cold Reg Sci Technol 53:102–114CrossRefGoogle Scholar
  28. Raza ZA, Rehman A, Khan MS, Khalid ZM (2007) Improved production of biosurfactant by a Pseudomonas aeruginosa mutant using vegetable oil refinery wastes. Biodegrad 18:115–121CrossRefGoogle Scholar
  29. Sheppard JD, Jumarie C, Cooper DG, Laprade R (1991) Ionic channels induced by surfactin in planar lipid bilayer membranes. Biochem Biophys Acta 1064:13–23CrossRefGoogle Scholar
  30. Shin KH, Cha DK (2008) Microbial reduction of nitrate in the presence of nanoscale zero-valent iron. Chemo 72:257–262CrossRefGoogle Scholar
  31. Spain JC, VanVeld PA (1983) Adaptation of natural microbial communities to degradation of xenobiotic compounds: effects of concentration, exposure time, inoculum and chemical structure. Appl Environ Microbiol 45:428–435Google Scholar
  32. Walter MV, Nelson EC, Firmstone G, Martin DG, Clayton MJ, Clayton S, Simpson S (1997) Surfactant enhances biodegradation of hydrocarbons: microcosm and field study. J Soil Contam A 6:61–77CrossRefGoogle Scholar
  33. Yakimov MM, Timmis KN, Wray V, Fredrickson HL (1995) Characterization of a new lipopeptide surfactant produced by thermotolerant and halotolerant subsurface Bacillus licheniformis BAS50. Appl Environ Microbiol 61:1706–1713Google Scholar
  34. Zhang X, Song Yan S, Tyagi R, Surampall R (2011) Synthesis of nanoparticles by microorganisms and their application in enhancing microbiological reaction rates. Rev Chemo 82:489–494Google Scholar

Copyright information

© Islamic Azad University (IAU) 2017

Authors and Affiliations

  1. 1.Egyptian Petroleum Research Institute (EPRI)CairoEgypt

Personalised recommendations