AIMS Microbiology, 2017, 3(3): 467-482. doi: 10.3934/microbiol.2017.3.467

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Hydrocarbon degradation abilities of psychrotolerant Bacillus strains

1 Department of Bioengineering, Faculty of Engineering, Ege University, 35100, İzmir, Turkey
2 Department of Biotechnology, Graduate School of Natural and Applied Sciences, Ege University, 35100, İzmir, Turkey
3 Department of Environmental Engineering, Engineering Faculty, Aksaray University, Aksaray, Turkey
4 Department of Environmental Engineering, Engineering Faculty, Dokuz Eylül University, Buca, Kaynaklar Campus, 35160, İzmir, Turkey

Biodegradation requires identification of hydrocarbon degrading microbes and the investigation of psychrotolerant hydrocarbon degrading microbes is essential for successful biodegradation in cold seawater. In the present study, a total of 597 Bacillus isolates were screened to select psychrotolerant strains and 134 isolates were established as psychrotolerant on the basis of their ability to grow at 7 °C. Hydrocarbon degradation capacities of these 134 psychrotolerant isolate were initially investigated on agar medium containing different hydrocarbons (naphthalene, n-hexadecane, mineral oil) and 47 positive isolates were grown in broth medium containing hydrocarbons at 20 °C under static culture. Bacterial growth was estimated in terms of viable cell count (cfu ml–1). Isolates showing the best growth in static culture were further grown in presence of crude oil under shaking culture and viable cell count was observed between 8.3 × 105–7.4 × 108 cfu ml–1. In the final step, polycyclic aromatic hydrocarbon (PAH) (chrysene and naphthalene) degradation yield of two most potent isolates was determined by GC-MS along with the measurement of pH, biomass and emulsification activities. Results showed that isolates Ege B.6.2i and Ege B.1.4Ka have shown 60% and 36% chrysene degradation yield, respectively, while 33% and 55% naphthalene degradation yield, respectively, with emulsification activities ranges between 33–50%. These isolates can be used to remove hydrocarbon contamination from different environments, particularly in cold regions.
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1. Kaczorek E, Chrzanowski L, Pijanowska A, et al. (2008) Yeast and bacteria cell hydrophobicity and hydrocarbon biodegradation in the presence of natural surfactants: Rhamnolipides and saponins. Bioresource Technol 99: 4285–4291.    

2. Toledo FL, Calvo C, Rodelas B, et al. (2006) Selection and identification of bacteria isolated from waste crude oil with polycyclic aromatic hydrocarbons removal capacities. Syst Appl Microbiol 29: 244–252.    

3. Solomons TWG, Fryhle CB (2000) Organic Chemistry, 7 Eds., New York: Wiley, 1–30.

4. Li H, Liu YH, Luo N, et al. (2006) Biodegradation of benzene and its derivatives by a psychrotolerant and moderately haloalkaliphilic Planococcus sp. strain ZD22. Res Microbiol 157: 629–636.

5. Chen HY, Teng YG, Wang JS (2013) Source apportionment for sediment PAHs from the Daliao River (China) using an extended fit measurement mode of chemical mass balance model. Ecotox Environ Safe 88: 148–154.    

6. Sawadogo A, Otoidobiga HC, Nitiema LW, et al. (2016) Optimization of hydrocarbons biodegradation by bacterial strains isolated from wastewaters in Ouagadougou, Burkina Faso: case study of SAE 40/50 used oils and diesel. J Agric Chem Environ 5: 1–11.

7. Lin C, Gan L, Chen ZL (2010) Biodegradation of naphthalene by strain Bacillus fusiformis (BFN). J Hazard Mater 182: 771–777.    

8. Deng MC, Li J, Liang FR, et al. (2014) Isolation and characterization of a novel hydrocarbon-degrading bacterium Achromobacter sp. HZ01 from the crude oil-contaminated seawater at the Daya Bay, southern China. Mar Pollut Bull 83: 79–86.

9. Yuan SY, Shiung LC, Chang BV (2002) Biodegradation of polycyclic aromatic hydrocarbons by inoculated microorganisms in soil. B Environ Contam Tox 69: 66–73.    

10. Perelo LW (2010) Review: In situ and bioremediation of organic pollutants in aquatic sediments. J Hazard Mater 177: 81–89.    

11. Delille D, Pelletier E, Rodriguez-Blanco A, et al. (2009) Effects of nutrient and temperature on degradation of petroleum hydrocarbons in sub-Antarctic coastal seawater. Polar Biol 32: 1521–1528.    

12. Männistö MK, Häggblom MM (2006) Characterization of psychrotolerant heterotrophic bacteria from Finnish Lapland. Syst Appl Microbiol 29: 229–243.    

13. Giudice AL, Casella P, Carusa C, et al. (2010) Occurrence and characterization of psychrotolerant hydrocarbon-oxidizing bacteria from surface seawater along the Victoria Land coast. Polar Biol 33: 929–943.    

14. Lechner S, Mayr R, Francis KP, et al. (1998) Bacillus weihenstephanensis sp. nov. is a new psychrotolerant species of the Bacillus cereus group. Int J Syst Bacteriol 48: 1373–1382.

15. El-Rahman HAA, Fritze D, Spröer C, et al. (2002) Two novel psychrotolerant species, Bacillus psychrotolerants sp. nov. and Bacillus psychrodurans sp. navo., which contain ornithine in their cell walls. Int J Syst Evol Microbiol 52: 2117–2133.

16. Vazquez SC, Coria SH, Cormack WPM (2004) Extracellular proteases from eight psychrotolerant antarctic strains. Microbiol Res 159: 157–166.    

17. Demir İ, Demirbağ Z, Beldüz AO (2000) Isolation and characterization of phenanthrene decomposing Pseudomonas sp. Fresen Environ Bull 9: 9–16.

18. Nievas ML, Commendatore MG, Estevas JL, et al. (2005) Effect of pH modification on bilge waste biodegradation by a native microbial community. Int Biodeter Biodegr 56: 151–157.    

19. Helmke E, Weyland H (2004) Psychrophilic Versus psychrotolerant bacteria occurrence and significance in polar and temperate marine habitats. Cell Mol Biol 50: 553–561.

20. Luigi M, Gaetano DM, Vivia B, et al. (2007) Biodegradative potential and characterization of psychrotolerant polychlorinated biphenyl degrading marine bacteria isolated from a coastal station in the Terra Nova Bay (Ross Sea, Antarctica). Mar Pollut Bull 54: 1754–1761.    

21. Okoh A, Ajisebutu S, Babalola G, et al. (2001) Potential of Burkholderia cepacia RQ1 in the biodegradation of heavy crude oil. Int Microbiol 4: 83–87.

22. Cooper DG, Goldenberg BG (1987) Surface-active agents from two Bacillus species. Appl Environ Microbiol 53: 224–229.

23. Christova N, Tuleva B, Nikolova-Damyanova B (2004) Enhanced hydrocarbon biodegradation by newly isolated Bacillus subtilis strain. Z Naturforsch C 59: 205–208.

24. Sponza DT, Gok O (2010) Effect of rhamnolipid on the aerobic removal of polyaromatic hydrocarbons (PAHs) and COD components from petrochemical wastewater. Bioresource Technol 101: 914–924.    

25. Sponza DT, Gok O (2012) Aerobic biodegradation and inhibition kinetics of poly-aromatic hydrocarbons (PAHs) in a petrochemical industry wastewater in the presence of biosurfactants. J Chem Technol Biotechnol 87: 658–672.    

26. Russell NJ, Cowan DA (2006) Handling of psychrophilic micro-organisms. Meth Microbiol 35: 371–374.

27. Deming JW (2009) Extremophiles: Cold Environments, In: Encyclopedia of Microbiology, 3 Eds., USA: Schaechter, 147–157.

28. Ruberto LAM, Vazquez SC, Cormack WPM (2008) Bacteriology of Extremely Cold Soils Exposed to Hydrocarbon Pollution, In: Dion P, Nautiyal CS, Ediors, Microbiology of Extrem Soils, USA: Springer, 247–274.

29. Brakstad OG, Booth AM, Fakness L (2010) Microbial Degradation of Petroleum Compounds in Cold Marine Water and Ice, In: Bej AK, Aislabie J, Atlas RM, Ediors, Polar Microbiology: the Ecology, Biodiversity and Bioremediation Potential of Microorganisms in Extremely Cold Environments, USA: CRC Press, 231–247.

30. Zhan Z, Gai L, Hou Z, et al. (2010) Characterization and biotechnological potential of petroleum-degrading bacteria isolated from oil-contaminated soils. Bioresource Technol 101: 8452–8456.    

31. Connaughton S, Collins G, O'Flaherty V (2006) Psycrophilic and mesophilic anaerobic digestion of brewery effluent; a comparative study. Water Res 40: 2503–2510.    

32. Rüger HJ, Fritze D, Spröer C (2000) New psychrophilic and psychrotolerant Bacillus marinus strains from tropical and polar deep-sea sediments and emended description of the species. Int J Syst Evol Microbiol 50: 1305–1313.    

33. Trejo-Hernandez MR, Ortiz A, Okoh AI, et al. (2007) Biodegradation of heavy crude oil Maya using spent compost and sugar cane bagasse wastes. Chemosphere 68: 848–855.    

34. Haddadin MSY, Arqoub AAA, Reesh IA, et al. (2009) Kinetics of hydrocarbon extraction from oil shale using biosurfactant producing bacteria. ‎Energy Convers Manage 50: 983–990.    

35. Dhote M, Juwarkar A, Kumar A, et al. (2010) Biodegration of chrysene by the bacterial strains isolated from oily sludge. World J Microbiol Biotechnol 26: 329–335.    

36. Al-Saleh ES, Drobiova H, Obuekwe C (2009) Predominant culturable crude oil-degrading bateria in the coast of Kuwait. Int Biodeter Biodegr 63: 400–406.    

37. Toledo FL, Gonzalez-Lopez J, Calvo C (2008) Production of emulsifiers by Bacillus subtilis, Alcaligenes faecalis and Enterobacter species in liquid culture Bioresourse Technol 99: 8470–8475.

38. Pathak H, Kantharia D, Malpani A, et al. (2009) Naphthalene degradation by Pseudomonas sp. HOB1: In vitro studies and assessment of naphthalene degradation efficiency in simulated microcosms. J Hazard Mater 166: 1466–1473.

39. Larter S, Wilhelms A, Head I, et al. (2003) The controls on the composition of biodegraded oils in the deep subsurface biodegradation rates in petroleum reservoirs. Org Geochem 34: 601–613.

40. Das N, Chandran P (2010) Bioremediation of petroleum hydrocarbon contaminants-an overview. Biotechnol Res Int 2011: 1–13.

41. Das K, Mukherjee AK (2007) Crude petroleum-oil biodegradation efficiency of Bacillus subtilis and Pseudomonas aeruginosa strains isolated from a petroleum-oil contaminated soil from North-East India. Bioresource Technol 98: 1339–1345.    

42. Margesin R, Feller G (2010) Biotechnological applications of psychrophiles. Environ Technol 31: 835–844.    

43. Zhuang WQ, Tay JH, Maszenan A, et al. (2002) Bacillus naphthovorans sp. nov. from oil-contaminated tropical marine sediments and its role in naphthalene biodegradation. Appl Microbiol Biotechnol 58: 547–554.

44. Swaathy S, Kavitha V, Pravin AS, et al. (2014) Microbial surfactant mediated degradation of anthracene in aqueous phase by marine Bacillus licheniformis MTCC 5514. Biotechnol Rep 4: 161–170.    

45. Ukiwe LN, Egereonu UU, Njoku PC, et al. (2013) Polycyclic aromatic hydrocarbons degradation techniques: a review. Int J Chem 5: 43–55.

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