Novel Bacterial Strains Pseudomonas sp. and Bacillus sp. Isolated from Petroleum Oil Contaminated Soils for Degradation of Flourene and Phenanthrene

Document Type : Original Research Paper


UIET, Biotechnology Branch, Panjab University, Chandigarh, P.O. Box 160014, India.


Flourene and phenanthrene are organic compounds with high hydrophobicity and toxicity. Being recalcitrant in nature they are accumulating in the environment at an alarming concentration, posing serious threat to living beings. Thus in the present study, microorganisms were screened for their ability to degrade these contaminants at high concentrations in least period of time. Two out of fifteen isolates screened showed growth in basal medium containing 25 mg/l of fluorene/phenanthrene as the only carbon source. These selected isolates were acclimatised with step wise increased concentrations of flourene/phenanthrene for 165 days in basal medium. The acclimatised strains were identified and characterised on the basis of their morphological and biochemical characteristics and 16S rRNA gene sequence analysis. Results showed close relatedness of the isolates to Pseudomonas aeruginosa sp. and Bacillus safensis sp. Biodegradation studies carried out with these acclimatised strains at optimum conditions (pH 7 and temperature 30°C) showed 62.44% degradation of fluorene and 54.21% of phenanthrene in 10 days by Pseudomonas sp. VB92, whereas, Bacillus sp. JK17 degraded 43.64% of fluorene and 59.91% of phenanthrene in 12 days, at an initial concentration of 200 mg/l, as determined by HPTLC. During fluorene degradation by Pseudomonas sp. VB92, one metabolite was identified as fluorene,1,4-dihydro. An anionic biosurfactant (emulsification index of 80%) produced by strain VB92 during growth with PAHs, improved its degradation rate. This showed strong potential of the acclimatised strains for bioremediation and reclamation of polyaromatic hydrocarbon contaminated sites.


Abdel-Shafy, H. I. and Mansour, M. S. M. (2016). A review on polycyclic aromatic hydrocarbons: Source, environmental impact, effect on human health and remediation. Egypt J Pet., 25(1); 107-123.
Agency for Toxic Substances and Disease Registry (ATSDR) (2009). Toxicity of Polycyclic Aromatic Hydrocarbons (PAHs). Case Stud Environ Med., 1-68.
Akdogan, H. A. and Pazarlioglu, N. K. (2011). Fluorene biodegradation by P. ostreatus - Part I: Biodegradation by free cells. Process Biochem., 46(4); 834-839.
Aparna, A., Srinikethan, G. and Smitha, H. (2012). Colloids and Surfaces B : Biointerfaces Production and characterization of biosurfactant produced by a novel Pseudomonas sp. 2B. Colloids and surfaces. B, Biointerfaces., 95; 23-29.
ATSDR. (1995). Toxicological profile for polycyclic aromatic hydrocarbons. U.S. Dep Heal Hum Serv, (August), 1-487.
Bamforth, S. M. and Singleton, I. (2005). Bioremediation of polycyclic aromatic hydrocarbons: Current knowledge and future directions. J Chem Technol Biotechnol., 80(7); 723-736.
Cappuccino, J. and Sherman, N. (2010). Microbiology: A Laboratory Manual, Pearson Education Limited.
Chauhan, A., Fazlurrahman., Oakeshott, J. G. and Jain, R. K. (2008). Bacterial metabolism of polycyclic aromatic hydrocarbons: Strategies for bioremediation. Indian J Microbiol., 48(1); 95-113.
Chen, B., Huang, J., Yuan, K., Lin, L., Wang, X., Yang, L., et al. (2015). Direct evidences on bacterial growth pattern regulating pyrene degradation pathway and genotypic dioxygenase expression. Mar. Pollut. Bull., 105(1); 73-80.
Chupungars, K., Rerngsamran, P. and Thaniyavarn, S. (2009). Polycyclic aromatic hydrocarbons degradation by Agrocybe sp. CU-43 and its fluorene transformation. Int Biodet Biodeg., 63(1); 93-99.
Drainas, C. and Koukkou, A. I. (2007). Taxonomic identification, phenanthrene uptake activity, and membrane lipid alterations of the PAH degrading. Appl Microbiol Biotechnol., 76; 709-717.
Fooladi, T., Moazami, N., Abdeshahian, P., Kadier, A., Ghojavand, H., Wan Yusoff, W. M. and Hamid, A. A. (2016). Characterization, production and optimization of lipopeptide biosurfactant by new strain Bacillus pumilus 2IR isolated from an Iranian oil field. J Pet Sci Eng., 145; 510-519.
Gupta, B., Puri, S. and Kaur, J. (2016). Isolation and characterization of pyrene degrading bacteria from petroleum oil contaminetd soil from Chandigarh. Int J Biol Pharm Allied Sci., 5(9); 2084-2096.
Haritash, A. K. and Kaushik, C. P. (2009). Biodegradation aspects of Polycyclic Aromatic Hydrocarbons (PAHs): A review. J Hazard Mater., 169(1-3); 1-15.
Holt, J. H., Krieg, N. R., Sneath, P. H. A., Staley, J. . and Williams, S. . (1 4). Bergey’s manual of determinative bacteriology ninth edition. European journal of paediatric neurology : EJPN : official journal of the European Paediatric Neurology Society., 13; 560.
Kasumba., J. and Holmen., B. A. (2018). Heterogeneous ozonation reactions of PAHs and fatty acid methyl esters in biodiesel particulate matter. Atmos. Environ.,175; 15-24.
Kuppusamy, S., Thavamani, P. and Singh, S. (2017). Polycyclic aromatic hydrocarbons (PAHs) degradation potential , surfactant production , metal
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resistance and enzymatic activity of two novel cellulose-degrading bacteria isolated from koala faeces. Environ Earth Sci., 76(1);1-12.
Ling, J., Zhang, G., Sun, H., Fan, Y., Ju, J. and Zhang, C. (2011). Isolation and characterization of a novel pyrene-degrading Bacillus vallismortis strain JY3A. Sci Total Environ., 409(10); 1994-2000.
Ma, J., Xu, L. and Jia, L. (2012). Degradation of polycyclic aromatic hydrocarbons by Pseudomonas sp. JM2 isolated from active sewage sludge of chemical plant. J Environ Sci., 24(12); 2141-2148.
Marchand, C., St-Arnaud, M., Hogland, W., Bell, T. H. and Hijri, M. (2017). Petroleum biodegradation capacity of bacteria and fungi isolated from petroleum-contaminated soil. Int Biodet Biodeg., 116; 48-57.
Marco-Urrea, E., Garca-Romera, I. and Aranda, E. (2015). Potential of non-ligninolytic fungi in bioremediation of chlorinated and polycyclic aromatic hydrocarbons. N Biotechnol., 32(6); 620-628.
Masakorala, K., Yao, J., Cai, M. and Chandankere, R. (2013). Isolation and characterization of a novel phenanthrene (PHE) degrading strain Psuedomonas sp. USTB-RU from petroleum contaminated soil. J Hazard Mater., 263; 493-500.
Mojarad, M., Alemzadeh, A., Ghoreishi, G. and Javaheri, M. (2016). Kerosene biodegradation ability and characterization of bacteria isolated from oil-polluted soil and water. J Environ Chem Eng., 4(4); 4323-4329.
Nie, M., Yin, X., Ren, C., Wang, Y., Xu, F. and Shen, Q. (2010). Novel rhamnolipid biosurfactants produced by a polycyclic aromatic hydrocarbon-degrading bacterium Pseudomonas aeruginosa strain NY3. Biotechnol Adv., 28(5); 635-643.
Oyehan, T. A. and Al-thukair, A. A. (2017) Isolation and characterization of PAH-degrading bacteria from the Eastern Province , Saudi Arabia. MPB., 115(1-2); 39-46.
Pedetta, A., Pouyte, K., Herrera, M. K., Babay, P. A., Espinosa, M., Costagliola, M., et al. (2013). Phenanthrene degradation and strategies to improve its bioavailability to microorganisms isolated from brackish sediments. Int Biodet Biodeg., 84; 161-167.
Rodrigues, E. M., Kalks, K. H. M., Marcos, R. T. (2015). Prospect, isolation, and characterization of microorganisms for potential use in cases of oil bioremediation along the coast of Trindade. J Environ Manage.,156; 1522.
Seo, J. S., Keum, Y. S. and Li, Q. X. (2009). Bacterial degradation of aromatic compounds. Int J Environ Res Public Health., 6.
Tsai, J., Kumar, M. and Lin, J. (2009). Anaerobic biotransformation of fluorene and phenanthrene by sulfate-reducing bacteria and identification of biotransformation pathway. J Hazard Mater., 164; 847-855.
Varjani, S. J., and Upasani, V. N. (2016). Biodegradation of petroleum hydrocarbons by oleophilic strain of Pseudomonas aeruginosa NCIM 5514. Bioresour Technol., 222; 195-201.
Waigi, M. G., Kang, F., Goikavi, C., Ling, W. and Gao, Y. (2015). Phenanthrene biodegradation by sphingomonads and its application in the contaminated soils and sediments: A review. Int Biodet Biodeg., 104; 333-349.
Williams, E. S., Mahler, B. J. and Metre, P. C. V. (2013). Cancer Risk from Incidental Ingestion Exposures to PAHs Associated with Coal-Tar-Sealed Pavement. Environ Sci Technol., 47; 1101-1109.
Zhang, G., Ling, J., Sun, H., Luo, J., Fan, Y. and Cui, Z. (2009). Isolation and characterization of a newly isolated polycyclic aromatic hydrocarbons-degrading Janibacter anophelis strain JY11. J Hazard Mater., 172; 580-586.
Zhou, L., Li, H., Zhang, Y., Han, S. and Xu, H. (2016). Sphingomonas from petroleum-contaminated soils in Shenfu, China and their PAHs degradation abilities. Braz J Microbiol., 47(2); 271-278.