STM Journals Peer-reviewed research from Consortium e-Learning Network

Research and Reviews : A Journal of Biotechnology Original Research

Investigations Into the Potential for Anthracene Decomposition of Novel Soil Bacterium, Pseudomonas putida P7

  1. Kushalatha M. Departmen of Biochemistry, Maharani Lakshmi Ammanni College for Women, Bangalore
  2. Umesh H.R. Department of Biochemistry, Maharani Lakshmi Ammanni College for Women, Bengaluru

Abstract

Bioremediation of hazardous contaminants holds immense importance for the preservation of a clean and healthy environment. Conversely, the accumulation of toxicants can be mitigated with the assistance of microorganisms capable of breaking down these harmful substances into benign molecules. Anthracene is frequently used as a model molecule for studies on PAH pollution because it is found in many carcinogenic PAHs. Therefore, the hunt for new microorganisms with the ability to break down anthracene is necessary. This work intended to the soil of a petroleum refinery near Bangalore was used in this study to isolate twenty different bacterial strains. These strains were cultured on solid nutrient agar and then purified by cultivation on mineral media supplemented with anthracene. Successful bacterial growth and anthracene degradation were confirmed through optical density measurements. The growth of bacterial isolate was observed in media at a pH of 7.0 and temperature of 28°C. However, the presence of certain nitrogen and carbon sources inhibited bacterial growth during anthracene breakdown. The efficacy of anthracene degradation by the isolate )1.8x1022 colony forming units/ml and optical density of 0.92) was determined to be 85.36% after one hundred twenty hours of incubation. Gas chromatography-Mass spectrometer analysis revealed the conversion of anthracene to anthraquinone and 9, 10-dihydroxy-anthracene. Through the biochemical tests and 16S rRNA analysis, the highly effective bacterial isolate was identified as P7, a novel strain of Pseudomonas putida, with 98% homology. High-performance liquid chromatography analysis demonstrated the substantial reduction in anthracene levels (up to 83.25%) and the formation of reaction products by the bacterial strain P7 sourced from the soil. The newly identified bacterial strain P7 shows promise as an effective tool for bio-remediating hydrocarbon pollutants to protect the environment from polyaromatic hydrocarbon contaminants.

Keywords

References (47)

  1. Jacques RJS, Santos EC, Bento FM, Peralba MCR, Selbach PA, Sá ELS, et al. Anthracene biodegradation by Pseudomonas sp. isolated from a petrochemical sludge landfarming site. International Biodeterioration & Biodegradation. 2005;56(3):143-150. doi:10.1016/j.ibiod.2005.06.005
  2. Bouchez M, Blanchet D, Vandecasteele JP. Degradation of polycyclic aromatic hydrocarbons by pure strains and by defined strain associations: inhibition phenomena and cometabolism. Applied Microbiology and Biotechnology. 1995;43(1):156-164. doi:10.1007/bf00170638
  3. Seg MY, Habib SA, Ali S. Evaluation of Corynebacterium variabilis Sh42 as a degrader for different poly aromatic compounds. Nour J Am Sci. 2010;6:343–56.
  4. Heitkamp MA, Cerniglia CE. Mineralization of polycyclic aromatic hydrocarbons by a bacterium isolated from sediment below an oil field. Applied and Environmental Microbiology. 1988;54(6):1612-1614. doi:10.1128/aem.54.6.1612-1614.1988
  5. Bamforth SM, Singleton I. Bioremediation of polycyclic aromatic hydrocarbons: current knowledge and future directions. Journal of Chemical Technology & Biotechnology. 2005;80(7):723-736. doi:10.1002/jctb.1276
  6. Hickey AM, Gordon L, Dobson ADW, Kelly CT, Doyle EM. Effect of surfactants on fluoranthene degradation by Pseudomonas alcaligenes PA-10. Applied Microbiology and Biotechnology. 2006;74(4):851-856. doi:10.1007/s00253-006-0719-5
  7. Wong JWC, Lai KM, Wan CK, Ma KK, Fang M. Isolation and optimization of PAH-degradative bacteria from contaminated soil for PAHs bioremediation. Water Air Soil Pollut. 2002;139:1–13.
  8. Sun R, Jin J, Sun G, Liu Y, Liu Z. Screening and degrading characteristics and community structure of a high molecular weight polycyclic aromatic hydrocarbon-degrading bacterial consortium from contaminated soil. Journal of Environmental Sciences. 2010;22(10):1576-1585. doi:10.1016/s1001-0742(09)60292-8
  9. Lily MK, Bhatt BA, Dangwal KK. Degradation of anthracene by a novel strain Brachybacterium paraconglomeratum BMIT637C [MTCC 9445]. Int J Environ Sci. 2013;3:1242–52.
  10. Dean-Ross D, Moody JD, Freeman JP, Doerge DR, Cerniglia CE. Metabolism of anthracene by aRhodococcusspecies. FEMS Microbiology Letters. 2001;204(1):205-211. doi:10.1111/j.1574-6968.2001.tb10886.x
  11. Shahsavari E, Poi G, Aburto-Medina A, Haleyur N, Ball AS. Bioremediation approaches for petroleum hydrocarbon-contaminated environments. In: Ball AS, editor. Enhancing Cleanup of Environmental Pollutants. Springer; 2017. p. 21–41.
  12. Shokrollahzadeh SG, SF. Study of Sphingopyxis isolates in degradation of polycyclic aromatic hydrocarbons. Chem Eng Trans. 2012;27:55–60.
  13. Haleyur N, Shahsavari E, Taha M, Khudur LS, Koshlaf E, Osborn AM, et al. Assessing the degradation efficacy of native PAH-degrading bacteria from aged, weathered soils in an Australian former gasworks site. Geoderma. 2018;321:110-117. doi:10.1016/j.geoderma.2018.02.004
  14. Poluyanov AM, Sokolova AY, Malashenko EA, Sergunova EV, Bobkova NV. Isolation, Identification and Quantitative Determination of Anthracene Derivatives by HPLC-UV Method in the Raw Materials of Some Representatives of the Genus Rumex of Three Vegetation Times. Drug development & registration. 2022;11(4):216-225. doi:10.33380/2305-2066-2022-11-4-216-225
  15. Koshlaf E, Shahsavari E, Aburto-Medina A, Taha M, Haleyur N, Makadia TH, et al. Bioremediation potential of diesel-contaminated Libyan soil. Ecotoxicology and Environmental Safety. 2016;133:297-305. doi:10.1016/j.ecoenv.2016.07.027
  16. Milic J, Beskoski V, Ilic M, Ali S, Gojgic-Cvijovic G, Vrvic M. Bioremediation of soil heavily contaminated with crude oil and its products: Composition of the microbial consortium. Journal of the Serbian Chemical Society. 2009;74(4):455-460. doi:10.2298/jsc0904455m
  17. Lee SH, Lee WS, Lee CH, Kim JG. Degradation of phenanthrene and pyrene in rhizosphere of grasses and legumes. Journal of Hazardous Materials. 2008;153(1-2):892-898. doi:10.1016/j.jhazmat.2007.09.041
  18. Joner EJ, Leyval C. Rhizosphere Gradients of Polycyclic Aromatic Hydrocarbon (PAH) Dissipation in Two Industrial Soils and the Impact of Arbuscular Mycorrhiza. Environmental Science & Technology. 2003;37(11):2371-2375. doi:10.1021/es020196y
  19. Alias SA, Omar M, Hussain NH. Degradation of 3-ring PAHs by S spiritovorum and C. urealyticum strain under non-indigenous condition. In: International Congress of Chemistry and Environment (ICCE); 2011. Port Dickson, Malaysia.
  20. Kumar G, Kumar R. Plasmid associated anthracene degradation by Pseudomonas sp. isolated from filling station site. Nat Sci. 2010;8:89–94.
  21. Khan K, M. Naeem, M. Javed Arshed, M. Asif. Extraction and Characterization of Oil Degrading Bacteria. Journal of Applied Sciences. 2006;6(10):2302-2306. doi:10.3923/jas.2006.2302.2306
  22. Sheng-wang P, Shi-qiang W, Xin Y, Sheng-xian C. The Removal and Remediation of Phenanthrene and Pyrene in Soil by Mixed Cropping of Alfalfa and Rape. Agricultural Sciences in China. 2008;7(11):1355-1364. doi:10.1016/s1671-2927(08)60185-6
  23. Ebrahimi MSM, Fallah R. Assessment of biodegradation efficiency of some isolated bacteria from oil contaminated sites in solid and liquid media containing oil-compounds. Int Res J Appl Basic Sci. 2012;3:138–47.
  24. Abdelhay A, Magnin JP, Gondrexon N, Baup S, Willison J. Optimization and modeling of phenanthrene degradation by Mycobacterium sp. 6PY1 in a biphasic medium using response-surface methodology. Applied Microbiology and Biotechnology. 2008;78(5):881-888. doi:10.1007/s00253-008-1365-x
  25. Leahy JG, Colwell RR. Microbial degradation of hydrocarbons in the environment. Microbiological Reviews. 1990;54(3):305-315. doi:10.1128/mr.54.3.305-315.1990
  26. Guo C, Dang Z, Wong Y, Tam NF. Biodegradation ability and dioxgenase genes of PAH-degrading Sphingomonas and Mycobacterium strains isolated from mangrove sediments. International Biodeterioration & Biodegradation. 2010;64(6):419-426. doi:10.1016/j.ibiod.2010.04.008
  27. Chulalaksananukul S, Gadd GM, Sangvanich P, Sihanonth P, Piapukiew J, Vangnai AS. Biodegradation of benzo(a)pyrene by a newly isolated Fusarium sp. FEMS Microbiol Lett. 2006;262:99–106. doi:10.1111/j.1574-6968.
  28. Rastogi G, Coaker GL, Leveau JHJ. New insights into the structure and function of phyllosphere microbiota through high-throughput molecular approaches. FEMS Microbiology Letters. 2013;348(1):1-10. doi:10.1111/1574-6968.12225
  29. Andleeb S, Shafique I, Naseer A, Abbasi WA, Ejaz S, Liaqat I, et al. Molecular characterization of plant growth promoting vermibacteria associated with Eisenia fetida gastrointestinal tract. PLOS ONE. 2022;17(6).
  30. Bhore SJ, Nithya R, Loh CY. Screening of endophytic bacteria isolated from leaves of Sambung Nyawa [Gynura procumbens (Lour.) Merr.] for cytokinin-like compounds. Bioinformation. 2010;5(5):191-197. doi:10.6026/97320630005191
  31. Urveg Ratsimamanga S, El Jaziri M, Rakotoniriana EF, Rafamantanana M, Randriamampionona D, Rabemanantsoa C. Study in vitro of the impact of endophytic bacteria isolated from Centella asiatica on the disease incidence caused by the hemibiotrophic fungus Colletotrichum higginsianum. Antonie Van Leeuwenhoek. 2013;103:121–33.
  32. Xie WY, Su JQ, Zhu YG. Phyllosphere Bacterial Community of Floating Macrophytes in Paddy Soil Environments as Revealed by Illumina High-Throughput Sequencing. Applied and Environmental Microbiology. 2015;81(2):522-532. doi:10.1128/aem.03191-14
  33. Upadhyay SK, Singh DP, Saikia R. Genetic Diversity of Plant Growth Promoting Rhizobacteria Isolated from Rhizospheric Soil of Wheat Under Saline Condition. Current Microbiology. 2009;59(5):489-496. doi:10.1007/s00284-009-9464-1
  34. Kamalraj S, Sridevi S, Gangodevi V, Venkatesan A, Muthumary J. Effect of NaCl on biochemical changes and endophytic assemblages in the leaves of a mangrove, Ceriopsrox Burghiana. J Sci Technol. 2008;1(1):1–7.
  35. Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Molecular Biology and Evolution. 2016;33(7):1870-1874. doi:10.1093/molbev/msw054
  36. Bhattacharya S, Das A, G M, K V, J S. Mycoremediation of Congo red dye by filamentous fungi. Braz J Microbiol. 2011;42:1526–36. doi:10.1590/S1517-838220110004000040. PubMed: 24031787.
  37. Rugha CL, Susilawati E, Kravchenko AN, Thomas JC. Biodegrader Metabolic Expansion during Polyaromatic Hydrocarbons Rhizoremediation. Zeitschrift für Naturforschung C. 2005;60(3-4):331-339. doi:10.1515/znc-2005-3-418
  38. Fernández-Luqueño F, Marsch R, Espinosa-Victoria D, Thalasso F, Hidalgo Lara ME, Munive A, et al. Remediation of PAHs in a saline–alkaline soil amended with wastewater sludge and the effect on dynamics of C and N. Science of The Total Environment. 2008;402(1):18-28. doi:10.1016/j.scitotenv.2008.04.040
  39. Sartoros C, Yerushalmi L, Béron P, Guiot SR. Effects of surfactant and temperature on biotransformation kinetics of anthracene and pyrene. Chemosphere. 2005;61(7):1042-1050. doi:10.1016/j.chemosphere.2005.02.061
  40. Doick KJ, Dew NM, Semple KT. Linking Catabolism to Cyclodextrin Extractability: Determination of the Microbial Availability of PAHs in Soil. Environmental Science & Technology. 2005;39(22):8858-8864. doi:10.1021/es0507463
  41. Li X, Li P, Lin X, Zhang C, Li Q, Gong Z. Biodegradation of aged polycyclic aromatic hydrocarbons (PAHs) by microbial consortia in soil and slurry phases. Journal of Hazardous Materials. 2008;150(1):21-26. doi:10.1016/j.jhazmat.2007.04.040
  42. Swaathy S, Kavitha V, Pravin AS, Mandal AB, Gnanamani A. Microbial surfactant mediated degradation of anthracene in aqueous phase by marine Bacillus licheniformis MTCC 5514. Biotechnology Reports. 2014;4:161-170. doi:10.1016/j.btre.2014.10.004
  43. Santos EC, Jacques RJS, Bento FM, Peralba MDCR, Selbach PA, Sá ELS, et al. Anthracene biodegradation and surface activity by an iron-stimulated Pseudomonas sp. Bioresource Technology. 2008;99(7):2644-2649. doi:10.1016/j.biortech.2007.04.050
  44. Somtrakoon K, Suanjit S, Pokethitiyook P, Kruatrachue M, Lee H, Upatham S. Enhanced Biodegradation of Anthracene in Acidic Soil by Inoculated Burkholderia sp. VUN10013. Current Microbiology. 2008;57(2):102-106. doi:10.1007/s00284-008-9157-1
  45. Simarro R, González N, Bautista LF, Molina MC. Biodegradation of high-molecular-weight polycyclic aromatic hydrocarbons by a wood-degrading consortium at low temperatures. FEMS Microbiology Ecology. 2012;83(2):438-449. doi:10.1111/1574-6941.12006
  46. Kafilzadeh F, Khosrobak A, Jamali H. Isolation and Identification of Phenanthrene Degrading Bacteria from the Soil around Oil Company of Andimeshk and Investigation of Their Growth Kinetics. Polycyclic Aromatic Compounds. 2015;36(1):58-71. doi:10.1080/10406638.2015.1037006
  47. Bibi N, Hamayun M, Khan SA, Iqbal A, Islam B, Shah F, et al. Anthracene biodegradation capacity of newly isolated rhizospheric bacteria Bacillus cereus S13. PLOS ONE. 2018;13(8):e0201620. doi:10.1371/journal.pone.0201620
Support