Isolation and Characterization of Alkane Hydrocarbons-Degrading Enterobacter ludwigii strain D8 from Petroleum-Contaminated Soils

Year 2022, Volume: 11 Issue: 2, 630 - 639, 30.06.2022

Ayşe Eren , Kemal Güven

https://doi.org/10.17798/bitlisfen.1071785

Abstract

A bacterial strain has been isolated from petroleum contaminated soil with in southeastern Turkey. This isolated strain was characterized to determine its hydrocarbon biodegradation potential. Phenotypic features and of 16 S gene sequence analysis of rRNA revealed that strain D8 belongs to the Enterobacter genus and most closely resembles Enterobacter ludwigii (100%). The optimum temperature and pH values for the growth of E. ludwigii D8 were found to be 30°C and 5.0, respectively. This bacterial strain grew in long and medium chain hydrocarbons such as 1% decane, pentadecane and squalene separately at the end of 3 day incubation in the basal medium (BM) under optimum conditions. It was shown that E. ludwigii strain D8 degrades about 27% of crude oil incubated for 5 days, while it degrades 29% of pentadecane after 3 days of incubation determined by Gas chromatography-MS analysis. The biodegradation potential of petroleum hydrocarbons of E. ludwigii strain D8 isolated and characterized in this study indicates that this strain may play a role in the bioremediation process.

Keywords

Oil pollution, bacterial characterization, 16S rRNA gene sequence analysis, GC-MS analysis, Bioremediation

Supporting Institution

Dicle University

Project Number

19.014

Alkan Hidrokarbonları Parçalayan Enterobacter ludwigii strain D8 Suşunun Petrolle Kirlenmiş Topraklardan İzolasyonu ve Karakterizasyonu

Abstract

Türkiye'nin güneydoğusundaki petrol bulaşmış topraktan bir bakteri türü izole edildi. Bu izole edilen bakteri, petrol hidrokarbonları açısından biyolojik bozunma potansiyelini belirlemek için karakterize edildi. Fenotipik özellikler ve 16 S rRNA geninin dizi analizi, suş D8'in Enterobacter cinsinin bir üyesi olduğunu ve en çok Enterobacter ludwigii ‘ ye (%100) benzediğini belirlendi. E. ludwigii suşu D8'in büyümesi için optimum sıcaklık ve pH değerleri sırasıyla 30°C ve 5.0 olarak bulunmuştur. Optimum koşullarda bazal ortamda (BM) uzun ve orta zincirli hidrokarbon olan %1’lik dekan, pentadekan ve skualen ile ayrı ayrı inkübasyon sonunda bu bakteri suşunun ürediği görülmüştür. E. ludwigii D8 suşunun, yapılan GC-MS analizlerinde, 5 günlük inkübasyon sonrasında ham petrolü yaklaşık % 27 oranında parçalarken, 3 günlük inkübasyonu sonrasında pentadekanı % 29 oranında parçaladığı belirlendi. Bu çalışmada, E. ludwigii strain D8 suşunun, petrol hidrokarbonlarını biyolojik olarak parçalama özelliğinden dolayı, biyoiyileştirme sürecinde rol alabileceğini göstermektedir.

Keywords

Petrol kirliliği, bakteriyel karakterizasyon, 16S rRNA gen dizisi analizi, GC-MS analizi, Biyoiyileştirme

Project Number

19.014

References

• [1]Acer Ö., Güven K., Matpan Bekler F., Gül-Güven R .2016. Isolation and characterization of long chain alkane-degrading Acinetobacter sp. BT1A from oil contaminated soil in Diyarbakır, in the Southeast of Turkey. Bioremediation Journal, 20:1.
• [2]Alikhani J., Shayegan J., Akbari A. 2015. Risk assessment of hydrocarbon contaminant transport in vadose zone as it travels to groundwater table: A case study. Advances in environmental technology, 2, 77-84.
• [3]Cerqueda-Garcíaa D.,García-Maldonadob J.Q., Aguirre-Macedoc L., García-Cruz U. 2020. A succession of marine bacterial communities in batch reactor experiments during the degradation of five different petroleum types. Marine Pollution Bulletin , 150:110775.
• [4]Panda S.K., Kar R.N., Panda C.R. 2013. Isolation and identification of petroleum hydrocarbon degrading microorganisms from oil contaminated environment. International journal of environmental sciences, 3(5): 1314-1321.
• [5]Ezeji U., Anyadoh S.O., Ibekwe V.I. 2007. Clean up of crude oil-contaminated soil, Terr. Aquat. Environ. Toxicol, 1(2) :54-59.
• [6]Liu G.H., Ye Z., Tong K., Zhang Y.H. 2013. Biotreatment of heavy oil wastewater by combined upflow anaer sludge blanket and immobilized biological aerated filter in a pilot-scale test, Biochem. Eng. J, 72: 48-53.
• [7]Moussavi G., Ghorbanian M. 2015. The biodegradation of petroleum hydrocarbons in an upflow sludge-blanket/fixedfilm hybrid bioreactor under nitrate-reducing conditions: Performance evaluation and microbial identification, Chem. Eng. J, 280: 121-131.
• [8]Awasthi M.K., Selvam A., Chan M.T., Wong J.W.C. 2018. Bio-degradation of oily food waste employing thermophilic bacterial strains. Bioresour. Technol, 248: 141-147.
• [9]Huang Y., Pan, H., Wang Q., Ge Y., Liu W., Christie P. 2019. Enrichment of the soil microbial community in the bioremediation of a petroleum-contaminated soil amended with rice straw or sawdust. Chemosphere ,224: 265-271.
• [10]Acer Ö., Güven K., Poli A., Donato P.D., Leone Buono L., Gül Güven R., Nicolaus B.,Finore I. 2020. Acinetobacter mesopotamicus sp. nov., petroleum-degrading bacterium, isolated from petroleum-contaminated soil in Diyarbakir, in the Southeast of Turkey. Current Microbiology,77(2):3192-3200.
• [11]Jyothi K., Surendra Babu K., Nancy Clara K., Kashyap A. 2012. Identification and isolation of hydrocarbon degrading bacteria by molecular characterization. Helix , 2:105-111.
• [12]Mahjoubi M, Jaouani A, Guesmi A, Ben Amor S, Jouini A .,Cherif H., Najjari A., Boudabous A., Koubaa N., Cherif A. 2013. Hydrocarbonoclastic bacteria isolated from petroleum contaminated sites in Tunisia: isolation, identification and characterization of the biotechnological potential. New Biotechnol, 30(6):723-733.
• [13]Xu X., Liu W., Tian S., Wang W., Qi Q., Jiang P., Gao X.,Li F., Li H., Yu H. 2018.Petroleum hydrocarbon-degrading bacteria for the remediation of oil pollution under aerobic conditions: A Perspective Analysis . Front. Microbiol, 9:2885.
• [14] Sajna K.V., Sukumaran R.K., Gottumukkala L.D., Pandey A. 2015. Crude oil biodegradation aided by biosurfactants from Pseudozyma sp. NII 08165 or its culture broth. Bioresour. Technol, 191: 133-139.
• [15]Wilkes H., Buckel W., Golding, B.T., Rabus R. 2016. Metabolism of hydrocarbons in nalkane utilizing anaerobic bacteria. J. Mol. Microbiol. Biotechnol, 26, 138-151.
• [16]Ledezma-Villanueva A., Adame-Rodríguez J.M., O'Connor-Sánchez I.A., Villarreal-Chiu J.F., Aréchiga-Carvajal E.T. 2016. Biodegradation kinetic rates of diesel-contaminated sandy soil samples by two different microbial consortia. Ann. Microbiol, 66 :197–206.
• [17]Rizzo C., Rappazzo A.C., Michaud L., De Domenico E., Rochera C., Camacho A., Giudice A.L. 2018. Efficiency in hydrocarbon degradation and biosurfactant production by Joostella sp. A8 when grown in pure culture and consortia. J. Environ. Sci, 67. 115-126.
• [18]Kumari S., Regar R.K., Manickam N. 2018. Improved polycyclic aromatic hydrocarbon degradation in a crude oil by individual and a consortium of bacteria. Bioresour. Technol, 254: 174-179.
• [19]Wanapaisan P., Laothamteep N., Vejarano F., Chakraborty J., Shintani M.,Muangchinda C., Morita T., Suzuki-Minakuchi C., Inoue K., Nojiri H., Pinyakong O. 2018. Synergistic degradation of pyrene by five culturable bacteria in a mangrovesediment-derived bacterial consortium. J. Hazard Mater, 342: 561-570.
• [20]Dussault H. P. 1955. An improved technique for staining red halophilic bacteria. Journal of Bacteriol, 70: 484-485.
• [21]Lányi B. 1988. 1 Classical and rapid identification methods for medically important bacteria. Method Microbiol , 19:1- 67.
• [22] Bergey D.H., Krieg N.R.,Holt J.G. 1989. Bergey's manual of systematic bacteriology. Williams & Wilkins Co, Baltimore, ISBN:0-683-04108-8.
• [23]Claus D., and Berkeley C.W. 1986. The genus Bacillus In: Bergey’s manual of systematic bacteriology, 2: 1105.
• [24]Abtahi H., Parhamfar M., Saeedi R., Villase~nor J., Majid S., Kumar V., Coulon F., Parhamfar M., Didehdar M., seifi H., Koolivand A. 2020. Effect of competition between petroleum-degrading bacteria and indigenous compost microorganisms on the efficiency of petroleum sludge bioremediation: Field application of mineral-based culture in the composting process. Journal of Environmental Management ,258 : 110013.
• [25]Dwivedi A., Chitranshi S., Gupta A., Kumar A., Lal Bhat J. 2019. Assessment of the Petroleum Oil Degradation Capacity of Indigenous Bacterial Species Isolated from Petroleum Oil‑Contaminated Soil. International Journal of Environmental Research ,13:735-746.
• [26]Gupta B.,Rajor A., K., Kaur J. 2018. Isolation, Characterisation of Novel Pseudomonas and Enterobacter sp. from contaminated soil of chandigarh for naphthalene degradation. In book: Utilization and Management of Bioresources , 175-186.
• [27] Zhang J., Cui1 T ., Li X. 2018. Screening and identification of an Enterobacter ludwigii strain expressing an active β-xylosidase. Annals of Microbiology, 68:261-271.
• [28]Wendy F.T., Abu Hena M.K.., Wong S. K., Idris M. H., Sharifuzzaman S. M, Ina-Salwany M. Y. 2014. Enterobacter ludwigii, a candidate probiont from the intestine of Asian seabass. Journal of Science and Technology in the Tropics, 10: 5-14.
• [29]Jerin Israt., Rahi M., Sultan T., Islam M.S., Sajib S.A., Hoque K.M.F., Reza M.A. 2021. Diesel degradation efficiency of Enterobacter sp., Acinetobacter sp., and Cedecea sp. isolated from petroleum waste dumping site: a bioremediation view point. Archives of Microbiology. 203:5075–5084.
• [30]Throne-Holst M., Wentzel A., Ellingsen T.E., Kotlar H.K., Zotchev S.B. 2007.Identification of Novel Genes Involved in Long-Chain n-Alkane Degradation by Acinetobacter sp. Strain DSM 17874. Applied and Environmental Microbiology, 73(10):3327-3332.
• [31]Muthukamalam S.,Sivagangavathi S.,Dhrishya D., Rani S.S. 2017. Characterization of dioxygenases and biosurfactants produced by crude oil degrading soil bacteria. Brazilian Journal of Microbiology, 48(4):637-647.
• [32]Simaria C., Pant G., Sibi G. 2015. Characterization and evaluation of polycyclic aromatic hydrocarbon (Pah) degrading bacteria isolated from oil contaminated soil. Applied Microbiology: Open Access, 1:1.
• [33]Hua X., Wu Z., Zhang H., Lu D., Wang M., Liu Y.,Liu Z.2010. Degradation of hexadecane by Enterobacter cloacae strain TU that secretes an exopolysaccharide as a bioemulsifier. Chemosphere 80: 951-956.
• [34]Kafilzadeh F and Khezri A. 2016. Biodegraditon of aniline by Enterobacter ludwigii KH-A5 isolated from the soil around shiraz refinery,iran. Global Nest Journal, 18(4): 697-707.
• [35]Wang D., Zheng G., Wang S., Zhang D. and Zhou L. 2011. Biodegradation of aniline by Candida tropicalis AN1 isolated from aerobic granular sludge, J Environ Sci, 23(12): 2063-2068.
• [36]Sarwade V. and Gawai K.2014. Biodegradation of aniline by alkaliphilic strain Bacillus badius D1., IOSR J Environ Sci Toxicol Food Technol, 8(5): 71-78.