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Isolation and Identification of Crude Oil Degrading and Biosurfactant Producing Bacteria from the Oil-Contaminated Soils of Gachsaran

Seyyedeh Zahra Hashemi, Jamshid Fooladi, Gholamhossein Ebrahimipour, Sajad Khodayari




Background and Objectives: Petroleum hydrocarbons are harmful to the environment, human health, and all other living creatures. Oil and its byproducts in contact with water block sunshine to phytoplanktons and thus break the food chain and damage the marine food source. This study aims to isolate the crude oil degrading and biosurfactant producing bacteria from the oil contaminated soils of Gachsaran, Iran.


Materials and Methods: Isolation was performed in peptone-water medium with yeast extract. Oil displacement area, emulsification index and bacterial phylogeny using 16S rRNA analysis were studied.


Results and Conclusion: Three isolates were able to degrade the crude oil. In the first day, there were two phases in the medium; after a few days, these three bacteria degraded the crude oil until there was only one phase left in the medium. One strain was selected as a superior strain by homogenizing until the medium became clear and transparent. This method confirmed that the strain produces biosurfactant. According to the morphological and biochemical tests, the strain isolated from the oil contaminated soils is a member of Bacillus subtilis, so to study the bacterial phylogeny and taxonomy of the strain, an analysis of 16S rRNA was carried out, and the phylogenic tree confirmed them. The results verified that oil contaminated soils are good source for isolation of the biosurfactant producing bacteria.


Bacillus, Bioremediation, Biosurfactant, Crude oil, Gachsaran


Ivshina IB, Kuyukina MS, Philp JC, Christofi N. Oil desorption from mineral and organic materials using biosurfactant complexes produced by Rhodococcus species. World J Microb Biot. 1998; 14: 711-717.

Souza EC, Vessoni-Penna TC, Oliveira PdS. Biosurfactant-enhanced hydrocarbon bioremediation: An overview. Int Biodeter Biodegr. 2014; 89: 88-94.

Abdel-Shafy I, Mansour SM. A review on polycyclic aromatic hydrocarbons: Source, environmental impact, effect on human health and remediation. Egypt J. 2015; In press.

Lai CC, Huanga Y, Wei YH, Chang J. Biosurfactant-enhanced removal of total petroleum hydrocarbons from. J Hazard Mater. 2009; 167: 609-614.

Singh AK, Cameotra SS. Efficiency of lipopeptide biosurfactants in removal of petroleum hydrocarbons and heavy metals from contaminated soil. Environ Sci Pollut Res. 2013; 20: 7367-7376.

Gojgic Cvijovic GD, Milic JS, Solevic TM, Beskoski VP, Ilic MV, Djokic LS, Naracic TM, Vrvis MM. Biodegradation of petroleum sludge and petroleum polluted soil by a bacterial consortium: a laboratory study. Biodegradation. 2012; 23: 1-14.

Asha A, Javarkar Sanjeev K, Singh Ackmez M. A comprehensive overview of elements in bioremediation. Rev Environ Sci Biotechnol. 2010; 9: 215-288.

Chandankere R, Yao J, Cai M, Masakorala K, Jain AK, Choi MF. Properties and characterization of biosurfactant in crude oil biodegradation by bacterium Bacillus methylotrophicus USTBa. Fuel. 2014; 122: 140-148.

Yemashova A, Murygina P, Zhukov V, Zakharyantz A, Gladchenko A, Appanna V , Kalyuzhnyi V. Biodeterioration of crude oil and oil derived products: a review. Rev Environ Sci Biotechnol. 2007; 6: 315-337.

Ferradji Z, Mnif S, Badis A, Rebbani S , Fodil D, Eddouaouda K, Sayadi S. Naphthalene and crude oil degradation by biosurfactant producing Streptomyces spp. isolated from Mitidja plain soil (North of Algeria). Int Biodeter Biodegr. 2014; 86: 300-308.

Duc LH, Hong HA , Barbosa M, Henriques O, Cutting M. Characterization of Bacillus Probiotics Available for Human Use. Appl Environ Microb. 2004; 70: 2161-2171.

Fujiya M , Musch W, Nakagawa Y, Hu S, Alverdy J, Kohgo Y, Schneewind O, Jabri B, Chang EB. The Bacillus subtilis Quorum-Sensing Molecule CSF Contributes to Intestinal Homeostasis via OCTN2, a Host Cell Membrane Transporter. Cell Host Microbe. 2007; 1: 299-308.

Afzal-Javan F, Mobibn-Dehkordi M. Amplification, Sequencing and Cloning of Iranian Native Bacillus subtilis Alpha-amylase Gene in Saccharomyces cerevisiae. Joundishapour Microb. 2013; 6(8): 1-7

Rufino RD, Luna JM, Marinho PHC, Farias CBB, Ferreira SRM, Sarubbo LA. Removal of petroleum derivative adsorbed to soil by biosurfactant Rufisan producedby Candida lipolytica. J Petrol Sci Eng. 2013; 109: 117-122.

Das P, Mukherjee S, Sen R. Improved bioavailability and biodegradation of a model polyaromatic hydrocarbon by a biosurfactant producing bacterium of marine origin. Chemosphere. 2008; 72: 1229-1234.

Zouari R, Besbes S, Chaabouni SE, Aydi D. Cookies from composite wheat–sesame peels flours: Dough quality and effect of Bacillus subtilis SPB1 biosurfactant addition. Food Chem. 2015; 194: 758-769.

Mnif I, Besbes S , Ellouze R, Ellouze Chaabouni S, Ghribi D. Improvement of Bread Quality and Bread Shelf-life by Bacillus subtilis Biosurfactant Addition. Food Sci Biotechnol. 2012; 21: 1105-1112.

Campos M, Stamford LM, Sarubbo A. Production of a Bioemulsifier with Potential Application in the Food Industry. Appl Biochem Biotechnol. 2014; 172: 3234-3252.

Makkar RS, Cameotra SS, Banat MI. Advances in utilization of renewable substrates for biosurfactant production. AMB Express. 2011; 1-5.

Morikawa M, Hirata Y, Imanaka T. A study on the structure function relationship of lipopeptide biosurfactants. Biochimica et Biophysica Acta. 2000; 1488: 211-218.

Gandhimathi R, Kiran G, Hema TA, Selvin J, Raviji T, Shanmughapriya S. Production and characterization of lipopeptide biosurfactant by a sponge-associated marine actinomycetes Nocardiopsis alba MSA10. Bioprocess Biosyst Eng. 2009; 32: 825-835.

Walter V, Syldatk C, Hausmann R. Screening Concepts for the Isolation of Biosurfactant Producing Microorganisms. Adv Exp Med Biol.2010; 672; 1-13.

Varjani SJ, Rana DP, Bateja S, Sharma MC, Upasani VN. Screening and identification of biosurfactant (bioemulsifier) producing bacteria from crude oil contaminated sites of Gujarat, India. Int J Innov Res Sci Eng Technol. 2014; 3(2): 9205-9213.

Donio MB, Ronica SF, Viji VT, Velmurugan S, Jenifer JA, Michaelbabu M, Cirarasu T. Isolation and characterization of halophilic Bacillus sp. BS3 able to produce pharmacologically important biosurfactants. Asian Pac J Trop M. 2013; 876-883.

Green DH, Wakerley PR, Page A, Barnes A, Baccigalupi L, Ricca E, Cutting SM. Characterization of Two Bacillus Probiotics. Appl Environ Microbial. 1999; 65(9): 4288-4291.

KV, Keharia H. Application of extracellular lipopeptide biosurfactant produced by endophytic Bacillus subtilis K1 isolated from aerial roots of banyan (Ficus benghalensis) in microbially enhanced oil recovery (MEOR). Biotech. 2014; 4; 41-48.

PPlaza GA, Turek A, Krol E, Szczyglowska R. Antifungal and antibacterial properties of surfactin isolated from Bacillus subtilis growing on molasses. Afr J Microbiol Res. 2013; 7(25); 3165-3170.

DOI: https://doi.org/10.22037/afb.v3i2.10286


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