Abstract
Environmental pollution from hydrocarbons and their derivatives is predominantly of anthropogenic origin, occurring mainly during mining, transportation, and storage. This often in an annual release of substantial amounts of pollutants into the environment. The effective restoration of petroleum-contaminated environments requires a series of interventions to remediate soil to its natural state. This study aimed to isolate and characterize biosurfactant-producing bacterial strains with potential bioremediation capabilities to assess the potential application for remediating soils polluted with spent engine oil in the Remo district, Ogun State. Bacterial isolates were recovered from soil samples collected from spent engine oil contaminated sites, using standard cultural techniques and characterized by biochemical and molecular techniques (16S rRNA amplification and sequencing). The isolates were further screened for biosurfactant activity, and the biosurfactant extract was characterized using gas chromatography-mass spectrometry (GC-MS) and Fourier-transform infrared (FTIR) spectroscopy. All 36 isolates recovered from the samples were positive for blood hemolysis, oil spread, and emulsification index assays. Pseudomonas sp. and Klebsiella sp. were identified as the most promising biosurfactant-producing isolates by 16S rRNA gene sequencing by the screening results. The GC-MS and FTIR spectroscopy of the biosurfactant extracts produced by Pseudomonas sp. revealed distinct peak values with various functional groups. FTIR analysis identified various functional groups, including aliphatic ether, ester, sulfone, alkene, allene, isothiocyanate, and alcohol, as the major compounds present in the biosurfactant. 20 bioactive compounds with different peak values were identified by GC-MS analysis. The rhlA gene associated with rhamnolipid production was detected in the two characterized P. aeruginosa strains. Subsequent laboratory-scale bioremediation of spent engine oil-contaminated soil demonstrated a significant (53%) removal of oil. The results highlight the potential applicability of eco-friendly biosurfactants for contaminated soil cleanup and suggest the need for further exploration, optimization, and elucidation of the genetic basis of biosurfactant production for improved efficacy.
References
Abdoli, S., Asgari Lajayer, B., Bagheri Novair, S., & Price, G. W. (2025). Unlocking the potential of biosurfactants in agriculture: novel applications and future directions. Sustainability, 17(5). https://doi.org/10.3390/su17052110
Abis, L. A., Hassan, O. M., & Hassan, K. T. (2026). Comparative bioremediation of petroleum-contaminated soil by live bacterial isolates and crude biosurfactant supernatants. Journal of Microbiological Methods, 107345. https://doi.org/10.1016/j.mimet.2025.107345
Albasri, H. M., Almohammadi, A. A., Alhhazmi, A., Bukhari, D. A., Waznah, M. S., & Mawad, A. M. (2024). Production and characterization of rhamnolipid biosurfactant from thermophilic Geobacillus stearothermophilus bacterium isolated from Uhud mountain. Frontiers in Microbiology, 15, 1358175. https://doi.org/10.3389/fmicb.2024.1358175
Al-Kanany, F. N., & Mohsen, N. (2023). New Record: Molecular Depiction of Rhamnolipids (rhlA) Gene in Locally Isolated Strains of Pseudomonas aeruginosa. Journal of Pure Applied Microbiology, 17(3), (1641–1649). https://doi.org/10.22207/JPAM.17.3.25
Al-Marri, S., Eldos, H. I., Ashfaq, M. Y., Saeed, S., Skariah, S., Varghese, L., … Raja, M. M. (2023). Isolation, identification, and screening of biosurfactant-producing and hydrocarbon-degrading bacteria from oil and gas industrial waste. Biotechnology Reports, 39, e00804. https://doi.org/10.1016/j.btre.2023.e00804
Angba, Y. J., Touré, A. O., Abro, K. D. M., Amadou Kiari, M. N., Drogui, A. P., & Yao, K. B. (2025). Optimization of biosurfactant production by Pseudomonas aeruginosa strain Pa using rubber tree seed oil as sole carbon source. Discover Applied Sciences, 7(9), 1020. https://doi.org/10.1007/s42452-025-07531-y
Apulu, O. G., Potravny, I. M., & Sukhorukova, I. V. (2021). Methods of Justification and Selection of Technologies for Remediation of Oil-contaminated Land. Ecology and Industry of Russia, 25, (38─43)126059. https://doi.org/10.18412/1816-0395-2021-6-38-43
Arabo, A. A., Bamanga, R. A., Abdulrazak, T. M., Ahmadu, J. H., Yakasai, H. M., Fadilu, M., ... Abdullahi, N. (2022). Isolation and Characterization of Biosurfactant Producing Pseudomonas aeruginosa YLA03 and its Diesel Degradation Potentials. Nigerian Journal of Biotechnology, 39(2), 64–73. https://dx.doi.org/10.4314/njb.v39i2.8
Babayola, N. M., & Adefisoye, M. A. (2025). Recent Advances and Prospects of Microbial Biosurfactant-Mediated Remediation of Engine Oil Pollution: A Comprehensive Review. Nature Environment and Pollution Technology, 24(1), 1–15. https://doi.org/10.46488/NEPT.2025.v24i01.D1655
Balakrishnan, S., Arunagirinathan, N., Rameshkumar, M. R., Indu, P., Vijaykanth, N., Almaary, K. S., ... & Chen, T. W. (2022). Molecular characterization of biosurfactant producing marine bacterium isolated from hydrocarbon-contaminated soil using 16S rRNA gene sequencing. Journal of King Saud University-Science, 34(3), 101871. https://doi.org/10.1016/j.jksus.2022.101871
Biktasheva, L., Gordeev, A., Usova, A., Kirichenko, A., Kuryntseva, P., & Selivanovskaya, S. (2024). Bioremediation of Oil-Contaminated Soils Using Biosurfactants Produced by Bacteria of the Genus Nocardiopsis sp. Microbiology Research, 15(4), 2575-2592. https://doi.org/10.3390/microbiolres15040171
Chang, J., Yang, W., Jin, Y., Zhou, Z., Zhao, W., Liang, S., & Ma, Y. (2026). Microbial Biosurfactants: A Bridge from Aquatic Environments to Subsurface Wettability Alteration in Oil Recovery. Life, 16(3), 484. https://doi.org/10.3390/life16030484
Chigede, N., Chikwambi, Z., Mpofu, I. D. T., & Madzimure, J. (2023). Isolation and characterization of biosurfactant-producing microbes isolated from the gastrointestinal system of broiler birds fed a commercial diet. Animal Biotechnology, 35(1). https://doi.org/10.1080/10495398.2023.2263771
Ciğeroğlu, Z., El Messaoudi, N., Şenol, Z. M., Başkan, G., Georgin, J., & Gubernat, S. (2024). Clay-based nanomaterials and their adsorptive removal efficiency for dyes and antibiotics: a review. Materials Today Sustainability, 26, 100735. https://doi.org/10.1016/j.mtsust.2024.100735
Darwesh, O. M., Mahmoud, M. S., Barakat, K. M., Abuellil, A., & Ahmad, M. S. (2021). Improving the bioremediation technology of contaminated wastewater using biosurfactants produced by novel bacillus isolates. Heliyon, 7(12). https://doi.org/10.1016/j.heliyon.2021.e08616
de Souza Araújo, L., Santana, L. A. R., Otenio, M. H., Nascimento, C. W., Cerqueira, A. F. L. W., & Rodarte, M. P. (2024). Biosurfactant production by Pseudomonas: a systematic review. Applied Biochemistry and Biotechnology, 1–15. https://doi.org/10.1007/s12010-024-05036-9
Deshmukh, N., & Kathwate, G. (2022). Biosurfactant production by Pseudomonas aeruginosa strain LTR1 and its application. Biointerface Research in Applied Chemistry, 13(10). https://doi.org/10.33263/BRIAC131.010
Dini, S., Bekhit, A. E. D. A., Roohinejad, S., Vale, J. M., & Agyei, D. (2024). The physicochemical and functional properties of biosurfactants: a review. Molecules, 29(11), 2544. https://doi.org/10.3390/molecules29112544
Elayaperumal, S., Sivamani, Y., Bhattacharya, D., Lahiri, D., & Nag, M. (2025). Eco-friendly biosurfactant solutions for petroleum hydrocarbon cleanup in aquatic ecosystems. Sustainable Chemistry for the Environment, 100207. https://doi.org/10.1016/j.scenv.2025.100207
El-Gebaly, E. (2020). Screening of biosurfactant production by bacterial strains isolated from oil contaminated sites near gas stations in Egypt. Al-Azhar Journal of Pharmaceutical Sciences, 61, 134–146. https://doi.org/10.21608/ajps.2020.86023
Faccioli, Y. E. D. S., De Oliveira, K. W., Campos-Guerra, J. M., Converti, A., Soares da Silva, R. D. C. F., & Sarubbo, L. A. (2024). Biosurfactants: Chemical properties, ecofriendly environmental applications, and uses in the industrial energy sector. Energies, 17(20), 5042. https://doi.org/10.3390/en17205042
Fei-Baffoe, B., Badu, E., Miezah, K., Sackey, L. N. A., Sulemana, A., & Amuah, E. E. Y. (2024). Contamination of groundwater by petroleum hydrocarbons: Impact of fuel stations in residential areas. Heliyon, 10(4). https://doi.org/10.1016/j.heliyon.2024.e25924
Ganiyu, S. A., Olobadola, M. O., & Adeyemi, A. A. (2023). Concentrations and health risk appraisal of heavy metals and volatile organic compounds in soils of automobile mechanic villages in Ogun State, Nigeria. Environmental Geochemistry and Health, 45(8), 6407-6433. https://doi.org/10.1007/s10653-023-01644-2
Garcia, K. L. M., Alemania, D. K. V., & Opulencia, R. B. (2026). Hydrocarbon degradation and genomic insights of Klebsiella pneumoniae from oil-contaminated soils in Guimaras Island, Philippines. Frontiers in Microbiology, 17, 1782430. https://doi.org/10.3389/fmicb.2026.1782430
Ge, H., Qiao, J., Zheng, J., Xu, H., Liu, R., Zhao, J., … Zheng, B. (2023). Emergence and clonal dissemination of KPC-3-producing Pseudomonas aeruginosa in China with an IncP-2 megaplasmid. Annals of Clinical Microbiology and Antimicrobials, 22(1), p.31. https://doi.org/10.1186/s12941-023-00577-z
Germer, A., Tiso, T., Müller, C., Behrens, B., Vosse, C., Scholz, K., ... Blank, L. M. (2020). Exploiting the natural diversity of RhlA acyltransferases for the synthesis of the rhamnolipid precursor 3-(3-hydroxyalkanoyloxy) alkanoic acid. Applied and Environmental Microbiology, 86(6), e02317-19. https://doi.org/10.1128/AEM.02317-19
Hamzah, A. F., Al-Mossawy, M. I., Al-Tamimi, W. H., Al-Najm, F. M., & Hameed, Z. M. (2020). Enhancing the spontaneous imbibition process using biosurfactants produced from bacteria isolated from Al-Rafidiya oil field for improved oil recovery. Journal of Petroleum Exploration and Production Technology, 10(8), 3767–3777. https://doi.org/10.1007/s13202-020-00874-9
Ja'afaru, M.I., Abbas, T., Ajunwa, O.M., & Olaifa, K. (2022). Characterization and statistical optimization of biosurfactant production using Bacillus subtilis isolated from automotive oil-contaminated soil in Yola, Nigeria. Scientific African, 17, p.e01357. https://doi.org/10.1016/j.sciaf.2022.e01357
Jain, S., Thomas, A., Zhuo, G. Y., & Mazumder, N. (2025). Fourier Transform Infrared (FTIR) Spectroscopy of Biomolecules. Recent Advances in Infrared Spectroscopy and Its Applications in Biotechnology, 101. https://doi.org/10.5772/intechopen.1010234
Jiang, M., Wang, H., Liu, J., Hou, X., Zhang, Y., Liu, X., ... Cui, Q. (2024). Isolation and Characterization of Biosurfactant-Producing Bacteria for Enhancing Oil Recovery. Processes, 12(11), p.2575. https://doi.org/10.3390/pr12112575
Jui, A. H., Bhuiyan, M. N. I., Bhowmik, B., Khatun, N., Chowdhury, A., Bhuiyan, R. H., ... & Afrin, S. (2024). Exploration and characterization of a newly isolated bacterium, Enterobacter quasihormaechei strain BDIFST24001, capable of producing rhamnolipid biosurfactant for oil remediation. Access Microbiology, 6(8), 000830-v4. https://doi.org/10.1099/acmi.0.000830.v4
Kaiyrmanova, G., Shaimerdenova, U., Assylbek, A., Amirgaliyeva, A., Yerzhan, A., & Yernazarova, A. (2025). Comprehensive Analysis of Formation Water Microorganisms for Their Biosurfactant Potential in MEOR Applications. Fermentation, 11(7), 367. https://doi.org/10.3390/fermentation11070367
Lavanya, M. (2024). Rhamnolipids: an insight to the overall characteristics of these extraordinary biomolecules. Green Chemistry Letters and Reviews, 17(1), 2371012. https://doi.org/10.1080/17518253.2024.2371012
Magri, M., & Abdel-Mawgoud, A. M. (2022). Identification of putative producers of rhamnolipids/glycolipids and their transporters using genome mining. Current Research in Biotechnology, 4, 152-166. https://doi.org/10.1016/j.crbiot.2022.02.002
Majeed, B. K., Shwan, D. M., & Rashid, K. A. (2025). A Review on Environmental Contamination of Petroleum Hydrocarbons, Its Effects and Remediation Approaches. Environmental Science: Processes & Impacts, 27, 526–548. https://doi.org/10.1039/D4EM00548A
Menkiti, N.A., Osuji, L.C. & Onojake, M.C. (2023). Hydrocarbon profile of oil-spill-impacted soils from Ogoni in Rivers State, Nigeria. Asian Journal of Applied Chemistry Research, 13(2), 1–1 15. https://doi.org/10.9734/ajacr/2023/v13i2238
Muvel, H., Jindal, M. K., Tewari, P. K., & Anand, V. (2025). Minimizing oil pollution: a review of current status and its treatment options. RSC Sustainability, 3(9), 3681-3723. https://doi.org/10.1016/j.totert.2023.100074
Nakano, Y., Domon, Y., & Yamagishi, K. (2023). Phylogenetic trees of closely related bacterial species and subspecies based on frequencies of short nucleotide sequences. Plos one, 18(4), p.e0268847. https://doi.org/10.1371/journal.pone.0268847
Ng, Y. J., Chan, S. S., Khoo, K. S., Munawaroh, H. S. H., Lim, H. R., Chew, K. W., ... & Show, P. L. (2023). Recent advances and discoveries of microbial-based glycolipids: Prospective alternative for remediation activities. Biotechnology Advances, 68, 108198. https://doi.org/10.1016/j.biotechadv.2023.108198
Nordin, A. R. R., Navarro, A. R., Reyes, J. C., Maragathavalli, S., Kristanti, R. A., Wulandari, R., & Bunrith, S. (2025). Microbial bioremediation of petroleum-contaminated soil: a sustainable approach. Tropical Aquatic and Soil Pollution, 5(1), 71-87. https://doi.org/10.53623/tasp.v5i1.683
Ozokolie, C. B., Amadi, C. C., Ezea, I. B., Iroha, N., Enyinnaya, I. C. N., Okeke, C. S., & Obashi, E. (2023). Contamination level of spent engine oil in the rhizosphere of Arachis Hypogea L. African Journal of Environmental Science and Technology, 17(5), 112–117. https://doi.org/10.5897/AJEST2023.3189
Patel, S., & Shah, G. (2023). Biodegradation of mixed hydrocarbon and eicosane by novel Enterococcus mundtii SS1 isolated from automobile service station soil. Sustainable Energy Technologies and Assessments, 55, 102993. https://doi.org/10.1016/j.seta.2022.102993
Ren, X. M., Chang, R. C., Huang, Y., Amorim Amato, A., Carivenc, C., Grimaldi, M., … Blumberg, B. (2023). 2,4-Di-tert-butylphenol induces adipogenesis in human mesenchymal stem cells by activating retinoid X receptors. Endocrinology, 164(4), bqad021. https://doi.org/10.1210/endocr/bqad021
Rice, E. W., Baird, R. B., Eaton, A. D. & Clesceri, L.S. (Eds.) (2012). Standard Methods for the Examination of Water and Wastewater (22nd ed.). American Public Health Association.
Rongsayamanont, W., & Phasukarratchai, N. (2025). Ultrasound-assisted extraction of biosurfactants from water hyacinth for enhanced soil washing of diesel-contaminated soils: performance evaluation and phytotoxicity assessment. Environmental Science and Pollution Research, 32(36), 21522-21542. https://doi.org/10.3390/pr8030322
Ropek, D. R., & Gospodarek, J. (2022). Entomopathogenic Nematode Steinernema feltiae as an Indicator of Soil Pollution with Oil Derivatives in Bioremediation Process. Agriculture, 12(12), 2033. https://doi.org/10.3390/agriculture12122033
Sałek, K., Euston, S. R., & Janek, T. (2022). Phase behaviour, functionality, and physicochemical characteristics of glycolipid surfactants of microbial origin. Frontiers in Bioengineering and Biotechnology, 10, 816613. https://doi.org/10.3389/fbioe.2022.816613
Shagufta, S. M. & Dharani, P. V. (2022). Screening and Isolation of Bacteria Producing Biosurfactants from Waste. Journal of Pure & Applied Microbiology, 16(1). http://dx.doi.org/10.22207/JPAM.16.1.55
Shu, Q., Lou, H., Wei, T., Liu, X., & Chen, Q. (2021). Contributions of glycolipid biosurfactants and glycolipid-modified materials to antimicrobial strategy: a re-view. Pharmaceutics, 13, 227. https://doi.org/10.3390/pharmaceutics13020227
Soltanighias, T., Singh, A. E., Satpute, S. K., Banpurkar, A. G., Koolivand, A., & Rahi, P. (2019). Assessment of biosurfactant-producing bacteria from oil-contaminated soils and their hydrocarbon degradation potential. Environmental Sustainability, 3, 285–296. https://doi.org/10.1007/s42398-019-00074-0
Stepanova, A. Y., Gladkov, E. A., Osipova, E. S., Gladkova, O. V., & Tereshonok, D. V. (2022). Bioremediation of soil from petroleum contamination. Processes, 10(6), 1224. https://doi.org/10.3390/pr10061224
Sui, X., Wang, X., Li, Y., & Ji, H. (2021). Remediation of petroleum-contaminated soils with microbial and microbial combined methods: Advances, mechanisms, and challenges. Sustainability, 13(16), 9267. https://doi.org/10.3390/su13169267
Yagoo, A., & Vilvest, J. (2023). Extraction of biosurfactant from Pseudomonas aeruginosa inhabiting oil-spilled soils. Future Journal of Pharmaceutical Sciences, 9(1), 58. https://doi.org/10.1186/s43094-023-00511-2
Zaman, S. A. U., Bhrdwaj, A., Nayarisseri, A., Khazanehdari, K. A., & Bhuyan, R. (2025). Isolation and characterization of novel hydrocarbon-degrading bacteria from oil polluted soil near Nacharam, Hyderabad, India. Scientific Reports, 15(1), 17219. https://doi.org/10.1038/s41598-025-01081-4
Zargar, A. N., Mishra, S., Kumar, M., & Srivastava, P. (2022). Isolation and chemical characterization of the biosurfactant produced by Gordonia sp. IITR100. Plos one, 17(4), e0264202. https://doi.org/10.1371/journal.pone.0264202
Recommended Citation
Babayola, Nafisa Mohammed; Lecky, Nasiru Bright; Onumaegbu, Chinweizu Obioma; Olajuyigbe, Olufunmiso Olusola; and Adefisoye, Martins Ajibade
(2026).
ASSESSMENT OF THE BIOREMEDIATION POTENTIALS OF BIOSURFACTANT-PRODUCING BACTERIAL STRAINS RECOVERED FROM SPENT ENGINE OIL-CONTAMINATED ENVIRONMENTS IN SOUTHWEST NIGERIA.
Journal of Environmental Science and Sustainable Development, 9(1), 111-128.
Available at: https://doi.org/10.7454/jessd.v9i1.1415