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ABSTRACTOil (or petroleum), consisting of a mixture of hydrocarbons, can be leaked from oil exploration, production, and use. Due to their complex mixture and interaction with the subsurface soil and water, they are hard to treat and can become a significant environmental concern. Rhamnolipid and sophorolipid biosurfactants, biologically produced surfactants, can be used to remove petroleum hydrocarbons. Nanoparticles have gained attention as promising materials for soil remediation. In many studies, nanoparticles have shown their effective degradation of organic contaminants in the soil, both at bench and field scales. In this study, suspensions of Fe-Cu nanoparticles and biosurfactants were employed for the remediation of oil-contaminated soil. The results showed that these suspensions displayed a high oil removal rate from contaminated soil, which followed the first-order reaction. For batch experiments, the oil remediation efficiency was up to 84%, based on the experimental conditions. Optimum conditions to achieve the highest oil remediation performance included a rhamnolipid biosurfactant: nanoparticle ratio of 10:1 (wt%: wt%), pH 7, room temperature, and shaking speed of 60 rpm for 60 minutes. The remediation rate was improved by higher temperature and lower ionic strength. In the presence and absence of nanoparticles, rhamnolipid biosurfactant demonstrated a higher remediation efficiency than sophorolipid biosurfactant and ultraplex surfactant. The presence of other surfactants decreased the treatment productivity by 9-14% compared to using only rhamnolipid biosurfactant. Nanoparticles were reused with a remediation efficiency of 59% after three cycles by rhamnolipid biosurfactant. These results suggested that biosurfactants/Fe-Cu nanoparticle suspension showed promise for the remediation of oil-contaminated soil. This work will significantly benefit the soil remediation field by providing an environmentally friendly and economical method for the remediation of oil-contaminated soil.