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Biosurfactants are the superb alternates for chemical surfactants due to non-toxicity, biodegradability and cost-effectiveness. The present study describes the biosurfactant production from two isolates Bacillus amyloliquefaciens SAS-1 and Bacillus subtilis BR-15. The produced biosurfactants were characterized by fourier transform infrared spectroscopy and mass spectrometry as lipopeptides, surfactins of molecular weight 1007, 1021, 1035 and 1049 Da. These molecules displayed an excellent emulsification (E₂₄ 60–78%) with hydrocarbons and stability at wide temperature (4–100 °C) and pH (4–10). The biosurfactants produced by SAS-1 and BR-15 accounted for 56.91 ± 1.52 and 66.31 ± 2.32% enhanced oil recovery respectively in sand pack column experiment on account of their high surface activity. Further, the biosurfactants efficiently augmented (75–94%) the engine oil degradation with microbial …