Biosurfactants plays a key role in tertiary recovery (EOR), and production, excessive minerals discharge from water during petroleum refinery process, environmental utilization and eco-friendly. Biosurfactant producing bacteria found to be sufficient in hydrocarbon-polluted soil samples; it is expected to more amounts of agrichemicals contemporary in the clay. These bacteria establish itself soil and region specific. So, in this study we made an attempt to identify and characterize biosurfactant produced by achromobacter xylos strain GSR21 from hydrocarbon polluted soil in Andhrapradesh, India. A battery of biosurfactant screening methods engaged were haemolytic activity, oil spreading technique, lipase activity, emulsification index (E24), emulsification assay, tilting glass slide, blood haemolysis test, drop-collapsed assay, and foaming activity. The organism isolate was studied based on molecular, phenotypic, and biochemical methods. Thin-layer chromatography I(TLC), Fourier transforms infrared spectroscopy (FTIR) and gas chromatography-mass spectrometry (GC–MS) investigates were used to identify and characterize the biosurfactant produced. The isolated biosurfactant was applied on chosen hydrocarbons to measure its emulsifying capacity.The phylogeny study of the 16S rRNA classified the isolate as A chromobacter xylos strain GSR21. The sequence secured from the isolate has been accumulated in GenBank covered by the accession number JQ746488. The result obtained from the study acknowledge high biosurfactant action with a maximum emulsification index (E24) of 62 % compared to emulsification index (E24) of 72% by sodium lauryl sulfate (SLS). Moreover, the biosurfactant appear emulsifying activity against the following hydrocarbons: diesel, methylbenzene, kerosene, dimethylbenzene, and petrol. The optimum cultural conditions (incubation time, carbon, pH, hydrocarbon, inoculum concentration, nitrogen, and temperature) for growth and biosurfactant produced by A chromobacter xylos GSR21 were analysed. The biosurfactant was characterized as a glycolipid using thin layer chromatography (TLC), while the gas chromatography-mass spectrometry (GC–MS) technique analyzed the glycolipid as dodecanoic acid-undecyl ester.Present study has exhibited the magnitude of Achromobacter xylos strain GSR21 isolated from hydrocarbon-polluted soil to produce biosurfactant and the effectiveness of the produced biosurfactant in emulsifying different hydrocarbons. Moreover, the biosurfactant produced was established to be held by the class, glycolipid based on the thin layer chromatography (TLC) and gas chromatography-mass spectrometry (GC–MS) analyses.