Biosurfactants are metabolites produced by microorganisms during their growth and reproduction. They are amphiphilic molecules capable of modifying surface and interfacial tension. A petrophilic, nitrogen-fixing, phosphorus-solubilizing, and biosurfactant-producing bacterial strain was isolated and characterized. The isolated strain was cultivated in Kim medium with a C:N ratio of 2:1, from which its growth kinetics was determined. Samples of the culture broth were taken at different times and the collapsed drop, oil dispersion and emulsification index tests were performed. Maximum culture growth occurred at 72 h. The biosurfactant was extracted from the crude cell-free broth using a mixture of solvents and a yield of 2.21 g/L was obtained. Thin-layer chromatography and FT-IR analyzes revealed the presence of a lipopeptide biosurfactant. Tensiometry tests showed that the biosurfactant was able to lower the surface tension of water from 72 mN/m to 40 mN/m and the critical micelle concentration was 0.058 (w/w). Probit analysis for toxicity determination showed that there was no weight loss in Eisenia foetida specimens in a range between 10000 and 50000 ppm.