In situ biosurfactant production by hydrocarbon degrader microorganisms is an attractive approach in the bioremediation of oil contamination because of their compatibility, biodegradability, environmental safety, and stability under extreme environmental conditions. Given the high efficiency of bacteria in degrading petroleum hydrocarbons, the present work studied the detection and characterization of a biosurfactant‐producing hydrocarbon degrader, Roseomonas aestuarii NB833. This strain was able to synthesize a biosurfactant during the biodegradation of crude oil, which reduced the surface tension of the aqueous system from 70 to 34 mN m−1, with a critical micelle concentration of 200 mg L−1. The emulsification ability of the biosurfactant was sustained at various temperatures, pH values, and salinities. The biosurfactant chemical structure was identified via FT‐IR, LC‐MS, and NMR analyses. These analyses confirmed the production of surfactin‐C14 with a molecular mass of 1007 g mol−1. These results revealed the high potential of R. aestuarii NB833 as an in situ surfactin‐producing bacteria for bioremediation applications under extreme environmental conditions.