Objective of the study was to isolate heavy metal resistant bacteria from chromiumcontaminated subsurface soil and investigate biosurfactant production and heavy metal bioremediation. Based on 16S rRNA gene sequence and phylogenetic analysis, the isolate was identified as Rahnella sp. RM. The biosurfactant production by heavy metal resistant Rahnella sp. RM was optimized using Box-Behnken design (BBD). The maximum emulsification activity was obtained 66 % at 6 % soybean meal in pH 7.0 and 33.5 °C. The biosurfactant was characterized using Field emission scanning electron microscopy (FE-SEM), Fourier transform infrared spectroscopy (FT-IR) and matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF). The highest metal removal rates using the biosurfactant were found 74.3, 72.5, and 70.1 %, respectively, at the 100 mg/L amended flasks at 48 h. This study indicated the biosurfactant from heavy metal resistant Rahnella sp. RM could be used as a potential tool to remediate the metals in contaminated environments.