Abstract The aim of this study is to assess the floatability of hematite using a crude biosurfactant (BS) extracted from Rhodococcus opacus. Throughout high-pressure ethanol extraction, unrefined surfactant was extracted from the bacteria with a yield of 0.3 g/L. An FTIR analysis on this extract, shows the presence of alcohol (–OH) and ketone (C=O) groups, as well as saturated and unsaturated carbon chains, which might originate from mycolates and trehalolipids moieties reported in literature. In addition, FTIR analysis on the modified mineral surface with the unrefined surfactant, confirms a strong interaction between hematite and the crude biosurfactant, showing similar vibrational peaks regarding the hydroxyl and carboxylate groups at 1631 and 3436 cm-1, respectively. Biosurfactant interaction with the mineral particles shifted the isoelectric point from 7.5 to 3.2, turning the hematite more hydrophobic in acidic environments. Surface tension measurements indicate that the unrefined surfactant is slightly more effective, decreasing the surface tension of water compared to the bacteria itself, with a variation of 4 mM/m. Finally, throughout microflotation tests in a Hallimond tube, it is shown that the surfactant doubles bacteria recovery efficiency, reaching a maximum recovery of around 95 % in acidic conditions, which is consistent with electrophoretic studies. The statistical models adjust well the experimental data, achieving R2 values of 93.6 and 91.1% for the hematite flotation using the biomass and the biosurfactant, respectively.