This work aimed to evaluate the electroflotation process of fine and ultrafine iron ore particles using biosurfactant extracted from Rhodococcus Opacus bacteria. Characterization studies of iron ore were carried out (particle size analysis, X-ray diffraction and X-ray fluorescence). Through infrared spectroscopy measurements it was possible to identify a possible interaction of the biosurfactant with the mineral, the hematite spectrum after the interaction showed characteristic functional groups of the biosurfactant (NH, CH2, C=O, COO-e PO2-).The measurements of zeta potential suggest a possible electrostatic interaction between the hematite and the biosurfactant, by shifting the isoelectric point from 5.3 to 3.5.Contact angle measurements suggest that after interaction, hematite may have become more hydrophobic, changing the contact angle from 40 degrees to 60 degrees.Surface tension measurements were performed to determine the physico-chemical properties of the biosurfactant. According to the results, the biosurfactant showed tensioactive characteristics, reducing the surface tension from 71 mN/m to 40 mN/m in 25 mg/L of biosurfactant, the formation of micelles occurred from 300mg/L of biosurfactant. Finally, electroflotation tests were performed with two size fractions of iron ore (-38+20 micrometre and -20 micrometre) to evaluate the pH (3-11), biosurfactant concentration (50-800 mg/L), current density (5.28-16mA/cm2) and solution agitation (300-700 rpm). According to the results obtained, the increase in the pH impaired the metallurgical recovery and iron grade. The best results occurred at pH 3, this behavior can be attributed to electrostatic interactions that occur in this pH range. The increase of the biosurfactant concentration favored metallurgical recovery and the iron grade, this behavior remained up to 300 mg/L, above this value occurred a decline probably caused by the formation of micelles. The increase in current density also favored metallurgical recovery and iron content. For the sample with particle size range from -38+20 micrometre the maximum metallurgical recovery was around 76 percent and 59 percent iron grade and the sample with particle size from -20 micrometre the maximum metallurgical recovery and grade of Fe were around 90 percent and 64 percent, respectively. The results obtained show a potential use of the electroflotation technique in the recovery of fine and ultrafine iron ore particles, using the biosurfactant extracted from the Rhodococcus opacus bacterium.