Previous research showed the effectiveness of bacterial strains as flotation reagents on Hematite beneficiation. The aim of this work is to study and evaluate Rhodococcus ruber as a biocollector. The sample was conditioned with the biomass suspension by stirring under specific conditions as particle size, biomass concentration, pH solution and conditioning time. The results showed a change in hematite zeta potential profile after interaction with R. ruber, and its adhesion onto the mineral surface was higher at pH 3 and at concentration of 0.60 g/L (109 cells/mL). Flotation studies were carried out in a 0.23 L modified Partridge-Smith cell flotation, and the highest floatability (84%) was achieved at size fraction -53 μm +38 μm under the conditions mentioned before. Complementary floatability studies were performed using the conventional frother Flotanol D24 combined with the R. ruber biomass, finding interesting results for the bigger particle size range. Thus, this research aims to evaluate the efficiency of bioflotation of minerals, particularly hematite, and the potential use of R. ruber as biocollector, projecting its future application in mineral flotation industry.