The recent literature has unveiled the potential use of microbial strains in mineral bioprocessing. Because of their affinity for different mineral systems, such microbial strains may modify the surface properties and in this way change the characteristics of a mineral surface. Mineral bioflotation uses microorganisms as flotation reagents. In the present work, the electrokinetic behavior of particles of quartz and hematite, before and after interaction with two strains of Bacillus subtilis, was studied. The experiments revealed a shift of the isoelectric point (IEP) which of hematite that changed from 4 to 2.5 after interaction with the strain B. subtilis BAM, suggesting a chemical adsorption mechanism, while the interaction between hematite and B. subtilis GLI presented itself much more pronounced in the alkaline pH range. The experimental measurements of the contact angle (sessile drop method) were taken for the surfaces of the mineral particles (hematite equal 27.4 degrees, 13.0 degrees equal quartz) and for the B. subtilis BAM (32.0 degrees) and B. subtilis GLI (41.0 degrees) strains. The B. subtilis GLI strain was capable of modifying the surface of the hematite (46.0 degrees), and to a lesser extent, the quartz (23.3 degrees). The contact angle values were used to calculate the interface free energy components of quartz, hematite and the bacterial strains. The microflotation tests on a modified Hallimond tube evidenced the application of B. subtilis GLI as bioreagent. The best isolated flotability of quartz and hematite conducted by a solution of B. subtilis GLI (600 mg.L(-1)) was obtained at pH 6, with a recovery of 40 and 80 percent, respectively. Subsequently, the flotation performance of a synthetic mixture, quartz and hematite (in ratio 1:1) in the presence of 600 mg.L(-1) of the strain B. subtilis GLI at pH 6, was evaluated and showed a concentrate with a content of 74 percent Fe2O3. The DLVO and X-DLVO theories were applied to assess the energies of interaction between strains and minerals depending on the distance. The X-DLVO theory was able to preview the interaction between B. subtilis GLI and hematite, justifying the results of the flotation tests. The results of this study indicated that the strain B. subtilis GLI is promising as a bioreagent in the selective flotation of hematite relative to quartz.