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Acidithiobacillus thiooxidanshas been used in bioleaching processes, being recognized for its high metal leaching ability from industrial wastes. In the present work, the sulfur-oxidizing activity of A. thiooxidansDSM 26636, adapted to increasing concentrations of ore was assessed, finding that at 28 days with 3% (w/v) of ore sample, the culture produces high amounts of the sulfate ion (SO4−2) and sulfuric acid (H2SO4) and causes a decrease in the pH value, compared to the negative control. Also, there was observed a net removal of As, Zn, Fe, Al, Cu, and Au in concentrations of 48.23 ± 3.11, 264.02 ± 26.27, 9055.81 ± 210.39, 605.29 ± 30.26, 2158 ± 359.04, and 0.99 ± 0.37 mg/kg, respectively. Thus, the present work shows the potential of A. thiooxidansDSM 26636 to be used for the extraction of metals contained in ores, especially for those containing Cu, Zn, As, Fe, and Al.