Abstract This study evaluates bioleaching treatments to remove copper from the Sarcheshmeh (Iran) molybdenite concentrate using a native strain of Acidithiobacillus ferrooxidans. Copper content of the concentrate was 0.83 wt.% as chalcopyrite. The tests showed selective dissolution of copper (chalcopyrite) from molybdenite concentrate. Up to 65% of copper content of molybdenite concentrate was removed via bioleaching with a native strain of A. ferrooxidans in less than 15 days. Ferrous sulfate, sulfur or pyrite was added to culture medium to enhance the activity of bacteria. Sulfur was the preferred additional source of energy for removing copper from molybdenite via bioleaching with A. ferrooxidans. In addition, 9K or Norris medium was also used as the culture medium in the experiments. The experiments showed that application of Norris medium would be better than that of 9K medium in order to remove copper from molybdenite via bioleaching. These results were backed up due to the fact that the cost of Norris medium was also less than that of 9K medium.