The bioleaching of pyrite and biosolubilization of rock phosphate (RP) in 9K basal salts medium were compared by the following strains of an autotrophic acidophilic bacterium, Acidithiobacillus ferrooxidans, a heterotrophic acidophilic bacterium, Acidiphilium cryptum, and mixed culture of At. ferrooxidans and A. cryptum. The results show that A. cryptum is effective in enhancing the bioleaching of pyrite and biosolubilization of RP in the presence of At. ferrooxidans, although it could not oxidize pyrite and solubilize RP by itself. This effect is demonstrated experimentally that A. cryptum enhances a decrease in pH and an increase in redox potential, concentration of total soluble iron and planktonic part bacterial number in the broth during pyrite bioleaching processes by At. ferrooxidans. The mixed culture of At. ferrooxidans and A. cryptum leads to the most extensive soluble phosphate released at 30 °C. Pulp density exceeding 3% is shown to adversely influence the release of soluble phosphate by the consortium of At. ferrooxidans and A. cryptum. It is essential to add pyrite to the 9K basal salts medium for the biosolubilization of RP by the mixed culture of At. ferrooxidans and A. cryptum, and the percentage of soluble phosphate released is the greatest when the mass ratio of RP to pyrite is 1:2 or 1:3.