The principle for the electro-generative simultaneous leaching (EGSL) is applied to simultaneous leaching of pyrite-MnO2 in this paper. A galvanic system for the bio-electro-generative simultaneous leaching (BEGSL) has been set up. The equation of electric quantity vs. time is used to study the effect of produced sulfur on electro-generative efficiency and quantity. It has been shown that the resistance decreased in the presence of Acidithiobacillus thiooxidans (A. thiooxidans) with the increase of electro-generative efficiency. The effects of temperature and grain size on rate of ferrous extraction from pyrite under the conditions of presence and absence of A. thiooxidans were studied, respectively. The changes in the extraction rate of Fe2+ as particle size in presence of A. thiooxidans were more evident than that in the absence, which indicated that the extraction in bio-electro-generative leaching was affected by particle size remarkably. Around the optimum culture temperature for A. thiooxidans, the bigger change in the conversion rate of Fe2+ was depending on temperature. The transferred charge in BEGSL including part of S0 to sulfate group in the presence of (A. thiooxidans) which is called as biologic electric quantity, and the ratio of biologic electric quantity reached to 58.10% in 72 h among the all-transferred charge.