The bioleaching mechanism of Co and Li from spent lithium-ion batteries by mixed culture of sulfur-oxidizing and iron-oxidizing bacteria was investigated. It was found that the highest release of Li occurred at the lowest pH of 1.54 with elemental sulfur as an energy source, the lowest occurred at the highest pH of 1.69 with FeS(2). In contrast, the highest release of Co occurred at higher pH and varied ORP with S + FeS(2), the lowest occurred at almost unchanged ORP with S. It is suggested that acid dissolution is the main mechanism for Li bioleaching independent of energy matters types, however, apart from acid dissolution, Fe(2+) catalyzed reduction takes part in the bioleaching process as well. Co(2+) was released by acid dissolution after insoluble Co(3+) was reduced into soluble Co(2+) by Fe(2+) in both FeS(2) and FeS(2) + S systems. The proposed bioleaching mechanism mentioned above was confirmed by the further results obtained from the experiments of bioprocess-stimulated chemical leaching and from the changes in structure and component of bioleaching residues characterized by XPS, SEM and EDX.