A defined mesophile consortium including Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans, and Leptospirilum ferriphilum was applied in bioleaching sediments contaminated with multiple heavy metals. Flask experiments showed that sulfur favored the acidification in the early stage while pyrite led to a great acidification potential in the later stage. An equal sulfur/pyrite ratio got the best acidification effect. Substrate utilization started with sulfur in the early stage, and then the pH decline and the community shift give rise to the utilization of pyrite. Solubilization efficiency of Zn, Cu, Mn, and Cd reached 96.1, 93.3, 92.13, and 87.65%, respectively. Bioleaching efficiency of other elements (As, Hg, Pb) was not more than 30%. Heavy metal solubilization was highly negatively correlated with pH variation. Logistic models were well fitted with the solubilization efficiency, which can be used to predict the bioleaching process. The dominant species in the early stage of bioleaching were A. ferrooxidans and A. thiooxidans, and the abundance of L. ferriphilum increased together with pyrite utilization and pH decline.