This study sought to elucidate the effect and mechanism of Galactomyces sp. Z3 in improving the bioleaching of heavy metals from sludge. Results showed that co-inoculation of Galactomyces sp. Z3 and two Acidithiobacillus strains (Acidithiobacillus ferrooxidans LX5 and Acidithiobacillus thiooxidans TS6) reduced the period required for sludge bioleaching by 4.5 days compared to Acidithiobacillus alone. Further, removal efficiencies of Cu, Zn and oxidation rate of Fe2+ and S-0 were higher in co-inoculation system than the Acidithiobacillus alone. Galactomyces sp. Z3 consumed the acetate, propionate, iso-butyrate, butyrate, and iso-valerate in sludge from the initial concentrations of 109.50, 28.80, 7.70, 34.30, and 18.40 mg/L to 10.20, 0.61, 0.63, 19.40 and 1.30 mg/L, respectively, after 12 h in the co-inoculation system, significantly lower than the concentrations observed in the Acidithiobacillus alone. Meanwhile, the surfactant properties of the extracellular polymeric substances produced by the Galactomyces accelerated the rate of sulfur oxidization by A. thiooxidans. (C) 2013 Elsevier Ltd. All rights reserved.