Bioleaching, a technologically and economically feasible technology, is considered as the high efficiency method to improve dewaterability in sewage sludge. The objective of this study was to investigate the effect of different sludge concentrations on bioleaching dewaterability and understand the mechanism of the effect of bioleaching on sludge dewaterability. Variation in pH, oxidation-reduction potential (ORP), capillary suction time (CST), specific resistance to filtration (SRF) and different fractions of extracellular polymeric substances (EPS) including slime EPS (S-EPS), loosely bound EPS (LB-EPS), and tightly bound EPS (TB-EPS) were determined. Different sludge concentrations (5, 10, 15, 20 and 30 g·L−1) were selected to investigate during bioleaching. Results indicated that sludge buffering capacity significantly inhibited bioleaching efficiency as sludge concentrations increased. Optimum enhancements in sludge dewaterability were observed during the 10 g·L−1 sludge concentration treatment, and reached a maximum when the pH was 2.11. The variation of different fractions of EPS revealed that the ratio of S-EPS/TB-EPS significantly affected sludge dewaterability. Principal component analysis and Pearson's correlation analysis both provided evidence that the higher TB-EPS followed by a very large reduction was positively correlated with sludge dewaterability. However, the increase of protein and DNA in S-EPS content was negatively correlated with sludge dewaterability.