The role of calcium (Ca) in enhanced biological phosphorus removal and its possible implications on the metabolic pathway have been studied. The experience has been carried out in an SBR under anaerobic-aerobic conditions for biological phosphorus removal during 8 months. The variations of influent Ca concentration showed a clear influence on the EBPR process, detecting significant changes in Y(PO4). These Y(PO4) variations were not due to influent P/COD ratio, pH, denitrification and calcium phosphate formation. The Y(PO4) has been found to be highly dependent on the Ca concentration, increasing as Ca concentration decreases. The results suggest that high Ca concentrations produce "inert" granules of polyphosphate with Ca as a counterion that are not involved in P release and uptake. Furthermore, microbiological observations confirmed that appreciable changes in PAO and GAO populations were not observed. This behaviour could suggest a change in the bacterial metabolic pathway, with prevailing polyphosphate-accumulating metabolism (PAM) at low influent Ca concentration and glycogen-accumulating metabolism (GAM) at high concentration.