Enhanced biological phosphorus removal (EBPR) is not always successfully achieved in activated-sludge processes by anaerobic-aerobic operation. It has been reported that the deterioration of EBPR has been caused by domination of glycogen-accumulating organisms (GAOs) in microbial sludge. After repeated operation of a sequencing batch reactor (SBR) supplied with acetate as a sole carbon source, it was found that an operational variable, pH, could be a tool to control the competition between GAOs and polyphosphate-accumulating organisms PAOs, which could mediate the success of EBPR. When anaerobic phase pH in the SBR was controlled at 7.0, GAOs dominated the culture, resulting in failure of EBPR. Without pH control, the pH during the anaerobic phase increased to 8.4 as a result of denitrification and acetate uptake. In this relatively high pH condition, almost complete EBPR was achieved. In this report, the mechanism of pH effect on the competition between GAOs and PAOs was elucidated through a characterization of activated sludge using previously proposed biochemical models.