Three activated sludges enriched with glycogen accumulating organisms (GAO) were acclimatized respectively with different ratios of propionic to acetic acid (i.e. biomass SBR-A, C and E) . The effect of different ratios of propionic/acetic acid on the metabolism of long-term cultivated GAO was investigated. Cultivated with high propionic/acetic acid ratio, GAO consumed less glycogen and synthesized less poly-beta-hydroxyalkanoates (PHA) in the anaerobic phase, and in the aerobic phase accumulated less glycogen and degraded less PHA, and at the same time the microbial growth was lower. When the carbon mole of acetic acid equaled that of propionic acid in the influent, GAO utilized acetic acid faster than propionic acid. Batch tests were carried out with biomass SBR-A and SBR-E to study the transient response of long-term cultivated GAO to short-term change of propionic/acetic acid ratio. The GAO cultivated with a high propionic/acetic acid ratio was able to utilize acetic acid immediately when the concentration of acetic acid in the feed suddenly increased. But when the biomass cultivated with a low propionic/acetic acid ratio was feed with high ratio propionic/acetic acid wastewater, the propionic acid uptake rate was only 41.1% of the rate of the GAO long-term cultivated with high propionic/acetic acid. The sudden increase of propionic/acetic acid ratio could effectively inhibit the metabolism of GAO.