SBR reactor was performed to incubate polyphosphate accumulating organism (PAO), and it was checked out of the system by fluorescence in situ hybridization. As PAO is a kind of ordinary heterotrophic bacteria, it was excluded the ability of phosphate release and uptake and it was considered only the capacity of denitrification of the target biomass. The results indicated that acetate and PHB can be the electron donors of PAO to denitrify. When fed with acetate, the denitrifying rate and PHB producing rate were independent of initial nitrate concentration. However, served as more nitrate in the reactor, it would be less PHB produced and fewer nitrate reducing when using same amount of acetate. In view of PHB stored as an internal carbon and energy source, it presented as a reaction of zero-order to the substrate by PAO to denitrify, such as nitrate, besides, the specific denitrifying rate was 0.9733 mg/(g x h) and the specific PHB consuming rate was 2.4626 mg/(g x h).