In order to control the water eutrophication, microorganism agents with high capability of denitrifying phosphorus accumulation are required. A denitrifying poly-phosphate accumulating organisms(DNPAOS), B8(Pseudomonas putida sp.)was isolated from activated sludge in oxidation ditch of Anhui Tianchang wastewater treatment plant. Intracellular polyphosphate granule(Poly-P)dyeing method was used to optimize the culture conditions of Poly-P-producing Pseudomonas putida strain B8 seed liquid. The results showed that the appropriate culture conditions were as follow: pH value and temperature ranged from 6.5 to 7.0 and 30~32 ℃m respectively. The saturation values of dissolved oxygen varied from 70% to 88%(when velocity was in range of 120~140 r/min), 15~20 h proved to be the cultivation time. The seed liquid produced by B8 showed high phosphorus uptake rate in the processes of anaerobic-anoxic and anaerobic-aerobic in wastewater(89.73% and 94.09% respectively)In the anaerobic-anoxic process, the highest nitrate removal efficiency was 53.48%. Extracellular polymeric substances(EPS)extracting method and analysis of phosphorus removal characterization revealed that phosphorus removal was mainly caused by intracellular uptake, rather than by extracellular biological adsorption.