In order to investigate effects of operation mode on the phosphorus removal and recovery performance of PAOs-biofilm, the continuous operating PAOs-biofilm reactor was carried out to recovery phosphorus from synthetic sewage wastewater with different operation modes. It showed that the operating modes resulted in significant differences of the environment for phosphorus uptake and release (phosphorus concentration in water). Under the “anaerobic-aerobic alternating” mode, the high phosphorus loading in aerobic phase strengthened the phosphorus uptake ability of the PAOs-biofilm, and the phosphorus recovery performance depended on the limitation of phosphorus uptake. In contrast, under the “aerobic-anaerobic alternating” mode, the high concentration of phosphorus in anaerobic phase depressed the phosphorus release ability of the PAOs-biofilm, which could be the main reason for the limited phosphorus recovery performance. Such differences in environment resulted from the operation modes led to different aerobic phosphorus uptake, anaerobic phosphorus release and carbon source utilization rate of biofilm. The average Prel/Cupt under “anaerobic-aerobic alternating mode” was 0.213 mmol P /mmol C, which was much higher than that of 0.046 mmol P/mmol C under “aerobic-anaerobic alternating” mode. In summary, the PAOs-biofilm showed higher efficiency in phosphorus removal under “anaerobic-aerobic alternating” mode, while it exhibited a more superior performance in phosphorus recovery under “aerobic-anaerobic alternating” mode, with phosphorus recovery liquid of 99.90 mg/L.