In this research, investigations were made on the effect of type and load of organic substrate on phosphorus release. Reactors of three different sizes were operated, being fed on five kinds of organic substrates. The quantitative analyses were made on phosphorus release and substrate utilization under anaerobic condition. The molar ratios of the uptaken organic substrate to the released phosphorus were 0.5 with acetate, 0.6 with glucose, 0.8 with glucose/acetate, and 1.2 with glucose/acids, respectively. The phosphorus release was inhibited at the higher organic load than the normal at stead state. Both acetate and acids/glucose enhanced phosphorus release- as well as uptake-rate, however, the complete phosphorus removal was achieved after the microbial adaptation to the new environment. In case with acetate, operation was hampered by the poor sludge settleability and phosphorus uptake was not enough although the phosphorus release was active. But with milk/starch, the phosphorus release and uptake was well developed even though phosphorus release was not comparatively high. From this study, it was concluded that organic substrates, such as glucose seemed to be converted fatty acids after fast bio-sorption, followed by concurrent uptake of these acids by excess phosphorus removing bacteria.