Phosphorus is considered the primary nutrient limiting eutrophication. The release of phosphorus from sediments not only provides hydrophytes with essential nutrients, but can also produce entrophication in lakes. In this study, the sediment characteristics, phosphorus speciation, and environmental factors affecting phosphorus release from sediments from Taihu Lake, Tien Lake, and East Lake were investigated. The results indicated that phosphorus levels of sediments ranged from 647 ± 67 to 2343 ± 122 mg/kg. Inorganic phosphorus comprised the majority of total phosphorus. The main phosphorus speciation of sediments obtained from Taihu Lake and Tien Lake was Residual-P, and the main phosphorus speciation of sediment obtained from East Lake was NaOH-P. Phosphorus release occurred in acidic and alkaline conditions but more frequently under alkaline conditions. The amount of phosphorus released from the sediments increased as the disturbing intensity rose. In addition, less phosphorus was released in natural than anaerobic conditions. The amount of phosphorus released was elevated as temperature increased.