Tunnel construction in watershed area of urban lakes would accelerate eutrophication by inputting nutrients into them, while mechanisms underlying the internal phosphorus cycling as affected by construction events are scarcely studied. Focusing on two main pathways of phosphorus releasing from sediment (enzymatic mineralization and anaerobic desorption), spatial and temporal variations in phosphorus fractionation, and activities of extracellular enzymes (alkaline phosphatase, β-1,4-glucosidase, leucine aminopeptidase, dehydrogenase, lipase) in sediment were examined, together with relevant parameters in interstitial and surface waters in a Chinese urban lake (Lake Donghu) where a subaqueous tunnel was constructed across it from October 2013 to July 2014. Higher alkaline phosphatase activity (APA) indicated phosphorus deficiency for phytoplankton, as illustrated by a significantly negative relationship between APA and concentration of dissolved total phosphorus (DTP). Noticeably, in the construction area, APAs in both sediment and surface water were significantly lower than those in other relevant basins, suggesting a phosphorus supply from some sources in this area. In parallel, its sediment gave the significantly lower iron-bound phosphorus (Fe(OOH)∼P) content, coupled with significantly higher ratio of iron (II) to total iron content (Fe(2+)/TFe) and dehydrogenase activities (DHA). Contrastingly, difference in the activities of sediment hydrolases was not significant between the construction area and other basins studied. Thus, in the construction area, subsidy of bioavailable phosphorus from sediment to surface water was attributable to the anaerobic desorption of Fe(OOH)∼P rather than enzymatic mineralization. Finally, there existed a significantly positive relationship between chlorophyll a concentration in surface water and Fe(OOH)∼P content in sediment. In short, construction activities within lakes may interrupt cycling patterns of phosphorus across sediment-water interface by enhancing release of redox-sensitive phosphate, and thereby facilitating phytoplankton growth in water column.