The regulatory effects of environmental factors on denitrification were studied in the sediments of Meiliang Bay, Taihu Lake, in a monthly sampling campaign over a one-year period. Denitrification rates were measured in slurries of field samples and enrichment experiments using the acetylene inhibition technique. Sediment denitrification rates in inner bay and outer bay ranged from 2.8 to 51.5 nmol N2/(g dw (dry weight)·hr) and from 1.5 to 81.1 nmol N2/(g dw·hr), respectively. Sediment denitrification rates were greatest in the spring and lowest in the summer and early autumn, due primarily to seasonal differences in nitrate concentration and water temperature. For each site, positive and linear relationships were regularly observed between denitrification rate and water column nitrate concentration. Of various environmental factors on denitrification that we assessed, nitrate was determined to be the key factor limiting denitrification rates in the sediments of Meiliang Bay. In addition, at the two sites denitrification rates were also regulated by temperature. The addition of organic substrates had no significant effect on denitrification rate, indicating that sediment denitrification was not limited by organic carbon availability in the sediments. Nitrate in the water column was depleted during summer and early autumn, and this suppressed effective removal of nitrogen from Taihu Lake by denitrification.