Wastewater treatment facilities in the Chesapeake Bay watershed are focusing their attention on nitrogen removal to meet future effluent limitations of about 3-4 mg N/L. External carbon is added during denitrification to facilitate the removal of nitrate. This study investigates the kinetics and stoichiometry of glycerol as an external carbon source for denitrification. Biodiesel-based glycerol acclimated mixed liquor was fed glycerol as an external carbon source in lab scale batch experiments to estimate specific denitrification rate (SDNR), maximum specific growth rate, C:N ratio, growth yield and half saturation constants for COD limiting conditions. The average SDNR for acclimated and unacclimated mixed liquor was 1.4 mg NO x -N /gVSS/hr. The maximum specific growth rate was estimated at 3.4 d -1 . Stoichiometric C:N ratio and growth yield were estimated as 4.2 mg COD/mg NO x -N and 0.32 mg biomass COD/mg NO x -N respectively. The half saturation constant was estimated between 5-8 mg COD/L.