An investigation of hydrolysis of particulate organic substrate by activated sludge has been made. Raw municipal wastewater was used as substrate. It was mixed with activated sludge from a high loaded activated sludge plant with pure oxygen aeration. During 4 days batch experiments under aerobic, anoxic and anaerobic conditions, the hydrolysis was following through the production of ammonia. The hydrolysis rate of nitrogeneous compounds is significantly affected by the electron donor available. The rate is high under aerobic conditions, medium under anaerobic conditions and low under anoxic conditions. The ratio between the hydrolysis rates under aerobic and under anoxic conditions are very similar to the respiration rates measured as electron equivalents.