A study was undertaken to investigate the production of amino acids from excess activated sludge (EAS) by enzymatic hydrolysis. Firstly, the protein was extracted from EAS. Secondly, the protein solution was further hydrolyzed under free enzyme or immobilized enzyme. The reversed phase high performance liquid chromatography (RP-HPLC) and inductively coupled plasma emission spectrometer (ICP) were applied to determine the contents of amino acids and heavy metals, respectively. The effects of enzyme/substrate(E/S), pH, temperature, and reaction time were investigated in detail. The results indicated that, the optimum conditions for protein hydrolysis were temperature 55℃, pH 10, E/S 9 g/L, and reaction time 8 h, and the highest yield of amino acids was more than 10 g/100 g dry sludge (DS) under free enzyme. Moreover, the security and nutrition were taken into consideration. There were seven kinds of essential amino acids and ten non-essential amino acids in the raw amino acid (RAA) solution, and the contents of heavy metals were lower, living up to Hygienical standard for feeds (China). This technology widens the source of amino acids and makes the extraction of amino acids from EAS more economic and effective.