Effective removal of aging biofilm deserves to receive more attention. This study aimed to characterized aging biofilm from a full-scale moving bed biofilm reactor treating pharmaceutical wastewater and evaluate the hydrolysis effects of biofilm by different enzymatic treatments. Results from FTIR and biochemical composition analyses showed that it was a predominately organic-based biofilm with the ratio of total protein (PN) to polysaccharide (PS) of 20.17. A reticular structure of extracellular polymeric matrix (EPM) with filamentous bacteria as the skeleton was observed on the basal layer through SEM-EDS test. Among the four commercial proteases and amylases from Genencor (R), proteases were shown to have better performances than amylases either on the removal of MLSS and PN/MLSS or on DOC (i.e., dissolved organic carbon)/MLSS raising of biofilm pellets. Difference of dynamic fluorescence characteristics of dissolved organic matters after treated by the two proteases indicated distinguishing mechanisms of the treating process. (C) 2013 Elsevier Ltd. All rights reserved.