In this study, the effects of lysozyme, protease and α-amylase pretreatments for improving the hydrolysis and biodegradability of waste activated sludge (WAS) were investigated. The results showed that lysozyme was more effective in increasing the soluble chemical oxygen demand (SCOD) concentration in the liquid phase of sludge and improving the release of protein and carbohydrate from sludge flocculation to enhance sludge hydrolysis. After 8 h hydrolysis, the net SCOD increase in a reactor with lysozyme was 2.23 times and 2.15 times that of the reactors with protease and α-amylase, respectively. Meanwhile, lysozyme and protease could improve the lysis of microorganism cells and the dissolution of extracellular polymeric substances (EPS) to a certain extent, and lysozyme was more effective. Furthermore, the compositional characteristics of dissolved organic matter (DOM) and EPS were analyzed by EEM fluorescence spectroscopy and fluorescence regional integration (FRI) analysis. Tryptophan-like protein was the main component of sludge, which accounted for 31% and 38% of DOM and EPS, respectively. Lysozyme could decrease the percentage of non-biodegradable materials in sludge, such as humic acid-like substances and fulvic acid-like substances, so it could improve the biodegradability of sludge. This study can provide valuable information for future studies about hydrolytic enzyme pretreatments for WAS disposal.