Abstract: In the extraction of chlorophyll from filter mud in sugar cane mill factories, volume fraction of 95% alcohol was used as extraction solvent.Chlorophyll extraction was investigated with respect to the effects of respective and synergic hydrolysis of filter mud by cellulase and pectinase. Three extraction parameters including enzymolysis temperature of mixed enzyme, enzymolysis time of mixed enzyme and ratio of material to liquid were optimized using central composite design and response surface methodology based on single factor investigations for achieving maximum the chlorophyll extraction quantity.The interaction of the respective variables and their influence on the chlorophyll extraction yield was studied by using box- behnken central composite design and response surface analysis theory, the simulated quadratic polynomial regression equation of prediction model was set up.The results indicated that more chlorophyll was extracted from filter mud by using both of the enzymes than using cellulase alone, the optimal conditions for extraction of chlorophyll were cellulase /pectinase ratio 2 ∶ 1, their total dosage 1.6g /100 g, enzymolysis temperature of 49℃, enzymolysis time of 3h and material to liquid ratio of1∶8 ( g /mL) . Under these conditions, the extraction yield of chlorophyll was 0.211mg·g- 1. Compared with the traditional extraction, enzymatic hydrolysis extraction revealed an enhanced yield of chlorophyll by 8%.