Abstract: In this study,the efficiency of enzymes(cellulase,pectinase and complex enzyme)on the hydrolysis of cell wall of H. pluvialis was studied. And response surface methodology was employed to optimize the hydrolysis conditions. Results indicated that when pectinase and cellulase mixed with the ratio of 1:1 (U/U),enzyme dosage 7000 U/mL,the optimal hydrolysis conditions were as follows:pH4.9,temperature 49 ℃,and time 6 h. Under these conditions,the theoretical and actual crude yield of astaxanthin were reached 70.21% and 71.08%±0.26%,with a relative error of 1.2%. This method of multi-enzymatic hydrolysis has the advantages of simple reaction,mild conditions,green security and high efficiency,which can apply in the practical extraction process of astaxanthin from H. pluvialis.