Abstract: This study aims to observe different factors which affected the bionic enzymatic hydrolysis of icariin into baohuoside Ⅰ and to optimize the reaction conditions in order to provide research foundation for building a novel bionic enzymolysis drug delivery system. To simulate the environment in vivo, 37℃ was set as the temperature and artificial intestinal juice and gastric juice were selected as the buffer solutions. Taking the conversion of baohuoside Ⅰ as index, the effects of the kinds of enzyme, enzyme activity, substrate concentration, reaction time, pancreatin in artificial intestinal juice and surfactant on the conversion of baohuoside Ⅰ were investigated. The results showed that cellulase, β-glucosidase and snailase were all inactive in artificial gastric juice and no baohuoside Ⅰ generated. Pancreatin in artificial intestinal juice couldn't significantly influence the activity of β-glucosidase or snailase (P > 0.05), but noticeably decrease the activity of cellulase (P <0.05). In artificial intestinal juice, the conversion of baohuoside Ⅰ was highest by using β-glucosidase, and the optimum reaction conditions were determined as follows: enzyme activity 10 U·mL^-1, substrate concentration 1 mg·mL^-1, 3 g·L^-1 rhamnolipid and reaction time 3 h. Under this condition, the conversion of baohuoside Ⅰ was 99.8%.