Abstract: Box-Benhnken’s central composite design and response surface method, polynomial regression model on the main influencing factors to enzymolysis of forest frog oil with papain such as substrate concentration, enzyme dosage,enzymolysis temperature, time and pH value was established and they were optimized. Meanwhile, its enzymatic hydrelysates were determined by mass sepectrometry. The optimal enzymatic hydrolysis conditions are determined as: substrate concentration 1%, enzyme dosage 1000 U/g, temperature 60 ℃, time 3.0 h, pH 6.5. According to determination results of laser desorption ionization time- of-fligh mass spectrometer, the hydrolysates of four different componetns, whose molecular weight are 712.4, 826.6, 1101.2 and 1326.1 D respectively.

Key words: response surface method, enzymolysis, molecular weight