Optimization of Hydrolysis Conditions for Obtaining Antithrombotic Peptides from Tenebrio Molitor Larvae

Tenebrio molitor larvae was hydrolyzed by pepsin and trypsin for preparing high antithrombotic activity hydrolysate. The significant variables were selected according to the Plackett-Burman design and further optimized by the Response Surface Methodology (RSM). Four factors, including substrate concentration, pepsin digestion time, pepsin amount and trypsin amount, respectively, influenced significantly on the hydrolysis effectiveness. The best hydrolysis conditions obtained using RSM experiments are as following: substrate concentration of 19.8 mg/mL, pepsin digestion time of 1.8 h, pepsin amount of 1634 U/mL and trypsin amount of 126 U/mL, respectively. Under the optimal hydrolysis conditions, the antithrombotic activity was 82.8% predicted by the RSM model, which was basically consistent with the 80.6% obtained in the three verification experiments. Moreover, the antithrombotic activity of the hydrolyzate had no linear relationship with the degree of hydrolysis.