关键词: 黄粉虫, 抗菌肽, 酶解, 响应面

Abstract: Objective: To determine the optimal conditions for enzymatic preparation of Tenebrio molitor antibacterial peptides. Methods: Antibacterial peptides were prepared by hydrolyzing the protein extracted from Tenebrio molitor by alkaline extraction and acid precipitation. The antibacterial activity of the peptides was evaluated by agar diffusion method. One-factor-at-a-time method and response surface analysis based on Box-Behnken design were used to explore the effects of hydrolysis temperature, pH, hydrolysis time, enzyme dosage and substrate concentration on antibacterial activity of Tenebrio molitor antibacterial peptides. Results: Alkaline protease was the most efficient protease for the preparation of Tenebrio molitor antibacterial peptides, and the optimum hydrolysis conditions were found to be hydrolysis at 54 ℃ and pH 9.5 for 4.4 h with an enzyme dosage of 440 U/g pro and a substrate concentration of 810 g/100 mL. The predicted maximum inhibition circle diameter was 15.17 mm, close to the actual value of 15.04 mm. Conclusion: Peptides with high antibacterial activity can be prepared by hydrolyzing Tenebrio molitor protein with alkaline protease, and the optimized process conditions allow accurate theoretical prediction.

Key words: Tenebrio molitor, antibacterial peptide, enzymatic hydrolysis, response surface methodology