超声波辅助酶解法提取鲫鱼鱼鳞抗菌肽工艺优化

Optimization of the extraction process of antibacterial peptides from Carassius auratus scales by ultrasound-assisted enzymatic hydrolysis

中文关键词:  鲫鱼鱼鳞  超声波  酶解  抗菌肽

英文关键词:Carassius auratus scales  ultrasonic wave  enzymatic hydrolysis  antimicrobial peptides

中文摘要:

      目的 优化鲫鱼鱼鳞抗菌肽的超声波辅助酶解提取工艺。方法 本研究以对大肠杆菌的抑菌圈直径和多肽回收率为评价指标, 对比分析胃蛋白酶、胰蛋白酶、木瓜蛋白酶和中性蛋白酶对鲫鱼鱼鳞的酶解效果, 筛选最佳酶解用酶。通过单因素实验, 系统分析底物浓度、酶活力、酶解时间和超声波功率对鱼鳞抗菌肽提取效果的影响, 在此基础上, 选择酶活力、酶解时间和超声波功率3个关键因素进行正交优化实验。结果 胃蛋白酶为最佳酶解用酶, 在底物质量浓度200 mg/mL、酶解时间4 h、超声波功率120 W、酶活力5000 U/g的酶解工艺下, 获得的抗菌肽抑菌效果最佳, 对大肠杆菌的抑菌圈直径可达(2.66±0.66) cm, 多肽回收率为22.52%±1.64%。结论 采用超声波辅助酶解得到鲫鱼鱼鳞抗菌肽液为后续的抗菌肽分离、纯化及提取提供了重要的理论基础。

英文摘要:

      Objective To optimize ultrasonic assisted enzymatic extraction of antimicrobial peptides from Carassius auratus scales. Methods In this study, the diameter of inhibition zone against Escherichia coli and peptide recovery rate were used as evaluation indicators. The enzymatic effects of pepsin, trypsin, papain, and neutral protease on Carassius auratus scales were compared and analyzed to screen the optimal enzyme for enzymolysis. Through single-factor experiments, the effects of substrate concentration, enzyme activity, enzymolysis time, and ultrasonic power on the extraction efficiency of antimicrobial peptides from crucian carp scales were systematically analyzed. On this basis, three key factors (enzyme activity, enzymolysis time, and ultrasonic power) were selected for orthogonal optimization experiments. Results Pepsin was identified as the optimal enzyme for enzymolysis. Under the enzymatic hydrolysis process with a substrate mass concentration of 200 mg/mL, enzymatic hydrolysis time of 4 h, ultrasonic power of 120 W, and enzyme activity force of 5000 U/g, the antibacterial effect of the obtained antimicrobial peptide was the best, with a inhibition zone diameter of (2.66±0.66) cm against Escherichia coli and a short peptide recovery rate of 22.52%±1.64%. Conclusion The ultrasonic assisted enzymatic extract of antimicrobial peptides from Carassius auratus scales provides a critical theoretical foundation for subsequent separation and purification.