响应面优化花生蛋白抗菌肽制备工艺

Optimization of Peanut Antibacterial Peptide Preparation by Response Surface Methodology

为了研究超声波辅助酶解制备花生抗菌肽工艺,以低温冷榨花生蛋白粉为原料,以蛋白酶解得到的抗菌肽复合物对枯草芽孢杆菌、大肠杆菌和米曲霉抑菌率为考察指标,在单因素试验基础上,通过响应面Box-Benhnken试验设计进行工艺优化。结果表明:超声波辅助酶解制备花生抗菌肽的最优工艺参数为底物浓度10.9%、初始pH值8.3、加酶量26.6 AU·L^(-1)、超声波功率210 W、超声波频率28 kHz、反应体系温度48℃、反应时间40 min;此工艺条件下的抗菌肽复合物对枯草芽孢杆菌、大肠杆菌和米曲霉抑菌率的验证试验值分别为132.9%±0.4%、128.6%±0.6%和84.1%±0.8%,与理论值的差异幅度分别为1.68%、1.38%和1.52%,说明模型拟合优度较好,预测精确度较高。本研究结果为花生抗菌肽的分离、纯化及应用等提供了理论基础。

The aim of this study was to optimize the preparation conditions of peanut antibacterial peptides by ultrasonic assisted enzymatic hydrolysis reaction. On the bases of single factor experiments, the response surface methodology(RSM) and Box-Behnken designs were used to determine the optimal technological conditions using the low-temperature pressed peanut protein powder as raw material and the inhibition rates of Bacillus subtilis, Escherichia coli and Aspergillus oryzae as main indexes. The results showed that the optimum conditions were substrate concentration, initial pH value, enzyme dosage, ultrasonic power, ultrasonic frequency, reaction temperature and time of 10.9%, 8.3, 26.6 AU·L^(-1), 210 W, 28 kHz, 48℃ and 40 min, respectively. Under these conditions, the inhibition rates of Bacillus subtilis, Escherichia coli and Aspergillus oryzae in verification experiment could reach 132.9%±0.4%, 128.6%±0.6% and 84.1%±0.8%, respectively, and their differences with predictive values were 1.68%, 1.38% and 1.52%. It could be concluded that the model fitted well with the actual situation and verified the correctness of the prediction model. Theses results could provide a theoretical basis for the isolation, purification and application of peanut antibacterial peptides.