响应面法优化超声波辅助酶解制备花生蛋白抗菌肽

Optimization of Peanut Antibacterial Peptides Preparation With Ultrasonic-assisted Enzymolysis by Response Surface Methodology

为了优化超声波辅助酶解制备花生蛋白抗菌肽工艺条件,以低温冷榨花生蛋白粉为原料,采用单因素试验和响应面Box-Benhnken试验设计方法,研究酶解得到的抗菌肽复合物对1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除率和对大肠埃希氏菌抑菌圈大小。结果表明,超声波辅助酶解制备花生蛋白抗菌肽复合物的最优工艺参数为底物浓度10%、初始pH值8.0、加酶量2.4 A·100 m L-1、超声波功率210 W、超声波频率45 k Hz、酶解温度47℃、酶解时间44 min;在此最佳工艺条件下,抗菌肽复合物对DPPH自由基清除率及大肠埃希氏菌的抑菌圈直径模型预测值分别为39.82%和1.77 cm,验证试验的DPPH自由基清除率(46.41%±2.10%)远大于模型预测值(39.82%),抑菌圈直径(1.70±0.04cm)与模型预测值(1.77 cm)相差不大。本研究结果为花生抗菌肽的分离、纯化、生物活性、构效关系及分子改造等方面的研究提供了理论依据。

In order to optimize the preparation process conditions of peanut protein antibacterial peptides with ultrasonic assisted enzymatic hydrolysis method,the low temperature-pressed peanut protein powder was chosen as the raw material,single factor experiments and response surface methodology based on Box-Behnken design were used to investigate the effects of antibacterial peptides complexes from enzymatic hydrolysis reaction on 1,1-diphenyl-2-picrylhydrazyl radical（DPPH） radical scavenging rate and the inhibiting bacterium circle of E. coli. The results showed that the optimum conditions obtained were as follows,substrate concentration,initial pH value,enzyme dosage,ultrasonic power,ultrasonic frequency,enzymolysis temperature and enzymolysis time of 10%,8. 0,2. 4 A·100 m L-1,210 w,45 k Hz,47℃ and 44 min,respectively. Under the optimum conditions,the model predictive DPPH radical scavenging rate and the inhibiting bacterium circle diameter of antibacterial peptides complexes were 39. 82% and 1. 77 cm,respectively. The DPPH radical scavenging rate in verification experiment could reach 46. 41% ± 2. 10%,which was much larger than the predictive value（39. 82%）. At the same time,the inhibiting bacterium circle diameter of verification experiment was 1. 70 ± 0. 04 cm,which was close to the model predictive value（1. 77 cm）. The results of this study provide a theoretical basis for the study of separation,purification,bioactivity,structure-function relationship and molecular modification of peanut antibacterial peptides.