超声预处理双酶分步水解制备滑菇抗氧化肽

Preparation of Antioxidant Peptides from Pholiota nameko Protein by Ultrasonic-pretreated Double Enzymatic Fractional Hydrolysis

研究超声预处理双酶分步水解滑菇蛋白制备抗氧化肽的工艺条件,为滑菇蛋白高附加值产品的开发提供理论依据。以羟自由基(·OH)清除率为指标,单因素试验研究功率、温度、时间等超声预处理参数对碱性蛋白酶水解效果的影响。从中性蛋白酶、胰蛋白酶、木瓜蛋白酶和风味蛋白酶等4种酶中筛选第二步水解的最适酶,通过正交试验优化其水解工艺。结果表明,超声波预处理的最佳工艺参数为:功率300 W、温度40℃、时间20 min,碱性蛋白酶水解产物的·OH清除率最高,为86.15%。风味蛋白酶作为第二步水解的最适酶,各因素对·OH清除率的影响程度依次为:水解时间>酶浓度>p H>温度,其最佳水解工艺条件为:酶浓度3 000 U/g、p H 7.0、温度55℃、水解时间1.5 h。在此条件下,水解产物·OH清除率为93.17%。超声波预处理双酶分步水解滑菇蛋白制备抗氧化肽的工艺可行。

To provide a theoretical basis for the development of high value added products from Pholiota nameko protein, the process condition of ultrasonic-pretreated double enzymatic fractional hydrolysis to produce antioxidant peptides were studied. According to the hydroxyl radical scavenging activity, the effects of ultrasonic pretreatment parameters such as power, temperature and time were analyzed through single factor experiment. The optimum protease for the second step hydrolysis was screened from neutral protease, trypsin, papain and flavourzyme, and the orthogonal test was used to optimize the factors affecting hydrolysis. The results showed that the optimum technology parameters of ultrasonic pretreatment were as follows： ultrasonic power 300 W, temperature 40 ℃ and ultrasonic time 20 min. Under this condition, the hydroxyl radical scavenging rate of the hydrolysate reached 86.15%. Flavourzyme was selected as the optimal enzyme for the second step hydrolysis, and the importance of the four factors affecting the hydroxyl radical scavenging activity was in the order of hydrolysis time, enzyme concentration, pH and temperature. The optimized process condition of ftavourzyme hydrolysis was as follows： enzyme concentration 3 000 U/g, pH 7.0, temperature 55℃ and hydrolysis time 1.5 h. Under this condition, the highest hydroxyl radical scavenging rate of the hydrolysate was 93.17%. The process technology for preparation of antioxidant peptides from Pholiota nameko protein by ultrasonic-pretreated double enzymatic fractional hydrolysis was feasible.