Zhongguo youzhi (Jan 2023)
双酶制备花生肽及其对NO·的清除能力Preparation of peanut peptide by double enzymes hydrolysis and its ability to scavenge NO·
Abstract
为对花生免疫活性肽的制备提供基础，采用蛋白酶水解花生蛋白制备花生肽，以花生蛋白水解度和NO·清除率为指标，优化酶种类组合及水解条件。结果表明：先采用Alcalase 2.4L碱性蛋白酶在花生蛋白质量浓度5 g/100 mL、超声功率400 W、超声时间5 min、pH 8.0、加酶量7 400 U/g、水解温度55 ℃条件下水解3 h，再加入2 480 U/g的中性蛋白酶，在pH 7.0、水解温度40 ℃条件下继续水解30 min，花生蛋白水解度可达到24.73%，花生肽NO·清除率的IC50为1.88 mg/mL（显著低于商品大豆肽的3.51 mg/mL和商品酪蛋白肽的11.71 mg/mL）;水解度在7.8%～39.1%范围内与NO·清除率有极显著线性关系，水解度超过22.6%时，与NO·清除率无显著线性关系。较高的水解度是花生肽拥有良好NO·清除能力的必要条件，但并非水解度越高越好。 In order to provide a basis for the preparation of peanut immune active peptide, the peanut peptide was prepared by hydrolyzing peanut protein with protease, and the combination of enzyme species and hydrolysis conditions were optimized with the degree of hydrolysis of peanut protein and NO·scavenging rate as indicators. The results showed that the peanut protein was first hydrolyzed for 3 h with Alcalase 2.4L alkaline protease under the conditions of mass concentration of peanut protein 5 g/100 mL, ultrasonic power 400 W, ultrasonic time 5 min, pH 8.0, enzyme dosage 7 400 U/g, and hydrolysis temperature 55 ℃, then 2 480 U/g neutral protease was added to hydrolyze for 30 min under the conditions of pH 7.0 and hydrolysis temperature 40 ℃, the degree of hydrolysis of peanut protein could reach 24.73%, and the IC50 of NO· scavenging rate of peanut peptide was 1.88 mg/mL (significantly lower than commercial soybean peptide 3.51 mg/mL and commericial casein peptide 11.71 mg/mL). The degree of hydrolysis in the range of 7.8%-39.1% had a very significant linear correlation with NO· scavenging rate. When the degree of hydrolysis was over 22.6%, there was no significant linear correlation between the degree of hydrolysis and NO·scavenging rate. Higher degree of hydrolysis is the necessary condition for peanut peptide to have good NO · scavenging ability, but the higher degree of hydrolysis is not the better.
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