羊肝蛋白酶解条件优化及酶解产物抗氧化活性研究

摘要

为优化羊肝蛋白酶解工艺条件及探讨其体外抗氧化活性,以水解度为评价指标,采用碱性蛋白酶酶解羊肝,在单因素试验基础上,通过响应面法优化羊肝蛋白的酶解工艺条件.结果表明,羊肝蛋白最佳酶解条件为酶解温度51℃、pH 8.5、酶解时间4.1h、加酶量0.40％.在此条件下,进行3次验证试验,测得羊肝酶解液实际水解度为(40.31±0.24)％.体外抗氧化试验结果表明,羊肝酶解产物具有一定的抗氧化活性,其清除羟自由基(·OH)、超氧阴离子自由基(O2-·)和DPPH自由基的IC50值分别为17.01 mg/mL、13.21 mg/mL和10.42 mg/mL,并具有一定的还原能力.%In order to optimize the sheep liver enzymolysis technology and explore itsin vitro antioxidant activity,using hydrolysis degree as evalua tion index,sheep liver was hydrolysed by alkaline protease.On the basis of single factor experiments,the enzymatic hydrolysis conditions of sheep liver proteins were optimized by response surface methodology.The results showed that the optimum enzymatic hydrolysis conditions were as follow:hydrolysis temperature 51 ℃,pH 8.5,hydrolysis time 4.1 h and enzyme addition 0.40％.Under the conditions,through three validation tests,the actu al hydrolysis degree of sheep liver hydrolysates was (40.31 ±0.24)％.The results ofin vitro antioxidant experiments showed that sheep liver hydrolysates had a certain antioxidant activities.The 1C50 of its scavenging hydroxyl radical,superoxide anion and DPPH free radical were 17.01 mg/ml,13.21 mg/ml and 10.42 mg/ml,respectively.Sheep liver hydrolysates had a certain reduction capacity.