鹿茸抗氧化多肽高压脉冲电场辅助酶法提取及纯化工艺优化

Process optimization for purification and preparation of hairy antler grounds antioxidant peptide assisted by high-intensity pulsed electric fields

以鹿茸渣为研究对象,利用高压脉冲电场辅助菠萝蛋白酶制备鹿茸抗氧化肽,并进行分离纯化。以水解度和DPPH·清除率为评价指标,通过单因素及响应面优化试验对鹿茸渣制备抗氧化活性多肽的酶解工艺进行优化。采用大孔吸附树脂对鹿茸抗氧化肽进行脱盐及纯化,并分析纯化后鹿茸肽的抗氧化活性。结果表明,抗氧化活性肽的最佳制备工艺为:底物浓度15%、酶添加量7.04 U/mg,电场强度17.3kV/cm,脉冲数7,该条件下制备的鹿茸蛋白水解液对DPPH·清除率达(74.76±2.03)%。75%乙醇溶液洗脱组分C3对DPPH·、·OH及O_2^-·的清除率最高,相比粗多肽的抗氧化活性大幅提高,鹿茸抗氧化肽具有较强的抗氧化活性,是制备抗氧化活性肽的良好原料。

Antioxidant peptides were prepared from hairy antler grounds by high-intensity pulsed electric fields（PEF）jointed bromelain,and then they were isolated and purified.The degree of hydrolysis and scavenging rates of DPPH· was considered as test indexes.According to the results of the single factor experiment,the response surface methodology（RSM）experiment was conducted on three factors at five levels to analyze the factors and interaction effects on the scavenging rates of DPPH·.Then the macroporous resin was used to desalt,separate and purify hairy antler antioxidant peptides,and the antioxidant activities of hairy antler peptides were also analyzed.The optimal conditions of enzymatic hydrolysis are as follows：the substrate concentration 15%,bromelain concentration 7.04 U/mg,electric filed intensity 17.3 kV/cm,pulse number 7,the scavenging rates of DPPH· by hairy antler antioxidant peptides was（74.76±2.03）%.The scavenging rates of DPPH·,·OH and O2^-· of 75%ethanol elution component C3 were the highest,and the antioxidant activity of elution component C3 sharply increased compared with crude peptide,when hairy antler peptides were featured with a considerable antioxidant activity,and hairy antler could be a potential source of preparing peptide with the strong antioxidant activity.