宽体金线蛭抗氧化活性肽的分离纯化及体外活性研究  

Separation,purification and in vitro activity of antioxidant peptides from Whitmania pigra

作　　者：尹娜[1] 林小清[2] 杨月伟[1] 徐海圣[2] 

机构地区：[1]曲阜师范大学生命科学学院,山东曲阜273165 [2]浙江大学动物科学学院,浙江杭州310058

出　　处：《浙江农业学报》2015年第3期348-353,共6页Acta Agriculturae Zhejiangensis

基　　金：浙江省重点科技创新团队项目(2011R50029);浙江省教育厅科研计划项目(Y201225771);湖州市科技计划项目(2011GN07);山东省自然基金(ZR2010CL013)

摘　　要：采用三水平四因素的正交试验,用木瓜蛋白酶酶解,以肽含量为指标确定了宽体金线蛭最佳酶解工艺,即加酶量7 500 U·g-1底物,酶解温度40℃,料液比1∶4（m/V）,酶解时间4 h,最初p H 8.5;测定了酶解原液及其经CM Sepharose FF分离纯化后活性较强部分清除ABTS自由基、羟基自由基、超氧阴离子自由基的活性。结果显示,宽体金线蛭短肽有较高的抗氧化活性,尤其经CM Sepharose FF分离纯化后的组分清除ABTS、羟基自由基活性明显强于抗坏血酸,但其清除超氧阴离子自由基活性较抗坏血酸弱。收集合并经CM Sepharose FF分离后活性强的部分,用Sephadex G-25脱盐,Q-TOF检测,抗氧化活性肽分子量大多为130.96~330.34 u。Separation of antioxidant peptides from enzymatic hydrolysates of Whitmania pigra and their antioxidation activity were studied. Ratio of solid to liquid,enzyme dose,temperature and hydrolysis time were selected as factors to optimize the hydrolysis parameters with four factors and three levels of the orthogonal experiment. Antioxidant peptides were separated and purified from the crude enzymatic hydrolysates by CM Sepharose Fast Flow and Sephadex G-25 gel chromatography. The antioxidant capability of each component and distribution of molecular weight were determined. Results showed the optimum condition for the enzymatic hydrolysis was as follows： ratio of solid to liquid1∶ 4,enzyme dose 7 500 U·g- 1,temperature 40℃,time 4 h and primary p H value 8. 5. The scavenging effects of antioxidant capability of each component on hydroxyl radical and ABTS radical were obvious and stronger than that of ascorbic acid. Whereas,their scavenging effects on superoxide anion radical were weaker than that of ascorbic acid.Moreover,the results indicated that molecular weight of these antioxidant peptides was mainly in the range of 130. 96-330. 34 u.

关 键 词：宽体金线蛭 寡肽 抗氧化活性 

分 类 号：S865.4[农业科学—野生动物驯养] Q74[农业科学—畜牧兽医]