浙江农业学报

• 动物科学 • 上一篇    下一篇

宽体金线蛭抗氧化活性肽的分离纯化及体外活性研究

  

  1. (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)
  • 出版日期:2015-03-25 发布日期:2015-03-31

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

  1. (1 College of Life Sciences, Qufu Normal University, Qufu 273165, China; 2 College of Animal Sciences, Zhejiang University, Hangzhou 310058, China)
  • Online:2015-03-25 Published:2015-03-31

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pH 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。

关键词: 宽体金线蛭, 寡肽, 抗氧化活性

Abstract: 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 liquid 1∶4, enzyme dose 7 500 U·g-1, temperature 40℃, time 4 h and primary pH 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.

Key words: Whitmania pigra Whitman, oligopeptides, antioxidant activity