中国畜牧兽医

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牛血Cu/Zn-SOD提取条件的研究

刘进,张峰,魏建民   

  1.  (内蒙古农业大学生命科学学院,内蒙古呼和浩特 010018)
  • 收稿日期:2013-07-08 出版日期:2014-02-20 发布日期:2014-03-27
  • 通讯作者: 张峰,男,教授,博士生导师,目前主要从事功能纳米复合材料制备、生物大分子自组装与人工操纵、蛋白质淀粉样纤维化机理、超灵敏检测、疏水药物包裹及靶向传送等研究工作。 魏建民,男,教授,硕士生导师,从事蛋白质化学研究。
  • 作者简介:刘进(1987—),男,内蒙古人,硕士生,研究方向:生物化学与分子生物学。
  • 基金资助:

    新型细胞内离子纳米传感器的开发;国家自然科学基金(医学部地区项目、81160213、30101-121152)。

Study on Extraction Conditions of Cu/Zn-SOD for Bovine Serum

LIU Jin,ZHANG Feng,WEI Jian-min   

  1.  (College of Animal Science,Inner Mongolia Agricultural University,Hohhot 010018,China)
  • Received:2013-07-08 Online:2014-02-20 Published:2014-03-27

摘要: 本研究以牛血为试验材料,研究了Cu2+浓度、Zn2+浓度、热变性温度和热变性时间4种单因素分别对牛血中Cu/Zn-SOD活性的影响。以4种单因素为基础设计了4因素5水平正交试验,通过邻苯三酚法测定了酶活性,考马斯亮蓝法测定了蛋白含量,聚丙烯酰胺凝胶电泳测定了蛋白纯度,确定了牛血Cu/Zn-SOD的最佳提取条件。结果表明:当Cu2+、Zn2+分别为6和12 mmol/L条件下,90 ℃热变性35 min,可得到较高活性和比活力的Cu/Zn-SOD。本研究意在为更简便、经济的牛血中Cu/Zn-SOD提纯方法提供参考。

关键词: 牛血; Cu/Zn-SOD; 提取条件; 活性

Abstract: In this assay, bovine blood as a sample, was studied the effect of four factors that concentrations of Cu2+ and Zn2+, denaturation temperature and thermal time on the activity of Cu/Zn-SOD in bovine blood. Four factors and five levels of orthogonal experiment was designed, and combined with methods of pyrogallol to measure the activity of enzyme, Coomassie brilliant blue method to determine the content of Cu/Zn-SOD protein, and polyacrylamide gel electrophoresis to detect the purity of the protein. We finally found the optimum extraction conditions of Cu/Zn-SOD in bovine blood. The results showed that when the concentrations of Cu2+ and Zn2+ were 6 and 12 mmol/L,respectively,at 90 ℃ thermal denaturation with 35 min, we could get Cu/Zn-SOD which had a higher activity. This study was aimed to provide a convenient and economical method for Cu/Zn-SOD extraction in bovine blood.  

Key words: bovine blood; Cu/Zn-SOD; extraction conditions; activity