食品科学 ›› 2013, Vol. 34 ›› Issue (9): 80-84.doi: 10.7506/spkx1002-6630-201309018

• 基础研究 • 上一篇    下一篇

磁性纳米粒子快速分离纯化超氧化物歧化酶

杨 林1,孙术国2,罗 章1,*   

  1. 1.西藏农牧学院食品科学学院,西藏 林芝 860000;
    2.中南林业科技大学食品科学与工程学院,稻谷及副产物深加工国家工程实验室,湖南 长沙 410004
  • 收稿日期:2012-11-14 修回日期:2013-04-11 出版日期:2013-05-15 发布日期:2013-05-07
  • 通讯作者: 罗章 E-mail:luozhangvip@163.com
  • 基金资助:

    西藏自治区科技厅自然科学基金项目

Fast Separation and Purification of Superoxide Dismutase Using Magnetic Nanoparticles

YANG Lin1,SUN Shu-guo2,LUO Zhang1,*   

  1. 1. College of Food Science, Tibet Agricultural and Animal Husbandry College, Linzhi 860000, China;
    2. National Engineering Laboratory for Rice and Byproduct Process, Faculty of Food Science and Engineering,
    Center South University of Forestry and Technology, Changsha 410004, China
  • Received:2012-11-14 Revised:2013-04-11 Online:2013-05-15 Published:2013-05-07
  • Contact: Zhang LUO E-mail:luozhangvip@163.com

摘要:

磁性纳米粒子快速分离纯化牦牛血液中超氧化物歧化酶(SOD),并建立吸附动力学模型,研究结合液(磷酸盐缓冲溶液)和解离液(醋酸钠缓冲溶液) pH值、NaCl浓度和吸附时间对SOD比活力和回收率的影响,以此确定最佳分离纯化条件。结果表明:随着结合液pH值增大,SOD比活力先升高后急剧降低,SOD回收率一直呈增大趋势,最佳pH值为7.5;随着结合液NaCl浓度增加,SOD比活力和回收率先升高,后降低,最佳NaCl浓度为0.5mol/L;随着解析液pH值增大,SOD比活力逐渐升高,而SOD回收率呈减小降趋势,最佳pH值为5.0;磁性纳米粒子对SOD最佳吸附时间为2h;随初始酶粗品浓度的增加,磁性纳米粒子对SOD吸附作用拟合Langmuir吸附模型,且获得的SOD比活力始终维持在较高水平(3000~3250U/mg),说明磁性纳米粒子对SOD分离效果良好。

关键词: 牦牛血液, 超氧化物歧化酶, 比活力, 吸附动力学

Abstract:

The magnetic nanoparticles used to quickly separate and purify superoxide dismutase (SOD) from yak blood was
evaluated and a kinetic model of SOD adsorption was established in our experiments. The effects of pH of binding buffer (sodium
phosphate buffer) and the dissociation buffer (sodium acetate buffer), NaCl concentration and adsorption time on the specific
activity and recovery of SOD were investigated to determine the optimal conditions for separation and purification of the
enzyme. The specific activity of SOD revealed an initial increase and a final decrease with the increase of pH, but the enzyme
recovery revealed a stable increase, suggesting that optimal pH of binding buffer was 7.5. The specific activity and recovery of
SOD exhibited an initial increase and following decrease with the increase of NaCl concentration. Meanwhile, the maximum
SOD adsorption was observed in the presence of 0.5 mol/L NaCl. When pH of dissociation buffer was increased from 4.0 to
5.5, the specific activity of SOD revealed an improvement, but the recovery of enzyme revealed a reduction. So the optimal
pH of dissociation buffer was determined to be 5.0. The experimental results also showed that under the optimal adsorption
conditions, the optimal static adsorption time was 2 h. As the concentration of crude enzyme extract was increased, SOD adsorption could be
expressed by Langmuir adsorption model. The specific activity of SOD was always maintained at a high level (3000—3250 U/mg), which
showed that the best separation selectivity for SOD can be achieved by using magnetic nanoparticles.

Key words: yak blood, superoxide dismutase, specific activity, adsorption kinetics

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