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化学学报
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2012 , Vol. 0 >Issue 05: 624 - 628

DOI: https://doi.org/10.6023/A1107232

研究论文

固定化乙酰胆碱酯酶的制备及其特性研究

  • 陈军辉 ,
  • 史倩 ,
  • 陈晨 ,
  • 李鑫 ,
  • 曹为 ,
  • 郑立 ,
  • 王小如
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  • a 国家海洋局第一海洋研究所 海洋生态研究中心 青岛 266061;
    b 厦门大学化学化工学院 厦门 361005;
    c 上海海洋大学水产与生命学院 上海 201306

收稿日期: 2011-07-23

  修回日期: 2011-11-22

  网络出版日期: 2011-12-09

基金资助

国家自然科学基金(No. 20905017)、国家海洋局青年基金(No. 2010140)和海洋一所基本科研业务专项(No. 2010G25)资助项目.

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Preparation and Characteristic Research of Immobilized Acetylcholinesterase

  • Chen Junhui ,
  • Shi Qian ,
  • Chen Chen ,
  • Li Xin ,
  • Cao Wei ,
  • Zheng Li ,
  • Wang Xiaoru
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  • a Research Center for Marine Ecology, The First Institute of Oceanography, State Oceanic Administration, Qingdao 266061;
    b College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005;
    c College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306

Received date: 2011-07-23

  Revised date: 2011-11-22

  Online published: 2011-12-09

Supported by

Project supported by the National Natural Scientific Foundation of China (No. 20905017), the Science Foundation for Youth Scholars of State Oceanic Administration (No. 2010140) and the Primary Scientific Foundation of the First Institute of Oceanography (No. 2010G25).

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摘要

本研究以期研制出能重复使用的固定化乙酰胆碱酯酶(AChE), 为天然产物复杂体系中AchE 抑制剂筛选新方法的发展奠定基础. 以氨基化硅胶(APS-Si)微球为载体, 戊二醛为交联剂对乙酰胆碱酯酶进行交联固定化, 并研究了酶的最佳固定化条件和固定化酶的性质. 结果表明, 0.05 g 氨基化硅胶微球载体, 用戊二醛溶液活化6 h 后, 在给酶量5 U, 28℃固定16 h 条件下, 得到固定化酶的活性最大. 固定化酶在常温(20～40 ℃), 以及较宽pH 范围内(pH 6～10)均具有较高的活性, 并且具有良好的保存稳定性和可重复利用率, 为基于固定化靶酶亲和-色谱质谱联用分析快速筛选乙酰胆碱酯酶抑制剂新方法的发展奠定了基础.

关键词： 乙酰胆碱酯酶; 固定化酶; 亲和分离材料; 氨基化硅胶

本文引用格式

陈军辉 , 史倩 , 陈晨 , 李鑫 , 曹为 , 郑立 , 王小如 . 固定化乙酰胆碱酯酶的制备及其特性研究[J]. 化学学报, 2012 , 0(05) : 624 -628 . DOI: 10.6023/A1107232

Abstract

A new method for preparation of a reusable immobilized acetylcholinesterase (AchE) was developed in order to lay a foundation for screening AchE inhibitors from natural products. AchE was immobilized on aminated silica gel microspheres with glutaraldehyde as cross-linking reagent. The optimum conditions and properties of immobilized enzyme were investigated. The optimal immobilization process was as follows: 0.05 g of aminated silica gel microspheres was activated by glutaraldehyde solution for 6 h, and mixed with AChE solution with the activity of 5 U. Then, the mixture was shook for 16 h at 28 . ℃ The immobilized enzyme produced by this method had the highest enzyme activity, and it could remain better activity with a wide range of pH (pH 6～10) at room temperature (20～40 ℃). The immobilized AChE showed higher storage stability and could be used repeatedly.

Key words： acetylcholinesterase; immobilized enzyme; affinity separation material; aminated silica gel

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