Acta Chimica Sinica ›› 2012, Vol. 0 ›› Issue (05): 624-628 .DOI: 10.6023/A1107232 Previous Articles     Next Articles

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陈军辉a, 史倩a, 陈晨a,c, 李鑫a, 曹为a, 郑立a, 王小如a,b   

  1. 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
  • 投稿日期:2011-07-23 修回日期:2011-11-22 发布日期:2011-12-09
  • 通讯作者: 陈军辉
  • 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.

Preparation and Characteristic Research of Immobilized Acetylcholinesterase

Chen Junhuia, Shi Qiana, Chen Chena,c, Li Xina, Cao Weia, Zheng Lia, Wang Xiaorua,b   

  1. 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:2011-07-23 Revised:2011-11-22 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).

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