SIOC 中文
ACS
Adv search

Acta Chimica Sinica >

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

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

Full Papers

Preparation and Characteristic Research of Immobilized Acetylcholinesterase

  • Chen Junhui ,
  • Shi Qian ,
  • Chen Chen ,
  • Li Xin ,
  • Cao Wei ,
  • Zheng Li ,
  • Wang Xiaoru
Expand
  • 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).

Fold

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

Cite this article

Chen Junhui , Shi Qian , Chen Chen , Li Xin , Cao Wei , Zheng Li , Wang Xiaoru . Preparation and Characteristic Research of Immobilized Acetylcholinesterase[J]. Acta Chimica Sinica, 2012 , 0(05) : 624 -628 . DOI: 10.6023/A1107232

References

1 Slon-Usakiewicz, J. J.; Ng, W.; Dai, J. R.; Pasternak, A.; Redden, P. R. Drug Discovery Today 2005, 10(6), 409.  

2 Espada, A.; Molina-Martin, M.; Dage, J.; Kuo, M. S. Drug Discovery Today 2008, 13(9～10), 417.

3 Slon-Usakiewicz, J. J.; Dai, J. R.; Ng, W.; Foster, J. E.; Deretey, E.; Toledo-Sherman, L.; Redden, P. R.; Pasternak, A.; Reid, N. Anal. Chem. 2005, 77(5), 1268.

4 Jin, N.-Z.; Lu, X.-H. Chin. J. Biochem. Pharm. 2003, 24(5), 1 (in Chinese). (金念祖, 陆晓和, 中国生化药物杂志, 2003, 24(5), 1.)

5 Huang, X. D.; Kong, L.; Li, X.; Chen, X. G.; Guo, M.; Zou, H. F. J. Chromatogr. B 2004, 812, 71.

6 Wang, S.; Sun, M.; Zhang, Y. M.; Du, H.; He, L. C. J. Chromatogr. A 2010, 1217(32), 5246.

7 Xin, G. Z.; Zhou, J. L.; Qi, L. W.; Li, P. Comb. Chem. High Throughput Screening 2011, 14(2), 93.

8 Huang, Y.; Tian, Q.-Q.; Li, T.-Y.; Chen, B.; Yao, S. Z. Sci. China, Ser. B: Chem. 2009, 39(8), 760 (in Chinese). (黄燕, 田清清, 李谭瑶, 陈波, 姚守拙, 中国科学B 辑: 化学, 2009, 39(8), 760.)  

9 Tang, Z. M.; Wang, Z. Y.; Kang, J. W. Electrophoresis 2007,28, 360.  

10 Hu, F. L.; Zhang, H. Y.; Lin, H. Q.; Deng, C. H.; Zhang, X. M. J. Am. Soc. Mass Spectrom. 2008, 19, 865.  

11 Hu, F. L.; Deng, C. H.; Zhang, X. M. J. Chromatogr. B2008, 871, 67.  

12 Rhee, I. K.; van de Meent, M.; Ingkaninan, K.; Verpoorte, R. J. Chromatogr. A 2001, 915(1～2), 217.

13 Zhang, X.-A.; Jia, H.-H.; Wang, X.-F.; Zhang, H.-L.; Yin, H.-W.; Chang, S.-L.; Wang, J.-F.; Wu, W.-J. Chin. Sci. Bull.2009, 54(12), 1701 (in Chinese). (张学骜, 贾红辉, 王晓峰, 张海良, 尹红伟, 常胜利, 王 建方, 吴文健, 科学通报, 2009, 54(12), 1701.)

14 Marinov, I.; Gabrovska, K.; Velichkova, J.; Godjevargova, T. Int. J. Biol. Macromol. 2009, 44(4), 338.  

15 Ellman, G. L.; Courtney, K. D.; Andres, V. Jr.; Feather-stone, R. M. Biochem. Pharmacol. 1961, 7, 88.  

16 Zhang, A.-J.; Meng, F.-P. Polym. Mater. Sci. Eng. 2011,27(4), 124 (in Chinese). (张爱静, 孟范平, 高分子材料科学与工程, 2011, 27(4),124.)  

17 Tumturk, H.; Sahin, F.; Demirel, G. Bioprocess Biosyst. Eng. 2007, 30, 141.  

18 Li, Y. R.; Gan, Z. Y.; Li, Y. F.; Liu, Q.; Bao, J. C.; Dai, Z. H.; Han, M. Sci. China Chem. 2010, 53(4), 820.  
Options
Outlines

/

〈 〉