|Table of Contents|

PURIFICATION AND CHARACTERIZATION OF CHOLINESTERASE FROM EISENIA FOETIDA(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
1997 03
Page:
246-251
Research Field:
Publishing date:

Info

Title:
PURIFICATION AND CHARACTERIZATION OF CHOLINESTERASE FROM EISENIA FOETIDA
Author(s):
Ding Shihua Li Qingyi
1. Dept Fisheries, SW Agricul Univ, Chongqing 630716;
2. Dept life Sci, SW Norm Univ, Chongqing 630715
Keywords:
cholinesteraseearthwormion-exchange chromatographygel filtration
CLC:
-
PACS:
DOI:
-
DocumentCode:

Abstract:
Cholinesterase from the Triton X-100 extract of Eisenia foetida was purified to electrophoretic homogeneity by extraction with PEG 2 000/potassium phosphate buffer, ion-exchange chromatography and gel filtration. The purified enzyme was found to be a kind of glycoproteins. The Mr determined by gel filtration was 59000 and by SDS-PAGE was 58000. The contents of acidic amino acid and basic amino acid were about 24% and 12%, respctively. Ultraviolet spectrum showed a maximum absorption at 272 um. The enzyme with optimum temperature of 39℃ was unstable when heated. It exhibited optimum activity at pH 7.8 and kept stable between pH 7.8~8.0. The Michaelis constant for acetylthiocholine iodide was 88 μmol/L. The enzyme activity affected slightly by Mg2+,Ca2+ and Mn2+ was inhibited by excess substrate and strongly by DFP.

References

1. 朱美财,王荫静,向培德.人脑及细胞膜乙酞胆碱醋酶的芳基酸胺酶活性.生物化学杂志.1994.10:180~184
2. Lockridge O. Substance P hydrolysis by human serum cholinesterase. J Neurochem. 1982.39:106~110
3. 黄雁,郭中英.简易、快速检测有机磷家药的酶片和生色基质片.环境科学.1995.18(3):180~184
4. Ellman GL. Courtney DK, Andres V et al. A new and rapid colorimetric determination of acetylcholinesterase activity. Bioclem Pharmacoi. 1961. 7:88~95
5. Wang CS,Smith RC. Lowry determination of protein in the presence of Triton X-100. Anal Biochem. 1975, 83:415~421
6. 张龙翔等.生化实验方法和技术.北京:人民教育出版社.1982
7. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T 4. Natare. 1970.227:680~685
8. 张翰,邢志勇,孙曼霁.乙酰胆碱酯酶的聚丙烯酞胺凝胶电泳.军事医学科学院院刊.1963.6:689~692
9. 彭浪,张龙翔.大熊猫乳酸脱氢酶同工酶H4的分离纯化及某些性质的研究.生物化学杂志.1995.11:85~90
10. 朱俭等.生物化学实验.上海:上海科学技术出版社,1981
11. Andersen RA,Aune T.Barstad JAB. Characteristics of cholinesterase of the earthworm Easeaea foelida. Comp Biachem Physeol. 1987,6IC:81~87
12. 施善平,张景园.黄姑鱼肌肉乙酞胆碱酷酶的多分子形式和亚基组成.生物化学与生物物理学报.1984,18:172~177
13. 屈贤铭.余微明,邱雪贞.浙江产眼镜蛇蛇毒胆碱醋酶的研究.生物化学与生物物理学报.1981.13:397~405
14. 孙曼霁,高天栋,邢志勇等.中国丁氏鳍电鳐乙酰胆碱酯酶的制备亲和层析.生物化学杂志.1985.1(1):47~55
15. Principato GB,Rosi G,Biagioni M et al. Kinetic studies on the reaction mechanism of the propionylcholinesterase from Hirudo medicinalis. Comb Btoclem Physiol. 1983,75C:185~192
16. 方允中.陈能乾.医学酶学.北京:人民卫生出版社,1984

Memo

Memo:
-
Last Update: 1900-01-01