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Received: 2012-04-08

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Journal of Zhejiang University SCIENCE B 2013 Vol.14 No.4 P.325-335


Purification and studies on characteristics of cholinesterases from Daphnia magna

Author(s):  Yan-xia Yang, Li-zhi Niu, Shao-nan Li

Affiliation(s):  Institute of Pesticide and Environmental Toxicology, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310029, China

Corresponding email(s):   snli@zju.edu.cn

Key Words:  Crustacea, Pseudocholinesterase, Cholinesterase, Substrate preference, Selective inhibitors

Yan-xia Yang, Li-zhi Niu, Shao-nan Li. Purification and studies on characteristics of cholinesterases from Daphnia magna[J]. Journal of Zhejiang University Science B, 2013, 14(4): 325-335.

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author="Yan-xia Yang, Li-zhi Niu, Shao-nan Li",
journal="Journal of Zhejiang University Science B",
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%T Purification and studies on characteristics of cholinesterases from Daphnia magna
%A Yan-xia Yang
%A Li-zhi Niu
%A Shao-nan Li
%J Journal of Zhejiang University SCIENCE B
%V 14
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%D 2013
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1200113

T1 - Purification and studies on characteristics of cholinesterases from Daphnia magna
A1 - Yan-xia Yang
A1 - Li-zhi Niu
A1 - Shao-nan Li
J0 - Journal of Zhejiang University Science B
VL - 14
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%@ 1673-1581
Y1 - 2013
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1200113

Due to their significant value in both economy and ecology, Daphnia had long been employed to investigate in vivo response of cholinesterase (ChE) in anticholinesterase exposures, whereas the type constitution and property of the enzyme remained unclear. A type of ChE was purified from Daphnia magna using a three-step procedure, i.e., Triton X-100 extraction, ammonium sulfate precipitation, and diethylaminoethyl (DEAE)-Sepharose™-Fast-Flow chromatography. According to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), molecular mass of the purified ChE was estimated to be 84 kDa. Based on substrate studies, the purified enzyme preferred butyrylthiocholine iodide (BTCh) [with maximum velocity (Vmax)/Michaelis constant (Km)=8.428 L/(min·mg protein)] to acetylthiocholine iodide (ATCh) [with Vmax/Km=5.346 L/(min·mg protein)] as its substrate. Activity of the purified enzyme was suppressed by high concentrations of either ATCh or BTCh. Inhibitor studies showed that the purified enzyme was more sensitive towards inhibition by tetraisopropylpyrophosphoramide (iso-OMPA) than by 1,5-bis(4-allyldimethylammoniumphenyl) pentan-3-one dibromide (BW284C51). Result of the study suggested that the purified ChE was more like a type of cholinesterase%29&ck%5B%5D=abstract&ck%5B%5D=keyword'>pseudocholinesterase, and it also suggested that Daphnia magna contained multiple types of ChE in their bodies.

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