Expression of the housefly acetylcholinesterase in a bioreactor and its potential application in the detection of pesticide residues |
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Authors: | Guo-Jun Lang Jin-Yan Shang Yan-Xia Chen Ying-Jun Cui Qiang Wang Zhen-Hua Tang Chuan-Xi Zhang |
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Institution: | (1) Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Science, Zhejiang University, 310029 Hangzhou, China;(2) Hangzhou Centre of Inspection and Testing for Agricultural Products, Ministry of Agriculture, 310021 Hangzhou, China; |
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Abstract: | Acetylcholinesterase is a key enzyme of the animal nerve system. The enzyme is the primary target of organophosphorous (OP)
and carbamate (CB) insecticides. The insect AChE is being extensively used in development of new insecticides or in vitro
selection of the new designed insecticides, and in pharmacological and toxicological field. Rapid assays using AChE-based
methods have been proposed as an efficient and rapid method for the detection of pesticides, especially in many Asian markets.
In this study, the acetylcholinesterase gene was cloned from housefly (Musca domestica) susceptible to organophosphate (OP) and carbamate (CB) insecticides, and expressed in baculovirus-insect cells system using
a bioreactor with oxygen supplementation. The recombinant housefly AChE was purified using ammonium sulfate precipitation
and procainamide affinity chromatography, and approximately 0.42 mg of the purified AChE with high biological activity (118.9 U/mg)
was obtained from 100 ml of culture solution. The purified AChE was highly sensitive to OP and CBs insecticides. In conclusion,
an efficient expression and purification system has been developed for large-scale production of recombinant housefly AChE.
The recombinant enzyme is potential to be used for the detection of pesticide residues. |
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