Preparing method for acetylcholinesterase

A technology of acetylcholinesterase and enzyme liquid, which is applied in the field of preparation of acetylcholinesterase, can solve the problems such as difficulty in popularization and application of rapid determination products, high price, complicated preparation process, etc., and achieves low production cost, high sensitivity and high product purity. Effect

Inactive Publication Date: 2004-02-04
GUANGDONG INST OF MICROORGANISM
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AI Technical Summary

Problems solved by technology

The rapid determination product currently used in China—pesticide rapid test paper, its main raw material acetylcholinesterase is imported from abroad. These acetylcholinesterases are expensive due to the complexity of product raw materials and preparation processes, which makes rapid detection of pesticide residues in fruits and vegetables. Determination products are difficult to popularize and apply

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The process steps of extracting acetylcholinesterase from chicken blood are as follows:

[0019] (1) After the fresh chicken blood is condensed at room temperature, place it in the refrigerator at 4°C for 2 hours, discard the precipitate, and take its supernatant;

[0020] (2) The supernatant was refrigerated and centrifuged at 8000 r.p.m at 4° C. for 15 minutes, and then the precipitate was discarded. The supernatant was the crude extract of acetylcholinesterase;

[0021] (3) Purification of acetylcholinesterase by ammonium sulfate precipitation method:

[0022] A, get acetylcholinesterase crude extract 10ml, add saturation and be 60% ammonium sulfate 3.61 grams (according to the ammonium sulfate saturation consumption table) under ice-bath slow stirring;

[0023] b. The solution obtained above was refrigerated and centrifuged at 11000 r.p.m at 4°C for 10 minutes, and the supernatant was poured off to obtain a precipitate with a saturation of ammonium sulfate of 60%; ...

Embodiment 2

[0033] The process steps of extracting acetylcholinesterase from pig blood are as follows:

[0034] (1) After the fresh pig blood is condensed at room temperature, place it in the refrigerator at 4°C for 2 hours, discard the precipitate, and take its supernatant;

[0035] (2) The supernatant was refrigerated and centrifuged at 8000 r.p.m at 4° C. for 15 minutes, and then the precipitate was discarded. The supernatant was the crude extract of acetylcholinesterase;

[0036] (4) Purification of acetylcholinesterase by ammonium sulfate precipitation method:

[0037] A, get acetylcholinesterase crude extract 10ml, add saturation and be 50% ammonium sulfate 2.91 grams (according to ammonium sulfate saturation consumption table) under ice bath slow stirring;

[0038] b. The solution obtained above was refrigerated and centrifuged at 11000 r.p.m at 4°C for 10 minutes, and the supernatant was poured off to obtain a precipitate with a saturation of ammonium sulfate of 50%;

[0039] c....

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Abstract

A process for preparing the acetylcholinesterase used to detect the content of residual agricultural chemical in fruit and vegetable includes such steps as extracting the coarse liquid extract from animal blood, purifying by ammonium sulfate deposition method, removing ammonium sulfate by dialysis method, concentrating and drying. Its advantages are widely available raw material, low cost and high purity and sensitivity of product.

Description

technical field [0001] The invention relates to a preparation method of acetylcholinesterase used for the determination of pesticide residues in fruits and vegetables. Background technique [0002] Pesticide residues in fruits and vegetables can cause acute poisoning and death in humans, and trace accumulation can cause chronic poisoning such as carcinogenicity, teratogenicity, and mutagenicity, which seriously endanger people's health. Vegetables currently on the market, in addition to relying on conventional methods such as gas chromatography for detection, rapid detection technology has also been applied. The rapid determination product currently used in China—pesticide rapid test paper, its main raw material acetylcholinesterase is imported from abroad. These acetylcholinesterases are expensive due to the complexity of product raw materials and preparation processes, which makes rapid detection of pesticide residues in fruits and vegetables. Determination products are d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/36C12N9/16C12Q1/46
Inventor 李盾陈仪本范怀忠陈娇娣
Owner GUANGDONG INST OF MICROORGANISM
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