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Process for preparing copolyester soybean esterase

A technology of soybean and esterase, which is applied in the direction of hydrolytic enzymes, etc., can solve the problems of detection sensitivity of residual pesticides and the minimum detection limit failing to meet the safety limit requirements, etc.

Inactive Publication Date: 2008-12-31
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing plant esterase sheet preparation report at present, as the patent office disclosed application number is 99114268.3 " the plant esterase sheet of rapid assay organophosphorus pesticide " patent of invention, and it is directly to use flour as enzyme source material, but prepared The sensitivity and minimum detection limit of esterase tablets to detect residual pesticides are far from the safety limit requirements stipulated in the national standard
In addition, there is no research report on the preparation method of plant esterase pure enzyme products

Method used

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Embodiment Construction

[0006] Concrete operation steps of the present invention are as follows:

[0007] a. Choose high-quality soybeans,

[0008] b. Grinding into fine powder with a pulverizer,

[0009] c. Phosphate buffer extraction,

[0010] c.1 Put the crushed soybean flour into a container of appropriate size, and add 0.3mol / L, pH 7.0-8.0 phosphate buffer solution at a ratio of 1:4-5 to liquid;

[0011] c.2 Shake or stir on a shaker or mixer for 10-30 minutes;

[0012] c.3 After shaking or stirring, continue to stand and extract in the above container

[0013] c.3.1 The temperature is 0-4°C

[0014] c.3.2 The time is 10-24h;

[0015] c.4 Filter the above extract with gauze, and take the supernatant, which is the crude extract;

[0016] d. Differential refrigerated centrifugation,

[0017] d.1 Centrifuge the crude extract at 5000-6000rpm for 10-15min, and collect the supernatant;

[0018] d.2 Dissolve the precipitate in the previous step with phosphate buffer solution of pH 7.0-8.0, then...

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PUM

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Abstract

The invention discloses a preparation method of soybean esterase used for the determination of pesticide residues in fruits and vegetables. The steps include: selecting high-quality soybeans, crushing, extracting with phosphate buffer solution, differential centrifugation, ammonium sulfate salting out and precipitation, ion exchange chromatography purification and freeze-drying. The preparation process of the invention is simple and stable, and the production cost is lower than the production cost of extracting pure enzyme products of acetylcholinesterase. The sensitivity of the soybean esterase purified by the invention is more than 19 times higher than that of the enzyme crude extract. The lowest detection limit (LDC) of monocrotophos, methamidophos, parathion, chlorpyrifos, dimethoate, malathion, trichlorfon and other pesticides can reach 0.03125ppm, and the LDC of chlorpyrifos and chlorpyrifos Even reached one thirty-second of its maximum residue limit (MRL), and the LDCs of parathion, parathion, thionon, etc. also reached one sixteenth of its MRL. The soybean esterase prepared by the invention can fully satisfy the rapid detection of various organophosphorus pesticide residues, and can be widely used as an enzyme marker for various methods of rapid detection of pesticide residues.

Description

technical field [0001] The invention relates to a method for detecting pesticide residues, in particular to a method for preparing soybean esterase for rapid detection of organic phosphorus and carbamate pesticide residues in fruits and vegetables. Background technique [0002] At present, organophosphorus pesticides and carbamate pesticides are still widely used in agricultural production, and they are very easy to cause residues in agricultural products, especially in fruits and vegetables, which are more prominent, causing harm to human health. Therefore, pesticide residues are important detection and control indicators for food quality and safety. In recent years, there have been many studies on rapid detection methods for pesticide residues. Among them, "enzyme inhibition-spectrophotometry" has been widely used because of its sensitivity, simplicity and speed, and has been listed as a national recommended standard (GB / T5009.199- 2003). The enzyme inhibition method is ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/16
Inventor 李建科温艳霞牛乐朱桂勤
Owner SHAANXI NORMAL UNIV
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