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Method for Degrading Organophosphorus Pesticides Using Bacterial Laccase

A technology of organophosphorus pesticides and laccase, which is applied in the field of environmental protection and achieves the effect of small usage

Active Publication Date: 2022-06-03
ZHONGKAI UNIV OF AGRI & ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there are research reports that Penicillium lilacinus, Trametes teneci fungus laccase, and Trichoderma can degrade organophosphorus pesticides. However, there are few reports on the degradation of organophosphorus pesticides by bacterial laccases, and their application conditions need to be further studied. Research

Method used

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  • Method for Degrading Organophosphorus Pesticides Using Bacterial Laccase
  • Method for Degrading Organophosphorus Pesticides Using Bacterial Laccase
  • Method for Degrading Organophosphorus Pesticides Using Bacterial Laccase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Enzyme production culture

[0038] 1) Preparation of medium: take yeast extract, peptone and sodium chloride and add water to prepare a yeast extract containing 0.5%, 1.0% peptone and 0.5% sodium chloride, adjust the pH to 7.0 to obtain an expansion medium; Copper sulfate pentahydrate was dissolved in water to make a 5% copper sulfate solution, and 8 mL of 5% copper sulfate solution was added to each L expansion medium to adjust the pH to 7.0 to obtain an enzyme production medium; the expansion medium and enzyme production culture The bases were sterilized in an autoclave at 121°C for 20 minutes.

[0039] 2) Inoculate the bacteria on the plate with the Lrr3S strain into the expansion medium on the ultra-clean workbench, and place it in a constant temperature incubator at 34°C for 1 day; when the culture medium is turbid, it is 1:3. The volume ratio was transferred to the enzyme production medium and cultured in a constant temperature incubator at 34°C for ...

Embodiment 2

[0042] Example 2: Detection of Organophosphorus Pesticide Degradation

[0043] 1) Standard curve drawing: use potassium dihydrogen phosphate as the standard, prepare the inorganic phosphorus standard solution group, use the phosphomolybdenum blue spectrophotometry, detect the OD value at 710nm, and draw the standard curve; the results are as follows figure 1 As shown, the phosphorus content has a good linear relationship with the absorbance value;

[0044] 2) Determination of total phosphorus content in pesticides: take 0.5mL of 0.2% pesticide diluent in a 50mL conical flask, add 5mL potassium persulfate solution, 1mL 1:35 sulfuric acid solution, add water to 25mL, and heat it to nearly Dry, and control the time within 30-40min, so that all the organic phosphorus of the pesticide is degraded into inorganic phosphorus. Take out to cool, and measure the phosphorus content by phosphomolybdenum blue spectrophotometry. Add an appropriate amount of water to dissolve the crystals...

Embodiment 3

[0047] Example 3: Single factor experiment

[0048] 1) The effect of temperature on the degradation of enzymatic pesticides

[0049] Prepare 6 groups of clean test tubes, add 2.5mL pesticide diluent, 2.3mL laccase, and 0.2mL 1mmol / L ABTS solution, respectively, take the inactivated laccase as control, at 20 ℃, 25 ℃, 30 ℃, 35 ℃ , 40℃, 45℃ for 6h. The degradation rate of the pesticide was measured, and a graph was drawn according to the relationship between the degradation rate and the temperature.

[0050] The result is as image 3 As shown, the temperature has an effect on the enzymatic degradation of the five organophosphorus pesticides, and the effect trend is basically the same. When it is between 20-30 ℃, the degradation rate increases with the increase of temperature, and between 30 and 45 ℃, it decreases with the increase of temperature. At 30℃, the degradation rates of five organic pesticides reached their peaks, among which dimethoate reached 8.43%, dichlorvos re...

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Abstract

The invention discloses a method for degrading organophosphorus pesticides by using bacterial laccase. In the method, Cu is added to the enzyme production medium. 2+ , at pH = 8‑10, temperature 25‑35 ° C, adding mediators for degradation, can degrade within a few hours at a concentration of about 0.2%, including dimethoate, dichlorvos, trichlorfon, chlorpyrifos, omethoate The degradation rate of organophosphorus pesticides can be as high as 15%.

Description

technical field [0001] The invention relates to the technical field of environmental protection, in particular to a method for degrading organophosphorus pesticides by using bacterial laccase. Background technique [0002] Laccase (Laccase, EC1.10.3.2) is a Cu-containing 2+ Polyphenoloxidases (PPO), the oxidation substrates of which include phenols and their derivatives, arylamines and their derivatives, carboxylic acids and their derivatives, steroid hormones, biological pigments, metal organic compounds and the like. [0003] Laccases can be divided into three categories according to their sources, including fungal laccases, bacterial laccases and plant laccases. At present, bacterial laccase has gradually attracted everyone's attention. Bacterial laccase often does not require glycosylation, has high thermal stability, has a wide optimal pH range for the enzyme, is stable in halogen solutions, has high resistance to extreme environments, and has a long growth cycle of ba...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A62D3/02A62D101/04A62D101/28A62D101/26
CPCA62D3/02A62D2101/04A62D2101/28A62D2101/26
Inventor 刘锐张晓柔陈欣强赵翾李南薇刘功良
Owner ZHONGKAI UNIV OF AGRI & ENG
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