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Application of glucose oxidase combined with peroxidase in mycotoxin detoxification

A technology of glucose oxidase and peroxidase, applied in food science and other fields, can solve the problem that manganese peroxidase cannot degrade toxins

Pending Publication Date: 2020-07-17
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] It has been reported that manganese peroxidase derived from white rot fungi can degrade mycotoxins, but it requires hydrogen peroxide as a substrate. If hydrogen peroxide cannot be introduced to detoxify mycotoxins in grain and feed, manganese Peroxidase cannot function to degrade toxins
There are relatively few reports on other peroxidases that degrade mycotoxins

Method used

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  • Application of glucose oxidase combined with peroxidase in mycotoxin detoxification
  • Application of glucose oxidase combined with peroxidase in mycotoxin detoxification
  • Application of glucose oxidase combined with peroxidase in mycotoxin detoxification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Combined application of glucose oxidase GOD and dye decoloring peroxidase DyP on the degradation effect of zearalenone

[0032] The dye decoloring peroxidase DyP is dissolved in sodium phosphate buffer (0.1M, pH 7.0), and 1 mL of dye decoloring peroxidase DyP solution (enzyme activity 5U / mL) is added to 0.0125, 0.025, 0.5, 1, 2, 4 and 8U / mL glucose oxidase GOD, then add 200μM glucose and ZEN, the concentration of ZEN reaches 10μg / mL; take the system without dye decoloring peroxidase DyP as control; react at 37℃ for 3h , 1mL methanol was added to terminate the reaction, and the ZEN content was detected by high performance liquid chromatography.

[0033] The result is figure 1 As shown, as the concentration of glucose oxidase GOD increases, the degradation rate of dye decoloring peroxidase DyP to ZEN increases. When the concentration of GOD is 1U / mL, the degradation rate of dye decoloring peroxidase DyP to ZEN reaches 68. %.

Embodiment 2

[0034] Example 2 Preparation method and application of a composite enzyme for degrading mycotoxins

[0035] Weigh a certain amount of glucose oxidase GOD and horseradish peroxidase HRP and mix them in a ratio of 1:3 by mass to obtain a composite enzyme that degrades mycotoxins; then use the prepared composite enzyme to degrade aflatoxin B1 (AFB1), dissolve 1mg of complex enzyme in 1mL sodium citrate buffer (0.1M, pH 6) reaction system, add 100μM glucose, then add AFB1, AFB1 concentration is 1μg / mL, react at 37℃ for 12h, add 1mL The reaction was terminated by methanol; the system without compound enzyme was used as a control; the content of AFB1 was detected by high performance liquid chromatography.

[0036] The results showed that the compound enzyme degrading mycotoxins reacted with 1μg / mL AFB1 for 12h, and the degradation rate of AFB1 was 82% ( figure 2 ).

Embodiment 3

[0037] Example 3 Preparation method and application of a composite enzyme for degrading mycotoxins

[0038] Weigh a certain amount of glucose oxidase GOD and lactoperoxidase LPO, mix them in a ratio of 1:4 by mass, and then add mediator ABTS that accounts for 1% of the amount of enzyme to prepare a composite enzyme that degrades mycotoxins; The obtained complex enzyme is used to degrade vomitin (DON). Dissolve 1 mg of complex enzyme in 1mL sodium citrate buffer (0.1M, pH5.0) reaction system, add 200μM glucose, and then add DON, the concentration of DON is 20μg / mL , React at 37°C for 24h, add 1mL methanol to terminate the reaction; take the system without adding complex enzyme as a control; use high performance liquid chromatography to detect the DON content.

[0039] The results showed that the compound enzyme degrading mycotoxins reacted with 20μg / mL DON for 24h, and its degradation rate of DON was 63% ( image 3 ).

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Abstract

The invention belongs to the technical field of agricultural biology, in particular relates to a combined application of glucose oxidase and peroxidase, and specifically relates to an application of glucose oxidase combined with peroxidase in the field of mycotoxin biological detoxification. A composite enzyme and a mycotoxin detoxification method provided by the invention are safe and efficient,and have a very wide application prospect.

Description

Technical field [0001] The invention belongs to the field of agricultural biotechnology, and specifically relates to the application of glucose oxidase combined with peroxidase to the field of mycotoxin biological detoxification. Background technique [0002] Peroxidase (Peroxidase, POD) is a type of oxidoreductase produced by microorganisms or plants. This type of enzyme uses hydrogen peroxide as an electron acceptor to catalyze the oxidation reaction of the substrate. It mainly exists in the peroxide of the carrier. In the enzyme body, iron porphyrin is used as a prosthetic group, which can catalyze hydrogen peroxide, oxidize phenol and amine compounds and hydrocarbon oxidation products, and have the ability to eliminate the toxicity of hydrogen peroxide and phenols, amines, aldehydes, and benzene. double effect. Peroxidase includes dye decolorizing peroxidase (Dye-decolorizing peroxidase, DyP), horseradish peroxidase (Horseradish Peroxidase, HRP), lactoperoxidase (Lactoperoxi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23L5/20
CPCA23L5/25Y02P60/87
Inventor 赵丽红郭永鹏马秋刚
Owner CHINA AGRI UNIV
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