CN111418756A - Application of glucose oxidase and peroxidase in mycotoxin detoxification - Google Patents
Application of glucose oxidase and peroxidase in mycotoxin detoxification Download PDFInfo
- Publication number
- CN111418756A CN111418756A CN202010014325.0A CN202010014325A CN111418756A CN 111418756 A CN111418756 A CN 111418756A CN 202010014325 A CN202010014325 A CN 202010014325A CN 111418756 A CN111418756 A CN 111418756A
- Authority
- CN
- China
- Prior art keywords
- peroxidase
- glucose oxidase
- complex enzyme
- mycotoxin
- oxidase god
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
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- 239000002636 mycotoxin Substances 0.000 title claims abstract description 68
- 108010015776 Glucose oxidase Proteins 0.000 title claims abstract description 51
- 239000004366 Glucose oxidase Substances 0.000 title claims abstract description 51
- 229940116332 glucose oxidase Drugs 0.000 title claims abstract description 51
- 235000019420 glucose oxidase Nutrition 0.000 title claims abstract description 51
- 238000001784 detoxification Methods 0.000 title claims abstract description 18
- 102000003992 Peroxidases Human genes 0.000 title claims abstract description 6
- 108010046301 glucose peroxidase Proteins 0.000 title abstract description 4
- 102000004190 Enzymes Human genes 0.000 claims abstract description 115
- 108090000790 Enzymes Proteins 0.000 claims abstract description 115
- 229940088598 enzyme Drugs 0.000 claims abstract description 115
- 108040007629 peroxidase activity proteins Proteins 0.000 claims abstract description 30
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/25—Removal of unwanted matter, e.g. deodorisation or detoxification using enzymes
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- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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Abstract
The invention belongs to the technical field of agricultural biology, in particular to combined application of glucose oxidase and peroxidase, and particularly relates to the field of application of the glucose oxidase and the peroxidase in biological detoxification of mycotoxin; the complex enzyme and mycotoxin detoxification method provided by the invention is safe and efficient, and has a very wide application prospect.
Description
Technical Field
The invention belongs to the technical field of agricultural biology, and particularly relates to the field of application of glucose oxidase and peroxidase in biological detoxification of mycotoxin.
Background
Peroxidases (PODs) are oxidoreductases produced by microorganisms or plants, which take hydrogen peroxide as an electron acceptor and catalyze substrates to perform oxidation reaction, mainly exist in peroxisomes of carriers, take iron porphyrin as a prosthetic group, can catalyze hydrogen peroxide, oxidize phenolic and amine compounds and hydrocarbon oxidation products, and have double effects of eliminating toxicity of hydrogen peroxide, phenols, amines and benzenes.
The method is characterized in that the method comprises the steps of taking hydrogen peroxide as a substrate, and if the mycotoxin in food and feed is detoxified, introducing the hydrogen peroxide to ensure that the manganese peroxidase cannot play a role in degrading the mycotoxin.
Disclosure of Invention
The invention aims to provide application of glucose oxidase GOD combined with peroxidase POD in mycotoxin detoxification.
The invention also aims to provide a compound enzyme containing the glucose oxidase GOD and the peroxidase POD.
The invention also aims to provide the application of the compound enzyme in mycotoxin detoxification.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a method for detoxifying mycotoxin, and particularly relates to a method for jointly applying glucose oxidase GOD and peroxidase POD to the field of mycotoxin detoxification.
In the detoxification method, glucose oxidase GOD is mixed with one or more of dye decolorizing peroxidase DyP, horse radish peroxidase HRP, lactoperoxidase L PO, lignin peroxidase L iP, manganese peroxidase MnP, chloroperoxidase CPO and soybean peroxidase SBP for preparation, and the method is used in the field of mycotoxin detoxification.
Preferably, glucose oxidase GOD and soybean peroxidase SBP, glucose oxidase GOD and horse radish peroxidase HRP, glucose oxidase GOD and dye decolorizing peroxidase DyP are jointly applied to mycotoxin detoxification;
more preferably, the glucose oxidase GOD and the dye decolorizing peroxidase DyP are used in combination for mycotoxin detoxification.
The dosage percentage of the glucose oxidase GOD and the peroxidase POD can be as follows: 1:1 or 1:2 or 1:3 or 1:4 or 1:5 or 1:6 or 1:7 or 1:8 or 1:9 or 1: 10.
In the above applications, the mycotoxins include but are not limited to zearalenone, α -zearalenol, β -zearalenol, α -zearalanol, β -zearalanol, zearalenone, aflatoxins B1, B2, G1, G2, M1 or M2, trichothecene toxoids (including vomitoxin, T-2 toxin, etc.), ochratoxins, fumonisins, patulin, gliotoxins and variegated aspergillons, and preferably zearalenone, aflatoxins B1 and vomitoxin.
In the application, the concentration of the mycotoxin is 0.001-500mg/kg, the concentration of the glucose oxidase GOD is 0.1U/m L-1U/m L, and the concentration of the peroxidase POD is 1U/m L-10U/m L.
The invention also provides a complex enzyme, which comprises glucose oxidase GOD and peroxidase POD.
The peroxidase POD includes, but is not limited to, one or more of dye decolorizing peroxidase DyP, horseradish peroxidase HRP, lactoperoxidase L PO, lignin peroxidase L iP, manganese peroxidase MnP, chloroperoxidase CPO and soybean peroxidase SBP.
The dosage percentage of the glucose oxidase GOD and the peroxidase POD can be as follows: 1:1 or 1:2 or 1:3 or 1:4 or 1:5 or 1:6 or 1:7 or 1:8 or 1:9 or 1: 10.
The compound enzyme also comprises various natural and synthetic mediators which assist peroxidase in degrading mycotoxin, the mediator includes but is not limited to one or more of acetosyringone, methyl syringate, acetovanillone, p-coumaric acid, syringaldehyde, cinnamaldehyde, citral, perillaldehyde, isovaleraldehyde, eugenol, carvacrol, thymol, vanillyl phenol, limonene, gamma-terpinene, allicin, anethole, menthone, linalool, zingerone, isothiocyanate, VIO, 2-hydrazine-bis (3-ethyl-benzothiazole-6-sulfonic acid) ABTS, 1-hydroxybenzotriazole HBT, 2 ', 6, 6' -tetramethylpiperidine oxide TEMPO or polyoxometallate, and the amount of the mediator is 1% -5% of the amount of the enzyme.
The invention also provides the application of the complex enzyme in mycotoxin detoxification.
In the above applications, the mycotoxins include but are not limited to zearalenone, α -zearalenol, β -zearalenol, α -zearalanol, β -zearalanol, zearalenone, aflatoxins B1, B2, G1, G2, M1 or M2, trichothecene toxoids (including vomitoxin, T-2 toxin, etc.), ochratoxins, fumonisins, patulin, gliotoxins and variegated aspergillons, and preferably zearalenone, aflatoxins B1 and vomitoxin.
In the application, the concentration of the mycotoxin is 0.001-500mg/kg, the concentration of the glucose oxidase GOD is 0.1U/m L-1U/m L, the concentration of the peroxidase POD is 1U/m L-10U/m L, and the dosage of the mediator accounts for 1% -5% of the dosage of the enzyme.
In the application, the glucose oxidase GOD and the peroxidase can be exogenously expressed by a genetic engineering technology or can be sourced from markets.
The invention has the advantages and beneficial effects that:
the invention provides the combined application of glucose oxidase GOD and peroxidase POD to the field of mycotoxin detoxification for the first time. Tests prove that the two enzymes can degrade various mycotoxins by composite application, and particularly, the detoxification efficiency on zearalenone, aflatoxin and vomitoxin is high. The two enzymes can be prepared by self, have low production cost and are suitable for biological detoxification of mycotoxin in feed and food.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.
FIG. 1 shows the effect of the complex enzyme of the present invention in degrading zearalenone;
FIG. 2 is a liquid chromatogram of the complex enzyme degradation AFB 1;
FIG. 3 is a liquid chromatogram of the complex enzyme for degrading vomitoxin;
FIG. 4 is a liquid chromatogram of the compound enzyme for degrading zearalenone according to the invention;
Detailed Description
The invention provides a method for detoxifying mycotoxin by jointly applying glucose oxidase GOD and peroxidase POD, and a person skilled in the art can use the contents for reference and appropriately improve the process parameters. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail below with reference to the accompanying drawings and embodiments. It will be apparent to those skilled in the art that the present technology can be implemented and applied by modifying or appropriately modifying or combining the methods and applications herein without departing from the spirit, scope, and content of the present invention. The material reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1 degradation Effect of glucose oxidase GOD and dye decolorizing peroxidase DyP on zearalenone
Dissolving dye decolorizing peroxidase DyP in sodium phosphate buffer solution (0.1M, pH7.0), taking 1M L dye decolorizing peroxidase DyP solution (enzyme activity 5U/M L), adding 0.0125, 0.025, 0.5, 1, 2, 4 and 8U/M L glucose oxidase GOD respectively, adding 200 μ M glucose and ZEN, wherein the ZEN concentration reaches 10 μ g/M L, taking a system without adding dye decolorizing peroxidase DyP as a control, reacting at 37 ℃ for 3h respectively, adding 1M L methanol to terminate the reaction, and detecting the ZEN content by high performance liquid chromatography.
As shown in the result of figure 1, the degradation rate of the dye-decolorizing peroxidase DyP on ZEN is increased with the increase of the concentration of glucose oxidase GOD, and the degradation rate of the dye-decolorizing peroxidase DyP on ZEN reaches 68% when the concentration of GOD is 1U/m L.
Weighing a certain amount of glucose oxidase GOD and horseradish peroxidase HRP, mixing according to a mass percentage of 1:3 to prepare a complex enzyme for degrading mycotoxin, then using the prepared complex enzyme for degrading aflatoxin B1(AFB1), dissolving 1mg of the complex enzyme in a 1M L sodium citrate buffer solution (0.1M, pH 6) reaction system, adding 100 mu M of glucose, then adding AFB1, wherein the concentration of the AFB1 is 1 mu g/M L, reacting for 12 hours at 37 ℃, adding 1M L of methanol to terminate the reaction, using the system without the complex enzyme as a control, and detecting the content of AFB1 by adopting high performance liquid chromatography.
The result shows that the compound enzyme for degrading the mycotoxin reacts with 1 mu g/m L AFB1 for 12 hours, and the degradation rate of the compound enzyme on AFB1 is 82% (figure 2).
Weighing a certain amount of glucose oxidase GOD and lactoperoxidase L PO, mixing according to a mass percentage of 1:4, adding a mediator ABTS accounting for 1% of the enzyme amount to prepare a complex enzyme for degrading mycotoxin, using the prepared complex enzyme for degrading vomitoxin (DON), dissolving 1mg of the complex enzyme in a 1M L sodium citrate buffer solution (0.1M, pH5.0) reaction system, adding 200 mu M glucose, adding DON with the concentration of 20 mu g/M L, reacting at 37 ℃ for 24h, adding 1M L methanol to terminate the reaction, using the system without the complex enzyme as a control, and detecting the content of DON by adopting high performance liquid chromatography.
The result shows that the compound enzyme for degrading the mycotoxin reacts with 20 mu g/m L DON for 24 hours, and the degradation rate of the compound enzyme on the DON is 63 percent (figure 3).
Weighing a certain amount of glucose oxidase GOD and dye decolorizing peroxidase DyP, mixing according to the enzyme activity ratio of 1:5 to prepare complex enzyme for degrading mycotoxin, using the prepared complex enzyme for degrading Zearalenone (ZEN), dissolving the complex enzyme containing 0.5U of glucose oxidase GOD and 2.5U of dye decolorizing peroxidase DyP in a 1M L sodium phosphate buffer solution (0.1M, pH8.0) reaction system, adding 300 mu M of glucose, adding ZEN with the concentration of 10 mu g/M L, reacting for 12h at 42 ℃, adding 1M L of methanol to terminate the reaction, using the system without the complex enzyme as a control, and detecting the content of ZEN by adopting high performance liquid chromatography.
The result shows that the complex enzyme for degrading the mycotoxin reacts with 10 mu g/m L ZEN for 12 hours, and the degradation rate of the complex enzyme on the ZEN is 96% (figure 4).
Example 5 preparation method and application of complex enzyme for degrading mycotoxin
Weighing a certain amount of glucose oxidase GOD and dye decolorizing peroxidase DyP, mixing according to the mass percentage of 1:3 to prepare complex enzyme for degrading mycotoxin, then using the prepared complex enzyme for degrading Zearalenone (ZEN), dissolving 1mg of complex enzyme in a 1M L sodium citrate buffer solution (0.1M, pH6.0) reaction system, adding 300 mu M glucose and 5mM ABTS, then adding ZEN, wherein the ZEN concentration is 10 mu g/M L, reacting for 6h at 42 ℃, adding 1M L methanol to terminate the reaction, using the system without the complex enzyme as a control, and detecting the ZEN content by adopting high performance liquid chromatography.
The result shows that the compound enzyme for degrading mycotoxin reacts with 10 mu g/m L ZEN for 6 hours in the presence of the mediator ABTS, and 100% of ZEN can be degraded.
Example 6 preparation method and application of complex enzyme for degrading mycotoxin
Weighing a certain amount of glucose oxidase GOD and soybean peroxidase SBP, mixing according to a mass percentage of 1:1, adding a mediator acetosyringone accounting for 2% of the enzyme amount to prepare a complex enzyme for degrading mycotoxin, then using the prepared complex enzyme for degrading Zearalenone (ZEN), dissolving 1mg of the complex enzyme in a 1M L sodium citrate buffer solution (0.1M, pH4.0) reaction system, adding 200 mu M glucose, adding ZEN with a ZEN concentration of 10 mu g/M L, reacting for 12 hours at 37 ℃, adding 1M L methanol to terminate the reaction, using the system without adding complex enzyme as a control, and detecting the content of ZEN by adopting high performance liquid chromatography.
The result shows that the complex enzyme for degrading the mycotoxin reacts with 10 mu g/m L ZEN for 12 hours, and the degradation rate of the complex enzyme on the ZEN is 86%.
Example 7 preparation method and application of complex enzyme for degrading mycotoxin
Weighing a certain amount of glucose oxidase GOD and lignin peroxidase L iP, mixing according to a mass percentage of 1:4, adding a mediator syringaldehyde accounting for 1.5% of the enzyme amount to prepare a complex enzyme for degrading mycotoxin, using the prepared complex enzyme for degrading Zearalenone (ZEN), dissolving 1mg of the complex enzyme in a reaction system of 1M L sodium phosphate buffer solution (0.1M, pH7.0), adding 300 mu M of glucose, adding ZEN with the concentration of 10 mu g/M L, reacting at 37 ℃ for 12h, adding 1M L of methanol to terminate the reaction, using the system without the complex enzyme as a control, and detecting the content of ZEN by adopting high performance liquid chromatography.
The result shows that the compound enzyme for degrading the mycotoxin reacts with 10 mu g/m L ZEN for 12 hours, and the degradation rate of the compound enzyme for the ZEN is 88%.
Weighing a certain amount of glucose oxidase GOD and manganese peroxidase MnP, mixing according to the enzyme activity ratio of 1:8 to prepare a complex enzyme for degrading mycotoxin, using the prepared complex enzyme for degrading aflatoxin B1(AFB1), dissolving the complex enzyme containing 0.3U of glucose oxidase GOD and 2.4U of manganese peroxidase MnP in a 1M L sodium malonate buffer solution (0.1M, pH4.5) reaction system, adding 100 mu M glucose and 1mM MnSO4Then adding AFB1 with AFB1 concentration of 1 μ g/m L, reacting at 37 deg.C for 12h, adding 1m L methanol to stop the reaction, using the system without complex enzyme as control, and detecting the content of AFB1 by high performance liquid chromatography.
The result shows that the compound enzyme for degrading the mycotoxin reacts with 1 mu g/m L AFB1 for 12 hours, and the degradation rate of the compound enzyme to AFB1 is 90%.
Example 9 preparation method and application of complex enzyme for degrading mycotoxin
Weighing a certain amount of glucose oxidase GOD and chloroperoxidase CPO, mixing according to a mass percentage of 1:5, adding a mediator syringaldehyde accounting for 0.8% of the enzyme amount and a mediator acetosyringone accounting for 0.7% of the enzyme amount to prepare a complex enzyme for degrading mycotoxin, then using the prepared complex enzyme for degrading vomitoxin (DON), dissolving 1mg of the complex enzyme in a 1M L sodium citrate buffer solution (0.1M, pH6.0) reaction system, adding 200 mu M glucose, adding DON with the concentration of 20 mu g/M L, reacting for 24h at 30 ℃, adding 1M L methanol to terminate the reaction, using the system without the complex enzyme as a control, and detecting the content of DON by adopting high performance liquid chromatography.
The result shows that the compound enzyme for degrading the mycotoxin reacts with 20 mu g/m L DON for 24 hours, and the degradation rate of the compound enzyme on the DON is 81 percent.
Example 10 preparation method and application of complex enzyme for degrading mycotoxin
Weighing a certain amount of glucose oxidase GOD, dye decolorizing peroxidase DyP and horseradish peroxidase HRP, mixing according to the mass percentage of 1:3:3 to prepare complex enzyme for degrading mycotoxin, then using the prepared complex enzyme for degrading aflatoxin B1(AFB1), dissolving 1mg of complex enzyme in a 1M L sodium citrate buffer solution (0.1M, pH6.0) reaction system, adding 100 mu M of glucose, then adding AFB1, wherein the concentration of AFB1 is 1 mu g/M L, reacting for 12h at 37 ℃, adding 1M L of methanol to terminate the reaction, using the system without the complex enzyme as a control, and detecting the content of AFB1 by adopting high performance liquid chromatography.
The result shows that the compound enzyme for degrading the mycotoxin reacts with 1 mu g/m L AFB1 for 12 hours, and the degradation rate of the compound enzyme to AFB1 is 85%.
Example 11 preparation method and application of Complex enzyme for degrading mycotoxin
Weighing a certain amount of glucose oxidase GOD, dye decolorizing peroxidase DyP and lignin peroxidase L iP, mixing according to the enzyme activity ratio of 1:5:1 to prepare complex enzyme for degrading mycotoxin, dissolving the complex enzyme containing 0.4U of glucose oxidase GOD, 2U of dye decolorizing peroxidase DyP and 0.4U of lignin peroxidase L iP in a 1M L sodium phosphate buffer solution (0.1M, pH8.0) reaction system, adding 150 mu M of glucose, adding ZEN with the ZEN concentration of 10 mu g/M L, reacting for 12 hours at 42 ℃, adding 1M L of methanol to terminate the reaction, using the system without the complex enzyme as a control, and detecting the ZEN content by adopting high performance liquid chromatography.
The result shows that the compound enzyme for degrading the mycotoxin reacts with 10 mu g/m L ZEN for 12 hours, and the degradation rate of the compound enzyme for the ZEN is 90%.
Example 12 preparation method and application of Complex enzyme for degrading mycotoxin
Weighing a certain amount of glucose oxidase GOD, dye decolorizing peroxidase DyP and lactoperoxidase L PO, mixing according to the enzyme activity ratio of 1:5:3 to prepare a complex enzyme for degrading mycotoxin, then using the prepared complex enzyme for degrading aflatoxin B1(AFB1), dissolving the complex enzyme containing 0.5U of glucose oxidase GOD, 2.5U of dye decolorizing peroxidase DyP and 1.5U of lactoperoxidase L PO in a 1M L sodium phosphate buffer solution (0.1M, pH7.0) reaction system, adding 400 mu M glucose, adding ABTS (1mM) according to 1% of the total mass, adding AFB1, wherein the concentration of AFB1 is 1 mu g/M L, reacting at 37 ℃ for 24h, adding 1M L methanol to terminate the reaction, using the system without adding the complex enzyme as a control, and detecting the content of AFB1 by adopting high performance liquid chromatography.
The result shows that the compound enzyme for degrading the mycotoxin reacts with 1 mu g/m L AFB1 for 12 hours, and the degradation rate of the compound enzyme to AFB1 is 93%.
Example 13 preparation method and application of Complex enzyme for degrading mycotoxin
Weighing a certain amount of glucose oxidase GOD, dye decolorizing peroxidase DyP, horseradish peroxidase HRP and soybean peroxidase SBP, mixing the materials according to the mass percentage of 1:2:1:2, mixing a mediator syringaldehyde according to the proportion of 1% of the total mass to prepare a complex enzyme for degrading mycotoxin, and then using the prepared complex enzyme for degrading α -zearalenol, β -zearalenol, α -zearalenol, β -zearalenol, zearalenone, aflatoxin B2, G1, G2, M1 or M2, T-2 toxin, ochratoxin, fumonisin, patulin, gliotoxin and variegated aspergillin.
α -zearalenol, β -zearalenol, α -zearalanol, β -zearalanol, zearalenone, AFB2, AFG1, AFG2, AFM1 or AFM2, T-2 toxin, ochratoxin, fumonisin, patulin, gliotoxin and variegated aspergillin are respectively dissolved in dimethyl sulfoxide, the test is carried out according to a reaction system that 1mg of complex enzyme is dissolved in 1M β sodium phosphate buffer (0.1M, pH8), 500 μ M glucose is added, and then toxin is added, wherein α -zearalenol, β -zearalenol, α -zearalanol, β -zearalenol and zearalenone are 10 μ g/M β, AFB2, AFG1, AFG2, 1 or 2, AFM2, fumonisin, fipronil and fustin are respectively used as a control system for detecting the concentration of the fumonisins, the degradation efficiency is not detected by adding methanol, the concentration of 8653-L g of the complex enzyme, the liquid phase, the degradation efficiency is not detected by adopting the same as the concentration of the liquid phase, and the concentration of the liquid phase, the.
TABLE 1 degradation efficiency of Complex enzymes for the degradation of mycotoxins on different toxins
Example 14 Effect of Complex enzymes for degrading mycotoxin on degradation of ZEN in mildew feed
Weighing a certain amount of glucose oxidase GOD, dye decolorizing peroxidase DyP, horseradish peroxidase HRP, soybean peroxidase SBP and chloroperoxidase CPO, mixing according to the mass percentage of 1:1:1:1:2, mixing with acetosyringone according to the proportion of 1.5% of the total mass to prepare a complex enzyme for degrading mycotoxin, dissolving 1g of the complex enzyme, 300 MuM of glucose and 1g of mildew feed (2 Mug/g of ZEN) in 10M L pH2.5 artificial gastric juice, placing in a 37 ℃ water bath, keeping warm and digesting for 3h in the dark, adjusting the pH value to 6.8 by 0.1 mol/L sodium hydroxide solution, adding 10M L artificial simulated intestinal juice, placing in a 37 ℃ water bath, keeping warm and digesting for 8h in the dark, adding 20M L of methanol to terminate the reaction after the digestion is completed, using a system without adding complex enzyme as a reference, and detecting the content of ZEN in the system by adopting an immunoaffinity column-high performance liquid chromatography method.
The result shows that the complex enzyme can degrade 73% of ZEN in the mildewed feed.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (11)
1. The combined use of glucose oxidase GOD and peroxidase POD in mycotoxin detoxification.
2. The use according to claim 1, wherein the glucose oxidase GOD is used in combination with one or more peroxidases selected from the group consisting of dye decolorizing peroxidase DyP, horseradish peroxidase HRP, lactoperoxidase L PO, lignin peroxidase L iP, manganese peroxidase MnP, chloroperoxidase CPO, soy peroxidase SBP;
preferably, the glucose oxidase GOD and soybean peroxidase SBP, the glucose oxidase GOD and horse radish peroxidase HRP, the glucose oxidase GOD and dye decolorizing peroxidase DyP are used in combination;
more preferably, the combination of glucose oxidase GOD and dye decolorizing peroxidase DyP is used.
3. The glucose oxidase GOD and peroxidase POD as claimed in claim 1 in a percentage of 1:1 to 1: 10.
4. The use of claim 1, wherein the mycotoxins include, but are not limited to, zearalenone, α -zearalenol, β -zearalenol, α -zearalanol, β -zearalanol, zearalenone, aflatoxins B1, B2, G1, G2, M1 or M2, vomitoxin, T-2 toxin, ochratoxin, fumonisin, patulin, gliotoxin and variegated aspergillotoxin, preferably zearalenone, aflatoxin B1 and vomitoxin.
5. A complex enzyme is characterized by comprising glucose oxidase GOD and peroxidase POD.
6. The peroxidase POD of claim 5 comprising but not limited to one or more of dye decolorizing peroxidase DyP, horseradish peroxidase HRP, lactoperoxidase L PO, lignin peroxidase L iP, manganese peroxidase MnP, chloroperoxidase CPO, soy peroxidase SBP.
7. The complex enzyme according to claim 5, wherein the dosage percentage of the glucose oxidase GOD and the peroxidase POD is 1:1-1: 10.
8. The complex enzyme of claim 5, further comprising various natural and synthetic mediators that assist the peroxidase in degrading mycotoxins, the mediator includes but is not limited to one or more of acetosyringone, methyl syringate, acetovanillone, p-coumaric acid, syringaldehyde, cinnamaldehyde, citral, perillaldehyde, isovaleraldehyde, eugenol, carvacrol, thymol, vanillyl phenol, limonene, gamma-terpinene, allicin, anethole, menthone, linalool, zingerone, isothiocyanate, VIO, 2-hydrazine-bis (3-ethyl-benzothiazole-6-sulfonic acid) ABTS, 1-hydroxybenzotriazole HBT, 2 ', 6, 6' -tetramethylpiperidine oxide TEMPO or polyoxometallate, and the amount of the mediator is 1% -5% of the amount of the enzyme.
9. Use of a complex enzyme according to any one of claims 5 to 8 for the detoxification of mycotoxins.
10. The use of claim 9, wherein the mycotoxins include, but are not limited to, zearalenone, α -zearalenol, β -zearalenol, α -zearalanol, β -zearalanol, zearalenone, aflatoxins B1, B2, G1, G2, M1 or M2, vomitoxin, T-2 toxin, ochratoxin, fumonisin, patulin, gliotoxin and variegated aspergillotoxin, preferably zearalenone, aflatoxin B1 and vomitoxin.
11. The use according to claim 1 or claim 9, wherein the concentration of mycotoxin is between 0.001 and 500mg/kg, the concentration of glucose oxidase GOD is between 0.1U/m L and 1U/m L, and the concentration of peroxidase POD is between 1U/m L and 10U/m L.
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| CN113699127A (en) * | 2021-08-05 | 2021-11-26 | 中国农业科学院北京畜牧兽医研究所 | Method for improving mycotoxin degradation rate of dye decolorizing peroxidase |
| CN114763540A (en) * | 2021-01-11 | 2022-07-19 | 中国农业大学 | Enzyme, preparation method and application thereof |
| CN117887681A (en) * | 2024-03-13 | 2024-04-16 | 潍坊新希望六和饲料科技有限公司 | Application of horseradish peroxidase in preparation of mildew-removed feed |
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| CN114763540A (en) * | 2021-01-11 | 2022-07-19 | 中国农业大学 | Enzyme, preparation method and application thereof |
| CN114763540B (en) * | 2021-01-11 | 2024-04-05 | 中国农业大学 | Enzyme and preparation method and application thereof |
| CN113699127A (en) * | 2021-08-05 | 2021-11-26 | 中国农业科学院北京畜牧兽医研究所 | Method for improving mycotoxin degradation rate of dye decolorizing peroxidase |
| CN113604447A (en) * | 2021-09-28 | 2021-11-05 | 中国农业科学院北京畜牧兽医研究所 | Application of manganese peroxidase in degradation of citrinin and method |
| CN113604447B (en) * | 2021-09-28 | 2021-12-24 | 中国农业科学院北京畜牧兽医研究所 | Application of manganese peroxidase in degradation of citrinin and method |
| CN117887681A (en) * | 2024-03-13 | 2024-04-16 | 潍坊新希望六和饲料科技有限公司 | Application of horseradish peroxidase in preparation of mildew-removed feed |
| CN117887681B (en) * | 2024-03-13 | 2024-05-24 | 潍坊新希望六和饲料科技有限公司 | Application of horseradish peroxidase in preparation of mildew-removed feed |
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