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Application and method of manganese peroxidase in degradation of patulin

A technology of manganese peroxidase and patulin, which is applied in the application field of degrading patulin, can solve the problems of less patulin degrading enzyme and the like, and achieves the effects of low cost and wide application range

Pending Publication Date: 2021-11-26
INST OF ANIMAL SCI CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, few patulin-degrading enzymes have been isolated and identified

Method used

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  • Application and method of manganese peroxidase in degradation of patulin
  • Application and method of manganese peroxidase in degradation of patulin
  • Application and method of manganese peroxidase in degradation of patulin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1 Preparation of recombinant manganese peroxidase MrMnP

[0017] The MrMnP gene sequence from Moniliophthora roreri was fully synthesized from Jinweizhi Company. The X33 / MrMnP Pichia pastoris engineering strain containing the recombinant plasmid was inoculated in 50 mL of YPD medium, shaken at 30 °C and 220 rpm for 48 h, and then transferred to 300 mL of BMGY medium at a ratio of 2%, at 30 °C. , 220 rpm shaking culture for 48 h, the BMGY yeast culture solution was centrifuged at 5,500 rpm for 5 min, and the supernatant was discarded. Add 200 mL of BMMY medium (with the addition of heme at a final concentration of 100 µM) to the fermentation flask. Cultivate on a shaker at 30°C for 48 hours at 200 rpm, centrifuge at 5,500 rpm for 5 minutes, collect the fermentation broth, and prepare the recombinant manganese peroxidase MrMnP.

Embodiment 2

[0018] Example 2 Degradation of patulin by manganese peroxidase

[0019] Dissolve patulin in acetonitrile to prepare a 5 g / L stock solution, and use the following reaction system: 250 μL malonic acid buffer (0.2 M, pH 5.0), 100 μL patulin solution (50 mg / L ), 250 μL manganese sulfate (40 mM), 250 μL manganese peroxidase (5000 U / L) prepared in Example 1, 200 μL hydrogen peroxide (5 mM). The system without adding manganese peroxidase was used as a control, and the reaction system was repeated three times. The reaction was carried out at 30°C. After 12 h, three times the volume of methanol was added to terminate the reaction. The degradation rate of patulin was analyzed by high performance liquid chromatography (HPLC). The liquid chromatography is Shimadzu Nexera UHPLC high-performance liquid chromatography analysis system, the chromatographic separation column is Zorbax SB-C18 (4.6×250 mm, 5 μm), mobile phase A (water with 0.1% acetic acid), mobile phase B (acetonitrile) ; Elu...

Embodiment 3

[0020] Example 3 Degradation of patulin in fruit juice by manganese peroxidase

[0021] Dissolve patulin in acetonitrile to prepare a 5g / L stock solution, and follow the following reaction system: 250 μL malonic acid buffer (0.2 M, pH 5.0, apple juice as solvent), 25 μL patulin solution ( 200 mg / L), 250 μL manganese sulfate (40 mM, apple juice as solvent), 125 μL manganese peroxidase (10000 U / L) prepared in Example 1, 100 μL hydrogen peroxide (10 mM), 250 μL apple juice. The system without adding manganese peroxidase was used as a control, and the reaction system was repeated three times. The reaction was carried out at 30°C. After 24 h, three times the volume of methanol was added to terminate the reaction. The degradation rate of patulin was analyzed by high performance liquid chromatography (HPLC). The liquid chromatography is Shimadzu Nexera UHPLC high-performance liquid chromatography analysis system, the chromatographic separation column is ZorbaxSB-C18 (4.6×250 mm, 5 μ...

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Abstract

The invention relates to the technical field of biology, in particular to application and a method of manganese peroxidase in degradation of patulin. The invention provides application of manganese peroxidase with an amino acid sequence as shown in SEQ ID NO:1 in degradation of patulin. The manganese peroxidase derived from Moniliophthora roreri efficiently degrades mycotoxin patulin, and the method is low in cost and wide in application range, and can be widely applied to the field of food toxin degrading enzymes.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to the application and method of manganese peroxidase for degrading patulin. Background technique [0002] Mycotoxins are toxic secondary metabolites produced by fungi that are potentially harmful to food safety. Patulin is one of the most widespread mycotoxins in agricultural products. Patulin is usually associated with apples and apple products, however, it is also found in other fruits and vegetables such as pears, figs, tomatoes, etc. Scientific investigations have also found patulin contamination in grains such as wheat, rice and corn. Patulin poses a number of health risks to humans and animals, including mutagenic, teratogenic and carcinogenic effects. [0003] Traditional methods of controlling mycotoxins include physical, chemical and biological methods. There have been reports that enzymes can degrade mycotoxins, for example, dye decolorization peroxidase degrades zearalenon...

Claims

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Application Information

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IPC IPC(8): A23L5/20A23L2/84C12N9/08
CPCA23L5/25A23L2/84C12N9/0065C12Y111/01013
Inventor 苏小运王帅张伟姚斌王晓璐秦星徐欣欣王苑张红莲罗会颖
Owner INST OF ANIMAL SCI CAAS
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