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Zearalenone toxin degradation enzyme and coding gene and application thereof

A technology of zearalenone and coding genes is applied in the field of enzyme engineering to achieve the effect of controlling food pollution

Active Publication Date: 2013-02-06
ACAD OF NAT FOOD & STRATEGIC RESERVES ADMINISTRATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, traditional physical and chemical methods have certain limitations. With the application of biotechnology, new progress has been made in the degradation of ZEN.

Method used

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  • Zearalenone toxin degradation enzyme and coding gene and application thereof
  • Zearalenone toxin degradation enzyme and coding gene and application thereof
  • Zearalenone toxin degradation enzyme and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Acquisition of zearalenone-degrading enzyme gene zlhy-6 and expression of zearalenone-degrading enzyme

[0030] 1. Construction of zearalenone degrading enzyme gene plasmid library

[0031] Break the mouth of the ampoule tube of the standard strain M. pink ACCC No.31535 (purchased from the Culture Collection Center, Institute of Microbiology, Chinese Academy of Sciences) and add 300-500 μL of liquid PDA medium (potato 200g / L, glucose 20g / L) , agar 20g / L, PH natural), and then the suspension of the strain was added to 100ml of the same medium, and the shaking conditions were 220 rpm, 30 ° C, and 72 hours. After the cultivation was completed, the cells were collected by centrifugation at 12,000 rpm in a centrifuge. RNA extraction (Trizol method);

[0032] (2) synthesizing a cDNA sequence by reverse transcription using RNA as a template;

[0033] 2. Obtaining the zearalenone degrading enzyme gene

[0034] Using the cDNA sequence obtained in step 1 as a templa...

Embodiment 2

[0045] Embodiment 2 Activity detection of zearalenone degrading enzyme

[0046] Sample processing: Recombinant Pichia strains (experimental group) and wild-type Pichia strains (control group, Pichia strains integrated with empty plasmids) were inserted into BMGY medium and continued to culture to the OD of the bacterial solution 600 = 2-3, the cells were collected by centrifugation at room temperature, resuspended in BMMY medium for 5 days, and 1 mL of culture medium was taken out every day, centrifuged at 12000 rpm for 5 min, and the supernatant was collected. Take 980 μL of the supernatant into a 1.5 mL centrifuge tube, and then add 20 μL of 1000 ppm ZEN to make the final concentration 20 ppm. After the addition, the reaction was performed at 30°C for 2h-10h, and the post-treatment was placed at 4°C for detection. Take 20 μL of the treated sample and put it into high performance liquid chromatography to detect the ZEN residue. The detection method is carried out according ...

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Abstract

The invention discloses a zearalenone toxin gradation enzyme and a coding gene and application thereof. The enzyme is derived from Gliocladiumroseum and is protein in (a) or (b): (a) protein consisting of an amino acid residue sequence of SEQIDNO: 1 in a sequence table; and (b) protein which is derived from the SEQIDNO: 1, has a zearalenone degradation activity and is formed through carrying out substitution and / or deletion and / or addition of one or a plurality of amino acid residues on the amino acid residue sequence of SEQIDNO: 1 in the sequence table. The zearalenone toxin degradation enzyme has highly-efficient degradation effect on zearalenone (ZEN).

Description

technical field [0001] The invention belongs to the field of enzyme engineering, and particularly relates to a zearalenone degrading enzyme and its encoding gene, and the application of the zearalenone degrading enzyme. Background technique [0002] Zearalenone (ZEN) is an estrogen-like mycotoxin produced by Fusarium. In 1999, D.Mello et al. found that ZEN can reduce the embryonic survival rate of pregnant animals and the birth weight of newborn fetuses. The effects of ZEN on the human body are mainly to induce tumors, induce DNA shrinkage, and cause chromosomal aberrations. In addition, ZEN can bind to 17β-estradiol receptors, leading to fat oxidation, cell apoptosis, and inhibition of protein and DNA synthesis. The biosynthesis of other macromolecules can also be inhibited. [0003] A large number of domestic and foreign literatures show that ZEN is poisonous to animals and humans, and that people eating food contaminated with ZEN can lead to diseases such as liver cance...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/18C12N15/55C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A62D3/02C12R1/84A62D101/28
Inventor 吴子丹孙长坡伍松陵路子显
Owner ACAD OF NAT FOOD & STRATEGIC RESERVES ADMINISTRATION