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: 2011-09-28
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 nozzle of the standard strain Glioma pink ACCC No.31535 (purchased from the Culture Collection Center of the 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 shaker condition was 220 rpm, 30°C, 72 hours. After the cultivation was completed, the bacteria were collected by centrifugation at 12000 rpm. RNA extraction (Trizol method);

[0032] (2) Using RNA as a template to reverse transcribe and synthesize a cDNA sequence;

[0033] 2. Acquisition of zearalenone degrading enzyme gene

[0034] Using the cDNA sequence obtained in step 1 as a template, a PCR reaction was performed ...

Embodiment 2

[0045] Example 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 until the OD of the bacterial liquid 600 =2-3, collect the bacteria by centrifugation at room temperature, resuspend in BMMY medium and culture for 5 days, take out 1mL of culture solution every day, centrifuge at 12000rpm for 5min, and collect the supernatant. Take 980 μL of the supernatant and put it in a 1.5 mL centrifuge tube, then add 20 μL of 1000ppm ZEN to make the final concentration reach 20ppm. After the addition, react at 30°C for 2h-10h and put it in 4°C for detection. Take 20 μL of the processed sample and enter it into high performance liquid chromatography to detect the residue of ZEN. The detection method is carried out according to the GB / T 23504-2009 method. HPLC detect...

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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, in particular to a zearalenone degrading enzyme and its coding gene, and the application of the zearalenone degrading enzyme. Background technique [0002] Zearalenone (Zearalenone, ZEN) is an estrogenic mycotoxin produced by Fusarium fungus. In 1999, D. Mello et al. found that ZEN can reduce the embryo survival rate and birth weight of newborn fetuses in pregnant animals. The impact of ZEN on the human body is mainly to induce tumors, induce DNA shrinkage, and cause chromosomal abnormalities. In addition, ZEN can bind to 17β-estradiol receptors, resulting in fat oxidation, cell apoptosis, and inhibition of protein and DNA synthesis. Also inhibits the biosynthesis of other macromolecules. [0003] A large number of documents at home and abroad show that ZEN is toxic to animals and humans, and people eating food contaminated by it will cause diseases such as liver cancer, testicular cancer, eso...

Claims

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Application Information

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Patent Type & Authority Applications(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
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