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Zearalenone degrading enzyme, gene, preparation method and application thereof, as well as zearalenone degradation method

A technology for degrading zearalenone and enzymes, applied in botany equipment and methods, biochemical equipment and methods, applications, etc., can solve problems such as long degradation time, degradation rate needs to be improved, and limit the application range of biological methods, etc. To achieve the effect of broad application prospects

Active Publication Date: 2018-07-06
COFCO NUTRITION & HEALTH RES INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be seen that the existing methods for microbial degradation of ZEN take a long time to degrade, and the degradation rate still needs to be improved.
[0006] In addition, most of the existing biological methods for degrading ZEN are carried out under mild conditions (such as 28 °C, pH around 7), however, under high temperature loads (such as during transportation in containers or in feed pellets) There is no better solution under harsh acidic and alkaline conditions, which limits the scope of application of biological methods in degrading ZEN

Method used

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  • Zearalenone degrading enzyme, gene, preparation method and application thereof, as well as zearalenone degradation method
  • Zearalenone degrading enzyme, gene, preparation method and application thereof, as well as zearalenone degradation method
  • Zearalenone degrading enzyme, gene, preparation method and application thereof, as well as zearalenone degradation method

Examples

Experimental program
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Effect test

Embodiment 1

[0074] This example is used to illustrate the zearalenone degrading enzyme provided by the present invention and its preparation method and application.

[0075] (1) Acquisition of genes

[0076] The following nucleotide sequence was synthesized by artificial chemical synthesis (Zuobao Bioengineering (Dalian) Co., Ltd., the same below): a protective base CGC was added to the 5' end of the nucleotide sequence shown in SEQ ID NO: 7 And NdeI restriction site, add protective base CCG and XhoI restriction site at the 3' end to obtain the corresponding gene fragment.

[0077] (2) Construction of recombinant plasmids

[0078] Use restriction endonucleases NdeI and XhoI (purchased from NEB Company) to carry out double enzyme digestion on the gene fragment obtained by step (1) and the PET30a plasmid (with His tag, purchased from Invitrogen Company, USA) respectively, and enzyme in a water bath at 37°C After cutting for 4 hours, the enzyme digestion system (50 μL) is as follows:

[0...

Embodiment 2

[0101] This example is used to illustrate the zearalenone degrading enzyme provided by the present invention and its preparation method and application.

[0102] Prepare zearalenone degrading enzyme according to the method of Example 1, the difference is that the nucleotide sequence as shown in SEQ ID NO: 8 synthesized by artificial chemical synthesis is used to replace the nucleotide sequence in step (1) of Example 1 The nucleotide sequence shown in SEQ ID NO: 7 to obtain the gene fragment.

[0103] Table 1 shows the effect of reaction time on enzyme activity, Table 2 shows the effect of temperature on enzyme activity, and Table 3 shows the effect of pH on enzyme activity.

Embodiment 3

[0105] This example is used to illustrate the zearalenone degrading enzyme provided by the present invention and its preparation method and application.

[0106] Prepare zearalenone-degrading enzyme according to the method of Example 1, the difference is that the nucleotide sequence shown in SEQ ID NO: 9 is used instead of the nucleotide sequence in Example 1 step (1) synthesized by artificial chemical synthesis. The nucleotide sequence shown in SEQ ID NO: 7 to obtain the gene fragment.

[0107] Table 1 shows the effect of reaction time on enzyme activity, Table 2 shows the effect of temperature on enzyme activity, and Table 3 shows the effect of pH on enzyme activity.

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PUM

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Abstract

Relating to the field of microorganisms, the invention discloses a zearalenone (ZEN) degrading enzyme, a gene, a preparation method and application thereof, as well as a zearalenone degradation method. Specifically, the invention discloses a zearalenone degrading enzyme, which has an amino acid sequence shown as (a) and / or (b) as follows: (a) an amino acid sequence shown as SEQ ID NO:2; and (b) anamino acid sequence that is obtained by substitution, deletion or adding of one or more of the 1st-262nd and 264th amino acid residues in the amino acid sequence shown as SEQ ID NO:2 and still has zearalenone degrading enzyme activity. The zearalenone degrading enzyme provided by the invention can achieve efficient and rapid degradation of ZEN, and has good industrial application prospects.

Description

technical field [0001] The present invention relates to the field of microorganisms, in particular to a zearalenone-degrading enzyme, a gene encoding the zearalenone-degrading enzyme, a recombinant vector and strain containing the gene, an additive, and grain and oil containing the additive Or feed, the method for expressing the zearalenone-degrading enzyme, and the application of the above-mentioned zearalenone-degrading enzyme, gene, recombinant vector, bacterial strain and additive in degrading zearalenone, and degrading zearalenone The keto approach. Background technique [0002] Zearalenone (Zearalenone, ZEN) is a mycotoxin produced by Fusarium fungus and released into the soil environment. It is the most widely polluted Fusarium toxin in the world. In 1999, D. Mello et al. found that ZEN can reduce the embryo survival rate and the birth rate of newborn fetuses in pregnant animals. The impact of ZEN on the human body is mainly to induce tumors, induce DNA shrinkage, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/16C12N15/55C12N15/63A23L5/20
CPCC12N9/16C12N15/63
Inventor 苏会波林海龙李文钊张子剑黄锦唐堂杨鑫李凡陈博王小艳张媛
Owner COFCO NUTRITION & HEALTH RES INST
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