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A kind of vomitoxin-degrading enzyme and its gene, preparation method and application, and method for degrading vomitoxin

A technology for detonating toxin and degrading enzyme is applied in the fields of deoxygenating toxin degrading enzyme and its gene, preparation and application, as well as degrading deoxygenation toxin. Efficient and rapid degradation, the effect of expanding the scope of application

Active Publication Date: 2021-10-22
COFCO NUTRITION & HEALTH RES INST +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patent application CN103243047A discloses a strain of Bacillus subtilis that efficiently degrades deoxynivalenol and its application. 900 μL of Bacillus subtilis ANSB471 fermentation broth is reacted with 100 μL of deoxynivalenol (100 μg / ml), and the degradation rate of deoxynivalenol for 2 hours is 25 %, the degradation rate of vomitoxin in 24 hours of reaction is 56%, and the degradation rate of vomitoxin in 48 hours of reaction is 80%, and the degradation rate still needs to be improved
[0005] In addition, most of the existing biological methods for degrading vomitoxin are carried out under mild conditions (such as a temperature of 25-37°C and not exceeding 40°C, and a pH of about 7), however, under higher temperature loads (such as During transport in containers or during feed pelleting) or harsh acidic and alkaline conditions, there is no better solution, which limits the scope of application of biological methods in the degradation of vomitoxin

Method used

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  • A kind of vomitoxin-degrading enzyme and its gene, preparation method and application, and method for degrading vomitoxin
  • A kind of vomitoxin-degrading enzyme and its gene, preparation method and application, and method for degrading vomitoxin
  • A kind of vomitoxin-degrading enzyme and its gene, preparation method and application, and method for degrading vomitoxin

Examples

Experimental program
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Effect test

Embodiment 1

[0071] This example is used to illustrate the deoxynivalenol-degrading enzyme provided by the present invention and its preparation method and application.

[0072] (1) Acquisition of genes

[0073] 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: 11 And NdeI restriction site, add protective base CCG and XhoI restriction site at the 3' end to obtain the corresponding gene fragment.

[0074] (2) Construction of recombinant plasmids

[0075] Use restriction endonucleases NdeI and XhoI (purchased from NEB Company) to carry out double enzyme digestion on the gene fragment of step (1) and PET30a plasmid (with His tag, purchased from Invitrogen Company, USA) respectively, and digest in a water bath at 37°C for 4h , the enzyme digestion system (50 μL) is as follows:

[0076]

[0077] After perf...

Embodiment 2

[0097] This example is used to illustrate the deoxynivalenol-degrading enzyme provided by the present invention and its preparation method and application.

[0098]The deoxynivalenol-degrading enzyme was prepared according to the method of Example 1, except that the nucleotide sequence shown in SEQ ID NO: 12 synthesized by artificial chemical synthesis was used to replace the SEQ ID NO in step (1) of Example 1 : Nucleotide sequence shown in 11.

[0099] 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

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

[0102] The deoxynivalenol-degrading enzyme was prepared according to the method of Example 1, the difference being that the nucleotide sequence shown in SEQ ID NO: 13 synthesized by artificial chemical synthesis was used to replace the SEQ ID NO in the step (1) of Example 1 : Nucleotide sequence shown in 11.

[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.

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Abstract

The invention relates to the field of microorganisms, and discloses a vomitoxin degrading enzyme, its gene, preparation method and application, and a method for degrading vomitoxin. Specifically, the present invention provides a deoxynivalenol-degrading enzyme having the following amino acid sequence (a) and / or (b): (a) the amino acid sequence shown in SEQ ID NO: 2; (b) an amino acid sequence in which one or more of the 155th and 430-450th amino acid residues in the amino acid sequence shown in SEQ ID NO: 2 still have deoxynivalenolase activity after substitution, deletion or addition. The deoxynivalenol degrading enzyme provided by the invention can efficiently and rapidly degrade the deoxynivalenol, and has good industrial application prospects.

Description

technical field [0001] The present invention relates to the field of microorganisms, in particular to a deoxynivalenol-degrading enzyme, a gene encoding the deoxynivalenol-degrading enzyme, a recombinant vector and strain containing the gene, an additive, grain oil or feed containing the additive, expressing the vomiting The method for degrading the toxin, and the application of the above-mentioned deoxynivalenolase, gene, recombinant vector, bacterial strain and additive in degrading the deoxynivalenol, and the method for degrading the deoxynivalenol. Background technique [0002] Vomitoxin (vomitoxin), also known as deoxynivalenol (deoxynivalenol, DON), the chemical name is 3α, 7α, 15-trihydroxy Fusarium-9-en-8-one, because it can cause pigs Named after vomiting, it is mainly trichothecene toxins produced by Fusarium graminearum and Fusarium culmorum infecting wheat, barley, oats, corn and other grains. Worldwide, vomitoxin is one of the main contaminating mycotoxins in g...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/00C12N15/52A23K20/189A23L5/20
CPCC12N9/00
Inventor 林海龙李文钊胡梦龙何景李慧苏会波李凡陈博王小艳张媛佟易李义李久仁樊维荣郭翠周勇
Owner COFCO NUTRITION & HEALTH RES INST