Vomitoxin degrading enzyme and gene thereof, preparation method and application, as well as vomitoxin degradation method

A technique for degrading vomitoxins and degrading enzymes, which is applied in the fields of detonating toxin degrading enzymes and their genes, preparation and application, and degrading vomitoxins.

Active Publication Date: 2018-07-06
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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  • Vomitoxin degrading enzyme and gene thereof, preparation method and application, as well as vomitoxin degradation method
  • Vomitoxin degrading enzyme and gene thereof, preparation method and application, as well as vomitoxin degradation method
  • Vomitoxin degrading enzyme and gene thereof, preparation method and application, as well as vomitoxin degradation method

Examples

Experimental program
Comparison scheme
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: 7 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 perfo...

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, the difference being that the nucleotide sequence shown in SEQ ID NO: 8 synthesized by artificial chemical synthesis was used to replace the SEQ ID NO in the step (1) of Example 1 : Nucleotide sequence shown in 7.

[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: 9 synthesized by artificial chemical synthesis was used to replace the SEQ ID NO in the step (1) of Example 1 : Nucleotide sequence shown in 7.

[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

Relating to the field of microorganisms, the invention discloses a vomitoxin degrading enzyme and a gene thereof, a preparation method and application, as well as a vomitoxin degradation method. Specifically, the invention provides the vomitoxin degrading enzyme having an amino acid sequence shown as the following (a) and/or (b): (a) an amino acid sequence shown as SEQ ID NO:2; and (b) an amino acid sequence that is obtained by substitution, deletion or adding of one or more of the 430th-450th amino acid residues in the amino acid sequence shown as SEQ ID NO:2 and still has vomitoxin degradingenzyme activity. The vomitoxin degrading enzyme provided by the invention can achieve efficient and rapid degradation of vomitoxin, 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 Applications(China)
IPC IPC(8): C12N9/00C12N15/52A23K20/189A23L5/20
CPCC12N9/00
Inventor 林海龙李慧胡梦龙何景李文钊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
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