Fumonisin degrading enzyme, coding gene, recombinant vector, cell, additive and application thereof

A fumonisin and recombinant cell technology, applied in the field of biotechnology, can solve the problems of long degradation time, unavailability, low removal efficiency, etc., and achieves the effects of good stability, low pH value and environmental friendliness.

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

At present, the use of mycotoxins to pollute grains as raw materials to ferment and produce ethanol is one of the most important methods, but the toxins are further concentrated in a large number of by-product distillers grains obtained from fermentation production, which greatly exceeds the national limit standard and cannot be used
[0009] However, the current bioremoval methods for fumonisins either have low removal efficiency, long degradation time, or poor thermal stability and acid-base stability. Therefore, it is urgent to develop an energy-saving, environmentally friendly and efficient Method for degrading fumonisins

Method used

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  • Fumonisin degrading enzyme, coding gene, recombinant vector, cell, additive and application thereof
  • Fumonisin degrading enzyme, coding gene, recombinant vector, cell, additive and application thereof
  • Fumonisin degrading enzyme, coding gene, recombinant vector, cell, additive and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] This example is used to illustrate the degrading enzyme provided by the present invention and its preparation method and application.

[0070] (1) Acquisition of genes

[0071] The following nucleotide fragments were synthesized by artificial chemical synthesis (commissioned Bao Bioengineering (Dalian) Co., Ltd., the same below); at the 5' end of the nucleotide sequence shown in SEQ ID NO: 1, the initiation codon ATG was added Add the Xho I restriction site before the Nde I restriction site and the 3' end codon TAG.

[0072] (2) Construction of recombinant plasmids

[0073] Use restriction endonucleases Nde I and Xho I (purchased from NEB Company) to perform double enzyme digestion on the PET30a plasmid (with His tag, purchased from Invitrogen Company, USA), and digest it in a water bath at 37°C for 4 h, and the enzyme digestion system (50 μL) as follows:

[0074]

[0075] After agarose gel electrophoresis, the digested product was purified and recovered. Then, a...

Embodiment 2

[0086] This example is used to illustrate the degrading enzyme provided by the present invention and its preparation method and application.

[0087] The degrading enzyme was prepared according to the method of Example 1, and the influence of temperature and pH on the enzyme activity was determined, except that SEQ ID NO: 3 was used instead of SEQ ID NO: 1.

Embodiment 3

[0089] This example is used to illustrate the degrading enzyme provided by the present invention and its preparation method and application.

[0090] The degrading enzyme was prepared according to the method of Example 1, and the influence of temperature and pH on the enzyme activity was determined, except that SEQ ID NO: 4 was used instead of SEQ ID NO: 1.

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Abstract

The invention relates to the biotechnology field, in particular to a fumonisin degrading enzyme, a coding gene, a recombinant vector, a cell, an additive and application thereof, and more specificallyrelates to the fumonisin degrading enzyme with a sequence shown as SEQ ID NO:2 or its mutant, the coding gene of the enzyme, the vector and cell containing the coding gene, the additive containing the enzyme and / or cell and / or the fermentation product thereof, application of the enzyme, the coding gene, the vector, the cell or the additive in degradation of fumonisins and / or other fungal toxins,as well as a method for degradation of fumonisin / or other fungal toxins. The enzyme provided by the invention has the advantages of: environmental friendliness, efficient and short time degradation offumonisins, and no production of harmful by-products. The enzyme can tolerate catalysis at high temperature up to 70DEG C, has low requirement for pH value and good stability, and also can degrade vomitoxin and T2 toxin to a certain degree.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a fumonisin degrading enzyme and its coding gene, a recombinant vector and a recombinant cell containing the coding gene, containing the degrading enzyme and / or the recombinant cell and / or its Additives for fermentation products, the application of the degrading enzyme, the coding gene of the degrading enzyme, the recombinant vector, the recombinant cell or the additive in degrading fumonisins and / or other mycotoxins, and a degradation Fumonisins and / or other mycotoxin methods. Background technique [0002] Mycotoxins are a class of secondary metabolites produced by toxin-producing fungi during the damage process, mainly including aflatoxin, deoxynivalenol (also known as vomitoxin, DON), zearalenone (ZEN) and Fumonisins (FUM), ochratoxins and T-2 toxins, etc. Among them, fumonisin is a kind of mycotoxin, which is a water-soluble metabolite produced by Fusarium moniliforme. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/18C12N15/55A23K20/189A23L5/20
CPCC12N9/18
Inventor 林海龙苏会波熊强唐堂谭剑黄锦张子剑李文钊李凡陈博杨鑫臧传刚王靖
Owner COFCO NUTRITION & HEALTH RES INST
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