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

A technology of ochratoxin and recombinant cells, applied in application, recombinant DNA technology, enzymes, etc., can solve the problems of unutilization, long degradation time, poor thermal stability, acid-base stability, etc., and meet the requirements of low and stable pH value Good performance and environment-friendly effect

Active Publication Date: 2020-11-17
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
[0008] However, the current methods for biological removal of ochratoxin 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 ochratoxin

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] This example is used to illustrate the degrading enzyme provided by the present invention and its preparation method and application.

[0069] (1) Acquisition of genes

[0070] The following nucleotide fragments were synthesized by artificial chemical synthesis (commissioned Bao Biological Engineering (Dalian) Co., Ltd., the same below); after the initiation codon ATG at the 5' end of the nucleotide sequence shown in SEQ ID NO:1 Add an Nde I restriction site, add an Xho I restriction site and a stop codon TAG at the 3' end.

[0071] (2) Construction of recombinant plasmids

[0072] 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:

[0073]

[0074]

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

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 field of biotechnology, in particular to an ochratoxin degrading enzyme, a coding gene, a recombinant vector, a cell, an additive and application thereof. The invention more specifically relates to the ochratoxin degrading enzyme with a sequence shown as SEQ ID NO:2 or a mutant thereof, a coding gene of the enzyme, a vector and a cell containing the coding gene, and anadditive containing the enzyme and / or cell and / or a fermentation product thereof, application of the enzyme, the coding gene, the vector, the cell or additive in degradation of ochratoxin and / or other fungal toxins, as well as a method for degradation of ochratoxin / or other fungal toxins. The ochratoxin degrading enzyme provided by the invention is environment-friendly, can efficiently degrade ochratoxin in a short time, and does not generate any harmful by-product. The enzyme can tolerate high temperature catalysis up to 80DEG C, also has low pH value requirement, has good stability, and achieve degradation of fumonisins and T2 toxin to a certain degree.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular, to an ochratoxin 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 degradative enzyme, the coding gene of the degradative enzyme, the recombinant vector, the recombinant cell or the application of the additive in degrading ochratoxin and / or other mycotoxins, and a degradation Ochratoxin 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, ochratoxin is another mycotoxin that has attracted worldwide attention after aflatoxin. It is an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/00C12N15/52A23K10/18A23K20/189A23L5/20
CPCC12N9/00
Inventor 林海龙苏会波熊强唐堂谭剑黄锦张子剑李文钊陈博李凡杨鑫臧传刚王靖
Owner COFCO NUTRITION & HEALTH RES INST
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