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Method for preparing and purifying DON toxin

A purification method and toxin technology, applied in the field of preparation and purification of DON toxin, can solve the problems of large amount of organic reagents, long production cycle, difficult purification, etc., and achieve the effects of convenient extraction and purification, less impurities and high purity

Inactive Publication Date: 2012-05-30
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The DON toxin obtained by this method has a long production period. Because wheat and rice contain rich components such as protein and sugar, there are often a large amount of impurities in the crude extract of the toxin, which leads to the extraction of the DON produced by this method. The steps of toxin are complicated, the operation time is long, the amount of organic reagent is large, and it will make the purification difficult in the later stage, and the rate of obtaining pure toxin is low

Method used

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  • Method for preparing and purifying DON toxin
  • Method for preparing and purifying DON toxin
  • Method for preparing and purifying DON toxin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Culture medium of the present invention:

[0025] The seed medium formula is mixed according to the volume percentage of the following components: 1% sucrose; 0.2% ammonium nitrate; 0.05% magnesium sulfate; 0.02% iron sulfate; 0.2% potassium dihydrogen phosphate; 0.4% peptone; The balance is water.

[0026] The fermentation medium is mixed according to the volume percentage of the following components: 0.1% of ammonium nitrate, 0.3% of potassium dihydrogen phosphate, 0.05% of magnesium sulfate, 0.4% of sodium chloride, 2% of sucrose, 1% of glycerin, 0.002% of zinc sulfate, and The amount is water.

Embodiment 2

[0028] Fusarium graminearum GZ3639 (the strain was donated by the Mycotoxin Research Center of the United States Department of Agriculture)

[0029] A) adopt the seed culture medium and fermentation medium of embodiment 1

[0030] Transfer Fusarium graminearum from the refrigerated slant to the PDA medium plate for activation, culture at 25-28°C for 4-5 days, pick a small amount of mycelium and inoculate it into the seed medium, and culture it on a shaker at 25-28°C for 3 days , to obtain the seed solution.

[0031] Then inoculate the seed liquid in the fermentation medium with an inoculation amount of 5%, and culture it in the dark at 28°C for 15-20 days in a triangular flask or a fermenter, take 1 liter of the fermentation culture liquid and centrifuge to get the supernatant, and use ethyl acetate Extract, and concentrate the extracted ethyl acetate to obtain a crude extract of DON toxin.

[0032] The crude extract of DON toxin was dissolved with cyclohexane: ethyl acetate...

Embodiment 3

[0042] Fusarium graminearum F10 (the strain was isolated and preserved by our laboratory)

[0043] A) adopt the seed culture medium and fermentation medium of embodiment 1

[0044] The preparation process is the same as the method A of Example 2, the only difference being that there is a difference in the source of Fusarium graminearum.

[0045] B) Using wheat grain culture medium

[0046] The preparation process is the same as the B method of Example 2, the only difference being that there is a difference in the source of Fusarium graminearum.

[0047] C) compare the crude toxins obtained by the two methods

[0048] The crude extract of DON toxin prepared and extracted by method A and the crude extract of DON toxin prepared and extracted by method B were detected by liquid chromatography, and the results are shown in figure 2 .

[0049] . Although the source of Fusarium graminearum used in this embodiment is different from that in Example 2, the same conclusion can still...

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Abstract

The invention relates to a method for preparing and purifying a DON toxin, which produces the DON toxin by using fusarium graminearum. A seed culture medium and a fermentation culture medium used in the production are both liquid culture mediums, wherein the seed culture medium is formed by mixing following components in percentage by volume: 1% of sucrose, 0.2% of ammonium nitrate, 0.05% of magnesium sulfate, 0.02% of iron sulfate, 0.2% of potassium dihydrogen phosphate, 0.4% of peptone, 0.2% of yeast extract and balance of water; and fermentation culture medium is formed by mixing followingcomponents in percentage by volume: 0.1% of ammonium nitrate, 0.3% of potassium dihydrogen phosphate, 0.05% of magnesium sulfate, 0.4% of sodium chloride, 2% of sucrose, 1% of glycerol, 0.002% of zinc sulfate and the balance of water. In the specific method, the purified DON toxin is obtained by the steps of activation, culture, fermentation, purification and the like.

Description

technical field [0001] The invention relates to a method for preparing and purifying DON toxin, in particular to a process for preparing and purifying DON toxin by using Fusarium graminearum through liquid seed medium and liquid fermentation medium. Background technique [0002] Mycotoxins are secondary metabolites produced by pathogenic bacteria infecting plants. Among all mycotoxins, trichothecenes are known to be one of the natural substances with high toxicity, carcinogenicity and pollution frequency. Among the mycotoxins, deoxynivalenol (DON) is the most common. DON, also known as vomitoxin (VT), is a mycotoxin produced after wheat and other grains are infected by scab. In 1970, it was first isolated from barley infected with scab in Kagawa Prefecture, Japan. Its structure is 3 , 7,15-trihydroxy-12,13-epoxytrichothecene-9-en-8-one (UenoY et al., 1984). DON contamination of grains will cause severe yield loss. Li Dawei (1989) reported that the loss of wheat due to toxi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/18C07D493/10C12R1/77
Inventor 徐剑宏史建荣祭芳林凡云
Owner JIANGSU ACAD OF AGRI SCI
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