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Eaglezella eggerthella sp. Application of D II‑9 in Degradation of Vomitoxin

A technology of Eaglezella and deoxynivalenol, which is applied in the field of mycotoxin degradation, can solve the problem of no microbial report, etc., and achieve the effect of wide range of metabolic temperature and pH value, stable activity and strong metabolic ability

Active Publication Date: 2018-03-06
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although there are purely cultured microorganisms that deepoxidize and metabolize DON in foreign countries, they are all registered as patents, and there are no reports of related microorganisms in China.

Method used

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  • Eaglezella eggerthella sp. Application of D II‑9 in Degradation of Vomitoxin
  • Eaglezella eggerthella sp. Application of D II‑9 in Degradation of Vomitoxin
  • Eaglezella eggerthella sp. Application of D II‑9 in Degradation of Vomitoxin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Analysis of DON deepoxidation metabolism capacity of chicken intestinal microbial mixed flora

[0037] 1. Method

[0038](1) A Wen’s chicken was randomly purchased from the market (the vegetable market around South China Agricultural University), and its carotid artery was cut with a scalpel, causing the chicken to die due to excessive blood loss.

[0039] (2) Dissect the abdomen and take out the cecum, large intestine and small intestine of the chicken, each 3-5cm, and mix the contents of the intestine with L10 medium.

[0040] (3) Stand still for 3-5 minutes, take 100 μl and add it to 900 μl L10 medium, add 10 μl of 10 mg / ml DON stock solution to make the final concentration 100 μg / ml.

[0041] (4) Put the inoculated sample and anaerobic bag into an anaerobic box, and incubate anaerobically at 37°C for 3 days.

[0042] (5) Take a sample of 100 μl, add three times the volume of acetonitrile to the bacterial solution to be extracted, vortex and shake to mix,...

Embodiment 2

[0046] Example 2 Isolation and identification of Eggerthella sp. D II-9

[0047] 1. Antibiotics, gradient dilution, 96-well plate screening

[0048] First, antibiotics, gradient dilution, and 96-well plate screening were performed on the chicken intestinal microbial flora described in Example 1 to obtain enriched flora with DON deepoxidation metabolism ability. The specific method is as follows:

[0049] (1) Use tetracycline, ampicillin, carbenicillin, tylomycin, lincomycin, cephalosporin, cefotaxime, streptomycin, chloramphenicol and other antibiotics to incubate with chicken intestinal microorganisms to screen out an antibiotic Combination: tetracycline + tylomycin + lincomycin + ampicillin, under the conditions of this combination of antibiotics, the growth of non-target bacteria can be greatly inhibited.

[0050] (2) Take the active flora after antibiotic screening as strains, and serially dilute 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 、10 -8 、10 -9 , HP...

Embodiment 3

[0068] Example 3 Optimum conditions for degradation of DON by Eggerthella sp. D II-9

[0069] 1. Temperature analysis

[0070] Set a series of temperature gradients of 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, and 50°C, and take samples at fixed points to detect the metabolic DON.

[0071] The results are attached Figure 5 It was shown that the optimum temperature range for the deepoxidation of DON is 30-45°C.

[0072] Low temperature (50 ℃) will make the bacteria Inactivation, completely loses the ability of deepoxidation to metabolize DON.

[0073] 2. pH analysis

[0074] Set a series of pH gradients of 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, and take samples at fixed points to detect the metabolic DON.

[0075] The results are attached Image 6 As shown, in the range of pH 5-10, DON is completely converted into DOM-1, but the optimum pH range for deepoxidation of DON is between 6.5-10.0.

[0076] Eaglezella will not lose the activity under acidic condition...

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Abstract

The invention discloses the application of a strain of Eggerthella sp.D II‑9 in degrading vomitoxin. The strain was deposited in the Guangdong Microbial Culture Collection Center on June 23, 2016, and its preservation number is GDMCC NO: 60049. The strain can efficiently and completely degrade DON into its attenuated product, deepoxyvoxime (DOM‑1), and the strain's maintenance of DON degradation activity does not depend on the persistence of DON. The strain has stable activity, strong metabolic ability, wide range of metabolic temperature and pH value, and can be used for the detoxification of DON in the field of feed processing and food processing, the preparation of DON detoxification preparations, the separation of enzymes related to DON deepoxidation metabolism, and the detoxification of DON. The preparation of epoxy metabolism engineering bacteria and the cultivation of DON-tolerant transgenic plants have good application prospects.

Description

technical field [0001] The invention belongs to the technical field of mycotoxin degradation. More specifically, it relates to the use of Eggerthella sp. D II-9 in degrading vomitoxin. Background technique [0002] Deoxynivalenol (DON) is also known as vomitoxin because it can cause vomiting in pigs, dogs, cats and other animals. DON contamination of grains is very common, and many grains are seriously polluted, such as wheat, oats, soybeans and rye. There have been reports of DON contamination of food around the world, including China, Brazil, the United States, Argentina, and South Africa. In 2016, the mycotoxin contamination status of 359 flour samples was investigated in Shandong, China, and it was found that the detection rate of DON was the highest, reaching 97.2%, and the average contamination concentration was 86.7 μg / kg (Li et al., 2016). DON pollution is not only serious in grain crops, but also in feed raw materials and complete compound feeds. From 2012 to 20...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L5/20C12R1/01
CPCC12N1/205C12R2001/01
Inventor 邓诣群母培强高小娟陈庆梅文继开
Owner SOUTH CHINA AGRI UNIV
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