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A strain of Eggerthella sp. that efficiently degrades vomitoxin D II‑9

A DON, DII-9 technology, applied in bacteria, microorganism-based methods, microorganisms, etc., to achieve the effect of stable activity, wide range of metabolic temperature and pH value, and strong metabolic capacity

Active Publication Date: 2017-07-11
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report of DON deepoxidation metabolizing microorganisms in China

Method used

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  • A strain of Eggerthella sp. that efficiently degrades vomitoxin D II‑9
  • A strain of Eggerthella sp. that efficiently degrades vomitoxin D II‑9
  • A strain of Eggerthella sp. that efficiently degrades vomitoxin D II‑9

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Analysis of DON deepoxidation metabolism capacity of chicken intestinal microbial mixed flora

[0033] 1. Method

[0034] (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.

[0035] (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.

[0036] (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.

[0037] (4) Put the inoculated sample and anaerobic bag into an anaerobic box, and incubate anaerobically at 37°C for 3 days.

[0038](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

[0042] Example 2 Isolation and identification of Eggerthella sp. D II-9

[0043] 1. Antibiotics, gradient dilution, 96-well plate screening

[0044] 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:

[0045] (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.

[0046] (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

[0064] Example 3 Optimal conditions for degradation of DON by Eggerthella sp. D II-9

[0065] 1. Temperature analysis

[0066] 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.

[0067] The results are attached Figure 5 It was shown that the optimum temperature range for the deepoxidation of DON is 30-45°C.

[0068] Low temperature (50 ℃) will make the bacteria Inactivation, completely loses the ability of deepoxidation to metabolize DON.

[0069] 2. pH analysis

[0070] 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.

[0071] The results are attached Figure 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.

[0072] Eaglezella will not lose its activity under acidic conditio...

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Abstract

The invention discloses a strain of Eggerthella sp. D II‑9 that efficiently degrades vomitoxin (DON). 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, deoxynivalenol (DOM‑1). The strain has the characteristics of a wide pH range (5-10) and wide temperature range (30-45°C) of DON degrading activity, and the maintenance of its activity does not depend on the continuous presence of DON in the culture system. The bacterial strain can be applied to the preparation of DON deoxygenase and DON detoxification preparation, and has a good application prospect in the detoxification of DON.

Description

technical field [0001] The invention belongs to the technical field of microbial screening and mycotoxin degradation. More specifically, it involves a strain of Eggerthella sp. D II-9, which is highly efficient 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 mater...

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 邓诣群高小娟母培强陈庆梅文继开
Owner SOUTH CHINA AGRI UNIV
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