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(Slackia sp.) D-G6 and applications in degrading of vomitoxin

A technology of Rickettsia bacteria and strains, which is applied in the field of mycotoxin degradation, can solve problems such as failure to resolve relevant proteins or enzymes, and achieve the effects of good promotion and application prospects, stable activity, and strong ability to metabolize DON.

Active Publication Date: 2019-10-22
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Regarding the deepoxidation metabolism of DON, only the related strains can carry out the deepoxidation metabolism reaction, but the related proteins or enzymes have not been analyzed yet.

Method used

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  • (Slackia sp.) D-G6 and applications in degrading of vomitoxin
  • (Slackia sp.) D-G6 and applications in degrading of vomitoxin
  • (Slackia sp.) D-G6 and applications in degrading of vomitoxin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Preliminary Screening of DON Deepoxidation Metabolism Primary Chicken Gut Microorganisms

[0043] Preliminary screening of primary chicken intestinal microbial DON deepoxidation metabolism using continuous subculture method, the specific methods and results are as follows:

[0044] 1. Experimental method

[0045] (1) Take 20 intestinal tracts of freshly slaughtered live chickens from the market, put them into an anaerobic box, and return to the laboratory as soon as possible for follow-up experiments;

[0046] (2) Take the contents of the large intestine, small intestine, and cecum from each intestinal tract, put them into a 50mL Corning centrifuge tube, mix evenly according to the ratio of 1g of contents plus 1mL of brain heart infusion (BHI) medium, and obtain a bacterial solution for later use;

[0047] (3) Use the bacterial solution prepared in step (2) as the seed solution, then inoculate the bacterial solution into BHI medium (200 μL system) at a ratio ...

Embodiment 2

[0053] The isolation and identification of embodiment 2 Shrekella D-G6

[0054] 1. Enrichment and culture of positive samples by serial serial dilution method

[0055] The 6 positive samples in Example 1 were serially diluted and screened on a 96-well plate to obtain enriched bacterial groups with DON deepoxidation metabolism ability. The specific methods and results are as follows:

[0056] 1. Experimental method

[0057] (1) 6 positive samples obtained in embodiment 1 are carried out gradient dilution 10 with 96 orifice plate -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 、10 -8 、10 -9 、10 -10 、10 -11 、10 -12 (If no instructions are given during the screening process, the medium used is BHI medium, and the final concentration of DON added is 25 μg / mL). The screening principle is: in the process of gradient subculture, if the gradient dilution -5 Active, serially diluted 10 -6 Inactive, choose 10 -5 Continue to screen the preserved species until a stable positive...

Embodiment 3

[0088] The optimization of embodiment 3 Shrekella D-G6 metabolism DON conditions

[0089] 1. Different media affect the ability of Shrekella D-G6 to metabolize DON

[0090] 1. Experimental method

[0091] (1) Inoculate the preserved strains 1:100 into different media, including basal media and improved media. The basal media include: NB, NB+1% L-Arg, BHI, BHI+0.3% L- Arg+0.3% L-His+0.05% L-Cys, WCA, GAM, TSB; the improved medium is based on the above basal medium, adding 50% chicken intestinal extract (Extract);

[0092](2) Add DON toxin so that the final concentration is 25 μg / mL, culture anaerobically at 37° C. for 3 days, and then use HPLC to detect the ability to metabolize DON.

[0093] The formula of WCA modified medium is: L-arginine 0.5g, tryptone 5.0g, glucose 0.5g, vitamin K 1 0.00025g, sodium chloride 2.5g, peptone 5.0g, hemin 0.0025g, yeast extract powder 0.5g, sodium pyruvate 0.5g, and then add 500mL filter-sterilized chicken intestinal extract.

[0094] 2. Ex...

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Abstract

The invention discloses (Slackia sp.) D-G6 and applications in the degrading of vomitoxin. The (Slackia sp.) D-G6 is firstly obtained through screening, the strain has been deposited on May 8th, 2019in the microbial species culture collection center of Guangdong province and assigned the accession number GDMCC NO : 60661. The strain can efficiently degrade DON into DOM-1, and is stable in activity and strong in ability to metabolize DON; the strain can be applied to the detoxification of vomitoxin in the field of feed processing or food processing, the preparation of DON detoxification preparations and DON de-epoxidation metabolic enzyme, the construction of DON detoxification engineering bacteria, the cultivation of DON tolerant transgenic plants; and the strain has important meanings onfurther screening detoxification genes and has good popularization and application prospects on controlling the pollution of mycotoxin.

Description

technical field [0001] The invention belongs to the technical field of mycotoxin degradation. More specifically, it relates to a strain of Shrekia and its use in degrading vomitoxin. Background technique [0002] Deoxynivalenol (DON), a mycotoxin mainly produced by Fusarium graminearum and Fusarium culmorum, is also known as vomitoxin because it can cause vomiting in pigs and other fungal animals . Food crops are very easy to breed fungi in humid and warm environments. Fungi such as Fusarium graminearum parasitize on crops and will inevitably accumulate secondary metabolites. DON is one of the secondary metabolites, and DON is present in all The detection rate is the highest (average about 95.8%) among the mycotoxins known to be detectable. When crop pollution is serious, the median content of DON can be as high as 933.0mg / kg, which far exceeds the limit standard of DON formulated by my country (1.0mg / kg, national standard GB 2761-2011). At present, the whole world is fa...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/00C12N15/82A01H5/00A23L5/20C12R1/01
CPCA23L5/28C12N9/00C12N15/8279C12N1/205C12R2001/01
Inventor 邓诣群高小娟母培强文继开祝勋花吴玉婷
Owner SOUTH CHINA AGRI UNIV
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