A strain that efficiently degrades aflatoxin b 1 Escherichia coli cg1061

An aflatoxin and Escherichia coli technology, applied in bacteria, microorganism-based methods, microorganisms, etc., can solve the problems of unclear toxicity, limited types and application limitations of degrading bacteria metabolites, and achieve good application prospects.

Active Publication Date: 2020-08-07
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present domestic and foreign AFB 1 Although there are many reports on degrading bacteria, the types are still limited, and the metabolites and toxicity of most degrading bacteria are not clear, so the practical application is limited

Method used

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  • A strain that efficiently degrades aflatoxin b <sub>1</sub> Escherichia coli cg1061
  • A strain that efficiently degrades aflatoxin b <sub>1</sub> Escherichia coli cg1061
  • A strain that efficiently degrades aflatoxin b <sub>1</sub> Escherichia coli cg1061

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Chicken cecum microorganism AFB 1 Degradative activity

[0026] 1. Method

[0027] (1) Randomly buy a live chicken from Wushan Vegetable Market in Guangzhou. After slaughtering, take out the cecum, cut open the cecum to get its contents, add 10ml of LB medium, mix well, let it stand for 10min, and the supernatant is the cecal mixed microorganisms.

[0028] (2) Add 100μl to 900μl LB medium, add 250μg / ml AFB 1 10 μl of the mother solution to make the final concentration 2.5 μg / ml.

[0029] (3) Culture at 37°C, 180r / min, and dark for 3 days.

[0030] (4) Take 200μl and add the same amount of dichloromethane to extract three times, centrifuge at 10000rpm for 1min, and wash the supernatant with N 2 blow dry. Add 150 μL of methanol to fully dissolve the residue, and filter it to the sample bottle with a 0.22 μm nylon needle filter.

[0031] (5) Use high performance liquid chromatography (HPLC) for AFB 1 Residues are tested.

[0032] (6) AFB 1 Metabolic rate...

Embodiment 2

[0034] Example 2 Isolation and identification of Escherichia coli CG1061

[0035] 1. Screening of AFB by plate separation method 1 Degrading bacteria

[0036] Carry out the dilution plate method separation AFB to the chicken gut microbiota described in embodiment 1 1 degrading bacteria. Pour the sterilized LB solid medium into a sterile plate while it is hot, and set it aside after solidification. Chicken intestinal microbial flora was serially diluted, and 10 -5 、10 -6 Draw 200 μL of the bacterial suspension from the two bacterial dilutions and put it in the center of the plate, do 3 repetitions for each dilution, spread evenly on the surface of the medium with a sterile glass coating stick, and let it stand at room temperature for 5 ~10min. All the plates were placed upside down in a constant temperature incubator at 37°C for 24 hours. According to the shape of the colonies, 50 single colonies were randomly picked, respectively inserted into 96-well cell culture plate...

Embodiment 3

[0040] Example 3 Escherichia coli CG1061 pathogenicity detection

[0041] 1. Extract Escherichia coli CG1061 genomic DNA according to the instructions of the DNA extraction kit, perform multiplex PCR according to the instructions of the Diarrhoeogenic E. coli PCR Kit (Statens Serum Institut, Denmark), and detect the ten pathogenic genes currently reported ipa H, aat A, elt A, vtx 2, eae , agg R, vtx 1, aai C, est A-porcine, est The expression of A-human.

[0042] 2. The pathogenicity test results of Escherichia coli CG1061 are attached Figure 4 shown. The results showed that Escherichia coli CG1061 did not carry the above-mentioned known pathogenic genes.

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Abstract

The invention discloses a high-efficiency degradation of aflatoxin B 1 Escherichia coli ( Escherichia coli ) strain CG1061. The strain was deposited in the Guangdong Microbial Culture Collection Center on February 1, 2018, with the preservation number GDMCC NO: 60324. The strain is isolated from chicken cecum and is a non-pathogenic Escherichia coli that can efficiently degrade AFB 1 , can be applied to the preparation of aflatoxin B 1 Detoxification bacteria agents, detoxification enzyme preparations, etc., have good application prospects in food, feed processing and other fields.

Description

technical field [0001] The invention belongs to the technical field of mycotoxin degradation. More specifically, it relates to a strain that efficiently degrades aflatoxin B 1 (AFB 1 ) of Escherichia coli ( Escherichia coli ) strain CG1061. Background technique [0002] Aflatoxin (AFT) is a class of secondary metabolites produced by fungi such as Aspergillus flavus and Aspergillus parasiticus, which have carcinogenic, teratogenic and mutagenic effects, among which aflatoxin B 1 (Aflatoxin B 1 , AFB 1 ) is the most toxic and harmful. AFB 1 The direct toxicity to animals is manifested as liver carcinogenicity, and the indirect toxicity is manifested as the reduction of feed conversion rate, immunity, and fecundity of animals. Aflatoxins mainly exist in peanuts, soybeans, corn and other food crops, and the processed food and feed enter the market, which seriously threatens the health of humans and livestock. Therefore, there is an urgent need for an efficient detoxific...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L5/20C12R1/19
CPCA23L5/28C12N1/205C12R2001/19
Inventor 邓诣群汪玲玲蒋珺吴骏母培强邓凤如
Owner SOUTH CHINA AGRI UNIV
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