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Microbial preparation for degrading aflatoxin and application

A technology for aflatoxin and microbial preparation, which is applied in the field of microbial preparations for degrading aflatoxin, can solve the problems of low efficiency and different detoxification efficiency, and achieves good degradation effect, reduced aflatoxin content and good degradation effect.

Active Publication Date: 2019-01-08
SHANDONG PEANUT RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Different microorganisms have significantly different detoxification efficiencies, and many microorganisms can remove AFB 1 The efficiency is low, and it is necessary to find strains with high detoxification efficiency

Method used

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  • Microbial preparation for degrading aflatoxin and application
  • Microbial preparation for degrading aflatoxin and application
  • Microbial preparation for degrading aflatoxin and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Sample Collection, Strain Isolation and Screening

[0032] 1. Sample collection time and place

[0033] On the morning of May 5, 2017, soil samples of peanut fields were collected from the experimental base of Shandong Peanut Research Institute (Wangcheng Town, Laixi City, Qingdao City).

[0034] 2. Sample processing, strain isolation and screening

[0035] Take 1 g of the collected soil sample and suspend it in 10 mL of sterile water in an ultra-clean bench, shake and dilute to prepare a soil suspension, and then dilute 100 times with sterile distilled water. Take 100 μL of the suspension and spread it on the LB solid plate, and place it at 37°C for cultivation. After 3 days, the grown colony is streaked and purified on the LB solid plate. After 3 times of streaking and purification, a strain is obtained, numbered A2025 .

Embodiment 2

[0036] Identification of embodiment 2 bacterial strain A2025

[0037] According to the method described in "Berger Bacteria Identification Handbook" (Eighth Edition), the morphological characteristics and physiological and biochemical characteristics of strain A2025 were identified, and the specific results were as follows:

[0038] 1. Morphological characteristics: The single colony of strain A2025 is raised on LB medium, milky white, opaque ( figure 1 ), after 2 days of culture at 37°C, the colony was about 4-6mm, and wrinkles appeared on the surface of the colony after 3 days of culture.

[0039] 2. Physiological and biochemical characteristics: Oxidase activity is negative, can grow at 4-54°C, Gram staining is positive, can hydrolyze and utilize casein, gelatin and starch, but cannot utilize Tween 40 and Tween 80.

[0040] 3. 16S rRNA gene analysis

[0041] The bacterial genomic DNA of A2025 was extracted, and PCR amplification was carried out using 16S rRNA gene univers...

Embodiment 3

[0043] The cultivation of embodiment 3 Bacillus siamese A2025

[0044] The medium of Bacillus siamese A2025 is LB medium: 10g peptone, 10g sodium chloride, 5g yeast extract, 1000mL water; it can grow at 4-54°C, the optimal culture temperature is 26-37°C, and it can grow at pH 5-9 growth, the optimal pH is 7-8. It can also grow in GY medium, GY liquid medium: glucose 20g, yeast powder 5g, water 1000mL; GY solid medium: glucose 20g, yeast powder 5g, agar 20g, water 1000mL.

[0045] Take out the frozen Bacillus siamica A2025 from the refrigerator at -80°C, inoculate 50 μL of the Bacillus siamica A2025 bacterial solution into a 5 mL LB liquid medium test tube in the ultra-clean workbench, and cultivate the activated strain for 12 hours. Insert 500 μL of activated bacterial solution into 100 mL of LB liquid medium, shake and culture at 37°C for 48 hours, and the concentration of the bacterial solution is 1.6×10 8 cfu / mL.

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Abstract

The invention discloses a microbial preparation for degrading aflatoxin and application, and belongs to the technical field of aflatoxin detoxification. An active ingredient of the microbial preparation for degrading the aflatoxin, which is disclosed by the invention, is a bacillus siamensis bacterium suspension or a culture of a bacillus siamensis whole culture solution or a crude extract of bacillus siamensis or an extracellular metabolite of the bacillus siamensis; and the bacillus siamensis is bacillus siamensis A2025 and is preserved in the China General Microbiological Culture CollectionCenter on April 12, 2018, the preservation number is CGMCC NO:15611, the address is No. 3, No. 1 Courtyard, Beichen West Road, Chaoyang District, Beijing, and the preservation requiring institution is Shandong Peanut Research Institute. The bacillus siamensis A2025 disclosed by the invention has a high-efficiency degrading effect on the aflatoxin.

Description

technical field [0001] The invention belongs to the technical field of aflatoxin detoxification, and in particular relates to a microbial preparation for degrading aflatoxin and its application. Background technique [0002] Aflatoxin is a class of secondary metabolites mainly produced by fungi such as Aspergillus flavus and A. parasiticus. Analyzed from the molecular structure, the toxoid is a structural analog composed of difuran ring and coumarin. Aspergillus toxins are highly teratogenic, carcinogenic, and mutagenic, and widely contaminate agricultural products and food such as peanuts, corn, cottonseed, rice, dried fruits, and milk. my country is a big exporter of peanuts. Every year, peanuts exported to Japan and the European Union are returned due to excessive aflatoxin, which is the main problem affecting my country's peanut exports. More than two dozen aflatoxins have been identified, of which aflatoxin B 1 (AFB 1 ) is the most widely distributed, and the most t...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P1/04A23L5/20C12R1/07
CPCA23L5/28C12N1/20C12P1/04
Inventor 王明清孙杰张初署于丽娜毕洁杨庆利刘宾赵善仓江晨
Owner SHANDONG PEANUT RES INST
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