Bacillus strain for degrading aflatoxin B1 and screening and application thereof

A kind of technology of aflatoxin and Bacillus, which is applied in the screening technology of new strains and the field of microorganisms, can solve the problems of difficult large-scale production, large loss of nutrients, and unstable effect, and achieve high efficiency and strong specificity

Active Publication Date: 2018-06-01
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Physical methods mainly include light, ultrasonic and ultra-high temperature, etc., but these processes may be affected by factors such as temperature and time, resulting in damage and loss of nutrients in food
The chemical method is to add some substances to degrade or destroy aflatoxins, such as adding oxidants, ammonium solution and sodium hydroxide method

Method used

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  • Bacillus strain for degrading aflatoxin B1 and screening and application thereof
  • Bacillus strain for degrading aflatoxin B1 and screening and application thereof
  • Bacillus strain for degrading aflatoxin B1 and screening and application thereof

Examples

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Effect test

example 1

[0022] Example 1: Rapid screening of microbial strains degrading aflatoxin according to the present invention

[0023] 1. Implementation steps

[0024] A number of soil samples were randomly collected from the Fruit and Vegetable Garden of the Institute of Landscape Architecture, Zhejiang University, Hangzhou City, Zhejiang Province, and placed in ziplock bags, and the collection name, location, time and other information were indicated. Weigh 5 g of the collected soil sample into 50 mL of 0.85% sterile physiological saline, stir and oscillate evenly to make a soil suspension. Transfer the soil suspension into a sterilized centrifuge tube and centrifuge at 3000-4000g for 3-5min, take 1mL of the supernatant of the soil suspension, and use the concentration gradient dilution method to gradually dilute 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 , draw 200 μL and spread evenly on the solid medium with coumarin as the only carbon source [ingredients: coumarin 1g / L, KH 2 PO 4 ...

Embodiment 2

[0027] Embodiment 2: Degradability determination of microbial strains of the present invention to aflatoxin B1

[0028] 1. Implementation steps

[0029]Inoculate the selected single colony into a 250mL Erlenmeyer flask filled with sterilized 50mL LB liquid medium (ingredients: 8-10g / L peptone, 5-8g / L yeast powder, 10-15g / L sodium chloride), Place it in a constant temperature shaker and culture it at 37°C and 180rpm for 18-24h, then take 900μL of fermentation broth into a sterilized 2mL centrifuge tube, add 100μL of AFB1 toxin and mix well to make the final concentration 2μg / mL , and placed in a 37°C incubator for 3-4 days in the dark. The control group was a sterile liquid medium containing the same concentration of AFB1, and the extraction and detection of aflatoxin B1 were carried out after the reaction was carried out at 0h, 6h, 12h, 24h, 48h and 96h, respectively.

[0030] Extraction of aflatoxin B1: Add 1mL of dichloromethane to the above reaction mixture and extract 3 ...

Embodiment 3

[0037] Embodiment 3: the identification of microbial strain of the present invention

[0038] 1. Implementation steps

[0039] According to the steps of Takara Bacteria Genomic DNA Extraction Kit (TakaraBacteria Genomic DNA Extraction Kit), the total DNA of the strains described in the present invention was extracted, the 16S rDNA gene of the bacteria was amplified with general primers 27F and 1492R, and the amplified products were recovered and sequenced , by determining the sequence of 16S rDNA for bacterial species identification. The obtained sequence results were compared by Blast at NCBI, and the recognized standard sequence data homologous to the 16S rDNA of the strain was obtained from the GenBank database. The sequence similarity was calculated using MEGA software and the Neighbor-Joining algorithm (Neighbor-Joining) was used as the system developmental analysis.

[0040] 2. Implementation result analysis

[0041] The strain of the present invention is identified t...

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Abstract

The invention discloses a bacillus strain for degrading aflatoxin B1 and screening and application thereof. The class of the strain is named bacillus aryabhattai DT and perserved in the China GeneralMicrobiological Culture Collection Center, with a perservation number of CGMCC 14949. A treatment method comprises: strain screening and separation by using a medium taking coumarin as a unique carbonsource, and 16S rDNA authentication. Meanwhile, the degradation effect of the fermentation culture solution of the strain on aflatoxin B1 is explored. A single colony of bacillus aryabhattai on a solid medium is picked and inoculated in 50mL of growth medium and then vibration is performed for 12-24h on a shaking table at 30-40 DEG C; the fermentation culture solution of the bacillus aryabhattaireacts with 2mu g/mL aflatoxin B1 for 3-4d in a dark place to realize a degradation rate of 82.98%. The screened bacillus aryabhattai realizes an obvious degradation effect on aflatoxin B1 and has thecharacteristics of high efficiency, energy saving, environmental friendliness and the like, and can be applied to the preparation of relevant aflatoxin detoxification agents.

Description

technical field [0001] The invention belongs to the field of new strain screening technology and microorganism technology, and particularly relates to a bacillus strain capable of degrading aflatoxin B1 and application thereof. Background technique [0002] Aflatoxin is the most toxic mycotoxin found so far, and its basic structure is difuran ring and coumarin. It is mainly a secondary heterocyclic metabolite produced by Aspergillus flavus and Aspergillus parasiticus. Aflatoxins are highly toxic substances with strong carcinogenicity. Long-term intake of toxins accumulate in the liver can cause liver cancer and cause cancer in other organs such as kidneys, lungs, and gastrointestinal tract. Among all aflatoxins, aflatoxins B1 (AFB1) is the most toxic and has been classified as a Class I human carcinogen by the International Agency for Research on Cancer (IARC). Aflatoxins are mainly found in moldy peanuts, grains, nuts and rice. According to the statistics of the Food and...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02A23L5/20C12R1/07
CPCA23L5/28C12N1/02C12N1/20C12N1/205C12R2001/07
Inventor 周文文汤曦郑晓冬张一鞠刘姝妤
Owner ZHEJIANG UNIV
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