Bacillus amyloliquefaciens capable of degrading ZEN (zearalenone) efficiently and application of bacillus amyloliquefaciens

A technology of zearalenone and amylolytic spores, which is applied to Bacillus amyloliquefaciens that can efficiently degrade zearalenone and its application field, and can solve the problem of chemical detoxification agent residue, unsatisfactory detoxification effect, and loss of nutrients and other issues, to achieve the effects of degradation safety, improvement of economic benefits of animal husbandry, and low production and use costs

Active Publication Date: 2015-11-25
ACAD OF NAT FOOD & STRATEGIC RESERVES ADMINISTRATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the physical and chemical methods of detoxification have a certain degree of effect, there are still many shortcomings, such as the unsatisfactory detoxification effect, which may cause the loss of some important nutrients, the residue of chemical detoxification agents, and the high cost, etc.

Method used

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  • Bacillus amyloliquefaciens capable of degrading ZEN (zearalenone) efficiently and application of bacillus amyloliquefaciens
  • Bacillus amyloliquefaciens capable of degrading ZEN (zearalenone) efficiently and application of bacillus amyloliquefaciens
  • Bacillus amyloliquefaciens capable of degrading ZEN (zearalenone) efficiently and application of bacillus amyloliquefaciens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Acquisition and Identification of Bacillus amyloliquefaciens ASAGF142

[0030] Soil samples were collected from cornfields in Wuhu, Anhui, and the 96-microwell plate was used as the culture carrier. Firstly, the enrichment culture method with 5 consecutive zearalenone toxin concentration gradients was used to obtain a bacterial suspension capable of degrading zearalenone. Then spread the bacterial suspension on the LB agar plate (yeast extract 0.5%, tryptone 1%, NaCl 1%, agar powder 1.6%, pH7.2, Bacteria 25min), pick colonies with good growth and separation degree and different colony morphological characteristics and colors, in LB liquid medium with zearalenone concentration of 10 μg / mL yeast extract 0.5%, tryptone 1%, NaCl 1% , agar powder 1.6%, pH 7.2, sterilized at 121°C for 25min) to carry out detoxification test, extract residual zearalenone with methanol and perform HPLC detection to verify the degradation effect of zearalenone in each pure culture, and...

Embodiment 2

[0037] Example 2 Detoxification effect of ASAGF142 in fermentation medium (Fermentation Medium, hereinafter referred to as "FM")

[0038] The activated ASAGF142 was inoculated into 50mL FM liquid medium containing 20μg / mL zearalenone at an inoculum amount of 1%, and detoxified culture was carried out under the shaking condition of 220 rpm, and it was taken out regularly and extracted with methanol. The residual zearalenone was extracted, and the remaining content of zearalenone in the final culture was detected by HPLC. The results showed that the zearalenone in FM was completely degraded, and the degradation rate of ZEN in ASAGF142 reached 100% after 4 hours. %(See figure 1 ). And draw under the liquid reaction system, the degradation effect figure (degradation curve) of above-mentioned bacterial strain (see figure 2 ).

Embodiment 3

[0039] The preparation of embodiment 3ASAGF142 liquid bacterial agent

[0040] Components and proportions of solid medium: 0.5% yeast extract, 1% tryptone, 1% NaCl, 1.6% agar powder, pH 7.2-7.4, high-temperature steam sterilization at 121°C for 25 minutes.

[0041] Components and proportions of seed medium: yeast extract 0.5%, tryptone 1%, NaCl 1%, pH7.2-7.4, high temperature steam sterilization at 121°C for 25 minutes; fermentation medium components and proportions: ASAGF142: lactose 2%, beef extract 1%, NaCl5mM / L, Tween400.005%, adjust the pH to 7.2.

[0042] Strain activation: ASAGF142 with the preservation number CGMCCNo.9464 was inoculated on solid medium, cultured at 37°C for 2 days, and its zearalenone degradation performance was measured, and then inoculated on the inclined surface of the test tube for later use.

[0043] Seed culture: Pick a single colony from the slant medium and insert it into the seed medium, and cultivate it to the logarithmic phase at 37°C to pr...

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Abstract

The invention discloses bacillus amyloliquefaciens capable of degrading ZEN (zearalenone) efficiently and an application of the bacillus amyloliquefaciens. The collecting number of the bacillus amyloliquefaciens ASAGF142 is CGMCC (China General Microbiological Culture Collection Center) No. 9464. The invention further provides an inoculant comprising the collected strain and a preparation method of the inoculant. Besides, the invention provides the application of the strain or the inoculant in degradation of ZEN. The strain can degrade 20 mu g/ml of ZEN completely in short time, and the degradation rate is 100%.

Description

technical field [0001] The invention relates to a strain of bacillus amyloliquefaciens and its application in degrading zearalenone. Background technique [0002] Zearalenone (Zearalenone, ZEN), also known as F-2 toxin, is a An estrogenic mycotoxin with a dihydroxybenzoic acid lactone structure, usually produced by Fusarium graminearum, Fusarium culmorum, Fusarium crookwellense and other Fusarium fungi and released into the environment middle. ZEN is the most widely polluted mycotoxin in the world. ZEN can be detected in grains and agricultural by-products all over the world, including Europe, South America, North America, Asia, Africa and Oceania. ZEN was first isolated from moldy corn by Baldwin et al. Now it is known that there are more than 11 derivatives. ZEN can be found in maize at a very high concentration of 11.8 mg / kg, and is also widely detected in oats, wheat, barley and sorghum. ZEN generally enters the food chain through contaminated plant feed, so dairy pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L1/015A23K1/16C12R1/07
Inventor 张小溪汪洋张晓琳赵晨韩伟
Owner ACAD OF NAT FOOD & STRATEGIC RESERVES ADMINISTRATION
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