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Bacillus subtilis and application thereof in degrading zearalenone

A Bacillus subtilis, zearalenone technology, applied in the direction of bacteria, microorganism-based methods, microorganisms, etc., can solve problems such as the inability to detect or determine the transformation of zearalenone or the toxicity of degradation products, so as to improve animal husbandry. Industrial economic benefits, low production and use costs, and bacterial strain safety effects

Active Publication Date: 2021-02-26
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are some drawbacks in the existing bacterial agents for removing zearalenone, such as the inability to detect or determine the toxicity of the conversion or degradation products of zearalenone
Therefore, although the existing bacterial agents remove zearalenone, they may still be transformed into derivatives with hidden toxicity

Method used

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  • Bacillus subtilis and application thereof in degrading zearalenone
  • Bacillus subtilis and application thereof in degrading zearalenone
  • Bacillus subtilis and application thereof in degrading zearalenone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1. Isolation, Identification and Preservation of Bacillus subtilis 816 CGMCC No.14854

[0062] 1. Isolation of zearalenone-degrading bacteria 816

[0063] 1. Add 1g of soil sample (collected from the corn field in Chifeng City, Inner Mongolia Autonomous Region, China) to a shaker flask (50mL) filled with 30mL LB liquid medium, mix well; then vibrate at 37°C and 180rpm for 12h to obtain cultured bacteria liquid.

[0064] 2. After completing step 1, use the 96-well plate as the culture carrier, take the culture liquid and adopt the enrichment culture method of 5 consecutive zearalenone toxin concentration gradients to obtain a bacterial suspension capable of degrading zearalenone.

[0065] 3. Take the bacterial suspension and dilute it with sterile water to obtain bacterial solutions with different dilutions. 0.1 mL of each dilution was evenly spread on the LB solid plate, and cultured overnight at 37°C.

[0066] 4. Inoculate colonies with good separation degre...

Embodiment 2

[0079] Example 2, Detection of the effect of Bacillus subtilis 816 on degrading zearalenone

[0080] 1. A single colony of Bacillus subtilis 816 was inoculated in 5 mL of LB liquid medium, and cultured at 37° C. and 180 rpm for 24 hours to obtain a seed solution.

[0081] 2. The seed liquid was inoculated into LB liquid medium containing 100 μg / mL zearalenone according to the inoculum amount of 10% (v / v) to obtain a culture system.

[0082] 3. Extract residual zearalenone in the culture system obtained in step 2 with methanol, and detect by HPLC.

[0083] 4. Take the culture system obtained in step 2, and culture it at 37° C. and 220 rpm for 2 hours, 3 hours, 4 hours or 5 hours to obtain a culture liquid. The residual zearalenone in the culture liquid was extracted with methanol, and detected by HPLC.

[0084] See the test results figure 1 . The results showed that Bacillus subtilis 816 could degrade zearalenone; when cultured for 5 hours, the degradation rate of zearalen...

Embodiment 3

[0085] Example 3, Detection of the conversion rate of Bacillus subtilis 816 to zearalenone at different temperatures

[0086] The concentration of ZEN in the ZEN toxin stock solution was 1 mg / mL, and the solvent was methanol.

[0087] 1. A single colony of Bacillus subtilis 816 was inoculated in 5 mL of LB liquid medium, and cultured at 37° C. and 200 rpm for 24 hours to obtain a seed liquid.

[0088] 2. The seed liquid was inoculated into 5 mL LB liquid culture medium according to the inoculum amount of 1% (v / v), and cultured at 37° C. and 200 rpm for 24 hours to obtain a fermentation liquid.

[0089] 3. Thoroughly mix 2 mL of fermentation broth and 40 μl of ZEN toxin stock solution, and then culture at 10°C, 20°C, 30°C, 40°C, 50°C or 60°C for 2.5 hours at 200 rpm to obtain culture broth. The content of zearalenone in the culture broth was detected by HPLC.

[0090] The fermentation broth was replaced with LB liquid medium, and other steps were unchanged. as a blank contro...

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Abstract

The invention discloses Bacillus subtilis and application thereof in degrading zearalenone. The Bacillus subtilis is specifically Bacillus subtilis 816, and the preservation number of the Bacillus subtilis 816 in the China General Microbiological Culture Collection Center is CGMCC No. 14854. Experiments prove that the Bacillus subtilis 816 CGMCC No. 14854 can degrade the zearalenone into zearalenone phosphate which is greatly reduced in biotoxicity and exists stably. The Bacillus subtilis has important application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a bacillus subtilis and its application in degrading zearalenone. Background technique [0002] Zearalenone (ZEN), also known as F-2 toxin, is a 2,4-dihydroxybenzoic acid lactone compound, which has extensive and serious pollution to grain, food and feed. ZEN itself is not toxic but has strong biological effects, because its chemical structure is similar to naturally occurring estrogens, resulting in ZEN's reproductive toxicity, immunotoxicity, genotoxicity and carcinogenicity. The pollution of ZEN leads to the reduction of animal husbandry production and human diseases, which brings huge economic losses. Therefore, countries all over the world have begun to pay attention to the pollution of ZEN. Pinotti et al. investigated and evaluated the prevalence of ZEN contamination in cereals and feeds worldwide in 2016, and pointed out that the incidence of ZEN contamination in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L5/20C12R1/125
CPCA23L5/28C12N1/20C12N1/205C12R2001/125
Inventor 付晓平郑宏臣宋诙杨世彬赵兴亚甄杰徐健勇
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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