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A strain of Rhodococcus and its application in degrading zearalenone

A technology of zearalenone and rhodococcus, applied in the direction of bacteria, microorganism-based methods, microorganisms, etc., can solve the problems of weak degradation ability and low ZEN degradation rate, and achieve easy operation, fast and efficient detoxification ability, and production The effect of low cost of use

Active Publication Date: 2018-06-01
ACAD OF NAT FOOD & STRATEGIC RESERVES ADMINISTRATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the degradation rate of these microorganisms to ZEN is not high, and the degradation ability is weak, and after the analysis of metabolites, some microorganisms convert ZEN into ZEN isomers and zearalenol, and the estrogen toxicity of these products Or lower than ZEN or stronger than ZEN

Method used

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  • A strain of Rhodococcus and its application in degrading zearalenone
  • A strain of Rhodococcus and its application in degrading zearalenone
  • A strain of Rhodococcus and its application in degrading zearalenone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Obtaining and Identification of Rhodococcus sp.

[0029] Soil samples were collected from the corn cultivation layer in the high-incidence area of ​​scab in Shangzhai Village, Linzhou, Henan Province, and the 96-well deep-well plate was used as the culture carrier, and the micro-enrichment culture method was adopted to first prepare the sample into a bacterial suspension in sterile water , inoculated in MM liquid medium (NH 4 NO 3 1g / L, (NH 4 ) 2 SO 4 0.5g / L, MgSO 4 ·7H 2 O 0.41g / L, KH 2 PO 4 0.5g / L, K 2 HPO 4 1.5g / L, NaCl 0.5g / L, FeSO 4 ·7H 2 (O 0.037g / L, pH 7.0-7.2, sterilized at 121°C for 30min), the final concentration of ZEN in the medium was 25μg / ml, and after 7 days of culture, transplant with 10% inoculum, and the concentration of ZEN in the medium remained unchanged. After 5 consecutive transplants, the ZEN content was detected by HPLC, and the MM medium containing the same ZEN concentration without inoculation was used as a negative c...

Embodiment 2

[0036] Example 2 Degradation effect of Rhodoccocus sp. on ZEN in MM liquid medium

[0037] Activate Rhodoccocus sp. stored at -80°C on LB agar medium, culture at 30°C, inoculate a single colony in LB liquid medium, culture at 30°C, 200 rpm for 24 hours; centrifuge at 4000 rpm for 10 minutes, Discard the supernatant, resuspend the bacteria with an equal volume of sterile water, discard the supernatant after repeating twice, add an equal volume of MM medium, and inoculate 10% of the inoculum in the MM medium containing 25 μg / ml ZEN , and at the same time, use the MM medium containing the same ZEN concentration without inoculation as the blank negative control, culture at 30°C, 200 rpm for 12 hours, take samples, add an equal volume of methanol, mix well, let stand for 1 hour, take the mixed solution at 14000 rpm Centrifuge for 10 min per minute, and take the supernatant for HPLC detection. The detection conditions are partially improved with reference to GB / T 23504-2009, the sp...

Embodiment 3

[0039] Example 3 Preparation of Rhodoccocus sp. liquid biological detoxification agent

[0040]Components and ratio of seed medium: 1% tryptone, 0.5% yeast extract, 1.0% NaCl, pH7.0-7.2. Fermentation medium components and ratio: yeast extract 0.8%, casein peptone 0.4%, glucose 1.0%, (NH 4 ) 2 SO 4 0.1%, K 2 HPO 4 0.2%, MgSO 4 0.02%, NaCl 0.01%, pH 7.0-7.2.

[0041] (1) Strain activation: Inoculate the Rhodoccocus sp. cryopreservation tube with the preservation number CGMCC No.7186 on the solid medium, culture it at 30°C for 24-36 hours, measure its toxin degradation performance, and then inoculate it on Set aside on the slant of the test tube.

[0042] (2) Seed culture: pick bacteria from the slant medium and insert them into the seed medium, and cultivate them to the logarithmic phase at 30°C to obtain first-class seeds; then, use a 500-liter seed tank to feed the seed medium Amount of 350 liters, 121 DEG C of high-pressure damp heat sterilization after feeding, coo...

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Abstract

The invention discloses a rhodococcus strain and its application for degrading zearalenone. The preservation number of the Rhodococcus sp. is CGMCC No.7186. The invention also provides a biological detoxification agent containing the preserved strain and a preparation method thereof. In addition, the present invention also provides the application of the strain or biological detoxification agent in degrading zearalenone.

Description

technical field [0001] The invention relates to a rhodococcus strain Rhodococcus sp. and its application in degrading mycotoxin zearalenone. Background technique [0002] Zearalenone (Zearalenone, ZEN) is an estrogenic mycotoxin with a dihydroxybenzoic acid lactone structure, and its chemical name is 6-(10-hydroxy-6-oxo-trans-1-deca -Carbene)-β-syringoic acid-lactone, which is widely present in moldy corn, sorghum, wheat, oats, barley and other cereal crops, mainly by Fusarium graminearum (Fusarium graminearum), yellow Fusarium (Fusarium culmorum), Crookwell Fusarium (Fusarium crookwellense) produced. ZEN is mainly reproductive toxicity. After entering the human and animal body, it will produce estrogen effect syndrome, cause excessive estrogen and estrous reaction in animals, including infertility or infertility, and can also cause testicular atrophy in male animals and poor semen quality. symptom. ZEN also has hematotoxicity, hepatotoxicity, immunotoxicity and genotoxic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L5/20C12R1/01
Inventor 张晓琳史競赵晨杨博磊汪洋韩伟
Owner ACAD OF NAT FOOD & STRATEGIC RESERVES ADMINISTRATION
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