Rhodococcus sp. and application of rhodococcus sp. to zearalenone degradation

A technology of Rhodococcus and mutant strains is applied to a strain of Rhodococcus and its application field of degrading zearalenone, which can solve the problems of low ZEN degradation rate and weak degradation ability, and achieve fast and efficient detoxification ability, easy Operation, the effect of solving the problem of toxin contamination

Active Publication Date: 2015-12-16
ACAD OF NAT FOOD & STRATEGIC RESERVES ADMINISTRATION
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  • 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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  • Rhodococcus sp. and application of rhodococcus sp. to zearalenone degradation
  • Rhodococcus sp. and application of rhodococcus sp. to zearalenone degradation
  • Rhodococcus sp. and application of rhodococcus sp. to zearalenone degradation

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Experimental program
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Effect test

Embodiment 1

[0028] Example 1 Rhodococcus Rhodococus sp. acquisition and identification

[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 O0.41g / L, KH 2 PO 4 0.5g / L, K 2 HPO 4 1.5g / L, NaCl0.5g / L, FeSO 4 ·7H 2 (00.037g / L, pH7.0-7.2, sterilized at 121°C for 30min), the final concentration of ZEN in the medium was 25 μg / ml, and transplanted with 10% inoculum after culturing for 7 days, the concentration of ZEN in the medium was constant . 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 co...

Embodiment 2

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

[0037] Activate the 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 For 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 Take the MM medium containing the same concentration of ZEN without inoculation as the blank negative control, incubate at 30°C and 200 rpm for 12 hours, take a sample, add an equal volume of methanol, mix well, let stand for 1 hour, take the mixed solution at 14000 rpm After centrifugation for 10 min, the supernatant was taken for HPLC detection. The detection conditions were partially improved with reference to GB / T23504-...

Embodiment 3

[0039] Embodiment 3Rhodoccocus sp. Preparation of 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 40.02%, NaCl0.01%, pH7.0-7.2.

[0041] (1) Strain activation: inoculate the Rhodoccocussp. cryopreservation tube with the preservation number CGMCCNo.7186 on the solid medium, culture it at 30°C for 24-36 hours, measure its toxin degradation performance, and then inoculate it on the slant of the test tube on standby.

[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, cooling t...

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Abstract

The invention discloses rhodococcus sp. and application of the rhodococcus sp. to zearalenone degradation. The preservation number of the rhodococcus sp. is CGMCC No.7186. The invention also provides a biological detoxication agent containing the preserved bacterial strain of the rhodococcus sp. and a preparing method of the biological detoxication agent. In addition, the invention also provides application of the bacterial strain or the biological detoxication agent to the zearalenone degradation.

Description

technical field [0001] The invention relates to a strain of 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 widely exists in moldy corn, sorghum, wheat, oats, barley and other cereal crops, mainly by Fusarium graminearum (Fusarium graminearum), yellow Fusarium (Fusarium culmorum ), produced by Fusarium crookwellense. 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 genotoxicity. [0003] my country's fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L1/015C12R1/01A23L5/20
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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