Microbacterium capable of completely degrading zearalenone and application thereof
A gibberellone, completely degradable technology, applied in the field of microorganisms
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Embodiment 1
[0020] Example 1: Screening, isolation and identification of zearalenone-degrading strains
[0021] Soil samples were collected from turf (N36.34.243 E100.32.285) near Qinghai Lake on the Qinghai-Tibet Plateau. Take 2 g of soil samples, add 10 mL of 0.15M sodium chloride aqueous solution, shake for 30 min to fully disperse the samples, let stand at room temperature for 1 min, immediately take 1 mL of supernatant, spread on LB plates, and incubate at 30 °C until single For clonal growth, single colonies of different shapes were picked and inoculated in liquid LB medium (adding zearalenone with a final concentration of 25 μg / ml dissolved in methanol), cultured at 30°C for 2 days, centrifuged to obtain the supernatant, and washed with an appropriate amount of ethyl acetate. Extract, collect ethyl acetate and evaporate to dryness, dissolve the eluate with a small amount of methanol, and use TLC method (petroleum ether: acetone: ethyl acetate = 5:4:1 as the developing solvent) to...
Embodiment 2
[0023] Example 2: Microbacterium sp. Fermentation of FY1538
[0024] Microbacterium sp. The fermentation supernatant of FY1538 strain is used to degrade erythralenone, which needs to be induced by methanol. After isolation and purification of the strains, a single colony was picked and inoculated in 50 mL of LB medium, and methanol at a final concentration of 10 μl / ml was added, shaken and cultured at 30 °C for 48 h, and the fermentation broth was centrifuged at 12,000 rpm to obtain the fermentation supernatant.
Embodiment 3
[0025] Example 3: Microbacterium sp. Analysis of the ability of FY1538 fermentation supernatant to degrade erythralenone
[0026]Take 500μl of the fermentation supernatant, add 25ug zearalenone, react in a water bath at 30°C for 24 hours, extract the reaction solution with 1000μl ethyl acetate, collect the ethyl acetate phase, evaporate to dryness and dissolve the eluate with a small amount of methanol, use TLC method ( MERCK company TLC silica gel plate 60F 254 , the developer is petroleum ether: acetone: ethyl acetate = 5:4:1) to check the residue of erythralenone. Such as image 3 as shown, Microbacterium sp. After 24 hours of catalytic reaction between FY1538 fermentation supernatant and zearalenone, no zearalenone in the reaction system could be detected by TLC method.
[0027] Further analysis by HPLC Microbacterium sp. The ability of FY1538 fermentation supernatant to degrade erythralenone at different reaction times. The HPLC chromatographic column was Agile...
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