Bacillus cereus for degrading aureomycin and application thereof
A technology of Bacillus cereus and aureomycin, which is applied in the direction of bacteria, microorganism-based methods, and treatment of microorganisms with electricity/wave energy, can solve the problems of toxicity and no research reports, and achieve excellent degradation effect of aureomycin
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Embodiment 1
[0022] Screening and identification of strains:
[0023] (1) Strain isolation:
[0024] Under sterile conditions, take 10g of cow dung raw material and add it to 100mL of sterile water, shake it at 30°C and 120r / min for 45min, remove the supernatant after standing still, and dilute according to the gradient multiple, and use the dilution coating plate method to determine the dilution 10 -6 ~10 -8 The supernatant was transferred to LB plates, and each dilution was repeated three times, and cultured in a 30°C incubator. After the flora grew out, they were repeatedly streaked and separated on the LB plate, and observed under a microscope to confirm that they were pure single colonies.
[0025] (1) Strain enrichment and domestication:
[0026] Add the isolated colony to 100mL inorganic salt basal medium containing 20mg / L aureomycin, place it in a shaker incubator, and culture it in the dark at 30°C and 180r / min for 7 days, then take 10mL of the enriched culture Inoculate agai...
Embodiment 2
[0031] Example 2: Mutagenesis and screening of original strains
[0032] Atmospheric room temperature plasma mutagenesis (ARTP) was adopted, and the mutagenesis steps were as follows:
[0033] (1) Add the above-mentioned colonies into sterile water to prepare a steamed stuffed bun suspension, and then dilute it to 10 -7 / mL concentration, take 10uL of the cell suspension and evenly spread it on the surface of the ARTP slide (sterilized) for mutagenesis treatment. The working gas for mutagenesis is helium, the gas flow rate is 10s / m, the power is 100W, and the distance is 4mm. Irradiate for 90s under the hood for mutagenesis, then transfer the slides to a centrifuge tube containing 1mL of normal saline, shake for 1min, and place in a shaking incubator for shaking for 1h.
[0034] (2) Obtain the bacterium suspension after the mutagenesis, and dilute the bacterium suspension after the mutagenesis to 10 -7 / mL concentration, take 100 μL of the diluted solution, spread it on a PD...
Embodiment 3
[0037] Degradation of aureomycin by Bacillus cereus LZ01 obtained by mutagenesis treatment:
[0038] In the degradation process of aureomycin by the original strain, it was found that pH had a great influence on the degradation efficiency. The original strain before mutagenesis treatment was inoculated into LB liquid medium, activated and cultivated at 30°C and 180rpm, and when the original strain grew to the logarithmic growth phase, the bacterial cells were collected by centrifugation, then washed and centrifuged, and 0.2 % (w / v) of the inoculum amount of the original strain was added to 50mg / L aureomycin without pH adjustment (the pH of the degradation system was weakly acidic at this time), and the removal of aureomycin in 12-48h The rate showed a rapid upward trend, rising from 31.75% to 55.21%.
[0039]Inoculate the Bacillus cereus LZ01 obtained by the mutagenesis treatment into LB liquid medium, and activate the culture at 30°C and 180 rpm. When the Bacillus cereus LZ0...
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