Composite microbial preparation for degrading quinolones antibiotics and preparation method and application
A composite microbial inoculum and quinolone technology, which is applied in the field of soil organic pollutant degradation, can solve the problems of difficult technical application, low efficiency, low degradation efficiency of quinolone antibiotics, etc.
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Embodiment 1
[0062] 1. Preparation of Quinolone Antibiotic Degrading Bacteria
[0063] Take degrading bacteria: Staphylococcus caprae (Staphylococcus caprae)-NOR-36 (already in the literature "Fu Boming, Chen Liwei, Cai Tianming, et al. Isolation and screening of norfloxacin degrading bacteria NOR-36 and research on degradation characteristics [J] .Journal of Environmental Science, 2017,37(2):576-584."Open), Candida parapsilosis (Candida parapsilosis) CICC 1973, Rhodococcus (Rhodococcus sp.) CGMCC 13024, Pseudomonas sp. ) ACCC 11715, Acinetobacter (Acinetobacter sp.) ACCC 11710, Sphingomonas (Sphingomonas sp.) ACCC 11703, the individual bacteria were cultured in LB liquid medium, and the single yeast were cultured in YPD liquid medium Cultivate to the logarithmic phase at 29°C and shake at 150r / min. Adjust the concentration of each obtained degrading bacteria seed stock solution so that the concentration of each bacteria solution is about 1×10 11 cfu / mL, then mixed evenly according to th...
Embodiment 2
[0071] 1. Preparation of Quinolone Antibiotic Degrading Bacteria
[0072] Individual degrading bacteria from Staphylococcus caprae-NOR-36 and Pseudomonas sp. ACCC 11715 were cultured in LB liquid medium to the logarithmic phase, cultured at 30°C and 130r / min shaking under conditions. Then, carry out acclimatization: A, respectively get 200 μ L of cultured degrading bacteria suspension and place them in 250 mL Erlenmeyer flasks, add sterilized 100 mL liquid selective medium (norfloxacin 440 mg / L, sulfuric acid) in Erlenmeyer flasks Ammonium 5.0g / L, Potassium dihydrogen phosphate 1.0g / L, Magnesium sulfate 0.5g / L, Yeast extract 0.2g / L), the degrading bacteria were wildly domesticated for 5 days (the domestication conditions were 30°C, 130r / min shaking conditions); B. Take 200 μL of wild domestication solution and repeat step A6 times, and then in LB liquid medium, shake and cultivate for 48 hours under the conditions of 37°C and 120r / min, to prepare and form a single degrading b...
Embodiment 3
[0080] 1. Preparation of Quinolone Antibiotic Degrading Bacteria
[0081] Individual degrading bacteria in Staphylococcus caprae-NOR-36, Pseudomonas sp. ACCC 11715, and Sphingomonas sp. ACCC 11703 were cultured in LB liquid medium until In the logarithmic phase, the culture was carried out at 29°C and 150r / min shaking. Then, carry out domestication: A, respectively get 250 μ L of the degrading bacteria suspension after enrichment culture and place respectively in 250 mL Erlenmeyer flask, add sterilized 100 mL liquid selective medium (ciprofloxacin 345 mg / L) in Erlenmeyer flask , Ammonium Sulfate 5.0g / L, Potassium Dihydrogen Phosphate 1.0g / L, Magnesium Sulfate 0.5g / L, Yeast Extract 0.2g / L), the degrading bacteria were wildly domesticated for 8 days (the domestication conditions were 29°C, 150r / Min shaking conditions); B. Take 250 μL of wild domestication solution and repeat step A6 times, and then in LB liquid medium, shake and cultivate for 60 hours under the conditions of 3...
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