Penicillium citrinum LJ318 for degrading chlortetracycline
A technique of chlortetracycline and penicillium citrinum, applied in the field of microorganisms, can solve the problems of few research reports and no research reports, and achieve the effects of convenient use, good degradation, tolerance and wide concentration range of use
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Embodiment 1
[0029] 1. Screening method for a strain of Penicillium citrinum LJ318 degrading aureomycin
[0030] 1.1 Material preparation
[0031] Source of bacteria samples: collected from the waste of aureomycin production enterprises.
[0032] Inorganic salt solid medium: (NH4) 2 SO 4 2.00g, K 2 HPO 4 0.50g, KH 2 PO 4 0.50g, MgSO 4 7H2O 0.50g, NaCl 0.20g, CaCl 2 0.1g, FeSO 4 0.01g, 0.015g EDTA, 13.00g agar, 2.00g / L glucose, dissolved in 1000ml distilled water. After sterilizing at 115°C for 20 minutes, add 2.00 g / L aureomycin to the ultra-clean workbench.
[0033] Martin's medium: peptone 6.0g, glucose 10.0g, KH 2 PO 4 1.0g, MgSO 4 ·7H2O 0.5g, dissolved in 1000ml of distilled water. Sterilize at 115°C for 20 minutes.
[0034] Select medium: (NH4) 2 SO 4 2.00,K 2 HPO 4 0.50, KH 2 PO 4 0.50, MgSO 4 7H2O 0.50, NaCl 0.20, CaCl 2 0.10, FeSO 4 0.01g, 0.015g of EDTA, dissolved in 1000ml of distilled water. Sterilize at 115°C for 20 minutes, after cooling, a...
Embodiment 2
[0074] Growth and degradation experiments of strain LJ318 in aureomycin as the only carbon source medium:
[0075] Prepare aureomycin as the only carbon source and the concentrations of aureomycin are 25mg / L, 50mg / L, 100mg / L, 1000mg / L, 2000mg / L, 3000mg / L, 4000mg / L, 5000mg / L, 10000mg / L Inorganic salt solid medium of L. Pick a few LJ318 colonies from the activated culture medium, streak on the above-mentioned inorganic salt solid medium plate, and culture in the dark at 28°C. It is known through observation that the concentration of aureomycin for bacterial strain LJ318 is 25mg / L, 50mg / L , 100mg / L, 1000mg / L, 2000mg / L, 3000mg / L, 4000mg / L, 5000mg / L, and 10000mg / L plates all grew hyphae. It shows that the strain LJ318 can grow in the medium with the concentration of aureomycin being 25mg / L-10000mg / L.
[0076] The activated strain LJ318 was inserted into the inorganic salt liquid medium with chlortetracycline as the sole carbon source and the concentration of chlortetracycline was...
Embodiment 3
[0078] Strain LJ318 treatment experiment on aureomycin residue:
[0079] The strain LJ318 was activated with Martin's medium, then inoculated into fresh chlortetracycline residue, cultured in the dark at 30°C, and samples were taken regularly to determine the residual amount of chlortetracycline in the residue and the degradation rate was calculated.
[0080] The method for measuring the pH value is as follows: take 1 g of fungus residue and add 10 ml of distilled water, stir it evenly, and measure it with an acidity meter.
[0081] The determination method of chlortetracycline is: accurately weigh 2.0000g of bacteria residue, extract with 20mL of acetone: hydrochloric acid solution: water = 13:1:6 extract, centrifuge the extract for 5min at 8000r / min, and take the supernatant , filter with a filter membrane of 22 μm, utilize high performance liquid chromatography to measure the aureomycin content in the filtrate and calculate the degradation rate, the HPLC conditions are the sa...
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