Mixed inoculant for degrading polycyclic aromatic hydrocarbon pollutants
A technology of mixed bacterial agents and polycyclic aromatic hydrocarbons, which is applied in the field of remediation of polycyclic aromatic hydrocarbon pollutants, can solve the problems of no practical scale biological treatment technology, achieve strong reproductive ability and adaptability, high degradation rate, improve activity and biological volume effect
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Embodiment 1
[0027] Screening of 7 highly efficient degrading bacteria:
[0028] Preparation of bacterial solution: In 90ml of enrichment medium containing PAHs pollutants, add 10g of PAHs-contaminated soil aged in the sewage irrigation area (Sanbaotun North, Fushun City) aged PAHs-contaminated soil, shake at 120rpm / min constant temperature (30°C) Shake the culture in the bed for 7 days and wait for use.
[0029] The enrichment medium containing PAHs pollutants is as follows: 2.5ml PAHs solution is added in a sterile 500ml Erlenmeyer flask, so that the total concentration of polycyclic aromatic hydrocarbons in the enrichment medium is 200mg / L; , and then pour 90ml of aliquoted sterile enrichment medium into the Erlenmeyer flask. The enrichment medium is calculated by weight percentage: glucose 0.1%, peptone 0.1%, yeast extract 0.1%, NaCl 0.5%, NH 4 NO 3 0.1%, K 2 HPO 4 0.5%, MgSO 4 ·7H 2 O 0.05%, the balance is tap water, pH7.2. Sterilization conditions: 0.1Mpa damp heat steriliz...
Embodiment 2
[0044] The above-mentioned bacterial solution with PAHs as the only carbon source is inserted into the enrichment medium at a 10% inoculum amount for expanded culture, and a microbial mixed bacterial agent for degrading PAHs is obtained, which is stored for future use.
[0045] Then, the experiment of the degradation ability of mixed bacterial agents on PAHs was carried out, and each treatment was repeated three times. Carried out in inorganic salt liquid medium. Adjust the pH of the inorganic salt medium to 7.0, pack in 150ml Erlenmeyer flasks, 30ml per bottle, autoclave for 30min after bottling, and after cooling, add 1.5ml of the above-mentioned mixed bacterial agent (5% inoculum) to each bottle ; Cast Garfield, pyrene and benzo (a) pyrene solution. The initial concentrations (measured values) of each pollutant were 72.84mg / l, 67.49mg / l and 40.34mg / l respectively, and the total amount of the three pollutants was 180.67mg / l. 130 rev / min, keep at 28°C. After shaking cultur...
Embodiment 3
[0049] Take the above bacteria agent out of the refrigerator and place it at room temperature for activation. The activation is to inoculate 10% into sterilized beef extract-peptone medium under aseptic conditions, culture at 28° C. for 2-3 days with shaking at 130 rpm.
[0050] The experiment was carried out by shake flask liquid culture method. Each treatment has 3 repetitions. Under sterile conditions, draw 0.60ml of PAHs solution into a sterilized 150ml empty Erlenmeyer flask, cover with a ventilated high-temperature plastic film, ventilate until it is nearly dry, then add 0.5ml of acetone, dissolve the pollutants by shaking, ventilate for at least amount, add 30ml of sterilized inorganic salt medium. Inoculate 3 ml bacterial agent (10% inoculation amount) in 30 ml culture medium, 130 revs / min, 28 ℃ shaking culture 10 days.
[0051] The PAHs solution is a mixture of phenanthrene, pyrene and benzo(a)pyrene, the concentration of phenanthrene and pyrene is 4mg / ml, the conc...
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