Denitrifying phosphate-accumulating organism with nitrogen and phosphorus removal functions and applications thereof
A technology for denitrifying phosphorus-accumulating bacteria and denitrifying and dephosphorizing, applied in bacteria, chemical instruments and methods, microorganism-based methods, etc. Active, stable effect, simple process effect
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Embodiment 1
[0032] The isolation and identification of embodiment 1 bacterial strain of the present invention:
[0033] (1) Medium:
[0034] A. Strain isolation, purification, preservation medium ( / L):
[0035] CH 3 COONa, 2g; Peptone, 15g; Yeast extract, 3g; Glucose, 1g; NaCl, 6g; Agar, 12g; KNO 3, 1.5g; the pH is controlled at 7.0-7.2.
[0036] B. Strain screening, denitrifying medium (DM: Denitrifying Medium) ( / L):
[0037] CH 3 COONa, 2g; KH 2 PO 4 , 0.4g; MgSO 4 ·7H 2 O, 0.6g; CaCl 2 2H 2 O, 0.07g; KNO 3 , 1g; Tris buffer 12mL; trace elements 2mL; pH controlled at 7.0-7.2.
[0038] C. LB medium ( / L):
[0039] Peptone, 10g; yeast extract, 5g; sodium chloride, 10g; pH is controlled at 7.5.
[0040] (2) Isolation and purification of Pseudomonas CL-1 strain:
[0041] Separation and purification were carried out by dilution coating method and plate streaking method. First, use a sterile pipette to take 10 mL of the sludge mixture solution at the end of SBR hypoxia, put it ...
Embodiment 2
[0060] The cultivation of embodiment 2 bacterial strains of the present invention
[0061] (1) Medium used
[0062] A. Strain preservation medium ( / L): peptone, 5g; yeast extract, 3g; glucose, 1g; NaCl, 6g; agar, 12g; KNO 3 , 1.5g; the pH is controlled at 7.0-7.2.
[0063] B. Strain denitrification medium (DM: Denitrifying Medium) ( / L): CH 3 COONa, 2g; KH 2 PO 4 , 0.4g; MgSO 4 ·7H 2 O, 0.6g; CaCl 2 2H 2 O, 0.07g; KNO 3 , 1g; Tris buffer 12mL; trace elements 2mL; pH controlled at 7.0-7.2.
[0064] The above medium was sterilized at 121°C for 20 minutes before use.
[0065] (2) Culture conditions
[0066] Use an inoculation loop to scrape 1 ring of bacterial lawn from the Pseudomonas CL-1 preserved on the slant of the preservation medium, inoculate it into a 250mL Erlenmeyer flask containing 100mL of sterilized LB medium, and cultivate it with constant temperature and shaking at 30°C After 12 hours, the seed liquid can be obtained. During the experiment, the seed li...
Embodiment 3
[0067] The optimum denitrification and dephosphorization conditions of the bacterial strain of the present invention in embodiment 3
[0068] Put 100mL of sterilized denitrification medium into a 250mL Erlenmeyer flask, insert 10% of the inoculum into the seed bacterial solution, and let it stand for cultivation. The optimal denitrification temperature is between 25-30°C, the optimum temperature is 25°C, and the optimum pH value is 7-8. The growth cycle of the bacteria is long, entering the logarithmic growth phase after 48h, entering the stable phase after 96h, basically in the stable phase from 96 to 120h, and entering the decline phase after 120h. The nitrogen and phosphorus removal rates of the strain were 56.93% and 43.56% in the first 24 hours, and the nitrogen and phosphorus removal rates were 75.6% and 57.5% during the whole experiment.
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