Facultative anaerobic denitrifying bacteria having complete denitrification enzyme systems and use thereof
A technology of denitrifying bacteria and nitrifying enzymes is applied in the field of facultative anaerobic denitrifying bacteria, which can solve the problems of inability to remove nitrogen, and achieve the effects of saving operating costs, thorough denitrification, and high denitrification activity.
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Embodiment 1
[0036] Embodiment 1 Isolation and identification of bacterial strains of the present invention:
[0037] (1) Medium:
[0038] A. Strain isolation, purification, preservation medium ( / L):
[0039] 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.
[0040] B. Strain screening, denitrifying medium (DM: Denitrifying Medium) ( / L):
[0041] CH 3 COONa, 2g; KH2 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.
[0042] C. LB medium ( / L):
[0043] Peptone, 10g; yeast extract, 5g; sodium chloride, 10g; pH is controlled at 7.5.
[0044] (2) Isolation and purification of Pseudomonas XP-2 strain:
[0045] Weigh 10g of rhizosphere soil samples of cattail plants and place them in a 250mL Erlenmeyer flask, add 100mL of sterile water and a few glass beads, and vibrate on a shaker for 15min to disperse the soil samples ...
Embodiment 2
[0064] Embodiment 2 The cultivation of bacterial strain of the present invention
[0065] (1) Medium used
[0066] 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.
[0067] B. Strain denitrification medium (DM: Denitrifying Medium) ( / L): CH3COONa, 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.
[0068] The above medium was sterilized at 121°C for 20 minutes before use.
[0069] (2) Culture conditions
[0070] Use an inoculation loop to scrape 1 ring of bacterial lawn from the Pseudomonas XP-2 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 liquid...
Embodiment 3
[0071] Example 3 The optimal denitrification conditions of the bacterial strain of the present invention
[0072] 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. In the range of 10-30°C, Pseudomonas XP-2 can remove more than 80% of total nitrogen, and the removal rate of total nitrogen is the highest at 25°C, reaching 95%. The adaptable pH value of the bacterial strain of the invention is between 6 and 9, can reduce the total nitrogen of about 140 mg / L to below 10 mg / L, and has no accumulation of nitrite. The optimum C / N (mol / mol) ratio of the strain of the invention is 3, and when C / N≥3, the removal rate of total nitrogen can reach over 90%. The slightly dissolved oxygen environment of static culture is most suitable for the denitrification activity of the strain of the present invention.
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