Heterotrophic nitrification and aerobic denitrification bacterium Y15 and application thereof
A technology of aerobic denitrifying bacteria and heterotrophic nitrification, applied in the field of environmental microorganisms, can solve problems such as limited practical application, and achieve high genetic stability, reduce nitrogen content, and high genetic stability
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Embodiment 1
[0034] Example 1 Isolation and identification of Pseudomonas putida Y15
[0035] 1. Isolation of Pseudomonas putida Y15
[0036] 1.1 Sample source
[0037] The samples were taken from the activated aerated sludge of a sewage treatment plant in Nanming District, Guiyang City.
[0038] 1.2 Enrichment, separation and purification of denitrification strains
[0039] Take 1mL activated sludge sample and inoculate it in the enrichment medium (formula: trisodium citrate 5.66g / L, MgSO 4 .7H 2 O0.02g / L, KH 2 PO 4 1.5g / L, Na 2 HPO 4 7.9g / L, KNO 3 0.84g / L, trace element solution 1mL / L, pH 7.41), enrichment culture at 28°C, 150rpm for 72h, repeated three times. Take 1mL enriched culture solution dilution in 9mL sterilized normal saline, dilute to 10 -1 . were diluted to 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 and 10 -7 isogradient. Under sterile conditions, take 10 -5 、10 -6 and 10 -7 200 μL each of the equal-gradient bacterial solution was evenly spread on the denitrificat...
Embodiment 2
[0056] Antibiotic susceptibility test of embodiment 2 Pseudomonas putida Y15
[0057] A single colony of Y15 was taken and transferred into LB liquid medium, and cultured at 37°C and 150rpm for 24h. Dilute the bacterial solution 10 times, take 200 μL of the diluted solution and spread it evenly on the LB plate, paste three same drug-sensitive paper sheets at equal distances on each plate, place it in a constant temperature incubator at 37°C for static culture for 24 hours, and count the results of the drug sensitivity test . The results are shown in Table 2, Y15 is only resistant to erythromycin, sensitive to other antibiotics, and has high biological safety in sewage treatment.
[0058] Table 2 Y15 antibiotic susceptibility test results
[0059] Types of antibiotics Average inhibition diameter (mm) Types of antibiotics Average inhibition diameter (mm) Norfloxacin 19.1±0.1 Cyclofloxacin 23.1±0.2 Chloramphenicol 25.2±0.2 Amikacin 19.5±0.1 ...
Embodiment 3
[0060] Example 3 Nitrogen removal performance test of Pseudomonas putida Y15
[0061] The slanted seeds of Pseudomonas putida Y15 were transferred to 100 mL LB liquid medium, and cultured at 30°C and 150 rpm for 24 hours to obtain seed liquid. The seed solution was taken out, centrifuged at 8000 rpm for 1 min, the supernatant was discarded, the bacteria were washed 3 times with sterilized PBS solution, and finally suspended in 10 mL of PBS solution to obtain the bacteria solution. Take 0.5mL of the bacterial solution and transfer it to 500mL NH 4 + -N, NO 3 - -N simulated sewage medium ((NH 4 ) 2 SO 4 0.72g / L; trisodium citrate dihydrate 3.50g / L; Vickers salt solution 50mL / L; trace element solution 1mL / L, pH 7.0. Wherein, Vickers salt solution formula is: K 2 HPO 4 .3H 2 O 6.50g / L; MgSO 4 .7H 2 O2.50g / L; NaCl 2.50g / L; FeSO 4 .7H 2 O 0.05g / L; MnSO 4 .H 2 O 0.04g / L. The formula of trace element solution is: EDTA 50g / L; ZnSO 4 2.20g / L; Ammonium molybdate 1.10g / L...
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