A strain of aerobic denitrifying bacteria
An aerobic denitrification and bacterial technology, applied in the field of microorganisms, can solve the problems of limited application range, low denitrification effect of related strains, etc., and achieve strong nitrogen reduction effect, strong environmental adaptability, and strong degradation ability Effect
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Embodiment 1
[0019] Isolation of strain (Klebsiella sp.) HLNR05:
[0020] (1) 10g of activated sludge sample was collected in the oxidation ditch of Xinyuan Wastewater Plant, Jishui County, Jiangxi Province. After the sample was collected, it was added to 100mL of sterile water filled with glass beads, mixed well and left for 30min, then 1mL was taken The supernatant was transferred to a 100mL Erlenmeyer flask containing nitrate-rich medium, and the medium was placed in a shaking incubator at a temperature of 28°C and a vibration frequency of 200rpm / min for 4 days. After 4 days, take 1 mL of the bacterial liquid and put it in 99 mL of sterilized enrichment medium containing nitrate (139 mg / L N), and culture it under the above culture conditions for 4 days, then repeat the same operation 3 times for 4 cycles of enrichment. set training.
[0021] (2) Take 1 mL of the bacterium liquid obtained in the 4th cycle in step (1), add it to the acclimatization medium in which the nitrate concentrati...
Embodiment 2
[0029] Denitrification activity detection of strain HLNR05:
[0030] (1) The strains obtained from the step (4) of the isolation of the above-mentioned strain (Klebsiella sp.) HLNR05 were respectively inoculated in 100 mL of fresh denitrification medium at a ratio of 1:100, at 28 °C, 200 rpm / min Cultivate in a shaking incubator for 1 day, and detect nitrate nitrogen (NO 3 -N) and the total N concentration; a blank control group is set at the same time. Among them, NO 3 - -N uses hydrochloric acid-naphthalene ethylenediamine spectrophotometry; TN uses potassium persulfate-ultraviolet spectrophotometry.
[0031] The above nitrates (NO 3 - -N) is the nitrification medium of nitrogen source: glucose 169mg, KNO 3 37.71mg, KCI63mg, MgSO 4 23mg, anhydrous CaCl 2 23mg, NaHCO 3 65mg, KH 2 PO 4 23mg, 2mL of trace element solution (the formula is the same as above), 1000mL of distilled water, pH7.2.
[0032] (2) Test results: When the strain HLNR05 is cultured and grown in a ...
Embodiment 3
[0034] Molecular characterization of the HLNR05 strain:
[0035] (1) The HLNR0516S rDNA PCR product was separated by 1% agarose gel electrophoresis and purified with an agarose gel separation and purification kit. The purified 16S rDNA fragment was transformed into E.coli DH5a by TA cloning technology, ampicillin and blue-white screening to pick positive clones and send them to Shanghai Sangong for sequence determination. The sequencing results were compared with the 16S rDNA sequences in the GenBank database to obtain the 16S rDNA sequences of the most similar strains. The phylogenetic tree was constructed with Neighbor-joining by MEGA5.0 software.
[0036] (2) RDP cluster analysis showed that the strain HLNR05 belonged to the β-proteobacteria subphylum Klebsiella sp., and the similarity was 99%. The strain was named as Klebsiella sp.HLNR05 (see attached figure 2 ).
[0037] buy example four
[0038] The test of removing nitrate nitrogen and total nitrogen in simulated eu...
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