A Heterotrophic Nitrifying-Aerobic Denitrifying Bacteria l1 and Its Application
A technology of aerobic denitrifying bacteria and heterotrophic nitrification, applied in the field of environmental microorganisms, can solve the problems of application limitations, difficulty in mass cultivation, slow growth in the growth environment, etc., and achieve the effect of great economic value
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Embodiment 1
[0015] Example 1: Isolation of heterotrophic nitrifying and aerobic denitrifying bacteria L1
[0016] Take 5 g of river sediment in Jizhou District, Tianjin, and add 100 ml enrichment medium (NH 4 Cl0.382g / L, KNO 3 0.36g / L, sodium acetate 2g / L, MgSO 4 ·7H 2 O 0.2g / L, K 2 HPO 4 0.2g / L, NaCl 0.12g / L, MnSO 4 4H 2 O 0.01g / L, FeSO 4 0.01g / L, the rest is water, pH 7.0-7.2), 30°C, 150rpm shaking aerobic culture for 3 days, then 30°C, static culture for 3 days, repeat this again. Then take 5ml of the above-mentioned enrichment solution and insert it into 100ml enrichment medium (NH 4 Cl 0.382g / L, KNO 3 0.36g / L, sodium acetate 2g / L, MgSO 4 ·7H 2 O0.2g / L, K 2 HPO 4 0.2g / L, NaCl 0.12g / L, MnSO 4 4H 2 O 0.01g / L, FeSO 4 0.01g / L, the rest is water, pH 7.0-7.2), 30°C, 150rpm shaking aerobic culture for 3 days, then 30°C, static culture for 3 days, repeat this again. Then take the enrichment solution of the above-mentioned secondary enrichment and use solid enrichment me...
Embodiment 2
[0017] Example 2 Identification of heterotrophic nitrifying and aerobic denitrifying bacteria L1
[0018] Morphological characteristics of L1: short rod shape, no spores, and form yellow colonies on solid plates. Physiological and biochemical characteristics of L1: Negative Gram stain, negative urease reaction. The activated L1, CGMCC No.15917 was inoculated in liquid LB medium (10 g / L peptone, 5 g / L yeast powder, 10 g / L NaCl), and cultured with shaking at 30° C. and 150 rpm for 24 hours. Take 1ml of fresh culture bacteria solution and centrifuge at 4°C and 8000rpm for 5min, collect the bacteria, use DNA extraction kit to extract DNA, and then use general primer 8F / 1492R for PCR amplification, after electrophoresis detection of PCR products, if figure 1 Swimming lane 2 electrophoresis band, and determine the 16S rDNA gene sequence (as shown in SEQ ID NO: 1), after comparison, it is determined that L1 belongs to Planomicrobium sp. The phylogenetic tree of the 16S rDNA sequence...
Embodiment 3
[0021] Example 3: Detection of L1, CGMCC No.15917 Heterotrophic Nitrification Function
[0022] The activated L1, CGMCC No.15917 was inoculated in liquid LB medium (10 g / L peptone, 5 g / L yeast powder, 10 g / L sodium chloride), and cultured with shaking at 30° C. and 150 rpm for 24 hours. According to 5% (V / V), take fresh bacterial solution and inoculate it in heterotrophic nitrification simulated wastewater (NH 4 Cl 0.382g / L, sodium acetate 2g / L, MgSO 4 ·7H 2 O 0.2g / L, K 2 HPO 4 0.2g / L, NaCl 0.12g / L, MnSO 4 4H 2 O 0.01g / L, FeSO 40.01g / L, pH7.0~7.2), 30°C, 150rpm shaking aerobic culture for 7 days. And there is an uninoculated blank control. After culturing for 7 days, use Griess reagent and diphenylamine reagent to detect whether nitrite or nitrate appears in the culture medium. If there is nitrite or nitrate in the culture medium, it indicates that heterotrophic nitrification occurs, and it is positive for heterotrophic nitrification; if there is no nitrite and nitra...
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