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Denitrifying phosphorus accumulation organism (DPAO) with function of efficiently removing nitrogen and phosphorus and application of DPAO

A denitrifying phosphorus-accumulating bacteria, nitrogen and phosphorus removal technology, applied in the field of environmental microbiology, can solve the problems of single species of bacteria, limited practical application research, etc., and achieve the effect of completely denitrifying air

Active Publication Date: 2015-06-24
龙岩水发环境发展有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the dominant populations of denitrifying phosphorus accumulating bacteria are mainly concentrated in the genus Pseudomonas and Bacillus. The strains are relatively single, and most of them focus on degradation genes and degradation mechanisms. The practical application research is limited.

Method used

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  • Denitrifying phosphorus accumulation organism (DPAO) with function of efficiently removing nitrogen and phosphorus and application of DPAO
  • Denitrifying phosphorus accumulation organism (DPAO) with function of efficiently removing nitrogen and phosphorus and application of DPAO
  • Denitrifying phosphorus accumulation organism (DPAO) with function of efficiently removing nitrogen and phosphorus and application of DPAO

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Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1 Enrichment culture, purification, screening and identification of bacterial strains of the present invention

[0027] (1) culture medium

[0028] ① Enrichment medium: CH 3 COONa·3H 2 O 5g / propionic acid 5mL; KNO 3 2.0g; MgSO 4 .7H 2 O 0.2g; K 2 HPO 4 2.0g; trace elements 2mL; double distilled water 1000mL; pH 7.2.

[0029] ②LB medium: (liquid) yeast extract 5g, peptone 10g, NaCl 5g, double distilled water 1000mL, pH7.0-7.2; (solid) liquid medium add 25g agar.

[0030] ③LB-P medium: add 2.5g K to LB medium 2 HPO 4 and 0.25gKH 2 PO 4 , pH7.0~7.2.

[0031] ④1% BTB-denitrification medium: CH 3 COONa·3H 2 O 5.0g; KNO 3 2.0g; MgSO 4 .7H 2 O 0.2g; Agar 20g; 1%-BTB 4mL; H 2O 1000 mL; pH 7.6.

[0032] ⑤Phosphorus deficiency medium for denitrification: KNO 3 2g / L;CH 3 COONa·3H 2 O 5g / L; K 2 HPO 4 0.05g / L; MgSO 4 ·7H 2 O 0.2g / L; CaCl 2 0.5g / L; trace elements 2mL / L.

[0033] ⑥Phosphorus-rich medium for denitrification: KNO 3 2g / L;CH 3 COON...

Embodiment 2

[0067] Embodiment 2 The cultivation of bacterial strain Klebsiella sp.N14 of the present invention

[0068] (1) culture medium

[0069] Selective medium stock solution: CH 3COONa·3H 2 O 5g / L; K 2 HPO 4 0.05g / L; KH 2 PO 4 0.2g / L; MgSO 4 ·7H 2 O 0.2g / L; CaCl 2 0.5g / L; trace element solution 2mL / L; pH 7.0~7.6.

[0070] Phosphorus-rich medium for denitrification: KNO 3 2g / L;CH 3 COONa·3H 2 O 5g / L; K 2 HPO 4 0.05g / L; KH 2 PO 4 0.2g / L; MgSO 4 ·7H 2 O 0.2g / L; CaCl 2 0.5g / L; trace elements 2mL / L.

[0071] (2) The culture condition optimization of bacterial strain of the present invention

[0072] ①Different pH values

[0073] Prepare 100mL of selective medium mother solution with pH values ​​of 5, 6, 7, 8, and 9 respectively, take 5mL of bacterial suspension to inoculate, and adjust OD 600 =0.1±0.001, culture on a shaker (26°C, 140r / min) for 24 hours, measure OD 600 and PO in the supernatant 4 3- -P, NO 3 - -N, NO 2 - -N concentration, calculate the denitr...

Embodiment 3

[0080] Embodiment 3 Application of bacterial strain Klebsiella sp.N14 of the present invention in sewage treatment

[0081] Take 200mL of sewage from the regulating tank of Xuzhou Guozhen Water Operation Co., Ltd., Jiangsu Province, and conduct a nitrogen and phosphorus removal test. The specific steps are as follows:

[0082] Take 10mL of bacterial suspension and inoculate in 200mL of sewage to be tested, adjust the OD 600 =0.1±0.001, under the conditions of pH 8, temperature 30°C, aerobic culture on a shaking table with a shaking frequency of 140r / min; test water samples were taken at 0, 4, 8, 12, 16, 20, and 24 hours respectively , take the supernatant after centrifugation, and measure the PO in the supernatant 4 3- -P, NO 3 - -N, NO 2 - -N concentration change, see the results Image 6 . Depend on Image 6 It can be seen that the content of phosphorus and nitrate nitrogen in the supernatant liquid gradually decreased during the aerobic culture process, the concent...

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Abstract

The invention relates to denitrifying phosphorus accumulation organism (DPAO) with a function of efficiently removing nitrogen and phosphorus and application of the DPAO, and belongs to the field of environmental microbiology. The bacterial strain is named Klebsiella sp. N14, belongs to Klebsiella species, and is preserved in China general microbiological culture center (CGMCC), the preservation number of the bacterial strain is CGMCC No. 10290. The bacterial strain is a Gram negative bacteria without spore and has thick capsule in a thick and short rod shape; the bacterial colony is milky white, the center of the bacteria is protruded, the edge is regular, and the surface is wet, transparent and glossy. The bacterial strain has a function of synchronously removing the nitrogen and the phosphorus, and can be applied to sewage biological sewage treatment process; after the bacterial strain is aerobically cultivated in nitrogen and phosphorus containing sewage for 24 hours, the phosphorous removal rate is 81.79%, and the nitrogen removal rate is 85.94%. The denitrifying final product of the bacteria strain is identified to be N2 with the full denitrifying capability; when the bacterial strain is used for treating the nitrogen and phosphorus containing sewage, inorganic nitrogen, such as nitrate nitrogen and nitrite nitrogen, in a water body is directly reduced to be harmless nitrogen gas to be discharged out of the water body; the effect is remarkable, the cost is low, and no secondary pollution exists.

Description

technical field [0001] The invention relates to the field of environmental microbiology, in particular to a denitrifying phosphorus-accumulating bacterium with high-efficiency nitrogen and phosphorus removal and its application. Background technique [0002] With the advancement of science and technology, the development of industry and agriculture, and the increase of population, countries all over the world are facing an unprecedented crisis of water shortage. my country's per capita water resources are less than 1 / 3 of the world's average level. The lack of water resources has seriously restricted the development of my country's industry and agriculture and has become the bottleneck of my country's sustainable economic development. [0003] Corresponding to the shortage of water resources is the pollution of the water environment and the eutrophication of the water body. Studies have shown that eutrophication is closely related to the content of nitrogen and phosphorus i...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/22
CPCC02F3/02C02F3/34C02F2101/105C02F2101/163C02F2101/166C12N1/205C12R2001/22
Inventor 孙玲
Owner 龙岩水发环境发展有限公司
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