Heterotrophic nitrifying aerobic denitrifying Acinetobacter and applications thereof

A bacillus and culture medium technology, applied in the field of environmental microorganisms, can solve the problems of lack of simultaneous resistance to multiple, lack of efficient denitrification, etc.

Active Publication Date: 2017-07-21
上海华强环境科技工程有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Therefore, the technical problem to be solved in the present invention is to overcome the deficiency of heterotrophic nitrification-aerobic denitrification bacteria that are simultaneously resistant to multiple antibiotics in the prior art, thereby providing a heterotrophic nitrification-aerobic denitrification bacterium that is simultaneously resistant to tetracycline and streptomycin. Aerobic denitrifying Acinetobacter, the heterotrophic nitrifying-aerobic denitrifying Acinetobacter has the function of efficient denitrification
[0006] Another technical problem to be solved by the present invention is to overcome the lack of high-efficiency denitrification methods in the prior art, thereby providing the application of the above-mentioned heterotrophic nitrification-aerobic denitrification Acinetobacter in nitrogen-containing water body denitrification

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  • Heterotrophic nitrifying aerobic denitrifying Acinetobacter and applications thereof
  • Heterotrophic nitrifying aerobic denitrifying Acinetobacter and applications thereof
  • Heterotrophic nitrifying aerobic denitrifying Acinetobacter and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037]Example 1 Screening and isolation of heterotrophic nitrification-aerobic denitrification strains

[0038] Take the activated sludge from the sewage treatment tank of the No. 1 Sewage Treatment Plant in Zhuyuan, Pudong, Shanghai, and take 5mL of mud-water mixture from the activated sludge into 45mL of sterilized NSM enrichment medium.

[0039] Then placed at 30°C, 200rpm air bath shaker enrichment culture for 12h. After gradient dilution of the enriched solution, spread evenly on NSM separation medium (agar 20g / L, other components are the same as NSM). After culturing in a constant temperature incubator at 30°C for 1 day, single clones with different shapes and sizes were picked, streaked and purified, and numbered and stored to obtain the strains for primary screening.

[0040] Wherein, the formula of NSM medium is, by every liter: (NH 4 ) 2 SO 4 0.945g, sodium citrate 16.34g, MgSO 4 ·7H 2 O 1.0g, NaCl 0.12g, MnSO 4 ·H 2 O 0.01g, FeSO 4 ·7H 2 O0.02g, phosphate ...

Embodiment 2

[0045] Example 2 Molecular biology identification

[0046] Genomic DNA of YN-3 was extracted using a kit from Axygen, and 16S rDNA was amplified using this as a template. Primers: 16S rDNA-8F: AGAGTTTGATCCTGGCTCA, 1492R: GGTTACCTTGTTACGACTT.

[0047] PCR reaction system (50 μL): template DNA 1 μL, PCR Taq mix 25 μL, upstream and downstream primers 1.5 μL, DMSO 2 μL, add ddH 2 O to 50 μL of the reaction system. PCR program: 94°C for 5min, 94°C for 30s, 50°C for 1min, 72°C for 2min, 72°C for 5min, 72°C for 10min; 30 cycles. The purification and sequencing of PCR products were completed by Meiji Biomedical Technology Co., Ltd., and the sequencing sequences were registered in NCBI for homologous sequence search (Blastn analysis), and the obtained sequences were imported into the bioinformatics software Mega6 for genetic tree analysis.

[0048] The effective sequence length of the 16S rDNA of the amplified strain YN-3 is 1437bp, as shown in SEQ ID NO.1 in the sequence table. Aft...

Embodiment 3

[0049] Embodiment 3 (Acinetobacter sp.) The growth adaptability of (Acinetobacter sp.) YN-3 and the effect analysis of removing ammonia nitrogen

[0050] NM medium was used to cultivate strain YN3, and YN3 was inoculated into LB medium, and cultured at 30° C. on an air-bath shaker at 200 rpm for 12 hours. Then get the centrifugal washing of bacterium liquid respectively, inoculate in the NM liquid nutrient solution of 50mL respectively, inoculum amount is 0.3g / L by thalline wet weight, and the acidity and alkalinity (pH) of nutrient solution is respectively 4.0,4.5,5.0,6.0, 7.0, 8.0, 9.0, 10.0, 10.5, 11.0. At 30°C, 200rpm air bath shake flask cultured for 24h, the results are as follows: image 3 shown.

[0051] And by image 3 It can be seen that YN3 can grow between pH 5.0 and 10.0. Although the growth slows down at pH 5.0 or 10.0, the growth of the strain still continues; There is no growth phenomenon or the growth is extremely slow, and when the pH value is 5.0-10.0, t...

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Abstract

The present invention discloses a heterotrophic nitrifying aerobic denitrifying Acinetobacter and applications thereof, wherein the heterotrophic nitrifying aerobic denitrifying Acinetobacter is Acinetobacter sp. YN-3, has the preservation number of CGMCC No.11366, and can be used for nitrogen-containing water body denitrification. According to the present invention, the Acinetobacter sp. YN-3 can simultaneously resist streptomycin and tetracycline; the heterotrophic nitrifying performance determination results show that the Acinetobacter sp. YN-3 can highly remove ammonia nitrogen while the accumulation of the intermediate products such as nitrate and nitrite is little; and the aerobic denitrifying performance determination results show that the Acinetobacter sp. YN-3 can well remove the nitrate nitrogen, the residual nitrite accumulation is low, and no secondary pollution is generated.

Description

technical field [0001] The invention belongs to the field of environmental microorganisms, and in particular relates to a heterotrophic nitrifying aerobic denitrifying Acinetobacter and application thereof. Background technique [0002] The rapid development of industry and agriculture and human production activities have brought serious pollution to the water environment. Excessive nitrogen source is the most common type of pollution in water pollution. It causes eutrophication in water, leading to algae blooms, reduced dissolved oxygen in water, and black water. It not only endangers human water safety, but also seriously affects the ecology of water. balance. Excessive discharge of nitrogen sources will destroy the nitrogen balance and cause the water body to lose its self-purification function. Therefore, nitrogen-rich sewage needs to be denitrified before it can be discharged into the natural environment. [0003] Nitrogen exists in water mainly in two forms: organic ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/38
CPCC02F3/34C02F2101/38C12N1/205C12R2001/01
Inventor 丁少华孙叶凤史吉平孙俊松颜薇芝
Owner 上海华强环境科技工程有限公司
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