A kind of heterotrophic nitrifying aerobic denitrifying Citrobacter and its application

A technology of Freund's citric acid and bacillus, applied in the field of applied microorganisms, can solve the problems of ammonia nitrogen cannot be processed in time, shorten the hydraulic retention time, and long hydraulic retention time, and achieve good biocatalytic function, wide temperature adaptability and growth adaptability strong effect

Active Publication Date: 2020-09-25
玉凰生态科技股份有限公司
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  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

However, the treatment efficiency of this method is not high, the required hydraulic retention time is long, and the high concentration of ammonia nitrogen in the water body will not be treated in time. The emergence of heterotrophic nitrification and aerobic denitrification microorganisms has updated the understanding of microbial biological nitrogen removal. This new metabolic pathway can be completed by a single microorganism, and allows nitrification and denitrification to be completed in the same reactor, so it can not only speed up the reaction process, shorten the hydraulic retention time, but also treat high-concentration ammonia nitrogen wastewater

Method used

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  • A kind of heterotrophic nitrifying aerobic denitrifying Citrobacter and its application
  • A kind of heterotrophic nitrifying aerobic denitrifying Citrobacter and its application
  • A kind of heterotrophic nitrifying aerobic denitrifying Citrobacter and its application

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Embodiment 1

[0028] Enrichment culture and isolation of embodiment 1 Citrobacter freundii S06

[0029] The activated sludge was taken from the treatment pond of Shanghai Pudong Zhuyuan No. 1 Sewage Treatment Plant, and 5mL of mud-water mixture was inoculated into 45mL of sterilized NRM enrichment medium (the formula of NRM medium was: (NH 4 ) 2 SO 4 2.0g / L, sodium citrate 20.0g / L, MgSO 4 ·7H 2 O 1g / L, 1.0mM phosphate buffer saline K-PBS, pH 6.8. ); 30°C, 200rpm, cultured on a constant temperature shaker for 24h; then the enriched seed solution was serially diluted, and coated onto an NRM solid culture plate (NRM, containing 2% agar); cultivated in a constant temperature incubator at 30°C After 24 hours, the grown microbial clones were cultured separately, inoculated into liquid bromothymol blue (BTB) medium, and cultured in a constant temperature incubator at 30° C. for 24 hours. The formula of BTB medium is (per liter): KNO 3 1g, sodium succinate 8.5g, MgSO 4 ·7H 2 O 1g, CaCl 2...

Embodiment 2

[0030]Identification and Genetic Tree Analysis of Example 2 Bacterial Strains

[0031] Collect 2ml of fresh strain culture solution, centrifuge at 10,000g for 2min, use a genomic DNA extraction kit, such as the kit from Axygen, to extract the genomic DNA of S06, dissolve and dilute to 100mg / L, and use this as a template to amplify 16SrDNA The primers used were 16S-F and 16S-R, and the sequences were 16S-F: AGAGTTTGATCCTGGCTCA, and 16S-R: GGTTACCTTGTTACGACTT. The PCR reaction mixture was prepared as follows: template DNA 1 μL, PCR Taq mix 25 μL, upstream and downstream primers 1.0 μL (20 μM), add ddH 2 0 to 50 μL. The PCR program was run as follows: 94°C for 5 min, 94°C for 30 s, 53°C for 1 min, 72°C for 2 min, 30 cycles, 72°C for 5 min, 4°C for 1 min. The PCR product was sequenced by a professional biological company, such as Shanghai Sangon Biotechnology Co., Ltd. The obtained sequence was analyzed by Blasm using a network tool, and the genetic tree analysis was completed b...

Embodiment 3

[0033] The cultivation and biological deamination analysis of embodiment 3Citrobacter freundii S06

[0034] Inoculate the monoclonal colony on the LB plate into 50ml of LB liquid medium, 30°C, 200rpm, shake and culture in a 250ml shake flask for 24h, take 3% (v / v) culture solution and centrifuge, 3,000g, 2min, to Wash with 1ml of NRM medium, then inoculate into 100ml of NRM liquid medium, shake and culture in a 500ml shaker flask for 72h at 30°C, 200rpm, regularly sample and measure ammonia nitrogen (NH 3 -N), nitrate (NO 3 -N) and nitrite (NO 2 -N) concentration, by analyzing the residual amount of total nitrogen (TN) to measure the removal rate of the strain to inorganic nitrogen, and thus to judge the heterotrophic nitrification and aerobic denitrification performance of the strain. Depend on figure 2 It can be seen that the biological denitrification performance of the isolated strain S06 in the synthetic medium is very high, and it has a good denitrification performan...

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Abstract

The invention discloses a heterotrophic nitrifying aerobic denitrifying Citrobacter freundii, which can be used in industrial and agricultural production or domestic sewage treatment. The said strain has been preserved in China General Microorganism Culture Collection and Management Center, numbered CGMCC No. 13434, and it can be applied to the biological denitrification of nitrogenous water bodies and the biodegradation of various antibiotics in various water body environments. Citrobacter freundii S06 in the present invention is not only capable of biological removal of organic or inorganic nitrogen, but also found to tolerate phenol in water and can effectively degrade it. The test results of heterotrophic nitrification performance show that the above-mentioned Acinetobacter Citrobacter freundii S06 has a high removal rate for organic and inorganic ammonia nitrogen, and there is no accumulation of intermediate metabolites nitrate and nitrite; The denitrification treatment of nitrate under certain conditions has low requirements for C / N nutrition, strong reproductive ability in the sewage environment, and the tolerance and degradation of phenol make it very good in the green treatment of industrial pollutants in rivers. Application prospect.

Description

technical field [0001] The invention belongs to the field of applied microorganisms, and in particular relates to a heterotrophic nitrifying aerobic denitrifying Citrobacter which can be applied to sewage treatment. Background technique [0002] The development of animal husbandry and modern planting has made agricultural pollution more than industrial sludge, becoming the largest industry pollution source in the country; in addition, the city's water environment is also facing challenges due to dense urban population and imperfect domestic garbage disposal schemes and facilities. Excessive nitrogen and phosphorus levels in the water environment are very common, which can lead to eutrophication and algal blooms, affecting the ecological balance of the water and its self-purification function. Therefore, nitrogen-rich domestic sewage needs to be denitrified. In addition, the water environment around cities and industrial areas often has serious phenolic pollution due to the p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/38
CPCC02F3/34C02F2101/38C12N1/205C12R2001/01
Inventor 邱建贺翁雯
Owner 玉凰生态科技股份有限公司
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