Heterotrophic nitrification and aerobic denitrification Citrobacter species and application thereof

A technology of citric acid and bacillus freundii, applied in the field of applied microorganisms, can solve the problems of shortening hydraulic retention time, ammonia nitrogen cannot be processed in time, and hydraulic retention time is long, and achieves good biocatalysis function, wide temperature adaptation range, and growth adaptability. strong effect

Active Publication Date: 2017-12-12
FOSHAN YUHUANG ECOLOGICAL ENVIRONMENT TECH
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  • Abstract
  • Description
  • Claims
  • 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

Method used

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  • Heterotrophic nitrification and aerobic denitrification Citrobacter species and application thereof
  • Heterotrophic nitrification and aerobic denitrification Citrobacter species and application thereof
  • Heterotrophic nitrification and aerobic denitrification Citrobacter species and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0029] Activated sludge is taken from the treatment tank of Shanghai Pudong Zhuyuan No. 1 Sewage Treatment Plant, and 5mL mud-water mixture is inoculated into 45mL sterilized NRM enrichment medium (the formula of NRM medium is: (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 (according to per liter): KNO 3 1g, sodium succinate 8.5g, MgSO 4 ·7H 2 O 1g, CaCl 2 0....

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 CFS06, dissolve and dilute to 100mg / L, and use this as a template for 16S rDNA extraction For amplification, the primer pairs used are 16S-F and 16S-R, and the sequences are respectively, 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 O 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 Blastn using a network tool, ...

Embodiment 3

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

[0034] Inoculate the monoclonal colony on the LB plate into 50ml of LB liquid medium, 30°C, 200 rpm, shake culture in a 250ml shake flask for 24h, take 3% (v / v) culture solution and centrifuge at 3,000g, 2min, Wash with 1ml of NRM medium, then inoculate into 100ml of NRM liquid medium, shake and culture in a 500ml shake 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 CFS06 in the synthetic medium is very high, and it has a good denitrification performance...

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Abstract

The invention discloses a heterotrophic nitrification and aerobic denitrification Citrobacter species applicable to the treatment of wastewaters from industrial and agricultural production or domestic wastewater. The species is already collected in China General Microbiological Culture Collection Center under CGMCC No. 13434 and is applicable to biological denitrification of nitrogen-bearing water and biological degradation of various antibiotics in various water environments. Citrobacter freundii CFS06 herein is capable of biologically removing organic or inorganic nitrogen and is discovered to tolerate phenols in water and effectively degrade the same. The results of heterotrophic nitrification performance measurement show that the above Acinetobacter, Citrobacter freundii CFS06, has high removal rate for organic and inorganic ammonia nitrogen, with zero accumulation of intermediate metabolites, nitrates and nitrites; the species is capable of denitrifying nitrates under aerobic condition, has low requirement on C/N nutrition, has great breeding capacity in wastewater environments, and has a promising application prospect in terms of green treatment of river channel industrial contaminants owing to its tolerance and degradation for phenols.

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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IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/38
CPCC02F3/34C02F2101/38C12N1/205C12R2001/01
Inventor 邱建贺翁雯
Owner FOSHAN YUHUANG ECOLOGICAL ENVIRONMENT TECH
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