Low-carbon and high-nitrogen resistant heterotrophic nitrification-aerobic denitrification sphingomyza sp. CY-10 and application thereof

A technology for nitrifying sphingosine box and heterotrophic nitrification, which is applied in the application field of heterotrophic nitrification-aerobic denitrifying bacteria, can solve problems such as cost increase, and achieve the effects of fast growth rate of bacteria and high removal rate

Pending Publication Date: 2021-11-26
CHONGQING UNIV OF TECH
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  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the heterotrophic nitrification-aerobic denitrification function of sphingosine bacteria in China
Moreover, currently reported heterotrophic nitrification-aerobic denitrification bacteria require higher C / N in the denitrification process, and often require an external carbon source, which increases the cost

Method used

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  • Low-carbon and high-nitrogen resistant heterotrophic nitrification-aerobic denitrification sphingomyza sp. CY-10 and application thereof
  • Low-carbon and high-nitrogen resistant heterotrophic nitrification-aerobic denitrification sphingomyza sp. CY-10 and application thereof
  • Low-carbon and high-nitrogen resistant heterotrophic nitrification-aerobic denitrification sphingomyza sp. CY-10 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1, strain enrichment and optimization

[0027] 1) Enrichment culture: Take 2ml of biogas slurry water sample from Chongqing Muan pig farm in Banan District, Chongqing, inoculate it into a 250ml Erlenmeyer flask containing 100ml of LB medium, put it into a shaker set at 30°C, 160r / min; After culturing for 72 hours, inoculate at 2% into a 250ml Erlenmeyer flask also equipped with 100ml LB medium, and subculture twice to obtain the enriched bacterial solution.

[0028] 2) Subculture and domestication: Take 2ml of the enriched bacteria liquid, inoculate it into a 250ml Erlenmeyer flask containing 100ml of heterotrophic nitrification, aerobic and denitrification selective medium, put it into a shaker set at 30°C, 160r / min, and test every 24h OD in culture flask 600 , NH 4 + -N, TN, subcultured three times to obtain a mixed flora with stable denitrification effect.

[0029] 3) Isolation of pure bacteria: use cooled sterile distilled water to carry out 10-fold s...

Embodiment 2

[0031] Embodiment 2, strain identification

[0032] Sichuan Qingke Company was entrusted to complete the 16SrDNA sequencing of the screened strains. After sequencing, the 16S rDNA sequence of the strain was obtained. According to the result, Blast comparison and identification was carried out in NCBI, and the strain with similar homology to the target gene sequence was selected, and the phylogenetic tree was constructed by Neighbor-Joining method with MEGA 7.0 software.

[0033] The identification results showed that the bacteria belonged to Gram-negative bacteria, the colony diameter was 2-4mm, aerobic, the surface was moist, shiny and opaque. The sequencing of the 16S rDNA of Sphingopyxis sp.CY-10 is shown in SEQ ID NO.1, and the base sequence was compared with the strains SAS22 and T2 of Sphingopyxis sp. in the GenBank nucleic acid sequence database. , SM 105 sequences are similar, and the phylogenetic tree of Sphingopyxis sp.CY-10 is shown in figure 2 .

Embodiment 3

[0034] Example 3, Sphingopyxis sp.CY-10 degrading effect on ammonia nitrogen under different C / N

[0035] Prepare heterotrophic nitrification, aerobic and denitrification selection media with C / N of 1, 2, 4, 6, 8, 10, and 20 respectively, in triplicate, and sterilize the above culture at 121°C, and after cooling Inoculate 2ml of sphingosine box bacteria CY-10 bacterial solution in the ultra-clean bench, cover the rubber stopper and shake well, put it into a shaker set at 30°C and 160 rpm for cultivation, and detect the concentration of the bacterial solution every 24 hours ( OD 600 ), ammonia nitrogen (NH 4 + -N), total nitrogen (TN) concentration. The detection method of ammonia nitrogen is Nessler's reagent spectrophotometry, and the cuvette and wavelength are respectively 20mm glass cuvette and 420nm. The reagents used in the detection process were Nessler's reagent and potassium sodium tartrate. The detection method of total nitrogen is alkaline potassium persulfate s...

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Abstract

The invention discloses a low-carbon and high-nitrogen resistant heterotrophic nitrification-aerobic denitrification sphingomyza sp. CY-10 and application thereof. The strain is preserved in China Center for Type Culture Collection, the preservation number is CCTCC NO: M 2021682, and the strain is named as sphingomyza sp. CY-10. The strain can remove high-concentration ammonia nitrogen under the condition of low C / N, has the heterotrophic nitrification-aerobic denitrification function for the first time, can keep higher activity under the environment of low C / N and high ammonia nitrogen, can be used for biological treatment of culture biogas slurry, high-speed service area wastewater, decentralized sewage and the like, and is expected to make a new breakthrough in the field of biological denitrification in extreme environments.

Description

technical field [0001] The invention relates to the field of environmental microorganisms, in particular to low-carbon and high-nitrogen-resistant heterotrophic nitrification-aerobic denitrification sphingosine bacteria, and also relates to the application of low-carbon and high-nitrogen-resistant heterotrophic nitrification-aerobic denitrification bacteria. Background technique [0002] Excessive ammonia nitrogen in the water body will cause eutrophication of the water body and lead to deterioration of water quality, which will not only cause poisonous effects on fish and aquatic organisms, but also seriously endanger the ecological balance and human health. The biological denitrification method is widely used in the treatment of ammonia nitrogen wastewater due to its economical, high efficiency and no secondary pollution. The traditional biological denitrification process includes aerobic nitrification and anaerobic denitrification, which requires autotrophic nitrifying ba...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/02C02F3/34C12R1/01
CPCC02F3/02C02F3/34Y02W10/10
Inventor 赵天涛陈沛沛刘毫李国戬张丽杰
Owner CHONGQING UNIV OF TECH
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