Heterotrophic nitrifying bacterium for low-temperature sewage denitrification and application thereof

A heterotrophic nitrification, low temperature technology, applied in bacteria, biological water/sewage treatment, microorganism-based methods, etc., can solve the problem of low denitrification efficiency of low temperature sewage, and achieve the effect of broad application prospects.

Inactive Publication Date: 2020-04-07
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a strain of low-temperature heterotrophic nitrifying bacteria to solve the problem of low denitrification efficiency of low-temperature sewage

Method used

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  • Heterotrophic nitrifying bacterium for low-temperature sewage denitrification and application thereof
  • Heterotrophic nitrifying bacterium for low-temperature sewage denitrification and application thereof
  • Heterotrophic nitrifying bacterium for low-temperature sewage denitrification and application thereof

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Experimental program
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Effect test

Embodiment 1

[0025] The screening method of low-temperature heterotrophic nitrifying bacteria BC-15 is as follows: take an appropriate amount of biological turntable mud sample and add it to a 250mL conical flask filled with 100mL sterile water, shake it on a shaker for 3 hours, and then use a pipette gun to draw 10mL liquid to inoculate In the sterilized enrichment medium, culture at 13°C and 150rpm for 5 days; then take the enrichment medium and transfer it to a new enrichment medium, transfer 3 times, and enrich for 15 days in total.

[0026] Take 1 mL of the enriched culture solution and put it into a test tube filled with 9 mL of sterile water, and dilute it according to the doubling ratio to obtain a concentration of 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 Gradients of bacterial suspensions. Then, using the plate coating method, draw 200 μL from each gradient bacterial suspension and spread it on the heterotrophic nitrification medium, place it in a constant temperature...

Embodiment 2

[0032] Molecular Biological Identification of Low Temperature Nitrifying Bacteria BC-15:

[0033] Physiological and biochemical tests of strain BC-15 were Gram-negative (see figure 2 ), milky white and round, the surface is moist and easy to provoke, and the bacteria are rod-shaped. Use a genome kit to extract the bacterial genome of the target strain; use universal primers 27F: 5'-AGAGTTTGATCMTGGCTCAG-3', 1492R: 5'-TACGGYTACCTTGTTACGACTT-3' for PCR amplification, and verify by agarose gel electrophoresis (1%) ; Electrophoresis detection, gel-cutting purification and sequencing, and sequence determination by Meiji Biotechnology.

[0034] PCR reaction conditions: pre-denaturation at 94°C for 3min; denaturation for 30s; annealing at 54°C for 30s; extension at 72°C for 1min30s; repeat 24 cycles from the second step.

[0035] The sequence length of strain BC-15 is 1316bp, and its gene sequence list is shown in SED ID NO:1.

[0036] The strain sequence results were uploaded to ...

Embodiment 3

[0038] Growth and denitrification performance of strain BC-15:

[0039] Add the low-temperature digested bacteria BC-15 to the activated medium for expanded culture, centrifuge at 6000rpm for 3 minutes, keep the bacteria, prepare the bacterial suspension with sterile water, and inoculate it in the heterotrophic nitrification medium according to the inoculum size of 3%. The initial ammonia nitrogen concentration was 100mg / L, and the OD after inoculation 600 It is about 0.4, and cultivated at 13°C, 150rpm, on a shaker for 72h. Regularly take samples to determine the middle NH in the supernatant 4 + -N, OD600 value, see the result image 3 .

[0040] It can be seen from the figure that the BC-15 strain reaches the logarithmic growth phase after 27 hours of growth in the nitrifying medium, and the OD600 at this time is about 1.1. The ammonia nitrogen concentration gradually decreases while the bacteria grow, and the ammonia nitrogen concentration is the lowest in the logarithm...

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Abstract

The invention discloses a heterotrophic nitrifying bacterium for low-temperature sewage denitrification and an application thereof, which belong to the field of environmental microorganism and water treatment biological denitrification. The invention aims to solve the problem of low denitrification efficiency of low-temperature sewage. The low-temperature nitrifying bacterium belongs to acinetobacter and is preserved in China General Microbiological Culture Collection Center of No.3, Yard 1, West Beichen Road, Chaoyang District, Beijing, the preservation date is October 25, 2019, and the preservation number is CGMCC No.18736. The strain provided by the invention is obtained by screening from a biological rotating disc mud sample of a sewage treatment plant in winter, can effectively removeammoniacal nitrogen in sewage under a low-temperature condition, can be directly used for denitrification of a water body in winter after amplification culture, and has relatively great application potential.

Description

technical field [0001] The invention relates to the fields of environmental microorganisms and water treatment biological denitrification, in particular to a low-temperature nitrifying bacterial strain used for sewage treatment, and simultaneously provides the denitrification performance of the bacterial strain. Background technique [0002] At present, in my country's water environment quality, nitrogen content and pollution are relatively serious. Nitrogen in water mainly comes from wastewater produced by industry and agriculture and sewage discharged from daily life, and the accumulation of nitrogen is one of the main sources of pollution causing eutrophication of water. my country's urban sewage treatment mainly adopts biological methods, that is, the use of microorganisms to decompose pollutants to achieve the purpose of purifying sewage. However, the growth of microorganisms is limited by many factors, among which temperature is an important factor. Macroscopically, ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/16
CPCC12N1/20C02F2101/16C12R2001/01C12N1/205
Inventor 王晓慧刘琳海热提·吐尔逊李媛
Owner BEIJING UNIV OF CHEM TECH
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