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Aerobic denitrification psychrotolerant bacterium and preparation method thereof

An aerobic denitrification and cold-resistant technology, applied in biochemical equipment and methods, chemical instruments and methods, aerobic process treatment, etc., can solve the problems of slow denitrification rate, slow process, and low denitrification efficiency, and achieve high value, the effect of a wide temperature range

Inactive Publication Date: 2012-02-01
JILIN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In the past, aerobic denitrifying bacteria isolated by people can simultaneously use oxygen molecules and nitrate ions as electron acceptors to carry out the denitrification process, but the denitrification rate is relatively slow, and the existence of oxygen molecules has certain disadvantages for the strains to use nitrate ions influences
At the same time, some studies have shown that under anaerobic conditions, microorganisms can use nitrate ions as the only electron acceptor to carry out the denitrification process under the catalysis of anaerobic denitrification enzymes, but the process is slow and the denitrification efficiency is low.

Method used

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  • Aerobic denitrification psychrotolerant bacterium and preparation method thereof
  • Aerobic denitrification psychrotolerant bacterium and preparation method thereof
  • Aerobic denitrification psychrotolerant bacterium and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] (1) When the ambient temperature is 10-15°C, water samples from the heavily eutrophic water body of Yitong River in Changchun City, Jilin Province and the sediment in the shallow water area were collected and mixed according to the volume ratio of 1:1 as the bacterial source;

[0030] (2) Take 20ml of the bacterial source and add it to a sterilized 250ml Erlenmeyer flask containing 80ml of selective medium, culture it with shaking (160r / min) for 48h at 10°C, and then centrifuge (40000r / min, 15min) , take 0.01g of the precipitate and add it to an iodine flask filled with fresh medium for static culture, centrifuge after 48h, then take 0.01g of the precipitate, add it to the medium for aerobic culture, and cultivate alternately 10 times in this way, each time Do 3 repetitions and the control treatment without bacterial source;

[0031] (3) Apply the last aerobic culture solution on the solid medium for culture, select a single colony, and prepare it with sterile water con...

Embodiment 2

[0035] Pick a ring of bacterial cells of the present invention from the plate, add 100 μl of sterile twice-distilled water, vortex and mix, boil in water bath for 5 minutes, centrifuge at 12000r / min for 5 minutes, and use the supernatant for PCR. The primers are a pair of universal primers:

[0036] Forward primer Pf: 5'-AGAGTTTGATCCTGGCTCAG-3';

[0037] Reverse primer Pr: 5'-ACGGCTACCTTGTTACGACT-3',

[0038] Corresponding to bases 8-27 and bases 1495-1514 of the 16S rRNA gene of Escherichia coli, respectively;

[0039] The PCR reaction system (100 μl) was: 10× Ex Taq buffer 10 μl, 5 U Ex Taq, 10 mmol / L dNTPs 4 μl, 50 pmol / L primers 1 μl, 1 μl template, supplemented to 100 μl with twice distilled water. The PCR reaction conditions are: 95°C for 3min; 95°C for 1min; 58°C for 1min; 72°C for 1min for 30s, 30 cycles; 72°C for 10min, store at 4°C;

[0040] 5 μl of the PCR product was taken for identification by 1% agarose gel electrophoresis. The PCR product was recovered and pu...

Embodiment 3

[0042] Use 0.1mol / L sodium hydroxide and 0.1mol / L hydrochloric acid solution to adjust the pH value of the liquid medium to 7.2, add 90ml of liquid medium to 28 250ml Erlenmeyer flasks, and sterilize under high pressure steam at 121°C for 30min After cooling, add 10ml of bacterial suspension of the present invention to 21 Erlenmeyer flasks among them respectively, shake and cultivate at a temperature of 5, 10, 15, 20, 25, 30 and 35° C., and repeat each temperature three times. And do the control treatment without adding bacteria respectively, the shaking speed is 160rpm. Samples were collected at 24, 48, and 72 hours after culturing to measure the growth status of the bacterial cells.

[0043] The growth status of the bacterial cells was characterized by the absorbance at 600nm. The results showed that the strain entered the logarithmic growth phase after a brief adaptation in the new culture medium.

[0044] When the temperature is 5°C, the bacterial strain of the present i...

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Abstract

The invention provides an aerobic denitrification psychrotolerant bacterium and a preparation method thereof. The collection number of the aerobic denitrification psychrotolerant bacterium is CGMCC No.4753. A strain can undergo denitrification at high dissolved oxygen level and low dissolved oxygen level, and has a wider appropriate growing temperature range. After the aerobic denitrification psychrotolerant bacterium is inoculated into lake water polluted by agricultural runoff, and is cultured at 10 DEG C for 7 days, the removing rate of nitrate ions is over 80 percent. The aerobic denitrification psychrotolerant bacterium has a higher treatment value on low-temperature eutrophic water bodies.

Description

technical field [0001] The invention discloses an aerobic denitrification psychrophilic bacterium, and also provides a preparation method of the psychrotolerant bacterium, which belongs to the technical field of bioremediation of water environment pollution. Background technique [0002] my country's per capita water resources are poor, and a large amount of nitrogen-containing nutrients entering lakes and reservoirs can easily cause water eutrophication, further aggravating the situation of water shortage in my country, and seriously threatening the safety of drinking water and the health of the ecosystem. In 2008, 22 of my country's 28 nationally controlled key lakes (reservoirs) reached Grade V and below water quality, accounting for 78.6%, of which total nitrogen was one of the main pollution indicators. In essence, the prevention and control of water eutrophication is mainly to solve the problem of nitrogen and phosphorus pollution, that is, to reduce or cut off the inp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34C02F3/02C12R1/01
CPCY02W10/10
Inventor 李明堂郝林琳刘梦洋曹国军
Owner JILIN AGRICULTURAL UNIV
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