Denitrification phosphorus removal bacterium H-hrb02 as well as screening method and application thereof

A technology for denitrifying phosphorus-accumulating bacteria and denitrifying phosphorus-accumulating bacteria, applied in chemical instruments and methods, biochemical equipment and methods, bacteria, etc. The effect of polyphosphorus properties

Active Publication Date: 2013-01-09
宜兴环保产业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, denitrifying and phosphorus-accumulating strains are mainly concentrated in Pseudomonas Bacillus and Bacillus, the strains are relatively single, and the research is mostly focused on the degradation gene and degradation mechanism, and the practical application research is limited

Method used

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  • Denitrification phosphorus removal bacterium H-hrb02 as well as screening method and application thereof
  • Denitrification phosphorus removal bacterium H-hrb02 as well as screening method and application thereof
  • Denitrification phosphorus removal bacterium H-hrb02 as well as screening method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment approach 1

[0034] Embodiment 1: In this embodiment, Pseudomonas aeruginosa H-hrb02 bacteria ( Pseudomonasaeruginosa strain H-hrb02) is measured by the following steps:

[0035] 1. Isolation, purification and screening of Pseudomonas aeruginosa H-hrb02 bacteria:

[0036] a. The activated sludge at the end of the anoxic period of a certain period when the SBR reactor is running stably is used as the separation sludge

[0037] b. Separation was performed using the dilution-mixed-plate method. Denitrifying bacteria are isolated using denitrifying medium, and phosphorus accumulating bacteria are isolated using phosphorus-rich medium.

[0038]The specific separation steps are as follows: take 10 mL of activated sludge from the end of the anoxic section of the SBR reactor into a conical flask filled with 90 mL of sterile water, and add sterile glass beads. Put the triangular flask into the shaker and oscillate fully to make the bacteria disperse in the water in a single-cell state. Then...

Embodiment approach 2

[0043] Embodiment 2: 16S rDNA sequencing of genes, the specific steps of PCR amplification of 16S rDNA genes are as follows:

[0044] a. PCR system establishment (25 μL):

[0045] 10×PCR Buffer 2.5μL

[0046] dNTPs (2.5mmol / L concentration) 2μL

[0047] Primer 1 0.5 μL

[0048] Primer 2 0.5 μL

[0049] DNA template 0.5 μL

[0050] rTaq DNA polymerase (5U / μL) 0.5μL

[0051] Add sterile deionized water to 25 μL

[0052] b. PCR program setting:

[0053] Pre-denaturation at 95°C for 3 minutes

[0054] Pre-denaturation at 95°C for 5 minutes, denaturation at 94°C for 1 minute, renaturation at 58°C for 30 seconds, extension at 72°C for 3 minutes, a total of 30 cycles, and finally extension at 72°C for 10 minutes;

[0055] c. Primer sequence:

[0056] Primer 1 5'-CGCCAGGGTTTTTCCCAGTCACGAC-3'

[0057] Primer 2 5'-AGCGGATAACAATTTCACACAGGA-3'

[0058] The gene sequence of Pseudomonas aeruginosa H-hrb02 is shown in SEQ ID No.1.

Embodiment approach 3

[0059] Embodiment 3: In this embodiment, Pseudomonas aeruginosa H-hrb02 can use a variety of organic matter as carbon sources, and can be used for biological denitrification and phosphorus removal of sewage. Detection of the biological nitrogen and phosphorus removal ability of Pseudomonas aeruginosa H-hrb02:

[0060] 1. Determination of the growth curve of Pseudomonas aeruginosa H-hrb02. Measure the growth curve of the bacterial strain (Pseudomonas aeruginosa H-hrb02) according to the standard method for determining the bacterial growth curve, take the culture solution every 2 hours, and use photoelectric turbidimetry to measure the OD of the bacterial solution at a wavelength of 600nm 600 (Optical Density), and then filtered through a 0.22μm microporous membrane to detect the filtrate PO 4 3- -P, pH value and other indicators. Obtain the growth curve of Pseudomonas aeruginosa H-hrb02 as image 3 shown. from image 3 It can be seen from the figure that the growth curve ...

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Abstract

The invention discloses a denitrification phosphorus removal bacterium as well as a screening method and application efficiency thereof. According to identification, the denitrification phosphorus removal bacterium belongs to pseudomonas, is pseudomonasaeruginosa and is named H-hrb02 and collected in the China General Microbiological Culture Collection Center with the collection number CGMCC NO.5940 on February 27, 2012. An H-hrb02 bacillus is a non-fermented Gram-negative bacterium as well as an obligate aerobe (except that in a nitrate culture medium), grows in the temperature range of 25-42 DEG C, preferably grows at the temperature of 25-30 DEG C, does not grow at the temperature of 4 DEG C particularly, but can grow at the temperature of 42 DEG C. The bacillus is 1.5-5.0 mu m in length and 0.5-1mu m in width, is cue-stick-shaped or linear, and is arranged pairwise or in the form of a short chain; and one end of the bacillus is monotrichous and is free from spores. In a denitrification phosphorus removal strain H-hrb02 provided by the invention, a plurality of organic matters can be taken as carbon sources. After being activated and cultured, the strain (H-hrb02) has excellent denitrification and phosphorus removal performances and can be applied to a sewage biological nitrogen and phosphorus removal process, the phosphorus absorption rate can be up to 92 percent, and the nitrogen removing rate is up to 87.7 percent.

Description

technical field [0001] The invention relates to a denitrifying phosphorus-accumulating bacterium H-hrb02 and its application efficiency, and simultaneously provides a screening method for the bacterium. Background technique [0002] At present, with the rapid development of the national economy and the continuous deepening of the urbanization process, the total discharge of urban domestic and industrial wastewater is also increasing year by year. The widespread use of pesticides, chemical fertilizers and synthetic detergents has increased the concentration of nutrients in water bodies, among which nitrogen and phosphorus are one of the main causes of eutrophication in water bodies. Conventional biochemical treatment process can effectively reduce BOD in sewage 5 and SS, but when there are N, P and other nutrients in the sewage, only 10%-20% of the phosphorus in the sewage can be removed, and a large amount of phosphorus-containing sewage will be directly discharged into the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/68C02F3/34C12R1/385C02F101/16C02F101/10
Inventor 李昂孙移鹿马放张晓昕庞长泷崔迪张斯杜丛
Owner 宜兴环保产业有限公司
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