Denitrification phosphorus removal bacterium H-hrb02 as well as screening method and application thereof
A technology of denitrification phosphorus accumulation and bacteria, applied in the direction of chemical instruments and methods, biochemical equipment and methods, bacteria, etc., can solve the problems of single strain of bacteria and limited practical application research, and achieve excellent denitrification and phosphorus accumulation performance Effect
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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. The...
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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