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A kind of Zhaodong Pseudomonas strain and its application

A technology of Pseudomonas strains and seeds, which is applied in bacteria, biological water/sewage treatment, biochemical equipment and methods, etc., can solve the problems such as immature products, staying in the research stage, and large amount of bacterial inoculation, etc. To achieve the effect of shortening the fermentation cycle, reducing costs, and strong bacterial vitality

Active Publication Date: 2022-06-14
QINGDAO VLANDSAIDE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1. Most of the research is only in the research stage and has not yet formed a mature product;
[0006] 2. In most studies, the inoculum amount of bacteria is too large, and the cost in practical application is relatively high;
[0007] 3. The minimum effective temperature is mostly above 10°C, and the application cost is relatively high

Method used

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  • A kind of Zhaodong Pseudomonas strain and its application
  • A kind of Zhaodong Pseudomonas strain and its application
  • A kind of Zhaodong Pseudomonas strain and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. strain screening and performance testing

[0032] 1. Enrichment culture

[0033] Collect sewage from a chemical plant, absorb 10mL of sewage and transfer to 100mL enrichment medium (potassium nitrate 2g / L, dipotassium hydrogen phosphate 0.5g / L, magnesium sulfate 0.2g / L, potassium sodium tartrate 200g / L, adjusted pH=7.20) in a 250mL Erlenmeyer flask, cultured statically at 8°C for 7 days for the first enrichment. Then, draw 10 mL of the primary enrichment solution, add it into a fresh enrichment medium, and culture it statically at 8° C. for 7 days to perform the second enrichment. Carry out the third enrichment according to the same enrichment method as above.

[0034] 2. Primary screening

[0035] The third enrichment was diluted to 10 by serial dilution -6 , draw 10 respectively -3 、10 -4 、10 -5 、10 -6Add 200 μL of each dilution to the separation medium (potassium nitrate 2 g / L, dipotassium hydrogen phosphate 0.5 g / L, magnesium sulfate 0.2 g / L, pota...

Embodiment 2

[0065] Example 2. Detection and identification of bacterial strain DB-LT01

[0066] 1. Experimental method

[0067] 1.1 Extraction of bacterial genomic DNA

[0068] Collect 1.0×10 in a 2ml centrifuge tube 9 (1ml bacterial solution OD600 is 1-1.5) bacterial culture, centrifuge at 12,000×g for 30s, and discard the supernatant. Suspend the pellet with 150 μl Buffer S to which RNase A has been added.

[0069] Add 20 μl lysozyme stock solution, mix well, and let stand at room temperature for 5 minutes.

[0070] Add 30 μl 0.25mol / L EDTA (pH 8.0), mix well, and ice-bath for 5 minutes.

[0071] Add 450μl Buffer G-A, vortex for 15s, and bathe in water at 65°C for 10min.

[0072] Add 400μl Buffer G-B and 1ml Buffer DV (pre-cooled at 4°C), mix vigorously, and centrifuge at 12,000×g for 2min.

[0073] Discard the upper phase as much as possible, save the interphase pellet and the lower phase. Add 1ml of 4°C pre-cooled Buffer DV, mix vigorously, and centrifuge at 12,000×g for 2min. ...

Embodiment 3

[0111] Embodiment 3: preparation and storage of microbial bacterial agent

[0112] 3.1 Preparation of microbial agent:

[0113] (1) First-level seed culture: Pick one ring of Pseudomonas Zhaodong strain DB-LT01 in a sterile environment and insert it into 100mL enrichment medium (potassium nitrate 2g / L, dipotassium hydrogen phosphate 0.5g / L, Magnesium sulfate 0.2g / L, potassium sodium tartrate 200g / L, adjust pH=7.20) in the 250mL Erlenmeyer flask that is placed in 30 ℃, 120rpm conditions and cultivate 24h, obtain the primary seed culture solution;

[0114] (2) Secondary seed culture: In a sterile environment, transfer 5mL of the primary seed culture solution to four 500ml enrichment media (potassium nitrate 2g / L, dipotassium hydrogen phosphate 0.5g / L, sulfuric acid Magnesium 0.2g / L, potassium sodium tartrate 200g / L, adjust pH=7.20) in the 1L Erlenmeyer flask, place 30 ℃, cultivate 24h under the condition of 120rpm, obtain secondary seed culture solution;

[0115] (3) Disinfect...

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PUM

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Abstract

The invention relates to a low-temperature denitrification Pseudomonas zhaodongensis strain (Pseudomonas zhaodongensis) and a microbial agent containing it. The strain is preserved in the General Microbiology Center of the China Microbiological Culture Collection Management Committee, and the preservation number is CGMCC No.23567. The strain can maintain an excellent total nitrogen removal effect at a temperature as low as 5°C. At 8°C, the degradation rate of total nitrogen can reach more than 96%, and the degradation rate of nitrate nitrogen can reach more than 97%. The degradation rate of state nitrogen reaches 100%, and the degradation rate of ammonia nitrogen can reach more than 99%, and the degradation efficiency gradually increases with the increase of temperature.

Description

technical field [0001] The invention relates to a Pseudomonas Zhaodong strain and a microbial agent containing the same, in particular to a Pseudomonas Zhaodong strain capable of degrading nitrogenous substances in water at a temperature as low as 5°C and its The invention belongs to the technical field of environmental microorganisms. Background technique [0002] With the development of national industrialization and the improvement of people's living standards, the increasing water consumption and the discharge of pollutants have made the problem of water pollution more and more serious. Environmental incidents, public safety incidents and even major social incidents caused by water pollution have seriously affected people's health and social harmony and stability, directly threatening the living space of human beings. [0003] In the process of sewage treatment, organic matter and nutrients such as nitrogen and phosphorus in sewage can be degraded and utilized by microo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/38C02F101/16
CPCC12N1/20C02F3/348C02F3/34C02F2101/163C02F2101/166C02F2101/16
Inventor 朱威刘圣鹏张大飞吴娜孔令迎
Owner QINGDAO VLANDSAIDE BIOTECHNOLOGY CO LTD
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