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Pseudomonas zhaodongensis strain and application thereof

A technology of pseudomonas strains and seeds, applied in the field of environmental microorganisms, can solve the problems of immature products, stay in the research stage, and large inoculation amount of strains, and achieve shortened fermentation cycle, reduced costs, and strong bacterial vitality Effect

Active Publication Date: 2022-03-29
青岛蔚蓝赛德生物科技有限公司
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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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  • Pseudomonas zhaodongensis strain and application thereof
  • Pseudomonas zhaodongensis strain and application thereof
  • Pseudomonas zhaodongensis strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Screening and performance detection of strains

[0032] Enrichment training

[0033] Collect a chemical plant sewage, draw 10 ml of sewage transfer to a 100ml enrichment medium (2 g / L of nitrate, 0.5 g / L of hydrogen phosphate, 0.2 g / L of magnesium sulfate 200g / L, adjustment In a 250 mL triangular bottle of pH = 7.20, it was allowed to cultivate 7 days at 8 ° C for the first time. Then, a 10 mL of a enriched liquid was then drawn, added to a fresh enrichment medium, and allowed to stand for 7 days under 8 ° C, and the second enrichment was performed. The third enrichment is performed in the same enrichment method described above.

[0034] 2. First Sieve

[0035] Dilute the third enrichment to 10 with gradient dilution -6 , Draw 10 separately -3 10 -4 10 -5 10 -6200 μl of the dilution to the separation medium (potassium nitrate 2g / L, 0.5 g / L of hydrogen phosphate, 0.2 g / L of magnesium sulfate 200g / L, agar powder 20g / L, pH = 7.20) After the coating...

Embodiment 2

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

[0066] 1, experimental method

[0067] 1.1 Bacterial genome DNA extraction

[0068] Collect 1.0 × 10 with 2ML centrifuge tube 9 (1ml bacterial OD600 is 1-1.5) bacteria culture, 12,000 × g centrifugation 30s, abandoned. A buffer S suspended by 150 μL has been added to the RNASe A.

[0069] 20 μl of lyse reservoir was added, mixed uniform, and allowed to stand for 5 min at room temperature.

[0070] 30 μL of 0.25 mol / l EDTA (pH 8.0) was added, mixed uniform, ice bath 5 min.

[0071] 450 μl of buffer G-A, vortex oscillation 15s, 65 ° C water bath 10 min.

[0072] 400 μL of buffer G-B and 1 ml buffer DV (pre-cooling at 4 ° C) were added, mixed with force, and centrifuged at 12,000 × g for 2 min.

[0073] As much as possible, the upper phase is discarded, and the phase precipitation and the lower phase are retained. 1 ml of pre-cooling buffer DV was added, mixed with force, and centrifuate 12,000 × g for 2 min.

[007...

Embodiment 3

[0111] Example 3: Preparation and storage of microbial agents

[0112] 3.1 Preparation of microbial agents:

[0113] (1) Level 1 seed culture: Aseptic environment to pick 1 ring Zudong pseudogen strain DB-LT01 access to 100 mL enrichment medium (2 g / L of nitrate, 0.5 g / L of hydrogen phosphate) The 250 ml triangular bottle of 0.2 g / L, potassium tartrate sodium sodium sodium sulfate was placed in a 250 ml triangular bottle of 30 ° C, 120 rpm, and obtained at 1000 mPM.

[0114] (2) Secondary seed culture: in aseptic environment, 5 ml of seed culture fluid is transferred from 5 ml to four in 500 ml of enrichment medium (2 g / L of nitrate, 0.5 g / L, sulfuric acid In a 1L triangular bottle of 0.2 g / L, potassium tartrate, pH = 7.20), placed under 30 ° C, 120 rpm, and cultured under secondary seed culture fluid;

[0115] (3) Disinfection sterilization: In addition to glucose alone ingredients and sterilization, other materials are ingredients in the ingredients, then in 1T ferme...

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Abstract

The invention relates to a Pseudomonas zhaodongensis for low-temperature denitrification and a microbial agent containing the Pseudomonas zhaodongensis, the Pseudomonas zhaodongensis is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC No.23567, the Pseudomonas zhaodongensis can keep an excellent total nitrogen removal effect at the temperature as low as 5 DEG C, the total nitrogen degradation rate reaches 96% or above at the temperature of 8 DEG C, and the Pseudomonas zhaodongensis has the advantages that the Pseudomonas zhaodongensis can be used for removing nitrogen from the Pseudomonas zhaodongensis and the microbial agent containing the Pseudomonas zhaodongensis; the degradation rate of nitrate nitrogen can reach 97% or above, the degradation rate of nitrite nitrogen can reach 100%, the degradation rate of ammonia nitrogen can also reach 99% or above, and the degradation efficiency is gradually improved along with temperature rise.

Description

Technical field [0001] The present invention relates to a Jidong pseudogen strain and a microbial bacterium comprising therethrough, in particular to a Jidong pseudomonas strain capable of still having a nitrogen-containing substance in a temperature as low as 5 ° C. Application, belongs to the field of environmental microbiological technology. Background technique [0002] With the development of national industrialization and the improvement of people's living standards, the increasing amount of water supplies with pollutants has become more and more serious. The environmental incidents, public safety incidents and even major social events due to water pollution, and seriously affect the people's physical health and social harmony and stability, directly threaten human living space. [0003] During the sewage treatment, nutrients such as organics and nitrogen, phosphorus in sewage can be degraded and utilized by metabolism. The main role in sewage treatment is a medium temperat...

Claims

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Application Information

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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 青岛蔚蓝赛德生物科技有限公司
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