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Pseudomonas aeruginosa strain separating, purifying and domesticating method and use

A technology for separation and purification of Pseudomonas aeruginosa, applied in chemical instruments and methods, methods based on microorganisms, biochemical equipment and methods, etc., to achieve high practical value

Inactive Publication Date: 2010-10-13
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to provide a microorganism for efficient biochemical treatment of phenols and other aromatic compound pollutants. The technical problem to be solved is to select the required microorganisms from the polluted sludge and domesticate them to improve the resistance of the strains to refractory substances. Degradation rate

Method used

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Experimental program
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Effect test

Embodiment 1

[0023] Embodiment 1: Isolation and purification of Pseudomonas aeruginosa (Pseudomonas aeruginosa) TBPY

[0024] A. Under sterile conditions, mix the sludge contaminated with tribromophenol with water and let it stand for 0.5-2 hours. Take 1 mL of the supernatant and dilute it with 0.85% normal saline for 10 -1 ~10 -6 , to obtain 6 parts of diluted bacterial liquid;

[0025] B. On the ultra-clean workbench, take the separated and purified solid medium that has been melted and cooled to about 45°C and pour it on six plates, inject 15-20mL of medium into each plate, and after solidification, take the above 6 parts Add 0.1-0.2mL of diluted bacterial solution to six plates respectively, and spread evenly with a sterile applicator;

[0026] C. Invert the six plates prepared above and place them in a biochemical incubator at 28-32°C for 20-60 hours;

[0027] D. Use an inoculation loop to pick a single colony with a clear shape, number it, mark it on the isolated and purified soli...

Embodiment 2

[0030] Embodiment 2: the strain identification of Pseudomonas aeruginosa bacterial strain (Pseudomonas aeruginosa) TBPY

[0031] The colonies of Pseudomonas aeruginosa strain (Pseudomonas aeruginosa) TBPY obtained by separation and purification were round in shape with neat edges; the color was milky white or milky yellow, moist and shiny; the surface morphology of the colonies was high convex. Observed under an optical microscope, the strain is a brevibacterium, single, in pairs or in short chains. According to the "Bergey's Bacteria Identification Manual", the results of the strain identification show that the strain is Gram-negative, respiratory metabolism, never fermented, positive for contact enzymes, and can use a variety of organic compounds as the only carbon source growth, and was initially identified as belonging to the genus Pseudomonas.

[0032] The 16S rDNA sequence of the strain was sequenced, and the resulting sequence was compared in the NCBI database. The res...

Embodiment 3

[0034] Embodiment 3: the domestication of Pseudomonas aeruginosa bacterial strain (Pseudomonas aeruginosa) TBPY

[0035]Take the purified strain and inoculate it in the inorganic salt medium according to the inoculation amount of 5%-10% (W / V), and domesticate it with the TBP concentration increasing pressure method, so that the maximum concentration of TBP is 100-200mg / L . It is cultivated in a shaker at 25-35°C and 120-170rpm, and inoculated into a new medium every 5-10 days at an inoculation amount of 5%-10%, and the acclimatization time is 2-5 months in total.

[0036] The formula of the above-mentioned inorganic salt liquid medium: every 1000mL of water contains K 2 HPO 4 4~8g, KH 2 PO 4 0.2~0.8g, MgSO 4 0.2~0.6g, (NH 4 ) 2 SO 4 1~5g, MnSO 4 ·H 2 O 0.04~0.08g, FeSO 4 ·7H 2 O 0.008~0.02g, CaCl 2 0.01~0.03g, TBP 50~200mg.

[0037] In the above-mentioned inorganic salt liquid medium, add TBP so that the concentration is 100 mg / L, insert the Pseudomonas aerug...

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Abstract

The invention discloses a Pseudomonas aeruginosa strain TBPY, which is screened from tribromophenol(TBP)-polluted sludge and purified. The purified strain is domesticated by a pressure type domesticating method with gradually increased TBP concentration in an organic salt culture medium, and the domesticated strain can be used for the biological control of waste water containing persistent organic pollutants such as phenol, pyrocatechol, resorcinol, benzoic acid, p-hydroxybenzoic acid, vanillin, 4-chlorophenol, 2,4-dichlorophenol, trichlorophenol and tribromophenol and has high research and application values.

Description

1. Technical field [0001] The present invention relates to a method for extracting microorganisms present in sludge and uses thereof, specifically to the separation and purification of a strain of Pseudomonas aeruginosa strains present in sludge polluted by tribromophenol and its domestication method and use. 2. Background technology [0002] Phenolic compounds, especially halogenated phenolic compounds, are a class of major substances that cause environmental pollution, and they are products or intermediate products in various links of industrial and agricultural production. These compounds exist in soil, air and water. Due to their poor natural degradability, they will remain for a long time after being discharged into the environment, and the cumulative pollution effect on the environment is significant. Due to their relatively high water solubility, these pollutants have a higher potential to contaminate water bodies and contaminate surface runoff and groundwater source...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/385C02F101/36
Inventor 姚日生程莎莎于莹李晓菊王淮孙玉栋
Owner HEFEI UNIV OF TECH
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