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Method for screening phenanthrene polycyclic aromatic hydrocarbon degrading bacteria

A polycyclic aromatic hydrocarbon and degrading bacteria technology, applied in the field of microbial degradation, can solve the problems of cumbersome operation process, poor solubility, pollution, etc., achieve high chemical stability, low biodegradability, and reduce toxicity

Pending Publication Date: 2021-09-17
CHONGQING UNIV OF POSTS & TELECOMM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to solve the above-mentioned technical problems, the present invention provides a method for screening phenanthrene-type PAH-degrading bacteria to solve the problem of conventional use of naphthalene, anthracene, phenanthrene and other substances in the current experimental process of screening PAH-degrading bacteria. The solubility in water is extremely poor, and it is volatile, and other organic co-solvents cannot be uniformly added to the culture medium. Some use spraying and sublimation methods, which not only pollute the surrounding organic solvents, but also have toxic effects on bacteria. , the operation process is cumbersome and difficult to control

Method used

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  • Method for screening phenanthrene polycyclic aromatic hydrocarbon degrading bacteria
  • Method for screening phenanthrene polycyclic aromatic hydrocarbon degrading bacteria
  • Method for screening phenanthrene polycyclic aromatic hydrocarbon degrading bacteria

Examples

Experimental program
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Effect test

Embodiment

[0033] 1. To screen phenanthrene-like PAH-degrading bacteria, the steps are:

[0034] (1) Take 1 g of the soil near the gas station and put it in 100 mL of LB liquid medium containing 300 mg / L crystal violet, and cultivate it with shaking at 30 °C and 170 r / min for 24 hours;

[0035] (2) Take 100 μL of the liquid in step (1) and dilute it according to the concentration gradient, then take 50 μL and spread it on an LB plate containing 300 mg / L crystal violet, and incubate it upside down at 30°C for 24 hours;

[0036] (3) Observe the LB plate of step (2), pick 2 single colonies with colorless circles, put each single colony into a liquid LB test tube, and culture on a shaker at 30°C and 170r / min for 16 hours;

[0037] (4) Detect decolorization rate, step is:

[0038] A1. Configure blank control sample 15mL, the concentration of PBS in this sample is 0.01M, the volume fraction of LB liquid medium is 10%, namely the LB liquid medium added is 1.5mL;

[0039] B1. Configure 15 mL o...

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Abstract

The invention provides a method for screening phenanthrene polycyclic aromatic hydrocarbon degrading bacteria. The method for the screening phenanthrene polycyclic aromatic hydrocarbon degrading bacteria mainly comprises the following steps of culturing petroleum-polluted soil in an LB liquid culture medium containing crystal violet, diluting according to a concentration gradient, coating on an LB plate containing the crystal violet, inversely culturing, selecting a colorless circle single colony, inoculating into a liquid LB test tube, culturing, and calculating the decolorization rate, and continuously screening strains with the crystal violet decolorization rate of more than 80% to enable the strains with the phenanthrene degradation rate of more than 45% to be the phenanthrene polycyclic aromatic hydrocarbon degrading bacteria. According to the method for the screening phenanthrene polycyclic aromatic hydrocarbon degrading bacteria, the crystal violet is utilized, the dye is a common triphenylmethane dye; when an observed sample can be converted into light purple from purple and finally colorless, the bacteria can degrade the crystal violet and has the effect of degrading polycyclic aromatic hydrocarbon, and the bacteria capable of efficiently degrading the crystal violet and obtained through the screening method can degrade the polycyclic aromatic hydrocarbon at the same time.

Description

technical field [0001] The invention belongs to the technical field of microbial degradation, and more specifically relates to a method for screening phenanthrene-like polycyclic aromatic hydrocarbon-degrading bacteria. Background technique [0002] Polycyclic aromatic hydrocarbons refer to aromatic hydrocarbons containing two or more benzene rings, also called PAHs. Due to their toxicity, genotoxicity, mutagenicity and carcinogenicity, they can cause various hazards to the human body, such as respiratory system, circulatory system, Nervous system damage, damage to liver and kidneys. Identified as the main organic pollutants affecting human health. Due to the good chemical stability of PAHs, it is difficult to decompose through chemical actions such as hydrolysis under natural conditions. Except for a small part of photochemical decomposition, the vast majority of PAHs mainly disappear from the environment through biodegradation. of. Therefore, the use of microbial degrad...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12R1/01
CPCC12N1/20C12N1/02
Inventor 谢永芳黄小容梁亦龙向浏欣谭权钦甘玉立李洋
Owner CHONGQING UNIV OF POSTS & TELECOMM
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