Bacteria separation method for efficiently degrading high molecular weight polycyclic aromatic hydrocarbons and application thereof

A polycyclic aromatic hydrocarbon and bacteria technology, applied in the field of microbial technology and environmental biology, can solve problems such as few reports and achieve the effect of high degradation efficiency

Active Publication Date: 2013-04-17
北京市科学技术研究院资源环境研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For high-concentration polluted soil, especially for highly polluted aged soil with tetracyclic and above polycy

Method used

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  • Bacteria separation method for efficiently degrading high molecular weight polycyclic aromatic hydrocarbons and application thereof
  • Bacteria separation method for efficiently degrading high molecular weight polycyclic aromatic hydrocarbons and application thereof
  • Bacteria separation method for efficiently degrading high molecular weight polycyclic aromatic hydrocarbons and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] This embodiment provides a separation method for screening high-efficiency PAH-degrading bacterial complexes, which is specifically as follows:

[0042] 1. Add 50ml of sterilized distilled water and 10g of fresh high-concentration PAHs-contaminated soil into a 100ml Erlenmeyer flask, culture at 25-30°C, 90rpm.

[0043] 2. Add the acetone solution of pyrene and benzopyrene to two new sterilized Erlenmeyer flasks respectively, and make the acetone solution volatilize completely in the ultra-clean workbench, then add bacterial culture medium I, so that the final concentration of pyrene in polycyclic aromatic hydrocarbons is 200mg / L, the final concentration of benzopyrene is 10mg / L. Insert the bacteria cultured in the above 1 with 1% volume content, culture on a shaker in the dark at 25-30°C and 90rpm for 10 days, and repeat the operation from step 2 to step 1 for the degradation rate of pyrene and benzopyrene by the bacterial enrichment culture solution Until the stabili...

Embodiment 2

[0056] Embodiment 2: the degradation performance of efficient degrading bacteria to pyrene and benzopyrene in bacterial culture medium I

[0057] 1. Get the sterilized Erlenmeyer flask and add the acetone solution containing pyrene, and make the acetone solution volatilize completely in the ultra-clean workbench, then add bacterial culture medium I, so that the final concentration of pyrene in polycyclic aromatic hydrocarbons is 200mg / L. Insert the three strains obtained in the above step 9 respectively, under the condition of room temperature 90rpm, culture in a dark shaker for 2, 4, 6, 8, and 10 days, set 3 repeated experiments and bacterial culture medium I+200mg without inoculation / L pyrene was used as a control, and the effect of degrading pyrene of a single bacterial strain was determined by GC-MS ( figure 2 ).

[0058] Result: by figure 2It can be seen that mycobacteria have a strong ability to decompose pyrene, and the degradation rate of the pyrene solution with ...

Embodiment 3

[0066] Example 3: Degradation effect in long-term polluted soil with high concentration and high content of polycyclic aromatic hydrocarbons and the tracking detection of the added bacteria

[0067] 1. Experimental design:

[0068] The experiment was divided into 4 groups, each with 500g soil samples. They are sample 1: add distilled water; sample 2: raw soil without any treatment; sample 3: trace element solution + three kinds of bacteria; sample 4: bacterial culture medium I + 1% glucose + three kinds of bacteria.

[0069] 2. Bacterial culture

[0070] The strains purified from the three bacteria in the ninth step of Example 1 are respectively Acinetobacter sp., Microbacterium sp. and Mycobacterium sp., inoculated with the following medium, and cultured on a shaker for 24-48 hours at room temperature at 90 rpm to obtain 5×10 8 ~5×10 10 CFU / ml concentration of bacteria liquid. Culture medium composition: Add 10g-20g glucose and 3-15g sodium acetate per 20L of the above-m...

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Abstract

The invention relates to a method for biologically restoring soil polluted by high molecular weight polycyclic aromatic hydrocarbons, concretely a method for separating and screening efficient degrading bacteria complexes and applications thereof in a field polluted by polycyclic aromatic hydrocarbons. The method, utilizing PCR-DGG, comprises: rapidly determining relative advantage flora for degrading pollutants in a soil sample, separating and purifying the flora, optionally regrouping the flora, enlargedly culturing the flora, and re-putting the flora in to the long-term pollution soil containing high density and high molecular weight polycyclic aromatic hydrocarbons. By periodically adding a proper amount of nutrient elements with different ratio and controlling the humidity (40-60%), the advantage of the high efficiency degrading flora can be enduringly maintained. The method has high degradation efficiency, especially for the soil polluted by high molecular weight polycyclic aromatic hydrocarbons.

Description

technical field [0001] The invention belongs to the field of microbial technology and environmental biotechnology, and in particular relates to the bioremediation of soil polluted by high-molecular-weight polycyclic aromatic hydrocarbons. Background technique [0002] Polycyclic aromatic hydrocarbons (PAHs) refer to organic compounds in which two or more benzene rings are connected in the form of fused or non-fused rings. PAHs are ubiquitous in the environment, and their extremely low water solubility and stable ring structure are the main reasons for the persistence of these compounds. 70%-90% of human and animal cancer lesions are caused by chemical substances in the environment, and PAHs are the largest category of environmental carcinogenic chemicals. Due to the carcinogenic, teratogenic and mutagenic properties of these compounds, they are potentially dangerous to human health and the ecological environment, and have attracted widespread attention from all over the wor...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/68C12Q1/04B09C1/10C12R1/32C12R1/01
Inventor 金京华程言君高振郭鹏罗霂宋云
Owner 北京市科学技术研究院资源环境研究所
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