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Polycyclic aromatic hydrocarbon degrading bacterium suitable for electric field condition and application thereof

A polycyclic aromatic hydrocarbon, degrading bacteria technology, applied in the direction of microorganism-based methods, bacteria, and the restoration of polluted soil, etc., can solve the problems of affecting the activity of degrading microorganisms, reducing the degradation effect, affecting the bioremediation process, etc., and achieve good social benefits. , the effect of enhancing the degradation activity and improving the degradation ability

Active Publication Date: 2013-04-10
SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a large number of studies have shown that the electric field will affect the activity of degrading microorganisms, reduce the degradation effect, and affect the bioremediation process.

Method used

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  • Polycyclic aromatic hydrocarbon degrading bacterium suitable for electric field condition and application thereof
  • Polycyclic aromatic hydrocarbon degrading bacterium suitable for electric field condition and application thereof
  • Polycyclic aromatic hydrocarbon degrading bacterium suitable for electric field condition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Isolation and Screening of Pseudomonas fluorescens PB4

[0021] CCTCC No: Isolation and Screening of M 2011260

[0022] The sample comes from the PAH-contaminated soil around Benxi Iron and Steel Company. Weigh 5.0g of soil, add 50ml of sterile water and shake for 30min, then absorb the supernatant and enrich and culture it in the liquid inorganic salt medium containing 100mg / L PAH for 3d Finally, streak and isolate on a solid inorganic salt medium containing 100 mg / L polycyclic aromatic hydrocarbons, select a well-growing single colony, streak, isolate and purify three times, and then store the bacterial strain on a slant medium.

[0023] The composition of described liquid inorganic salt culture medium: MgSO 4 .7H 2 O 0.2g; CaCl 2 .2H 2 O 0.01g; FeSO 4 .7H 2 O 0.005g; K 2 HPO 4 0.4g; MnSO 4 .H 2 O 0.02g; NH 4 NO 3 1.0g; Na 2 HPO 4 0.6g; distilled water 1000ml; pH 7.2-7.4, sterilized at 121°C for 30min.

[0024] The solid inorganic salt medium: MgSO ...

Embodiment 2

[0027] Example 2 PCR amplification and sequence determination of the 16S rDNA gene of Pseudomonas fluorescens (Pseudomonas fluorescens) PB4

[0028] The total DNA of the above strain Pseudomonas fluorescens PB4 was extracted, and the 16S rDNA of the strain was amplified. The primers for PCR amplification were universal primers for bacteria, 27f: 5'-AGAGTTTGATCCTGGCTCAG-3', 1492R: 5'-GGTTACCTTGTTACGACT-3'. The PCR reaction conditions were: 94°C for 3min; 94°C for 1min, 55°C for 1min, 72°C for 3min, 35 cycles; 72°C for 10min. After the PCR amplification product is purified, it is sequenced, and the result and the sequence on Genbank are subjected to Blast analysis. The sequence length is 1424bp, and the phylogenetic tree of its 16S rDNA gene sequence (see figure 1 ), it can be seen that the above-mentioned isolated bacterial strain has great homology with Pseudomonas fluorescens.

Embodiment 3

[0029] Example 3 Determination of biological activity of Pseudomonas fluorescens PB4.

[0030] The above Pseudomonas fluorescens (Pseudomonas fluorescens) PB4 was inoculated in beef extract peptone liquid medium, and then shaken and cultured at 28°C and 150r / min constant temperature shaker for 3 days; Phosphate buffer was diluted, and the resulting diluent (phosphate buffer composition: KH 2 PO 4 34.0g; 175ml of 1mol / L NaOH solution; 825ml of distilled water; pH7.2; preparation of diluent: take 1.25ml of phosphate buffer, dilute to 1000ml with distilled water, and autoclave at 121°C for 30min) to prepare the bacteria into a suspension. The prepared bacterial suspension of Pseudomonas fluorescens (Pseudomonas fluorescens) PB4 was added to sterilized clean sandy loam, and the concentration of Pseudomonas fluorescens in the soil was 1.7×10 6 CFU / g dry soil. At the same time, unidirectional and switching electric fields were applied to the test soil, the voltages were 0.5V / cm,...

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Abstract

The invention belongs to the technical field of environmental organic pollutant biological treatment and particularly relates to a polycyclic aromatic hydrocarbon degrading bacterium suitable for an electric field condition and application of the polycyclic aromatic hydrocarbon degrading bacterium. The polycyclic aromatic hydrocarbon degrading bacterium is pseudomonas fluorescens PB4, and has been already collected in China Center for Type Culture Collection (CCTCC) on July 19th, 2011, and the collection register number of the polycyclic aromatic hydrocarbon degrading bacterium is CCTCC No: M2011260. The pseudomonas fluorescens PB4 is used for repairing contaminated soil. A bacterial strain obtained by separating the polycyclic aromatic hydrocarbon degrading bacterium is higher in activity under the action of the electric field condition, and thus, the ability of degradation of single polycyclic aromatic hydrocarbon and mixed polycyclic aromatic hydrocarbon can be improved. After an electric field of 0.5-2.0V / cm is applied, the degradation activity of the pseudomonas fluorescens is enhanced, and the removal rate of the single polycyclic aromatic hydrocarbon and the removal rate of the mixed polycyclic aromatic hydrocarbon are respectively increased by 12.46-30.62 percent and 8.11-20.20 percent after 28 days.

Description

technical field [0001] The invention belongs to the technical field of biological treatment of environmental organic pollutants, and in particular relates to a polycyclic aromatic hydrocarbon degrading bacterium suitable for electric field conditions and an application thereof. Background technique [0002] Polycyclic aromatic hydrocarbons (PAHs) refer to a class of hydrocarbons with two or more benzene rings, which are carcinogenic, teratogenic, and mutagenic, and are ubiquitous in the environment. These substances will bring potential long-term harm to the ecological environment and human health. The US Environmental Protection Agency (US EPA) has listed 16 of them as "priority control pollutants" in the 1980s. Therefore, the treatment and remediation of PAHs pollution in the environment has become a research hotspot. [0003] Soil is a complex multiphase system and a dynamic open system. Because the soil is rich in organic matter, and PAHs are lipophilic organic pollutan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/00B09C1/10C12R1/39
Inventor 李凤梅郭书海李刚张玲妍李婷婷吴波
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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