Screening and application of polybrominated diphenyl ethers (PBDEs) degrading bacterium

A technology of polybrominated diphenyl ethers and BDE209, which is applied in the direction of bacteria, microorganism-based methods, water/sludge/sewage treatment, etc., to achieve good degradation ability, simple operation process, and low requirements for management conditions

Active Publication Date: 2017-01-04
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are relatively few studies on the degradation of PBDEs, mainly focusing on chemical degradation, photodegradation, microbial degradation and some combined degradation technologies.
Among them,

Method used

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  • Screening and application of polybrominated diphenyl ethers (PBDEs) degrading bacterium
  • Screening and application of polybrominated diphenyl ethers (PBDEs) degrading bacterium
  • Screening and application of polybrominated diphenyl ethers (PBDEs) degrading bacterium

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Embodiment 1, the screening of Stenotrophomonas

[0037] Weigh 5-10g of soil samples contaminated by polybrominated diphenyl ethers (PBDEs) and add them to 50-100mL of high-temperature sterilized 0.85% normal saline, vortex fully, let stand for 10-30 minutes, and take 5mL of soil supernatant Pass through a 0.45μm filter membrane, transfer 1mL of the filtrate to an inorganic salt medium with 10mg / L BDE 209 as the sole carbon source, and culture in a constant temperature shaker at 30°C, 150r / min, and transfer once a week to make BDE 209 The final concentration reached 30mg / L. After 3 weeks of cultivation, the bacterial strains were isolated and purified on the LB solid medium by streaking method, and the dominant colony with faster growth was selected, and the purification was repeated 3 times to obtain a single pure colony. After expanded culture in liquid LB medium, the whole gene of the strain was extracted, PCR amplified, 16rDNA sequencing was performed, and the seque...

Embodiment 2

[0040] Determination of Growth of Stenotrophomonas in Different Concentrations of LB

[0041] After the bacterial strain screened in Example 1 was inoculated into the LB liquid medium to expand the culture, 5 × 10 4 cells / mL were transferred to new LB medium, samples were taken at 0, 4, 12, 23, 30, and 36 hours respectively, and the uninoculated LB medium was used as the blank control, and the bacterial growth was detected by flow cytometry, and the obtained Growth of Stenotrophomonas in different concentrations of LB. image 3 Stenotrophomonas in different concentrations of LB, respectively LB, 10-fold diluted LB (1 / 10LB), 100-fold diluted LB (1 / 100LB) and 1000-fold diluted LB (1 / 1000LB) for 36 hours It can be seen that the strain has good utilization of different concentrations of LB, and within 36 hours, the maximum growth in different concentrations reached 5.0×10 10 ,4.3×10 9 ,3.5×10 8 ,4.3×10 8 cells / mL.

Embodiment 3

[0043] Optimal Conditions for Degradation of BDE209 by Stenotrophomonas

[0044] Orthogonal experiment was used to select 5 factors pH, temperature, salinity, medium volume, BDE209 concentration, and set 5 levels respectively, pH (5, 7, 8, 9, 11), temperature (20°C, 25°C, 30℃, 35℃, 40℃), salinity (0%, 0.5%, 1%, 2%, 4%), medium volume (50mL, 100mL, 150mL, 200mL, 250mL), BDE209 concentration (50μg / L , 100μg / L, 250μg / L, 500μg / L, 750μg / L), optimize the degradation conditions of BDE209 by Stenotrophomonas. The experiment consisted of 25 treatments, each treatment was repeated 3 times, and the degradation time was 7 days. The results showed that the optimal degradation conditions for BDE209 by Stenotrophomonas were pH 5, temperature 25°C, and salinity 0.5%. The volume of the medium was 150 mL, and the concentration of BDE209 was 500 μg / L.

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Abstract

The invention discloses screening and application of a polybrominated diphenyl ethers (PBDEs) degrading bacterium, and belongs to the technical field of biological treatment of environmental pollutants. The PBDEs degrading bacterium comes from soil contaminated by PBDEs and is obtained through artificial enrichment, separation and purification. The strain is a stenotrophomonas pavanii strain WZN-1 and is preserved in the China General Microbiological Culture Collection Center, the preservation date is August 30, 2016, and the preservation serial number is CGMCC No.12918. The WZN-1 strain is a Gram-negative bacterium, is faint yellow and presents a milk white and non-transparent circular bacterial colony with the surface raised; in a scanning electron microscope, the bacterium is in a rod shape, and the diameter is (0.5-0.7) micrometer*(1-2) micrometer. In a liquid culture medium, the strain has the good degrading capability on both decabromodiphenyl ether (BDE 209) and tetrabromodiphenyl ether (BDE 47).

Description

technical field [0001] The invention belongs to the technical field of microorganisms and their application in remediating PBDEs pollution. It relates to a screening method of a PBDEs-degrading bacterium Stenotrophomonas and its degradation method and application to PBDEs (BDE 47 and BDE 209). Background technique [0002] Polybrominated diphenyl ethers (PBDEs) are commonly used brominated flame retardants. Due to their high flame retardant efficiency, good thermal stability, small amount of addition, small impact on material properties and low price, they are widely used in electronic , electrical appliances, chemicals, construction, transportation, textiles, petroleum and mining and other fields. The general chemical formula of PBDEs is C 12 h (0-9) Br (10-1) O, where the sum of hydrogen atoms and bromine atoms is 10, can be divided into 10 series according to the number of bromine atoms, with a total of 209 homologues. At present, the PBDEs products produced and used...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02C02F3/34A62D3/02C12R1/01C02F101/36A62D101/28
CPCA62D3/02A62D2101/28C02F3/34C02F2101/36C12N1/02C12N1/205C12R2001/01
Inventor 王莹莹吴志能谢苗苗
Owner NANKAI UNIV
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