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Method for screening polychlorinated biphenyl degrading bacterium and polychlorinated biphenyl degrading bacterium

A technology for polychlorinated biphenyls and screening methods, applied in microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problem of microbial toxicity, low bioavailability of polychlorinated biphenyls, and difficulty in obtaining polychlorinated biphenyls. Benzene degrading bacteria and other problems, to achieve the effect of promoting degradation, simple method, and promoting complete degradation

Active Publication Date: 2014-05-14
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Due to the low bioavailability of PCBs and their toxic effects on microorganisms, it is difficult to obtain PCB-degrading bacteria by conventional microbial enrichment and separation methods.
However, there is still a lack of reliable methods for screening PCB-degrading bacteria from the environment.

Method used

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  • Method for screening polychlorinated biphenyl degrading bacterium and polychlorinated biphenyl degrading bacterium
  • Method for screening polychlorinated biphenyl degrading bacterium and polychlorinated biphenyl degrading bacterium
  • Method for screening polychlorinated biphenyl degrading bacterium and polychlorinated biphenyl degrading bacterium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Screening of PCB-degrading bacteria

[0019] The present invention screens out PCB-degrading bacteria from the environment through enrichment and domestication, separation and purification, and PCB degradation experiments. The specific method is as follows:

[0020] (1) Enrichment and domestication. The soil samples used to isolate microorganisms were taken from a PCB-contaminated area in Luqiao District, Taizhou City, Zhejiang Province. Inorganic salt medium including KH 2 PO 4 1.00g / L, K 2 HPO 4 ·3H 2 O3.00g / L, MgSO 4 0.15g / L, FeSO 4 0.01g / L, CaCl 2 0.005g / L, NaCl1.00g / L, (NH 4 ) 2 SO 4 0.50g / L, trace element solution 0.1% (v / v), pH7.2; trace element solution formula is Na 2 Mo 4 ·H 2 O6.70g / L, ZnSO 4 ·5H 2 O28.00g / L, CuSO 4 ·5H 2 O2.00g / L, H 3 BO 4 4.00g / L, MnSO 4 ·5H 2 O4.00g / L, CoSO 4 ·7H 2 O4.70g / L, pH7.2. Fill 100 mL of the above culture medium into a 300 mL Erlenmeyer flask, and sterilize at 121 °C for 20 min before use. Befo...

Embodiment 2

[0027] Embodiment 2: Degradation of bacterial strain HC3 to benzoic acid and chlorobenzoic acid

[0028] Degradation of benzoic acid and chlorobenzoic acid. The degradation system is the same as in Example 1 (4), and the degradation substrates are benzoic acid, 2-chlorobenzoic acid, 3-chlorobenzoic acid and 4-chlorobenzoic acid, and the concentration of each substrate is 20-30 mg / L. After incubating at 180r / min and 30°C in the dark for 3 days, add 3mol / L HCl to each tube to adjust the pH to 2.5, then add 15mL ethyl acetate and seal the tube opening. Vortex for 5 minutes, sonicate for 15 minutes, centrifuge at 2500r / min for 5 minutes, take the upper organic phase and dry it with nitrogen, redissolve the sample in methanol, and finally transfer it to a sample vial for HPLC analysis. The high-performance liquid chromatograph is Agilent1100, and the DAD detector is selected, and the chromatographic column is Zorbax Eclipse XDB-C18 (4.6mm×250mm, 5μm). The ratio of mobile phase me...

Embodiment 3

[0032] Example 3: Degradation of polychlorinated biphenyls in soil by bacterial strain HC3

[0033] The soil (with a PCB content of 18.60 mg / kg) taken from a PCB-contaminated site in Taizhou City, Zhejiang Province was air-dried naturally, ground finely, passed through a 20-mesh sieve, mixed evenly, and divided into 200 g in each beaker. Then inoculate the bacterial strain HC3 (the preparation method of the bacterial solution is the same as the method described in Example 1) so that the bacterial concentration in the soil reaches 1.0×10 8 cfu / g (the same amount of heat sterilization was added to the control group), and an appropriate amount of water was added to make the soil moisture content reach 40%. The test group was placed in an artificial climate room, protected from light, and the soil temperature was maintained at 22-23°C during the test. On the fifth and tenth day of the test, soil samples were taken from each test group by the five-point method for the determinatio...

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PUM

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Abstract

The invention discloses a method for screening a polychlorinated biphenyl degrading bacterium and the polychlorinated biphenyl degrading bacterium. The method is applied to separation of polychlorinated biphenyl aerobic degradation bacteria from polychlorinated biphenyl contaminated soil. The polychlorinated biphenyl degrading bacterium disclosed by the invention is identified to be Sphingobium fuliginis HC3. The bacterium is capable of effectively degrading chlorodiphenyl, dichlorodiphenyl and trichlorodiphenyl, and can also degrade benzoic acid, 3-chlorobenzoic acid and 4-chlorobenzoic acid. After the strain HC3 is inoculated to the polychlorinated biphenyl contaminated soil, the degradation of chlorodiphenyl...tetrachlorophenyl can be remarkably prompted, and the total degradation rate of polychlorinated biphenyl can be 44.91% within 10 days; and the degradation rate is one time higher than that of a control group. The strain HC3 can degrade low-chloro multi-chlorophenyl and low-chloro benzoic acid under the aerobic condition, so that the strain can be applied to bioremediation engineering of polychlorinated biphenyl contaminated soil so as to prompt thorough degradation of multi-chlorophenyl and improve the remediation efficiency.

Description

technical field [0001] The present invention relates to a method for screening polychlorinated biphenyl degrading bacteria and a strain of polychlorinated biphenyl degrading bacteria Sphingobium fuliginis HC3, in particular to enriching, isolating and purifying polychlorinated biphenyl degrading bacterial strains from polychlorinated biphenyl contaminated soil The method and the bacterial strain Sphingobium fuliginis HC3 which can degrade low-chlorinated polychlorinated biphenyls and low-chlorinated benzoic acid under aerobic conditions obtained by using the method belong to the field of environmental microbial engineering. Background technique [0002] Polychlorinated biphenyls (polychlorinated biphenyls, PCBs) is a large class of compounds formed by replacing the hydrogen atoms in biphenyls with chlorine atoms to varying degrees, with a total of 209 species [Cao Xianzhong, Chen Huaguo, Shen Songmei, etc. Properties of PCBs and its harm to the environment. China Science and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/02C12N1/20C12R1/01
Inventor 胡金星沈超峰崔静岚
Owner ZHEJIANG UNIV
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