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Halogenated phenol-degrading strain and microbial agent prepared from halogenated phenol-degrading strain

A technology of halogenated phenol and degrading bacteria, which is applied in the field of biotechnology, and can solve the problems of non-point source pollution, strong toxicity, and high stability of compounds such as soil and water bodies

Active Publication Date: 2020-07-07
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the extremely strong electronegativity of halogen elements, it is easy to combine with the enzyme system in living cells, resulting in high stability, strong toxicity, and easy migration in the environment, resulting in serious non-point source pollution such as soil and water bodies.

Method used

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  • Halogenated phenol-degrading strain and microbial agent prepared from halogenated phenol-degrading strain
  • Halogenated phenol-degrading strain and microbial agent prepared from halogenated phenol-degrading strain
  • Halogenated phenol-degrading strain and microbial agent prepared from halogenated phenol-degrading strain

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, isolation and identification of bacterial strain

[0033] The invention provides a bacterial strain capable of efficiently degrading halogenated phenols and the bacterial agent produced therefrom. The bacterial strain used is a Gram-negative bacterial strain DCP-2, which is isolated from the soil of a pesticide factory in Nanjing, Jiangsu. The specific isolation and screening methods for strains are as follows:

[0034] Take 10.0g contaminated soil sample and add to 100mL containing 50mg L -1 2,4-DCP inorganic salt (hereinafter referred to as MM) culture medium, 30 ℃, 150rpm shaker culture 7d, with 15% inoculum size (v / v) transferred to fresh culture medium treated the same, continuous enrichment Set cultured four times. Ultraviolet spectrophotometer was used to detect the degradation of the fifth generation enrichment solution. The effective fifth-generation enrichment solution containing 50mg L -1 2,4-DCP was diluted and coated on MM solid medium, ...

Embodiment 2

[0036] Embodiment 2, laboratory degradation experiment

[0037] 2.1 Growth utilization and degradation of 2,4-DCP by strain DCP-2

[0038] Detection of DCP-2 by high-performance liquid chromatography: take 3mL sample, centrifuge at 12,000rpm for 5min, carefully absorb the supernatant, add 25% hydrochloric acid, and adjust the pH to about 3.0. Then, extract with an equal volume of dichloromethane, remove water with anhydrous sodium sulfate, take 2mL of dichloromethane and blow dry, then redissolve in 0.5mL of methanol, filter through an organic phase filter with a pore size of 0.22 μm, and detect by HPLC. Detection conditions: Shimadzu RID-10A is the high-performance liquid chromatograph; the chromatographic column is a C18 reversed-phase column with a size of 250mm×4.6 mm; the column temperature is 40°C; the mobile phase is methanol:water:acetic acid (80:20:0.5, V :V:V), the flow rate is 1.0mL min L -1 ; The detection wavelength is 220nm and 235nm.

[0039] The bacterial st...

Embodiment 3

[0051] Embodiment 3 bacterial agent preparation

[0052] The original seed of the halogenated phenol-degrading strain DCP-2 of the present invention is activated on a petri dish, and inoculated on the inclined surface of a test tube for later use. The test tube was inoculated in 200mL LB medium (LB medium formula: peptone 10g L -1 , yeast powder 5g L -1 , NaCl 5g L -1 , pH 7.4) in a 1,000mL shake flask, cultured with constant temperature shaking until the logarithmic phase, ready to inoculate the primary seed tank. The first-level seed tank is 50L, the feeding volume is 40L, and the medium formula is: glucose 8g L -1 , yeast extract 5g L -1 , K 2 HPO 4 1g L -1 , NaCl 5g L -1 , CaCO 3 2g L -1 , MgSO 4 0.2gL -1 , soybean oil 0.1% (v / v), pH value 7.2-7.5; After feeding, high-pressure steam sterilization, after cooling to 30 ° C, the above-mentioned cultured shake flask strains were inoculated into 50L- Grade seed tanks, cultivated to the logarithmic growth phase, ...

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Abstract

The invention discloses a halogenated phenol-degrading strain and a microbial agent prepared from the halogenated phenol-degrading strain. The strain is a Gram staining reaction negative strain DCP-2and is Bosea sp. through identification. The strain is preserved in the China Center for Type Culture Collection on December 23, 2019 and has a strain preservation number of CCTCC M20191083. The halogenated phenol-degrading strain DCP-2 can be applied to degradation of 2,4-dichlorophenol (2,4-DCP), o-chlorophenol, p-chlorophenol or m-chlorophenol, and is preferentially applied to degradation of 2,4-DCP in soil. A degradation microbial agent can be used for reducing the residual amount of 2,4-DCP in soil to be 95 percent or more by direct application, and the pollution problem of 2,4-DCP in soil can be solved.

Description

technical field [0001] The invention belongs to the field of biological high technology, and relates to a halogenated phenol degrading bacterial strain and the bacterial agent produced therefrom. [0002] technical background [0003] Most of the halogenated aromatic compounds have the advantages of stable chemical properties and superior chemical properties. They can be used as pesticides, drugs and various intermediates. They are widely used in industrial and agricultural production, resulting in huge economic and social benefits and greatly improving human health. Life. However, halogenated aromatic hydrocarbons also have the characteristics of high toxicity and high stability, can exist stably in the environment for a long time, and have certain irritation, carcinogenicity, teratogenicity, and neurological and reproductive toxicity. Halogenated aromatics pollution poses a serious threat to the ecological environment and human health, so the treatment of halogenated aroma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C12R1/01
CPCB09C1/10C12N1/20C12N1/205C12R2001/01
Inventor 蒋建东张龙乔文静
Owner NANJING AGRICULTURAL UNIVERSITY
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