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Halogenated benzoic acid degradation strain, inoculant produced by strain and application

A technology of halogenated benzoic acid and degrading bacteria, which is applied in the field of biotechnology, can solve problems such as threats to the environment and human health, and achieve the effects of maintaining human health, low production and use costs, and good repair effects

Inactive Publication Date: 2018-02-09
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, DBHB is the main intermediate metabolite of the herbicide bromoxynil. Due to the extensive use of bromoxynil, DBHB has been widely detected in the environment, which poses a greater threat to the environment and human health.

Method used

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  • Halogenated benzoic acid degradation strain, inoculant produced by strain and application
  • Halogenated benzoic acid degradation strain, inoculant produced by strain and application
  • Halogenated benzoic acid degradation strain, inoculant produced by strain and application

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Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1, isolation and identification of bacterial strain

[0027] The present invention provides a kind of bacterial strain capable of efficiently degrading halogenated benzoic acid and the bacterial agent produced thereof, the bacterial strain used is Gram-negative bacteria H8, which is isolated from the soil of a certain pesticide factory in Suzhou, Jiangsu. The specific isolation and screening methods for strains are as follows:

[0028] Take 5.0 g of the soil sample and add it to 100 ml of inorganic salt (hereinafter referred to as MM) medium containing 0.2 mM 3,5-dibromo-4-hydroxybenzoic acid (hereinafter referred to as MM) culture medium at 30 ° C and 150 rpm for 5 days, with 5% inoculum size (v / v) transfer to the same fresh medium, and enrich and culture continuously for four times. Dilute the fifth-generation enrichment solution on MM solid medium containing 0.2mM 3,5-dibromo-4-hydroxybenzoic acid, culture at 30°C for 4 days, pick a single colony on the ...

Embodiment 2

[0030] Embodiment 2, laboratory degradation experiment

[0031] 2.1 Growth utilization and degradation of 3,5-dibromo-4-hydroxybenzoic acid by strain H8

[0032] Detection of 3,5-dibromo-4-hydroxybenzoic acid by high-performance liquid chromatography: take 1mL sample, centrifuge at 12000rpm for 5min, carefully absorb the supernatant, and filter the supernatant through a 0.22μm aqueous membrane filter for detection by HPLC. Detection conditions: Shimadzu RID-10A is the high-performance liquid chromatograph; the chromatographic column is a C18 reversed-phase column, the specification is 250mm×4.6mm; the column temperature is 40°C; the mobile phase is acetonitrile:water:acetic acid (50:50:0.5, V :V:V), the flow rate is 1.0mL / min; the detection wavelengths are 221nm and 250nm.

[0033] The strain H8 was adjusted to a final concentration of 0.06-0.08 (OD 600The inoculum amount) was connected to 100mL MM containing 0.2mM 3,5-dibromo-4-hydroxybenzoic acid, cultured on a shaker at 3...

Embodiment 3

[0045] Embodiment 3, soil degradation experiment

[0046] The vegetable garden soil was taken as the test soil sample. Pass the soil sample through a 2mm sieve, and take a certain amount of 3,5-dibromo-4-hydroxybenzoic acid, 3-bromo-4-hydroxybenzoic acid, 3,5-dichloro-4-hydroxybenzoic acid and 3- Chloro-4-hydroxybenzoic acid powder was dissolved in 100mL of methanol, and then soaked in diatomaceous earth, so that the pesticide was completely absorbed. Diatomite after soaking is placed in fume hood to dry, and it is mixed in the soil, so that the concentration of pesticide in the soil is about 50mg / kg. Take 500g of each soil sample, insert the seed solution according to 10% inoculum amount, and cultivate it in a constant temperature incubator at 30°C. The soil sample with the same amount of sterile MM liquid is used as a control, and the water holding capacity of the soil is maintained at 60%. Samples were taken at 0, 1, 3 and 5 days respectively, and 3 samples were taken fo...

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Abstract

The invention discloses a halogenated benzoic acid degradation strain, an inoculant produced by the strain and an application. The halogenated benzoic acid degradation strain H8 is preserved in ChinaCenter for Type Culture Collection on April 28, 2017 with preservation number of CCTCC NO.M2017222. The invention provides the halogenated benzoic acid degradation inoculant which is produced by the halogenated benzoic acid degradation strain H8. The bacterium H8 provided by the invention can be used for effectively and rapidly degrading halogenated benzoic acid. The degradation strain H8 has a relatively wide degradation spectrum and is capable of degrading such halogenated aromatic hydrocarbon as 3,5-dibromo-4-hydroxybenzoic acid, 3-bromo-4-hydroxybenzoic acid, 3,5-dichloro-4-hydroxybenzoicacid, 3-chloro-4-hydroxybenzoic acid and the like. The halogenated benzoic acid degradation strain can be used for conditioning halogenated benzoic acid pollution.

Description

technical field [0001] The invention belongs to the field of biological high technology, and relates to a halogenated benzoic acid degrading bacterial strain and the bacterial agent produced therefor and application thereof. technical background [0002] Most of the halogenated aromatic compounds have the advantages of stable chemical properties and superior chemical properties. They can be used as pesticides, flame retardants, dyes, drugs and various intermediates, and are widely used in industrial and agricultural production, resulting in huge economic and social value. , greatly improving human 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, neurological and reproductive toxicity, etc. The pollution of halogenated aromatic hydrocarbons poses a serious threat to the ecosystem and human health...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34B09C1/10C12R1/01C02F101/36
CPCB09C1/10C02F3/34C02F2101/36C12N1/20C12N1/205C12R2001/01
Inventor 陈凯蒋建东李顺鹏简珊珊
Owner NANJING AGRICULTURAL UNIVERSITY
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