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Triclocarban metabolic plasmid pDCA-TCC and application thereof

A triclocarban and plasmid technology, applied in the field of applied environmental microorganisms, can solve problems such as carcinogenicity, genotoxicity, and endocrine disorders

Pending Publication Date: 2022-07-29
NANJING AGRICULTURAL UNIVERSITY
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  • Claims
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Problems solved by technology

In addition, triclocarban can accumulate in mammalian and human tissues through the food chain, leading to endocrine disorders, potential carcinogenic and genotoxic effects

Method used

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  • Triclocarban metabolic plasmid pDCA-TCC and application thereof
  • Triclocarban metabolic plasmid pDCA-TCC and application thereof
  • Triclocarban metabolic plasmid pDCA-TCC and application thereof

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Embodiment 1 3

[0023] Example 1. Construction of Triclocarban Metabolism Plasmid

[0024] 1.1 Plasmid pDCA extraction

[0025] The original host of plasmid pDCA, Achromobacter sp.ANB-1 (Zhang Long. Isolation of herbicide Ligulong and its metabolite-degrading strains, degrading enzyme genes and bacterial group synergistic metabolism research [D]. Nanjing: Nanjing Agricultural University, 2017) After a large amount of culture in LB medium, the DNA of the plasmid pDCA with higher purity was extracted by alkaline lysis method and dissolved in Tris-EDTA (pH 8.0) buffer. Compilation of Molecular Biology Experiment Guide. The quality and concentration of DNA samples were detected by agarose gel electrophoresis and NanoDrop 2000 (Thermo Fisher Scientific, Shanghai, China), respectively.

[0026] 1.2 Construction of recombinant strain Pseudomonas putida KT2440 (pDCA)

[0027] Since the original host of plasmid pDCA, Achromobacter sp. ANB-1, contains multiple antibiotic resistance, it is difficult ...

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Abstract

The invention discloses a triclocarban metabolic plasmid pDCA-TCC and an application of the triclocarban metabolic plasmid pDCA-TCC. The invention provides a plasmid pDCA-TCC for coding a triclocarban degradation pathway and a preparation method of the plasmid pDCA-TCC. The strain Pseudomonas putida KT2440 carrying a metabolic plasmid pDCA-TCC has a wide chloroaniline degradation substrate spectrum, not only can efficiently degrade an antibacterial agent triclocarban, but also can degrade toxic and harmful compounds such as a herbicide aniline and chlorpropham, a plant regulator forchlorfenuron, an insecticide diflubenzuron and the like. In urban domestic wastewater, the degrading bacterium KT2440 (pDCA-TCC) can be used for completely degrading 50 [mu] M of triclocarban within 3 days. The invention has good application potential and value in biologically enhanced remediation of triclocarban residue pollution, and has important significance in the aspects of protecting ecological environment safety, human health and the like.

Description

technical field [0001] The invention belongs to the field of applied environmental microorganisms, and relates to a triclocarban metabolizing plasmid pDCA-TCC and its application. technical background [0002] Trichlorocarbanilide (3,4,4'-trichlorocarbanilide) is a synthetic broad-spectrum halogenated aromatic antibacterial agent, which mainly inhibits Gram-positive bacteria and has partial antibacterial activity against Gram-negative bacteria and fungi. Over the past 60 years, triclocarban has been widely used in pharmaceutical and personal care products such as cosmetics, body washes, detergents, deodorants, and more. Large amounts of triclocarban are consumed worldwide every year and discharged to sewage treatment plants. Although the U.S. Food and Drug Administration (FDA) banned the use of triclocarban in daily necessities such as hand sanitizers and soaps in 2013, triclocarban still maintains huge market demand in China and India. Due to its poor water solubility and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/78C12N15/55C12N1/21C02F3/34C12R1/40C02F101/36C02F101/38
CPCC12N15/78C12N9/80C12Y305/01C02F3/34C02F3/342C02F3/348C02F2101/38C02F2101/36
Inventor 陈凯乔文静朱闻起董浩然张乐宁罗子睿柯壮蒋建东
Owner NANJING AGRICULTURAL UNIVERSITY
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