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Triclocarban degrading bacterial strain and application thereof

A technology of triclocarban and degrading bacteria, applied in bacteria, biological water/sewage treatment, microorganisms, etc., can solve the problem of affecting the reproductive ability and endocrine system of animals, inducing the generation and transfer of microbial resistance genes, affecting the activity of aquatic organisms, etc. question

Pending Publication Date: 2017-11-03
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to its stable chemical properties and strong hydrophobicity, triclocarban discharged with domestic sewage cannot be completely removed through the traditional sewage treatment process, but is mainly adsorbed on the remaining sludge and subsequently discharged into the environment
The farmland reuse or landfill of excess sludge has caused triclocarban to be further transferred to soil and sediment, and has been frequently detected in water bodies and sediment environments in various places, which has brought various harms to human health and the environment. harm
Triclocarban will not only affect the activity of aquatic organisms, but also affect the reproductive ability and endocrine system of animals. Triclocarban and its various transformation products are potential carcinogens. At the same time, as an antibacterial agent, triclocarban also has Potential risk of inducing microbial resistance gene production and transfer
Based on the characteristics of wide pollution range and low relative concentration of triclocarban, other treatment methods are difficult and costly. Therefore, the use of microbial degradation to remove triclocarban is an economical and effective way to reduce the environmental pollution caused by it. It is of great significance to reduce the ecological risk caused by it
At present, there are few reports on the screening and degradation pathway of triclocarban-degrading bacteria. Only two strains, Sphingomonas sp.YL-JM2C and Ochrobactrumsp.MC22, have been reported to have the ability to degrade triclocarban, but their degradation efficiency is relatively low. And the concentration of triclocarban that can be tolerated is low
However, there is no report about the degradation gene of triclocarban

Method used

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  • Triclocarban degrading bacterial strain and application thereof
  • Triclocarban degrading bacterial strain and application thereof
  • Triclocarban degrading bacterial strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 3

[0034] Example 1 Isolation and identification of triclocarban-degrading bacteria TCC-1 and its aerobic degradation of triclocarban

[0035] 1.1 Strain isolation and purification

[0036]The source of the bacteria was collected from the remaining sludge of a sewage treatment plant in Beijing. 20mg / L TCC was added to a 150mL Erlenmeyer flask as a mother liquor. First, the acetone was volatilized to remove, and then 50mL of sterilized basal salt medium (containing 0.05 % yeast extract), respectively insert about 5g of mud-water mixture samples, and place them in a constant temperature shaker at 30°C for cultivation, with a rotation speed of 150rpm. After culturing for 7-8 days, the culture medium was prepared in a similar manner, and 10% of the volume of the enriched solution was transferred to fresh culture medium. After 5 consecutive transfers, the degradation effect of the enrichment solution was measured. The strains were isolated and purified by dilution plate method and s...

Embodiment 2 3

[0046] Example 2 Triclocarban-degrading bacteria TCC-1 anaerobically degrades triclocarban, 4,4'-dichlorophenylurea and diphenylurea

[0047] Preparation of anaerobic medium: Supplement 1.5mM potassium nitrate and 15mM sodium acetate into the basal salt medium. After fully dissolving, inject high-purity nitrogen gas to remove oxygen, seal the culture bottle with anaerobic plug, and sterilize at high temperature and high pressure.

[0048] In the anaerobic medium prepared above, 5 mg / L of triclocarban, 4,4'-dichlorophenylurea and diphenylurea were respectively added in the form of mother liquor, and the bacterial strain TCC- 1 Inoculate into anaerobic basal salt medium, the initial OD600 value is about 0.035, each group has 3 repetitions, and observe the change of biomass and the degradation of the above substrates.

[0049] Continuously monitor the conversion of triclocarban, 4,4'-dichlorophenylurea and diphenylurea Figure 5 and 6 As shown, the results show that the tricloc...

Embodiment 3 3

[0050] Example 3 Cloning of the degrading gene tccA of the triclocarban-degrading bacteria TCC-1

[0051] 3.1 Extraction of bacterial genome total DNA

[0052] After the triclocarban-degrading bacterium TCC-1 was cultured in large quantities, the CTAB method was used to extract high-purity, large-segment genomic DNA. The extraction method was referred to the "Refined Molecular Biology Experiment Guide" edited by F. Osper et al.

[0053] 3.2 Digestion of total DNA

[0054] The total DNA of Ochrobactrum sp.TCC-1 was partially digested with Sau3AI.

[0055] 3.3 Recovery of DNA fragments

[0056] The digested DNA fragments were separated and purified by electrophoresis (TAE buffer), and recovered using a DNA recovery kit (Tiangen, Beijing). The recovered DNA was dissolved in TE buffer (pH 8.0) and placed at -20°C preservation.

[0057] 3.4 Enzyme linkage

[0058] Establish the following reaction system: pUC118(BamHI) (TaKaRa, Code: D3321) 0.5μL

[0059] Recover 5 μL of DNA f...

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Abstract

The invention discloses a triclocarban degrading bacterial strain and the application thereof. The triclocarban degrading bacterial strain is Ochrobactrum sp., and has the collection number of CGMCC No.14087. The invention further provides the application of the triclocarban degrading bacterial strain to the field of microbial degradation. The triclocarban degrading bacterial strain has the effective degradation temperature range of 20-40 DEG C and the effective pH range of 5.2-9.0, has the degradation ability under both aerobic and anaerobic conditions, can be well adapted to engineering application under a natural condition, and can degrade multiple types of amido bond-containing substrates.

Description

technical field [0001] The present invention relates to the application field of microorganisms and biodegradation thereof, specifically, the present invention relates to a triclocarban degrading bacterium and its application. Background technique [0002] The chlorinated aromatic hydrocarbon triclocarban (chemical name N-(4-chlorophenyl)-N-(3,4-dichlorophenyl)urea) is an antibacterial agent widely added to people’s daily necessities. It has good antibacterial effect on Gram-positive bacteria, negative bacteria and fungi. Due to its stable chemical properties and strong hydrophobicity, triclocarban discharged with domestic sewage cannot be completely removed through the traditional sewage treatment process, but is mainly adsorbed on the remaining sludge and subsequently discharged into the environment. The farmland reuse or landfill of excess sludge has caused triclocarban to be further transferred to soil and sediment, and has been frequently detected in water bodies and s...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/36C02F101/38
CPCC02F3/34C02F2101/36C02F2101/38C12N1/205C12R2001/01
Inventor 王爱杰梁斌云慧孔德勇李智灵
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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