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Escherichia coli (escherichia coli) LHM10-1 strain and its application

A technology of Escherichia coli and gdmccno, which is applied in the field of biological treatment of environmental pollutants, can solve the problems such as the degradability that remains to be studied and the environmental pollution of tetracycline degradation efficiency, and achieves a good degradation effect.

Active Publication Date: 2021-01-26
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Biodegradation includes phytodegradation and microbial degradation, but phytodegradation mostly focuses on using a certain plant to repair a specific polluted environment, and the degradation ability of compound pollution caused by multiple antibiotics remains to be studied
There are relatively few reports on microbial degradation of tetracyclines, but the effect is remarkable. For example, Pseudomonas hibiscus DT1 can degrade 83.21% of tetracyclines within 72 hours (patent publication number: CN104403965A) and Paleobacterium bacterium KSS10 within 96 hours Degrade 63.33% of oxytetracycline (Sicheng Shao et al., 2019, Ecotoxicology and Environmental Safety), the current tetracycline degradation efficiency is still not enough to solve the increasingly serious environmental pollution problem

Method used

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  • Escherichia coli (escherichia coli) LHM10-1 strain and its application
  • Escherichia coli (escherichia coli) LHM10-1 strain and its application
  • Escherichia coli (escherichia coli) LHM10-1 strain and its application

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Experimental program
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Effect test

Embodiment 1

[0022] A strain of Escherichia coli LHM10-1. The above-mentioned Escherichia coli LHM10-1 strain was deposited in the Guangdong Microbial Culture Collection Center (GDMCC) on March 28, 2019, and the preservation number is GDMCC No: 60622. The preservation address is: 5th Floor, Building 59, Compound, No. 100 Xianlie Middle Road, Guangzhou City, Guangdong Province.

[0023] Isolation and purification of the above-mentioned E. coli LHM10-1

[0024] In July 2017, a sample of pig manure was collected from a pig farm in Zhangshu City, Jiangxi Province, and 0.1g of fresh sample was weighed and added to 1ml of 0.9% normal saline, and vortexed to mix. Pipette 20 μl of the suspension into 4 ml of intestinal bacteria enrichment broth (Guangdong Huankai Microbiology Technology Co., Ltd.), and culture at 37° C. and 180 rpm for 12-16 hours. Subsequently, the above-mentioned culture solution was divided and divided into MacConkey agar medium containing 64 μg / ml tetracycline (Guangdong Hua...

Embodiment 2

[0031] Determination of minimum inhibitory concentration (MIC)

[0032] Using the micro broth dilution method, referring to the relevant standards of the American Clinical and Laboratory Standards Association (CLSI: M100-S26), the Escherichia coli LHM10-1 was tested for tetracycline, chlortetracycline, oxytetracycline, doxycycline, minol The MICs of cyclocycline, tigecycline and iravacycline (all purchased from Sigma-Aldrich, USA). Escherichia coli ATCC 25922 was used as the quality control strain.

[0033] The results showed (Table 1) that Escherichia coli LHM10-1 exhibited different levels of drug resistance to tetracycline, chlortetracycline, oxytetracycline, doxycycline, minocycline, tigecycline, and iravacycline. The sensitivity was 256 μg / ml, 128 μg / ml, 256 μg / ml, 16 μg / ml, 32 μg / ml, 8 μg / ml and 4 μg / ml, respectively.

[0034] Table 1 MICs of Escherichia coli LHM10-1 to tetracyclines

[0035]

[0036] Note: TC, tetracycline; CTC, chlortetracycline; OTC, oxytetracyc...

Embodiment 3

[0038]Pick a single colony of purified Escherichia coli LHM10-1, inoculate it into 4ml of LB broth (Guangdong Huankai Microbiology Technology Co., Ltd.), and cultivate it at 37°C and 180rpm for 4-6 hours. Subsequently, draw 200 μl of the above-mentioned E. coli culture solution to 200ml containing tetracycline (100mg / L), aureomycin (100mg / L), oxytetracycline (100mg / L), doxycycline (50mg / L), minol Incubate in LB broth of cyclocycline (10 mg / L), tigecycline (10 mg / L) and eravacycline (2.5 mg / L) for 48 hours at 37° C., 220 rpm and in the dark. And the culture medium without adding tetracycline was used as the control group. Finally, a standard digital camera was used to take images, and the degradation ability of Escherichia coli LHM10-1 to tetracyclines was preliminarily judged according to the color change.

[0039] the result shows( figure 1 ), for tetracycline, chlortetracycline, oxytetracycline and doxycycline, Escherichia coli LHM10-1 all changed the medium from yellow to...

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Abstract

The invention belongs to the technical field of biological treatment of environmental pollutants, in particular to Escherichia coli ( Escherichia coli ) LHM10‑1 strain and its application in degrading tetracycline family antibiotics, Escherichia coli ( Escherichia coli ) LHM10‑1 strain, which was deposited in Guangdong Microbial Culture Collection Center (GDMCC) on March 28, 2019, with the preservation number GDMCC No: 60622. The Escherichia coli LHM10‑1 strain of the present invention has good degradation effect on tetracycline family antibiotics. Further, the Escherichia coli LHM10‑1 provided by the present invention can degrade 83.91% of tetracycline and 92.24% of ilavacycline after 16 hours, providing a safe, efficient and environmentally friendly solution to the residual problem of tetracycline antibiotics .

Description

technical field [0001] The invention belongs to the technical field of biological treatment of environmental pollutants, and in particular relates to Escherichia coli (Escherichiacoli) LHM10-1 strain and its application in degrading tetracycline antibiotics. [0002] technical background [0003] Tetracycline antibiotics are a class of broad-spectrum antibiotics, including tetracycline, oxytetracycline, chlortetracycline, doxycycline, minocycline, tigecycline and ilavacycline, etc., which are widely used in humans and animals Treatment of bacterial infectious diseases and growth promotion in animals. At present, tetracycline antibiotics are one of the most widely used antibiotics in my country, and the usage in 2013 has reached 12,000 tons (Qian-Qian Zhang et al., 2015, Environmental Science & Technology). However, due to the low absorption rate of these drugs, a large number of tetracycline antibiotics exist in the environment in the form of parent or active metabolites. S...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12Q1/18A62D3/02C12R1/19
CPCA62D3/02C12Q1/18C12N1/205C12R2001/19
Inventor 孙坚陈冲崔超月吴小亭张燕廖晓萍刘雅红
Owner SOUTH CHINA AGRI UNIV
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