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Screening method of aureomycin degrading strains

A screening method and technology of chlortetracycline, which is applied in the field of microbial degradation of antibiotic residues, can solve the problems of long cycle, no consideration of the screening of low-concentration chlortetracycline degrading strains, cumbersome operation of the screening method, etc., and achieves high accuracy and precision. Effect

Inactive Publication Date: 2012-07-25
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The screening method described in this document is cumbersome to operate and has a long cycle. It needs to be domesticated for more than 60 days before strain screening; meanwhile, this method focuses on the screening of bacterial strains that are resistant to degradation at higher concentrations of aureomycin, and does not consider low concentrations of aureomycin. Screening of aureomycin-degrading strains

Method used

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  • Screening method of aureomycin degrading strains
  • Screening method of aureomycin degrading strains
  • Screening method of aureomycin degrading strains

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1) Bacteria sample collection and pretreatment

[0026] Collect bacteria residue and wastewater samples from aureomycin production enterprises, weigh 10 g of bacteria residue, put it in a 250 mL Erlenmeyer flask, add 100 ml of distilled water, shake it on a shaker, and use the leachate for later use.

[0027] 2) Strain isolation

[0028] Aspirate 400 μl of waste water and leaching solution of bacteria residues, spread them on inorganic salt solid medium plates respectively, and place them under the condition of 28°C-30°C for 2-7 days in shade. The inorganic salt solid medium consists of: (NH 4 ) 2 SO 4 2.0g, K 2 HPO 4 0.5g, KH 2 PO 4 0.5g, MgSO 4 ·7H 2 O 0.5g, NaCl 0.2g, CaCl 2 0.1g, FeSO 4 0.01g, EDTA 0.015g, glucose 2.0g, agar 13.0g. Dissolve the above substances in 1000ml of distilled water, and sterilize with damp heat at 115°C for 15-20min. Before pouring the plate, add aureomycin to the medium, the concentration is 0.1g / L, 1.0g / L, 2.0g / L, 3.0g / L, 4.0g...

Embodiment 2

[0052] Enrichment, Isolation and Purification of Chlortetracycline Degrading Strains

[0053] Compared with Example 1, this method first carries out strain enrichment, and then carries out separation and purification. include:

[0054] 1) Bacteria enrichment

[0055] Collect the fungus residue and waste water, weigh 10g of the waste residue, place it in a 250mL Erlenmeyer flask, add 100ml of distilled water, shake well on a shaker, and extract the leaching solution. Draw 10mL of waste water sample and bacteria residue leachate, put them into 250mL Erlenmeyer flasks with 100ml enrichment medium, respectively, and culture in dark for 2-3 days at 28°C-30°C, shaking at 170r / min. Then take 5ml of the culture solution and transfer it into a new same culture medium, and continue the enrichment culture once under the same culture conditions.

[0056] Inorganic salt liquid enrichment medium consists of: glucose 2g, (NH 4 ) 2 SO 4 2.0g, K 2 HPO 4 0.5g, KH 2 PO 4 0.5g, MgSO 4 ...

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Abstract

The invention relates to a screening method of aureomycin degrading strains, and belongs to the technical field of degrading antibiotic residues by microorganisms. The screening method comprises the following particular steps: isolating the aureomycin degrading strains; purifying the strains; verifying the degrading functions of the strains; and determining the degrading activities of the strains. The screening method adopted by the invention is quick, feasible, economic and low-priced, and is suitable to obtain strains capable of degrading aureomycin residues of different concentrations, especially the efficient strains capable of degrading low-concentration or oligotrophic-grade aureomycin. The obtained strains can be used for preparing an aureomycin degrading agent. According to the invention, the aureomycin-producing microbe culture residue and waste water are utilized as the original materials for screening; and the obtained strains and the prepared agents can be directly applied in treating waste of aureomycin production enterprises, and can be possibly applied in remediation of peripheral soil and water environment polluted by aureomycin. According to the invention, high performance liquid chromatography is used for determining the degrading activities of the strains, and the obtained results have high accuracy and precision.

Description

technical field [0001] The invention relates to a method for screening aureomycin-degrading bacterial strains, and belongs to the technical field of microbial degradation of antibiotic residues. Background technique [0002] Aureomycin (Chlortetracycline; Aureomycin), also known as chlortetracycline, belongs to tetracycline antibiotics. The chemical name is 6-methyl-4-(dimethylamino)-3,6,10,12,12a-pentahydroxy-1,11-dioxo-7-chloro-1,4,4a,5,5a , 6,11,12a-octahydro-2-tetracenecarboxamide, the molecular formula is C 22 h 23 ClN 2 o 8 , the molecular weight is 478.88, and the chemical structural formula is: [0003] [0004] Aureomycin is a broad-spectrum antibiotic that can inhibit Gram-positive bacteria, Gram-negative bacteria, spirochetes, rickettsia, mycoplasma, chlamydia, etc. Wide range of uses. [0005] my country is a big country producing aureomycin, and a large amount of bacteria residue and waste water will be produced in the production process of aureomycin....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00
Inventor 李艳菊蒋耘晨肖思颖董晓霞袁聪颖
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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