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Method for effectively increasing yield of tetraene macrolide antibiotics

A technology of macrolides and antibiotics, applied in the field of microorganisms, to achieve the effect of increasing production

Inactive Publication Date: 2016-03-16
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Amylase Streptomyces chromogenes ( Streptomyces diastatochromogenes ) D can synthesize three kinds of tetraenyl macrolide antibiotics: tetramycin P, tetramycin B and tetramycin A, and the amylase chromogenous Streptomyces chromogenes D can be improved to increase the four There is no report on the research on the synthesis ability of ene macrolide antibiotics

Method used

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  • Method for effectively increasing yield of tetraene macrolide antibiotics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Obtaining rifampicin-resistant mutant strains (the first round of screening of drug-resistant mutant strains)

[0024] A GYM solid plate with a final rifampicin concentration of 20 μg / mL was prepared, that is, the rifampicin concentration in the GYM solid plate was 20 μg / mL. Use a pipette to draw 100μL of amylase-producing Streptomyces chromogenes D spore suspension (1×10 6 Pieces / mL) on the GYM solid plate, spread evenly with a sterile coating rod, and place it in a 28°C incubator for 7 days. Pick a single colony to a new GYM solid plate containing 20μg / mL rifampicin (one A single colony is divided into a plate), placed in an incubator at 28°C for 5 days, and the strain that can grow again on the new GYM solid plate is the rifampicin-resistant mutant. A total of 2 rifampicin-resistant mutant strains were obtained, numbered f31 and f32, respectively. The two mutant strains were cultured in liquid fermentation according to step (4) in the technical scheme. The fe...

Embodiment 2

[0025] Example 2: Obtaining of low-concentration streptomycin resistant mutant strains (the second round of screening of resistant mutant strains)

[0026] A GYM solid plate with a final concentration of streptomycin of 30 μg / mL was prepared, that is, the concentration of streptomycin in the GYM solid plate was 30 μg / mL. Use a pipette to pipette 100μL of rifampicin-resistant mutant f31 spore suspension (1×10 6 Pieces / mL) on the GYM solid plate, spread evenly with a sterile coating rod, place it in an incubator at 28°C for 7 days, pick out single colonies to a new GYM solid plate containing 30μg / mL streptomycin (one A single colony is drawn on a plate), placed in an incubator at 28°C for 5 days, and the strain that can grow again on the new GYM solid plate is a low-concentration streptomycin resistant mutant. A total of 2 low-concentration streptomycin resistant mutant strains were obtained, numbered G2-13 and G2-18 respectively. The two mutant strains were cultured in liquid ferm...

Embodiment 3

[0027] Example 3: Obtaining high-concentration streptomycin-resistant mutant strains (the third round of screening of drug-resistant mutant strains)

[0028] Prepare a GYM solid plate with a final concentration of 300 μg / mL of streptomycin, that is, the concentration of streptomycin in the GYM solid plate is 300 μg / mL. Use a pipette to pipette 1000μL of low-concentration streptomycin resistant mutant strain G2-13 spore suspension (1×10 12 Pcs / mL) on the GYM solid plate, spread evenly with a sterile coating rod, and place it in a 28℃ incubator for 10-15 days. Pick a single colony to a new GYM solid plate containing 300μg / mL streptomycin (One single colony draws one plate), place it in a 28°C incubator for 7-10 days, and the strain that can grow again on the new GYM solid plate is a high-concentration streptomycin resistant mutant. A total of 2 high-concentration streptomycin resistant mutant strains were obtained, numbered G3-13 and G3-15 respectively;

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Abstract

The invention discloses a method for effectively increasing the yield of tetraene macrolide antibiotics, and belongs to the field of microbial technologies. According to the method, by screening joint drug-resistant mutant strains, mutant strains of streptomyces diastatochromogenes D with drug resistance to rifampicin, low-concentration streptomycin and high-concentration streptomycin are obtained, and compared with streptomyces diastatochromogenes D, the capability of the obtained mutant strains for synthesizing tetraene macrolide antibiotics is significantly improved.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a method for effectively increasing the production of tetraenyl macrolide antibiotics. Background technique [0002] At present, many self-developed new agricultural antibiotics are only in the laboratory stage, and the key reason is that the production level of the producing bacteria has not met the requirements of industrialization. Therefore, a major problem facing researchers is to improve the production levels of existing and developing agricultural antibiotic-producing bacteria, and to speed up the industrialization process of agricultural antibiotics. Conventional microbial mutation breeding is time-consuming, labor-intensive, and highly random. Genetic engineering can achieve directed evolution and overexpression of enzymes encoded by single genes, but the production of natural products such as antibiotics that require clustered gene encoding can be increased. It ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/62C12R1/525
CPCC12P19/62
Inventor 申屠旭萍俞晓平许益鹏汤谷刘楠楠
Owner CHINA JILIANG UNIV
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