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Bacterial strain for increasing yield of tetramycin Z and method for preparing tetramycin Z by using same

A technology of tetramycin and strain, applied in the field of microorganisms, can solve the problems of high randomness, time-consuming and laborious, difficulty in increasing the yield of natural products such as antibiotics, etc., and achieve the effects of increasing yield and strong inhibitory effect.

Active Publication Date: 2020-04-24
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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. is difficult

Method used

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  • Bacterial strain for increasing yield of tetramycin Z and method for preparing tetramycin Z by using same
  • Bacterial strain for increasing yield of tetramycin Z and method for preparing tetramycin Z by using same
  • Bacterial strain for increasing yield of tetramycin Z and method for preparing tetramycin Z by using same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Acquisition of low-concentration streptomycin-resistant mutants (first round of screening of drug-resistant mutants)

[0028] A GYM solid plate with a final concentration of 30 μg / mL of low-concentration streptomycin was prepared, that is, the concentration of streptomycin in the GYM solid plate was 30 μg / mL. Pipette 100 μL of Streptomyces chromogenes D spore suspension (1×10 6 colonies / mL) on the GYM solid plate, spread evenly with a sterile spreader bar, placed in a 28°C incubator for 7 days, and pick a single colony to a new GYM solid plate (one colony) containing 20 μg / mL streptomycin. A single colony was drawn on a plate), placed in a 28°C incubator for 5 days, and the strain that could grow again on the new GYM solid plate was the streptomycin-resistant mutant. A total of 15 low-concentration streptomycin-resistant mutant strains were obtained, numbered fz1 to fz15, and the 15 mutant strains were subjected to liquid fermentation culture according to st...

Embodiment 2

[0029] Example 2: Obtainment of high-concentration streptomycin-resistant mutants (second round of screening of drug-resistant mutants)

[0030] A GYM solid plate with a final streptomycin concentration of 300 μg / mL was prepared, that is, the streptomycin concentration in the GYM solid plate was 300 μg / mL. Pipette 1000 μL of low-concentration streptomycin-resistant mutant fz9 spore suspension (1×10 12 colonies / mL) on the GYM solid plate, spread evenly with a sterile coating rod, and culture in a 28°C incubator for 10 to 15 days. Pick a single colony to a new GYM solid plate containing 300 μg / mL streptomycin. (One single colony is drawn on one plate), and cultured in a 28°C incubator for 7 to 10 days. The strains that can grow again on the new GYM solid plate are high-concentration streptomycin-resistant mutants. A total of 3 high-concentration streptomycin-resistant mutant strains were obtained, numbered sz2-1, sz2-2 and sz2-3 respectively, and the three mutant strains were s...

Embodiment 3

[0031] Example 3: Acquisition of rifampicin-resistant mutants (the third round of screening of drug-resistant mutants)

[0032] A GYM solid plate with a final concentration of rifampicin of 20 μg / mL was prepared, that is, the concentration of rifampicin in the GYM solid plate was 20 μg / mL. Pipette 100 μL of the mutant sz2-3 spore suspension (1×10) obtained from the second round of screening with a pipette. 10 colonies / mL) on a GYM solid plate containing 20 μg / mL rifampicin, spread evenly with a sterile coating rod, and culture in a 28°C incubator for 7 days. The GYM solid plate of Fuping (a single colony is drawn on a plate), placed in a 28°C incubator 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 mutants were obtained in this screening, numbered tz3-1 and tz3-2.

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Abstract

The invention discloses a bacterial strain for increasing the yield of tetramycin Z and a method for preparing tetramycin Z by using the bacterial strain. The strain is classified and named as Streptomyces diastatochromogenes tz3-1 and is preserved on December 3, 2019, with an accession number being CGMCC No. 19069. The invention further discloses a preparation method of the streptomyces diastatochromogenes tz3-1. According to the streptomyces diastatochromogenes tz3-1 disclosed by the invention, the content of tetramycin Z in fermentation liquor obtained by fermenting the Streptomyces diastatochromogenes tz3-1 is 20.6 times higher compared with control Streptomyces diastatochromogenes D, so a foundation is laid for industrial production of tetramycin Z in the future.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a strain for improving the yield of tetramycin Z and a method for preparing tetramycin Z. 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 production bacteria does not meet the requirements of industrialization. Therefore, a major problem facing researchers at present is to improve the toxin-producing level of existing and developing agricultural antibiotic-producing bacteria, and to speed up the industrialization process of agricultural antibiotics. Conventional microbial mutagenesis breeding is time-consuming, labor-intensive, and highly random. Through genetic engineering, the directed evolution and overexpression of enzymes encoded by a single gene can be achieved, but the yield of natural products such as antibiotics that need to be encode...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/18C12R1/525
CPCC12N1/20C12P17/181C12N1/205C12R2001/525
Inventor 申屠旭萍赵若颖宋阳俞晓平刘光富许益鹏
Owner CHINA JILIANG UNIV
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