Method for increasing yield of erythromycin through saccharopolyspora erythraea SACE_1906 gene pathway

A technology of Saccharopolyspora erythromycetes and erythromycin, which is applied in the field of genetic engineering and can solve the problem of the absence of regulatory genes in synthetic gene clusters

Active Publication Date: 2020-05-26
ANHUI UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The biosynthetic genes of erythromycin are clustered in the genome of Saccharopolyspora erythromycetes, but there are no regulatory genes in the synthetic gene cluster

Method used

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  • Method for increasing yield of erythromycin through saccharopolyspora erythraea SACE_1906 gene pathway
  • Method for increasing yield of erythromycin through saccharopolyspora erythraea SACE_1906 gene pathway
  • Method for increasing yield of erythromycin through saccharopolyspora erythraea SACE_1906 gene pathway

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Embodiment

[0031] 1 Materials and methods:

[0032] 1.1 Strains, plasmids and growth conditions

[0033] The bacterial strains and plasmids used in the examples are shown in Table 1. Escherichia coli was cultured in liquid LB medium at 37°C or LB solid plate with no resistance or corresponding resistance added with 1.25% agar. The erythromycin-producing bacterium Erythropolyspora saccharopolyspora and its engineered strains were cultured on tryptone soybean broth (TSB) medium at 30°C or R3M solid plates containing 2.2% agar without resistance or corresponding resistance.

[0034] 1.2 Materials, DNA manipulation and sequencing

[0035] PEG3350, lysozyme, TES, casamino acids, thiostrepton, and apramycin were purchased from Sigma. TSB, yeast extract, and peptone were purchased from Oxoid. Glycine, agar powder, sodium chloride, and other biological reagents were purchased from reagent companies. The general operating techniques for Escherichia coli and Saccharopolyspora red mold follow ...

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Abstract

The invention discloses a method for increasing the yield of erythrocin through a saccharopolyspora erythraea SACE_1906 gene pathway. The SACE_1906 gene is inactivated in the saccharopolyspora erythraea through a genetic engineering approach, the target gene SACE_1905 of the saccharopolyspora erythraea is overexpressed to obtain the saccharopolyspora erythraea erythromycin high-yield engineering strain, the erythromycin is produced by fermenting the strain, and the yield of the erythromycin can be increased by fermenting the strain obtained by the technology.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for increasing the yield of erythromycin through the gene pathway of Saccharopolyspora erythromycetes SACE_1906. Background technique [0002] There are a variety of family transcriptional regulators in prokaryotic genomes, such as MarR, TetR, AsnC, Lrp, GntR and other family transcriptional regulators. Among them, TetR family transcriptional regulators (TetR family transcriptional regulators, TFRs) are common single-component prokaryotic signal transduction systems, which exist in the genomes of various bacteria, among which soil-dwelling bacteria encode the most. TetR, a transcriptional regulator of this family, was first discovered in Escherichia coli and is the founding member of this family. Regulators with structures and functions similar to it discovered later are classified into the TetR family. TFRs can regulate many cellular physiological processes...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12P19/62C12R1/01
CPCC07K14/195C12P19/62
Inventor 张部昌林浩吴杭
Owner ANHUI UNIVERSITY
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