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Method for increasing yield of erythromycin by SACE_5754 gene pathway of Saccharopolyspora erythraea

A technology of Saccharopolyspora erythromycin and erythromycin, which is applied in the field of genetic engineering and can solve problems such as uneconomical, time-consuming erythromycin A, and unsuitability for wide application

Active Publication Date: 2019-01-04
ANHUI UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional method of optimizing fermentation conditions to increase the yield of erythromycin A is time-consuming and uneconomical, and is not suitable for wide application

Method used

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  • Method for increasing yield of erythromycin by SACE_5754 gene pathway of Saccharopolyspora erythraea
  • Method for increasing yield of erythromycin by SACE_5754 gene pathway of Saccharopolyspora erythraea
  • Method for increasing yield of erythromycin by SACE_5754 gene pathway of Saccharopolyspora erythraea

Examples

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Embodiment

[0038] The bacterial strains and plasmids used in the examples are shown in Table 1. Escherichia coli were cultured in liquid LB medium at 37°C or on solid LB plates supplemented with 1.25% agar. The erythromycin-producing bacterium Erythropolyspora saccharopolyspora and its engineered strains were cultured in tryptone soybean broth (TSB) medium at 30°C or on R3M plates containing 2.2% agar.

[0039]

[0040]

[0041]

[0042] Bacteria and plasmids used in the present embodiment of table 1

[0043] In the examples, 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 standard operations. The synthesis of primers and DNA sequencing were completed by Sangon Bioengineering ...

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Abstract

The invention discloses a method for increasing yield of erythromycin by a SACE_5754 gene pathway of Saccharopolyspora erythraea, and belongs to the technical field of genetic engineering. According to the method, SACE_5754 is deleted from the Saccharopolyspora erythraea by the genetic engineering pathway, target genes SACE_0388 and SACE_6149 of the Saccharopolyspora erythraea are overexpressed, ahigh-yield erythromycin engineering strain is obtained, and when the obtained strain is applied to fermented production of erythromycin, yield can be substantially increased, and therefore, a new technical support is provided for erythromycin yield increase in industrial production.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a method for increasing the yield of erythromycin through the gene pathway of Saccharopolyspora erythromycetes SACE_5754. Background technique [0002] Erythromycin is a class of broad-spectrum macrolide antibiotics produced by the secondary metabolism of Saccharopolyspora erythromycetes. It has good activity against Gram-positive bacteria and low gastrointestinal side effects. Since it was first isolated from Saccharopolyspora erythraea in 1952, the utilization rate of erythromycin has been at the forefront of antibiotics in the world. Among the components of the secondary metabolite erythromycin of Saccharopolyspora erythromycetes, erythromycin A is widely used, and its antibacterial activity is the highest, while azithromycin, clarithromycin, and erythromycin Sodium and telithromycin are also the mainstream medical antibiotics today. It can be seen that improving the outpu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/80C12P19/62C12R1/645
CPCC12N15/80C12P19/62
Inventor 张部昌储祖灵吴杭汪焰胜
Owner ANHUI UNIVERSITY
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