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Recombinant vector and recombinant bacterium for improving yield of macrolide antibiotics and application of recombinant vector and recombinant bacterium

A recombinant vector and macrolide technology, applied in the field of genetic engineering, can solve the problems affecting the growth and yield of strains, and the accumulation of antibiotics soaring, so as to reduce feedback inhibition and improve yield.

Pending Publication Date: 2022-07-19
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, a variety of strategies have been used to construct industrial strains with high antibiotic production. However, the increase in the unit fermentation yield makes the accumulation of antibiotics soar in the cells. This phenomenon seriously affects the growth of the strain itself and the further improvement of the yield.

Method used

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  • Recombinant vector and recombinant bacterium for improving yield of macrolide antibiotics and application of recombinant vector and recombinant bacterium
  • Recombinant vector and recombinant bacterium for improving yield of macrolide antibiotics and application of recombinant vector and recombinant bacterium
  • Recombinant vector and recombinant bacterium for improving yield of macrolide antibiotics and application of recombinant vector and recombinant bacterium

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Embodiment 1

[0022] 1. Construction of macrolide efflux module

[0023]Using the genome of Streptomyces glaciformis BC-101-4 as the template (the accession number on GenBank is Genome ID: CP002047.1), all related promoters and efflux genes were amplified, among which the macrolide efflux module was Components include promoters, ribosomal recognition sites, and efflux genes. The combined fragment of described promoter and ribosome recognition site is PiRj, wherein Pi is promoter P1 (sequence as shown in SEQ ID NO:1), P2 (sequence as shown in SEQ ID NO:2), P3 (sequence as shown in SEQ ID NO:2) As shown in SEQ ID NO: 3), P4 (sequence as shown in SEQ ID NO: 4), P5 (sequence as shown in SEQ ID NO: 5), Rj is the ribosome recognition site Rn (Rn is each start (5'-GGAG-3'), Rb (5'-GAAAG-3'), Rc, (5'-GAGGA-3'), Rd (5') -GGGGAG-3'), using the genome of Streptomyces glaciformis BC-101-4 as a template, using the PiRj-F / PiRj-R primer pair to amplify the PiRj fragment (i represents 1-5, j represents n...

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Abstract

The invention discloses a recombinant vector and recombinant bacteria for improving the yield of macrolide antibiotics and application, and belongs to the field of gene engineering. The purpose of the invention is to improve the yield and titer of macrolide antibiotics. A starting vector of the recombinant vector for efflux of macrolide antibiotics is pSET152T, a streptomyces bingchengensis BC-101-4 genome is used as a template to amplify a promoter and an efflux gene, and a sequence obtained by connecting the promoter and the efflux gene is integrated into the starting vector to obtain the recombinant vector. The promoter is located at the upstream of the efflux gene. Recombinant bacteria are constructed through genetic engineering transformation, in fermentation production, the feedback inhibition effect of a final product is relieved, and the yield of milbemycins, abamectin and nemadectin is increased.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a recombinant vector and recombinant bacteria for improving the yield of macrolide antibiotics and application. Background technique [0002] Macrolide antibiotics are a class of biologically active secondary metabolites that can be produced by Streptomyces, many of which have been used in the large-scale production of biopesticides, such as avermectin, milbemycin, nimox Ding et al. At present, a variety of strategies have been used to construct industrial strains with high production of antibiotics. However, the increase of the unit fermentation yield makes the accumulation of antibiotics in the cell soar, which seriously affects the growth of the strain itself and the further improvement of the yield. It is well known that an increase in the accumulation of end-products can have a negative feedback inhibitory effect on the entire synthesis process. In particular...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/76C12N15/31C12N1/21C12P17/18C12R1/465
CPCC12N15/76C07K14/36C12P17/181
Inventor 向文胜李珊珊褚丽阳张艳艳董倬旭
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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