Genetic engineering high-yield strain streptomyces diastatochromogenes and production method and application of epsilon-polylysine

A technology for producing Streptomyces chromogenes and polylysine, applied in the biological field, can solve problems such as less reports, and achieve the effects of increasing concentration, increasing yield, and improving fermentation level

Active Publication Date: 2022-01-07
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] As mentioned above, although there are reports on the use of genetic engineering technology for the transformation of bacterial strains, there are few reports on the construction of engineered bacteria through gene knockout to improve the production of ε-polylysine

Method used

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  • Genetic engineering high-yield strain streptomyces diastatochromogenes and production method and application of epsilon-polylysine
  • Genetic engineering high-yield strain streptomyces diastatochromogenes and production method and application of epsilon-polylysine
  • Genetic engineering high-yield strain streptomyces diastatochromogenes and production method and application of epsilon-polylysine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] A genetic engineering high-yield amylase strain Streptomyces chromogenes ΔfabF1 (Streptomyces diastatochromogenes ΔfabF1), the construction steps are as follows:

[0085] (1) Obtaining the knockout component: the knockout component contains the allelic site of the fabF1 gene, that is, the upstream homologous fragment and the downstream homologous fragment of the gene, as well as the resistance fragment (apramycin resistance) as a selection marker.

[0086] Using the genome of S.diastatochromogenes 6#-7 as a template, the upstream and downstream homologous fragment primer sequences fabF1-L-F / fabF1-L-R and fabF1-R-F / fabF1-R-R were designed according to the fabF1 gene; The primer sequence fabF1-apr-F / fabF1-apr-R was designed for the mycin resistance gene (Apr).

[0087] Add 8 nucleotides at the upstream and downstream ends of the knockout module to form the restriction endonuclease EcoRI cutting site.

[0088] The sequence of the primers is:

[0089] fabF1-L-F: SEQ No.2...

Embodiment 2

[0105] A genetic engineering high-yield amylase strain Streptomyces chromogenes ΔfabF2 (Streptomyces diastatochromogenes ΔfabF2), the construction steps are as follows:

[0106] (1) Obtaining the knockout component: the knockout component contains the allelic site of the fabF2 gene, that is, the upstream homologous fragment and the downstream homologous fragment of the gene, and the resistance fragment (apramycin resistance) as a selection marker.

[0107] Using the genome of S.diastatochromogenes 6#-7 as a template, the upstream and downstream homologous fragment primer sequences fabF2-L-F / fabF2-L-R and fabF2-R-F / fabF2-R-R were designed according to the fabF2 gene; The primer sequence fabF2-apr-F / fabF2-apr-R was designed for the mycin resistance gene (Apr).

[0108] Add 8 nucleotides at the upstream and downstream ends of the knockout module to form the restriction endonuclease EcoRI cutting site.

[0109] The sequence of the primers is:

[0110] fabF2-L-F: SEQ No.13, name...

Embodiment 3

[0126] A genetic engineering high-yield amylase strain Streptomyces chromogenes ΔfabF3 (Streptomyces diastatochromogenes ΔfabF3), the construction steps are as follows:

[0127] (1) Obtaining the knockout component: the knockout component contains the allelic site of the fabF3 gene, that is, the upstream homologous fragment and the downstream homologous fragment of the gene, and the resistance fragment (apramycin resistance) as a selection marker.

[0128] Using the genome of S.diastatochromogenes6#-7 as a template, the upstream and downstream homologous fragment primer sequences fabF3-L-F / fabF3-L-R and fabF3-R-F / fabF3-R-R were respectively designed according to the fabF3 gene; using the pSET152 plasmid as a template, according to Apramycin Primer sequence fabF3-apr-F / fabF3-apr-R was designed for the protein resistance gene (Apr).

[0129] Add 8 nucleotides at the upstream and downstream ends of the knockout module to form the restriction endonuclease EcoRI cutting site.

[...

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Abstract

The invention relates to a streptomyces diastatochromogenes strain producing epsilon-polylysine in high yield in genetic engineering. The streptomyces diastatochromogenes strain is obtained by respectively knocking a key fatty acid ACP synthase gene FabF in a fatty acid anabolism pathway out of Streptomyces diastatochromogenes 6#-7. A genetic engineering recombinant strain is obtained by knocking out the key gene in the fatty acid anabolism pathway. Experiments prove that the genetic engineering strain of streptomyces has higher epsilon-polylysine production capacity than an original strain streptomyces diastatochromogenes TUST under the same condition, According to the invention, an excellent strain is provides for epsilon-polylysine production.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a genetic engineering strain and a construction method thereof, in particular to a production method and application of a genetic engineering high-yield amylase chromogenic strain Streptomyces chromogenes and ε-polylysine. Background technique [0002] ε-polylysine is a homotype monomer polymer containing 25-30 lysine residues, which is a polypeptide with antibacterial effect. It was first described by Japanese scholars Shima and Sakai in the early 1980s. This biological preservative is used in food preservation. Compared with traditional chemical preservatives and biological preservatives, ε-polylysine has good water solubility, high thermal stability and wide antibacterial spectrum, and has inhibitory effect on most bacteria, some fungi and some viruses. ε-polylysine belongs to biopolymer material, which has the advantages of being edible, degradable and non-toxic to humans and the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/21C12N15/76C12P13/02C12R1/525
CPCC12N9/93C12N15/76C12P13/02C12Y602/0102
Inventor 谭之磊董天宇贾士儒侯颖唐昆鹏周东浩闫佳佳许倍铭
Owner TIANJIN UNIV OF SCI & TECH
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