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Application of Bacillus licheniformis dw2δphop in the production of bacitracin

A technology of Bacillus licheniformis and bacitracin, which is applied in the field of Bacillus licheniformis strains with phoP gene knockout and its construction, to achieve the effect of increasing the yield of bacitracin and increasing the yield of bacitracin

Active Publication Date: 2021-06-25
LIFECOME BIOCHEM
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] There are many genes that are closely related to the synthesis of metabolites in Bacillus licheniformis, but it is still unknown which gene is related to the production of bacitracin, and the engineering bacteria with high production of bacitracin need to be further studied through genetic modification. Research

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  • Application of Bacillus licheniformis dw2δphop in the production of bacitracin
  • Application of Bacillus licheniformis dw2δphop in the production of bacitracin
  • Application of Bacillus licheniformis dw2δphop in the production of bacitracin

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Embodiment approach

[0029] The construction method of Bacillus licheniformis (Bacillus licheniformis) knocking out phoP comprises the following steps:

[0030] (1) Using the genomic DNA of Bacillus licheniformis DW2 as a template, PCR amplifies the upstream homology arm and the downstream homology arm of the phoP gene;

[0031] (2) connecting the upstream homology arm of the phoP gene and the downstream homology arm of the phoP gene by overlap extension PCR to form the target gene fragment;

[0032] (3) Double digestion of the target gene fragment with XbaI and SacI restriction endonucleases to obtain a knockout fragment;

[0033] (4) Prepare the plasmid T2(2)-ori, and use XbaI and SacI restriction endonucleases to double-enzyme digest the plasmid T2(2)-ori to obtain a linear plasmid fragment;

[0034] (5) Ligate the enzyme-cut knockout fragment obtained in step (3) and the linearized plasmid fragment obtained in step (4) through DNA ligase to obtain the knockout plasmid T2(2)-ΔphoP;

[0035] (...

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Abstract

The invention provides an application of Bacillus licheniformis DW2ΔphoP in the production of bacitracin. The invention successfully knocks out the gene of carbon metabolism transcription factor PhoP—phoP in the genome of Bacillus licheniformis DW2 through genetic engineering, in order to increase the production of bacitracin A new strategy is provided. Compared with the Bacillus licheniformis DW2, the bacitracin yield of the Bacillus licheniformis DW2ΔphoP constructed by the present invention is increased by more than 15%.

Description

technical field [0001] The invention relates to the field of bacillus licheniformis strain transformation, in particular to a bacillus licheniformis strain with phoP gene knockout and its construction method and application. Background technique [0002] Bacillus licheniformis is recognized as an important industrial microbial strain with biological safety (GRAS), which widely exists in nature. Due to its clear genetic background and high industrial application value, Bacillus licheniformis has been extensively studied in recent years. At present, Bacillus licheniformis is mainly used for the fermentation and production of poly-γ-glutamic acid, bacitracin, acetoin, 2,3-butanediol, lichenin and other biochemical products. [0003] Bacitracin, also known as subtilisin, is an unstable polypeptide composed of 12 amino acids. It can inhibit or kill certain pathogenic bacteria, strongly inhibit Gram-negative bacteria, and has a synergistic effect with other antibiotics such as p...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/75C07K7/58C12P21/04C12R1/10
CPCC07K7/58C07K14/32C12P21/02
Inventor 陈守文蔡冬波朱杉朱江李俊辉陈晓斌楼丽君
Owner LIFECOME BIOCHEM