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Application of Bacillus licheniformis dw2δlrpc in the production of bacitracin

A technology of Bacillus licheniformis and bacitracin, applied in the field of strain transformation of Bacillus licheniformis, can solve the problems that the regulatory mechanism of LrpC has not been analyzed and cannot be deduced

Active Publication Date: 2020-12-29
LIFECOME BIOCHEM
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current research has not resolved the specific regulatory mechanism of LrpC, therefore, it is impossible to deduce the relationship between LrpC and leucine synthesis and bacitracin production

Method used

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  • Application of Bacillus licheniformis dw2δlrpc in the production of bacitracin
  • Application of Bacillus licheniformis dw2δlrpc in the production of bacitracin
  • Application of Bacillus licheniformis dw2δlrpc in the production of bacitracin

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Experimental program
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Embodiment Construction

[0025] The method for constructing bacillus licheniformis by knocking out the lrpC gene comprises the following steps:

[0026] (1) Using the genomic DNA of Bacillus licheniformis DW2 as a template, the upstream homology arm of the lrpC gene and the downstream homology arm of the lrpC gene were amplified by PCR; The downstream homology arms of the gene are spliced ​​together to obtain the fusion gene sequence A;

[0027] (2) Carry out double enzyme digestion to the fusion gene sequence A obtained in step (1) using restriction endonucleases Xba I and Sac I to obtain the enzyme-cut gene sequence A;

[0028] (3) Prepare the plasmid T2(2)-ori, and use restriction endonucleases Xba I and Sac I to perform double digestion on the plasmid T2(2)-ori to obtain the restriction plasmid T2(2)-ori;

[0029] (4) connecting the enzyme-cut gene sequence A obtained in step (2) to the enzyme-cut plasmid T2(2)-ori obtained in step (3), to obtain the lrpC gene knockout plasmid T2(2)-ori-lrpC;

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Abstract

The invention provides a method for constructing bacillus licheniformis by knocking out an lrpC gene, a strain and an application of the strain. The bacillus licheniformis DW2 delta lrpC is obtained by knocking out the lrpC gene in a bacillus licheniformis genome with a genetic engineering method, and the yield of bacitracin of the strain in a fermentation liquid in the bacitracin fermentation process is increased by 10% or above.

Description

technical field [0001] The invention relates to the field of Bacillus licheniformis strain transformation, in particular to a method for constructing Bacillus licheniformis by knocking out the lrpC gene, a strain and an application thereof. Background technique [0002] Bacillus licheniformis belongs to Gram-positive bacteria, which is recognized as an important industrial microbial strain with biological safety (GRAS). Because of its clear genetic background and high industrial application value, Bacillus licheniformis has been widely used for research in recent years. [0003] At present, Bacillus licheniformis is mainly used to ferment and produce biochemical products such as poly-γ-glutamic acid, bacitracin, acetoin, 2,3-butanediol, and lichenin. Bacitracin, also known as subtilisin, can inhibit or kill certain pathogenic bacteria, strongly inhibit the growth of Gram-negative bacteria, and has a synergistic enhancement effect with other antibiotics (such as penicillin, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/75C12N15/90C12N15/66C12N15/31C12N1/21C12P21/04C12R1/10
CPCC07K7/58C07K14/32C12N15/66C12N15/75C12N15/902C12P21/02
Inventor 陈守文蔡冬波朱江李阳朱杉李俊辉楼丽君陈晓斌
Owner LIFECOME BIOCHEM
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