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Preparation method of bacillus licheniformis with enhanced YvbW expression and obtained strain and application thereof

A technology of Bacillus licheniformis and Bacillus licheniformis, which is applied in biochemical equipment and methods, botany equipment and methods, and microbial-based methods, and can solve the problem of unclear transfer-in or transfer-out, and failure to analyze the regulation mechanism of YvbW, etc. question

Active Publication Date: 2018-09-21
LIFECOME BIOCHEM
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Problems solved by technology

However, the current research has not resolved the specific regulatory mechanism of YvbW, and it is not clear about which amino acids YvbW will specifically affect the transport of amino acids and whether BcaP is responsible for the transfer of target amino acids. In addition, the amino acids in cells Has a strict and complex regulatory mechanism

Method used

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  • Preparation method of bacillus licheniformis with enhanced YvbW expression and obtained strain and application thereof
  • Preparation method of bacillus licheniformis with enhanced YvbW expression and obtained strain and application thereof
  • Preparation method of bacillus licheniformis with enhanced YvbW expression and obtained strain and application thereof

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Embodiment Construction

[0025] A kind of preparation method of bacillus licheniformis that strengthens YvbW expression, comprises the following steps:

[0026] (1) Using the genomic DNA of Bacillus licheniformis DW2 as a template, PCR amplified wxya gene fragments, and then wxya The gene fragment is connected upstream to the P43 promoter and downstream to the amylase terminator to form a complete wxya expression element;

[0027] (2) At the same time, using the genomic DNA of Bacillus licheniformis DW2 as a template, PCR amplified wxya The upstream homology arm of the gene and wxya The downstream homology arm of the gene;

[0028] (3) by overlap extension PCR wxya The upstream homology arm of the gene, wxya expression elements and wxya The downstream homology arms of the genes are connected together to form the target gene fragment, and the arrangement order of the target gene fragment is: wxya The upstream homology arm of the gene - wxya expression element- wxya The downstream h...

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Abstract

The invention provides a preparation method of bacillus licheniformis with enhanced YvbW expression and an obtained strain and application thereof. By adopting a genetic engineering method, the bacillus licheniformis DW2-yvbW is obtained by inserting an amino acid permease YvbW gene into a bacillus licheniformis genome, and the yield of bacitracin in a fermentation liquor in the bacitracin fermentation process is improved by 13% or above by adopting the strain.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering bacteria transformation, in particular to a preparation method of bacillus licheniformis which strengthens the expression of amino acid permease YvbW and the application of the obtained strain. Background technique [0002] Bacillus licheniformis is recognized as an important industrial microbial strain with biological safety (GRAS), which widely exists in nature. In view of the characteristics of clear genetic background and high industrial application value of Bacillus licheniformis, it has been widely used for research in recent years. Bacillus licheniformis is a Gram-positive bacterium, which is mainly used to ferment and produce biochemical products such as poly-γ-glutamic acid, bacitracin, acetoin, 2,3-butanediol, and lichenin. [0003] Bacitracin is a class of cyclic peptide antibiotics composed of 12 amino acid residues. The constituent amino acids of bacitracin include ornith...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/10C12N15/53C12N15/75C12N15/66C12P21/00C12R1/10
CPCC12N9/0022C12N15/66C12N15/75C12P21/00C12Y104/03002
Inventor 陈守文朱江蔡冬波朱杉刘紫薇李俊辉段茂华楼丽君陈晓斌
Owner LIFECOME BIOCHEM
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