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Bacillus licheniformis strain capable of realizing high production of bacitracin as well as preparation method and application thereof

A technology of Bacillus licheniformis and Bacillus licheniformis, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problem of unpredictable isoleucine production

Active Publication Date: 2018-05-11
LIFECOME BIOCHEM
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Problems solved by technology

Moreover, the synthesis of isoleucine is regulated by many factors. It is unpredictable whether simply strengthening one of the key enzymes in the synthesis pathway of isoleucine can increase the yield of isoleucine
In addition, the synthesis of bacitracin is also regulated and affected by many factors, and it is unpredictable whether simply increasing the content of isoleucine can increase the production of bacitracin

Method used

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  • Bacillus licheniformis strain capable of realizing high production of bacitracin as well as preparation method and application thereof
  • Bacillus licheniformis strain capable of realizing high production of bacitracin as well as preparation method and application thereof
  • Bacillus licheniformis strain capable of realizing high production of bacitracin as well as preparation method and application thereof

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Embodiment Construction

[0029] A high bacitracin-producing Bacillus licheniformis ( Bacillus licheniformis ) construction method, including the following steps:

[0030] A method for constructing a high-yield bacitracin bacillus licheniformis comprising the following steps:

[0031] (1) Using the genomic DNA of Bacillus licheniformis DW2 as a template, PCR amplified ilvB Gene fragment;

[0032] (2) in ilvB The gene fragment is connected upstream to the P43 promoter and downstream to the amylase terminator to form a complete ilvB expression element;

[0033] (3) Using the genomic DNA of Bacillus licheniformis DW2 as a template, PCR amplified ilvB The upstream homology arm of the gene and ilvB The downstream homology arm of the gene;

[0034] (4) by overlap extension PCR will ilvB The upstream homology arm of the gene, ilvB expression elements and ilvB The downstream homology arms of the genes are connected together to form the target gene fragment, and the arrangement order of the t...

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Abstract

The invention provides a bacillus licheniformis strain capable of realizing high production of bacitracin as well as a preparation method and application thereof. Bacillus licheniformis DW2-ilvB is obtained through a genetic engineering method and by inserting ilvB genes in a genomic DNA (Deoxyribonucleic Acid) sequence of bacillus licheniformis DW2; relative to the bacillus licheniformis DW2, theyield of the bacitracin in a fermentation solution during a bacitracin fermentation process of the strain is increased by 11 percent or more.

Description

technical field [0001] The invention relates to a bacillus licheniformis strain with high bacitracin production, a preparation method and application thereof. Background technique [0002] Bacitracin is a class of cyclic peptide antibiotics composed of 12 amino acid residues. The constituent amino acids of bacitracin include ornithine (Orn), D-phenylalanine (D-Phe), isoleucine (His ), D-Aspartic Acid (D-Asp), Asparagine (Asn), Lysine (Lys), D-Glutamic Acid (D-Glu), Cysteine ​​(Cys), Leucine (Leu), isoleucine (Ile) and valine (Val) 11 amino acids. Bacitracin is a broad-spectrum antibiotic that inhibits bacterial growth mainly by inhibiting cell wall synthesis. Bacitracin can effectively inhibit Gram-positive bacteria, such as Clostridium, Staphylococcus, Streptococcus, Corynebacterium and Neisseria, etc., and some Gram-negative bacteria such as meningococcus, influenza Bacillus and actinomycetes also have good inhibitory effect. Bacitracin is hardly absorbed in the intest...

Claims

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Application Information

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IPC IPC(8): C12N15/75C12N15/66C12N1/21C12P21/00C12R1/10
CPCC12N9/1022C12N15/66C12N15/75C12P21/00C12Y202/01006
Inventor 陈守文李阳蔡冬波吴非李俊辉陈晓斌楼丽君
Owner LIFECOME BIOCHEM
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