Low-peculiar smell bacitracin production bacillus licheniformis and construction method and application thereof

A technology of Bacillus licheniformis and Bacillus licheniformis, which is applied in the direction of bacitracin, chemical instruments and methods, and methods based on microorganisms, and can solve the problems of fermentation broth contamination, increased input costs, and economic losses

CN106755047AInactive Publication Date: 2017-05-31LIFECOME BIOCHEM
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2017-05-31
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a low-peculiar smell bacitracin production bacillus licheniformis and a construction method and an application of the low-peculiar smell bacitracin production bacillus licheniformis. A ptb gene in a genomic DNA sequence of the bacillus licheniformis DW2 is knocked out through a genetic engineering method to obtain the bacillus licheniformis DW2deltaptb, and compared with the bacillus licheniformis DW2, the synthetic amounts of isobutyric acid and isovaleric acid of the bacterial strain are respectively reduced to 33% and 37% in the fermentation process of the bacitracin, and the peculiar smell is remarkably reduced.
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Description

technical field

[0001] The invention relates to the field of microbial genetic engineering, in particular to a low-odor bacitracin-producing bacillus licheniformis and its construction method and application. Background technique

[0002] Bacitracin is a light yellow or off-white powder. It is a polypeptide antibiotic composed of 10 amino acids and 12 amino acid residues. It can effectively inhibit Gram-positive pathogenic bacteria and some Gram-negative bacteria. It is widely added to feed.

[0003] At present, bacitracin is produced by microbial fermentation, and the main production strain is Bacillus licheniformis. The bacitracin fermentation process has a strong odor, which seriously affects the environment, and the odorous substances are attached to the surface of the bacitracin product, which affects the product quality. In view of the low yield of bacitracin produced by most of the existing Bacillus licheniformis, it is not conducive to putting into industrialized pr...

Examples

Embodiment Construction

[0018] A method for constructing a low-odor bacitracin-producing bacillus licheniformis, comprising the following steps:

[0019] 1) According to the upstream and downstream sequences of the ptb gene in the Bacillus licheniformis (Bacillus licheniformis) DW2 genomic DNA sequence, design the upstream and downstream homology arm primers of the ptb gene; and use the genomic DNA of Bacillus licheniformis DW2 as a template, respectively The upper and lower homology arm primers of the ptb gene are amplified by PCR to obtain the upper homology arm of the ptb gene (usually 450-550bp) and the lower homology arm of the ptb gene (usually 450-550bp), and then use overlap extension PCR Methods The upstream homology arm of ptb gene and the downstream homology arm of ptb gene are spliced ​​together to obtain fusion gene sequence A;

[0020] 2) Use restriction endonucleases XbaI and BamH I to double-enzyme digest the fusion gene sequence A obtained in step (1) to obtain the restriction endonu...