Bacterial strain of bacillus licheniformis with vitreoscilla hemoglobin gene as well as construction method and application of bacterial strain

A technology of Bacillus licheniformis and hemoglobin, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, bacteria, etc., to achieve the effects of improving oxygen supply problems, improving oxygen uptake capacity, and improving physiological state

Active Publication Date: 2014-03-19
LIFECOME BIOCHEM
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  • Claims
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Problems solved by technology

[0004] At present, the main production strains of bacitracin are Bacillus licheniformis and Bacillus subtilis, which are produced by microbial fermentation. In the process of microbial fermentation, oxygen is particularly important for the growth and reproduction of aerobic microorganisms and energy and substance metabolism. Important, but microorganisms can only use dissolved oxygen in the medium, making oxygen supply a major bottleneck limiting fermentation yields

Method used

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  • Bacterial strain of bacillus licheniformis with vitreoscilla hemoglobin gene as well as construction method and application of bacterial strain
  • Bacterial strain of bacillus licheniformis with vitreoscilla hemoglobin gene as well as construction method and application of bacterial strain
  • Bacterial strain of bacillus licheniformis with vitreoscilla hemoglobin gene as well as construction method and application of bacterial strain

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specific Embodiment

[0023] Specific examples of the method for constructing Bacillus licheniformis carrying the Vitreoscilla hemoglobin gene are as follows:

[0024] (1) Plasmid pDG1730-vgb + The specific construction method

[0025] ① Plasmid pEG491-vgb + The 1246bp hemoglobin gene vgb fragment was obtained by double digestion with BamHI and HindIII;

[0026] ② Plasmid pDG780 was digested with HindIII to obtain 1360bp kanamycin resistance gene (Kan r ) Fragment;

[0027] ③The two fragments were inserted into the corresponding multiple cloning sites in plasmid pDG1730 to obtain plasmid pDG1730-vgb + .

[0028] (2) Plasmid pDG1730-vgb + Double digestion

[0029] ① Enzyme digestion system:

[0030] ② Enzyme digestion conditions: temperature 30℃, time 8 hours.

[0031] BamH I and Hind III are DNA restriction endonucleases, and 10×K buffer is the matching restriction enzyme buffer. BamH I, Hind III and 10×K buffer were all purchased from Bao Biological Engineering (Dalian) Co., Ltd.

[0032] After the above-ment...

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Abstract

The invention relates to a bacillus licheniformis with a vitreoscilla hemoglobin gene. The bacterial strain is the bacillus licheniformis (PHY300-vgb), wherein the bacterial strain is stored in China Typical Model Cultivation Center of Wuhan, and the storage number is CCTCC NO: M2011345. The invention aims to provide the bacterial strain of the bacillus licheniformis with vitreoscilla hemoglobin gene, which can produce bacitracin in high yield, as well as a construction method and an application of the bacterial strain.

Description

Technical field [0001] The invention relates to an engineered Bacillus licheniformis strain carrying a Vitreoscilla hemoglobin gene. Background technique [0002] Bacitracin is a pale yellow or white powder, odorless and bitter. It is a polypeptide antibiotic composed of 10 amino acids and 12 amino acid residues. It can effectively inhibit Gram-positive pathogens and some Gram-negative bacteria. Bacitracin interacts with various metal ions such as Cu through the imidazole group and thiazole ring of histidine. 2+ , Ni 2+ , Co 2+ , Zn 2+ And Mn 2+ The antibacterial activity of bacitracin in the form of zinc salt is enhanced to a certain extent, and the stability is the best. Bacitracin has the advantages of almost no absorption in the intestine, rapid excretion, and no residue in the body, so it is widely added to feed. [0003] Bacitracin's mechanism of action is: 1. Its complex can be adsorbed on the cell wall of bacteria, hindering dephosphorylation, affecting the operation of ph...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12P21/04C12R1/10
Inventor 李俊辉楼丽君赖建平
Owner LIFECOME BIOCHEM
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