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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, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., to achieve the effect of improving oxygen supply, oxygen uptake capacity, and physiological state

Active Publication Date: 2013-06-12
LIFECOME BIOCHEM
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  • Abstract
  • Description
  • Claims
  • Application Information

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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] The specific embodiment of the construction method of Bacillus licheniformis carrying Vitiligo hyaline hemoglobin gene is as follows:

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

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

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

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

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

[0029]

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

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

[0032] Af...

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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 engineering bacterial strain of Bacillus licheniformis carrying the hemoglobin gene of Vitella hyaline. Background technique [0002] Bacitracin is a light yellow or off-white powder, odorless and bitter in taste. It is a polypeptide antibiotic composed of 10 amino acids and 12 amino acid residues, which can effectively inhibit Gram-positive pathogenic bacteria and some Gram-negative bacteria. Bacitracin interacts with various metal ions such as Cu through the imidazolyl and thiazole rings of histidine 2+ 、Ni 2+ 、Co 2+ , Zn 2+ and Mn 2+ And so on to form a complex, in which bacitracin in the form of zinc salt, the antibacterial activity 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] The mechanism of action of bacitracin is as follows: 1. Its comple...

Claims

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Application Information

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