Method for preparing deoxidized violacein and special recombinant bacterium

A purple bacteriocin and deoxyviolet technology, applied in the field of production of deoxyviolet bacteriocin, can solve the problems of low yield, difficult separation, less research on properties and biological activities, etc., and achieve convenient extraction, simple separation and purification, and easy industrial production. Effect

Inactive Publication Date: 2008-12-10
TSINGHUA UNIV
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Problems solved by technology

[0005] Deoxyviobacterin is a by-product of the violacein synthesis pathway, the yield is very low, and it is difficult to separate, so there are few studies on its properties and biological activities

Method used

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  • Method for preparing deoxidized violacein and special recombinant bacterium
  • Method for preparing deoxidized violacein and special recombinant bacterium
  • Method for preparing deoxidized violacein and special recombinant bacterium

Examples

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Embodiment 1

[0025] Embodiment 1, the recombinant bacterium BL-DV that produces deoxyviolet violacein

[0026] 1) Construction of recombinant expression vectors for enzymes related to the synthesis pathway of deoxyviolet bacterin

[0027] Transfer Duganella sp. B2CGMCC №2056 into liquid medium (starch 15g / L, ferrous sulfate 0.03g / L, potassium nitrate 1g / L, dipotassium hydrogen phosphate 0.7g / L, sulfuric acid Magnesium 0.5g / L, tryptophan 0.5g / L, pH adjusted to 7.0), 25°C, 200rpm cultured for 36 hours, according to the instructions of the Shanghai Sangon Genomic DNA Extraction Kit, the Genomic DNA of Duchenne B2 was extracted.

[0028] According to the sequence of the violacein gene cluster, three pairs of primers were designed using Oligo 7.10 software. The primer sequences are shown in Table 1, wherein P1 and P2 amplify the partial sequence of vioA and part of the vioB gene, and the amplified product is named Fragment A; P3 and P4 amplify Part of the vioB and vioC genes, the amplified pro...

example 2

[0048] Example 2, recombinant bacterium BL-DV CGMCC No.2557 producing deoxyviolet violacein

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Abstract

The invention discloses a method for producing deoxidized violacein and special recombination strains thereof. The method comprises the following steps of: obtaining the recombination strains through inducting a deoxidized violacein synthesis associated gene cluster into bacillus coli BL21-CodonPlus(DE3)-RIL, producing the deoxidized violacein through the recombination strains which take tryptophan as a substrate to ferment, wherein the deoxidized violacein synthesis associated gene cluster is a recombination gene cluster which is obtained through getting rid of a VioD gene from a violacein synthetic gene cluster consisting of VioA, VioB, VioC, VioD and VioE, and the recombination strains are obtained through inducting the gene cluster into bacillus coli BL21-CodonPlus(DE3)-RIL and are capable of producing the deoxidized violacein. The method for producing the deoxidized violacein has a yield as high as 0.71g / L fermentation liquor with convenient extraction and simple separation and purification.

Description

technical field [0001] The invention relates to a method for producing deoxyviorin and its special recombinant bacteria. Background technique [0002] [0003] Violacein and deoxyviolacein are metabolites of microorganisms, which belong to indole derivatives and are formed by oxidative condensation of two tryptophan molecules. Since violacein was discovered at the end of the 19th century, people have carried out a lot of exploration on its biological function, and found that it has strong biological function, and it has attracted more and more attention. Studies have shown that violacein has strong biological activity: (1) has broad-spectrum antibacterial activity, such as anti-staploylococcus aureus, Bacillus sp, streptococcus sp, mycobacterium, Neisserig, pseudomonas (Sanchez et al., Reevaluation of the Violacein Biosynthetic Pathway and its Relationship to Indolocarbazole Biosynthesis.Journal 2006.7, 1231-1240); (2) anti-oxidation (Konzen et al., Antioxidant propertie...

Claims

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Application Information

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IPC IPC(8): C12N15/31C12N1/21C12N15/70C12P17/16C12R1/19
Inventor 邢新会王海胜蒋培霞赵洪新姜瑞波张翀娄恺魏东
Owner TSINGHUA UNIV
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