Myxobacterium for producing methsadazole compound and application of myxobacterium

A technology for mesadazole and compounds is applied in the application field of fermenting and preparing mesadazole compounds, and can solve the problems of high cost, high difficulty, low yield and the like

Active Publication Date: 2021-08-06
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The synthesis of multi-chiral centers and isoxazole-dimethylbenzimidazole mother nucleus is very difficult. At present, methadazole compounds can only be obtained by biosynthesis, which is costly, difficult and low yield.
Based on this, the development of strains producing methadazole compounds is of great significance for the production of methadazole compounds by fermentation. After searching, there are no reports on the myxobacteria producing methadazole compounds and their applications.

Method used

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  • Myxobacterium for producing methsadazole compound and application of myxobacterium
  • Myxobacterium for producing methsadazole compound and application of myxobacterium
  • Myxobacterium for producing methsadazole compound and application of myxobacterium

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Experimental program
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Effect test

Embodiment 1

[0039] The isolation of embodiment 1 bacterial strain

[0040] The separation method is as follows: after the WCX medium is solidified, spread Escherichia coli. Sprinkle fresh or air-dried samples (soil samples from the bottom of Dongcuo Lake in Gaize County, Tibet Autonomous Region) evenly on the E. coli mat. After culturing to the third day, check the plates under a stereomicroscope every day for the appearance of fruiting bodies or pellicles of myxobacteria. If there is, carefully pick the top of the fruiting body or scrape the edge of the bacterial film, and transfer to a new WCX medium plate or VY / 2 medium plate with E. coli bacteria pad for purification. In the experiment, a yellow bacterium was obtained, which was named strain SDU36.

[0041] Among them, the formula of the above-mentioned WCX medium: CaCl 2 2H 2 O 0.1g / 100ml, agar 1.5g / 100ml, pH 7.2. After sterilization, add cycloheximide to a final concentration of 25 μg / mL. Formula and preparation method of VY / 2...

Embodiment 2

[0045] The fermentation of embodiment 2 bacterial strain SDU36 and the detection of produced methadazole compound

[0046] The fermentation steps of bacterial strain SDU36 are as follows:

[0047] Inoculate Myxococcus sp. SDU36 on a fresh VY / 2 plate and culture at 30°C for 3–4 days to obtain activated bacteria; scrape the bacteria and inoculate them into a 100mL shake flask containing VY / 2 liquid seed medium , cultured at 30°C for 4–5 days, reached the logarithmic growth phase, transferred to fresh VY / 2 fermentation medium with 10% inoculum volume, continued to culture at 30°C, 200rpm for 7 days, and filtered out the bacteria , add 2% (w / v) adsorption resin HP-20 to the culture medium, adsorb for 20 hours at 30°C and 200rpm, collect the resin, elute with methanol, concentrate the eluate, take 10 mg of the crude product fermented by SDU36 strain, add 100 μL Chromatographic methanol was ultrasonically dissolved, centrifuged at 15,000 rpm for 30 min to take the supernatant, and ...

Embodiment 3

[0051] Example 3 Preparation of methadazole compounds by fermentation of Myxococcus sp. SDU36

[0052] (1) Inoculate Myxococcus sp. SDU36 on a fresh VY / 2 plate and culture at 28°C for 4 days to obtain activated bacteria;

[0053] (2) Inoculate the activated thallus (10 μL inoculation loop full circle) prepared in step (1) into a shake flask containing VY / 2 liquid seed medium, and culture at 28° C. for 4 days to obtain a seed liquid;

[0054] (3) Inoculate the seed liquid prepared in step (2) into the VY / 2 fermentation medium in an amount of 10% by volume, ferment for 8 days at 28° C., filter out the mycelium, and obtain methadazole-containing compound fermentation broth.

[0055] Wherein, above-mentioned various culture medium formulations are the same as embodiment 2.

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Abstract

The invention discloses a myxobacterium for producing a mesadazole compound. The myxobacterium is named as myxococcus sp. SDU36, and is preserved in the China Center for Type Culture Collection on May 12, 2021, and the preservation number is CCTCC NO: M 2021520. The invention also discloses an application of the strain to fermentation preparation of the methadazole compound. Experiments prove that the myxococcus culture process is simple, the period is short, pollution is not prone to occurring, a mesadazole compound product is stable, the myxococcus is a mesadazole compound producing strain with great research value, and social benefits and economic values are expected to be generated due to discovery of the myxococcus.

Description

technical field [0001] The present invention relates to a myxococcus sp. SDU36 strain producing methadazole compounds and its fermentative production of methadazole compounds, in particular to a myxococcus sp. application in compounds. The invention belongs to the technical field of microbial technology and its products and applications. Background technique [0002] Mesadazole compounds are a new alkaloid framework formed by the hybridization of fatty acid chains of isoxazole rings from myxobacteria and N-ribityl-5,6-dimethylbenzimidazole. The isoxazole ring is very rare in nature, and only five natural products containing isoxazole fragments have been found from actinomycetes and fungi: cycloserine (M.-L.Svensson, S.Gatenbeck, Arch.Microbiol. 1982,131,129-131), acivicin and 4-hydroxyacevicin (S.J.Gould, S.Ju, J.Am.Chem.Soc.1993,114,10166-10172), muscimol (M .Onda, H.Fukushima, M.Akagawa, Chem.Pharm.Bull.1964,12,751-751), ibotenic acid (K.Bowden,A.C.Drysdale,G.A.Mogey,Na...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/16C07D413/06C12R1/01
CPCC12N1/20C12P17/16C07D413/06
Inventor 李越中吴长生李岳兰吴姝鸽王晶晶
Owner SHANDONG UNIV
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