A myxobacterium producing methadazole compounds and its application

A technology for methadazole and compounds, which is applied in the field of fermentative preparation of methadazole compounds, and can solve the problems of difficulty, difficulty in synthesis, and high cost

Active Publication Date: 2022-04-12
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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  • A myxobacterium producing methadazole compounds and its application
  • A myxobacterium producing methadazole compounds and its application
  • A myxobacterium producing methadazole compounds and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Isolation of strains

[0040] The isolation method is as follows: After the WCX medium has solidified, smeared with E. coli. The fresh or air-dried samples (the soil samples from the lake bottom of Dongcuo Lake, Gaize County, Tibet Autonomous Region) were evenly spread on the E. coli bacteria pads. After culturing to day 3, the plates were examined daily under a stereo microscope for the appearance of fruiting bodies or biofilms of myxobacteria. If there is, carefully pick the top of the fruiting body or scrape the edge of the pellicle, and transfer it to a new WCX medium plate with E. coli pads or VY / 2 medium for purification. In the experiment, a yellow bacterium was obtained and named as strain SDU36.

[0041] Wherein, the formula of above-mentioned WCX medium: CaCl 2 ·2H 2 O 0.1g / 100ml, agar 1.5g / 100ml, pH 7.2. After sterilization, cycloheximide was added to a final concentration of 25 μg / mL. The formula and preparation method of VY / 2 medium: Angel y...

Embodiment 2

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

[0046] The fermentation steps of 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 cells; scrape the cells and inoculate them into 100 mL shake flasks containing VY / 2 liquid seed medium , cultured at 30°C for 4–5 days to reach the logarithmic growth phase, transferred to fresh VY / 2 fermentation medium with 10% of the inoculum volume, continued to culture at 30°C, 200rpm for 7 days, and filtered off the bacteria , add 2% (w / v) adsorption resin HP-20 to the culture solution, adsorb at 30°C for 20 hours at 200 rpm, collect the resin, elute with methanol, concentrate the eluate, take 10 mg of the crude fermentation product of SDU36 strain, add 100 μL Chromatography Dissolved by methanol ultrasonically, centrifuged at 15,000 rpm for 30 min to take the supernatant, and detected methad...

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 cells;

[0053] (2) inoculate the activated thalline (10 μL inoculation ring full) prepared in step (1) in a shake flask containing VY / 2 liquid seed medium, and cultivate at 28° C. for 4 days to obtain seed liquid;

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

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

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Abstract

The invention discloses a myxobacterium producing methadazole compounds. The myxobacterium is named Myxococcus sp. SDU36, and has been preserved in the China Center for Type Culture Collection on May 12, 2021, with the preservation number CCTCC NO: M 2021520. The invention also discloses the application of the bacterium in fermenting and preparing methadazole compounds. Experiments have confirmed that the myxococcus culture process of the present invention is simple, the cycle is short, it is not easy to pollute, and the methadazole compound product is stable. It is a methadazole compound producing bacterium with great research value. Its discovery is expected to produce social benefits and Economic Value.

Description

technical field [0001] The present invention relates to a Myxococcus sp. SDU36 strain and its fermentative production of methadazole compounds, in particular to a Myxococcus sp. Compound applications. It belongs to the technical field of microbial technology and its products and applications. Background technique [0002] Mesadazoles are a new type of alkaloid skeleton composed of a fatty acid chain of isoxazole ring derived from Myxobacteria and N-ribitol-5,6-dimethylbenzimidazole. Isoxazole rings are very rare in nature. At present, 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), Acevexin and 4-Hydroxyacevisine (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), Amanita (K. Bowden, A.C. Drysdale, G.A. Mogey, Nature 1965, 206, 1359-1360), Mesadazole...

Claims

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Application Information

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Patent Type & Authority Patents(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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