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Hexacyclic alkaloid compound, preparation method and application thereof

A technology of alkaloids and compounds, applied in the biological field

Inactive Publication Date: 2015-01-21
YUNNAN MINZU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention separates a hexacyclic alkaloid compound with significant anti-tobacco mosaic virus activity from the solid fermentation product of the Penicillium Cladoides group; the compound has not been reported so far

Method used

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  • Hexacyclic alkaloid compound, preparation method and application thereof
  • Hexacyclic alkaloid compound, preparation method and application thereof
  • Hexacyclic alkaloid compound, preparation method and application thereof

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preparation example Construction

[0031] The preparation method of the hexacyclic alkaloid compound of the present invention comprises the following steps:

[0032] A. Solid fermentation: Cultivate the strain in potato dextrose agar medium at 28°C for 7 days, inoculate it into 100ml potato dextrose liquid medium (cut 200 g potatoes into small pieces, add water and boil them until rotten (boil for 20-30 minutes, Potato cubes can be pierced by a glass rod), filter with eight layers of gauze, heat, add 5 g of agar, continue to heat and mix well, after the agar is dissolved, add 20 g of glucose, stir evenly, and cool down slightly Make up water to 1000ml, divide into test tubes or Erlenmeyer flasks, stopper and bandage, (121°C) for about 20 minutes after sterilization, take out the test tubes and place them on an inclined plane or shake well, cool down and store them in 250ml Erlenmeyer flasks, at Cultivate for 5 days at 28°C under the condition of rotation and shaking at 180rpm to obtain strains for fermentation,...

Embodiment 1

[0057] ------Solid fermentation and extract extraction:

[0058] The strain was cultured in potato dextrose agar medium for 7 days at a culture temperature of 28 °C; the agar plug was inoculated into a 250 mL Erlenmeyer flask containing 100 mL of potato dextrose, and cultured at 28 °C under the condition of rotating and shaking at 180 rpm After 5 days, the strains for fermentation were obtained. Solid fermentation was carried out in 500 mL Erlenmeyer flasks, each containing 100 g of rice and 120 mL of water, and inoculated with strains after high-temperature sterilization; 50 bottles were fermented in each batch, and fermented at 25 °C for 45 days . The fermentation product was extracted with 70% methanol, and then extracted with ethyl acetate, and the ethyl acetate extracted part was concentrated to obtain 210 g of extract.

Embodiment 2

[0060] ------compound separation

[0061] The extract was dissolved in pure methanol with a weight ratio of 2.0 times, and then mixed with 350 g of 100-mesh crude silica gel, and 1.2 kg of 160-mesh silica gel was packed into a column for silica gel column chromatography, and the volume ratio was 1:0, 9:1 , 8:2, 7:3, 6:4, 1:1, 0:1 chloroform-acetone gradient elution, TLC monitoring and merging the same parts to obtain 8 parts, wherein the volume ratio of 9:1 chloroform - The acetone eluted part was separated by Agilent 1100 semi-preparative high performance liquid chromatography, using 60% methanol as mobile phase, Zorbax SB-C18 (21.2 × 250 mm, 5 μ m ) The preparative column is a stationary phase, the flow rate is 12 ml / min, and the detection wavelength of the ultraviolet detector is 254 nm. Each injection is 200 μL, and the chromatographic peaks collected for 22.5 min are evaporated to dryness after repeated accumulation; Dissolved, and then using pure methanol as mobile phas...

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Abstract

The invention discloses a hexacyclic alkaloid compound, a preparation method and an application thereof. The hexacyclic alkaloid compound is obtained through fermentation, extract extraction, chromatography and purification on a Penicilliumramigena (YNCA0361) strain, a molecular formula is C19H20N2O6, a structural formula is shown as the following, the compound is named as Penicilliumramigena A; a bacterial strain preservation number of the Penicilliumramigena (YNCA0361) strain is CGMCC No.4824, and the preservation date is 2nd, May, 2011. The preparation method comprises the steps of solid fermentation, extract extraction, slica column chromatography, and separation and purification by high pressure liquid chromatography. The application of the present invention is the application of the hexacyclic alkaloid compound for preparing tobacco mosaic virus medicines. The compound has good application prospect for preparing the tobacco mosaic virus medicines. The compound has good activity, and can be obtained through a biological fermentation approach, and can be used as the guiding compound for the tobacco mosaic virus medicines.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a hexacyclic alkaloid compound and its preparation method and application. Background technique [0002] There are many types of microbial metabolites, and the products produced in the logarithmic growth phase of the bacteria, such as amino acids, nucleotides, proteins, nucleic acids, sugars, etc., are necessary for the growth and reproduction of the bacteria. These products are called primary metabolites; in the stationary phase of bacterial growth, some bacteria can synthesize some products with specific functions, such as antibiotics, alkaloids, bacterial toxins, plant growth factors, etc. These products have no obvious relationship with the growth and reproduction of bacteria, and are called secondary metabolites. Secondary metabolites are mostly small molecular compounds, but their chemical structure types are various. Secondary metabolites with different structures...

Claims

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Application Information

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IPC IPC(8): C07D491/22C12P17/18A01P1/00C12R1/80
CPCA01N43/90C07D491/22C12P17/182
Inventor 高雪梅黄相忠胡秋芬李银科叶艳清杜刚
Owner YUNNAN MINZU UNIV
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