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Amorocoelophoma sp. and application thereof in preparation of spirolactone derivative

A technology of plant endophytic fungi and derivatives, which is applied to the application field of plant endophytic fungi and the preparation of spirolactone derivatives, which can solve the problem that Amorocoelophoma has no secondary metabolites and the like.

Inactive Publication Date: 2021-06-15
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are no reports on secondary metabolites of fungi of the genus Amorocoelophoma

Method used

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  • Amorocoelophoma sp. and application thereof in preparation of spirolactone derivative
  • Amorocoelophoma sp. and application thereof in preparation of spirolactone derivative
  • Amorocoelophoma sp. and application thereof in preparation of spirolactone derivative

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] (1) Rinse the freshly collected, non-conditioned Yunnan Stone Skin with tap water, remove the soil and dust of the surface. The dried plant material was placed in an ultra-clean workbench through UV-sterilized, and 75% alcohol and 2% sodium hypochlorite were selected as surface disinfectants. The plant materials were first soaked with 75% alcohol for 30-60S, rinsed 2 times in sterile water; then treated with 2% sodium hypochlorite, 1.5-3 min, sterile water rinsing 3 times. Sterile filter paper suction moisture.

[0033] (2) The Yunnan Stone Timberry, which is disinfected with step (1), has a small piece of a small piece of about 1 cm, and the small piece of plant tissue is inoculated on the PDA medium, and the temperature is cultured at 28 ± 2 ° C. The PDA medium includes 200 g of peel potatoes, 20 g of glucose, grease 15g, distilled water 1000ml, pH nature.

[0034] (3) Observe the growth of the surface of the plant tissue block, and use a sterile bamboo stick to pick up t...

Embodiment 2

[0039] Amorocoelophoma sp.YE3351 Method for fermenting helium lysate derivatives, including the following steps:

[0040](1) PDB medium: weighing 200g, cut into small pieces, add 1000 mL of distilled water to boil, gauze filtrate, gauze, pH 20g, pH, 121 ° C Sterilization for 30 min, produce PDB medium Near, where the concentration of potatoes in the PDB medium is 0.2 g / ml, and the concentration of glucose is 0.02 g / ml.

[0041] (2) AmorocoElophoma sp.YE3351 strain is incorporated into a PDB medium, and 4D is cultured at 28 ± 2 ° C, 200R / min shaker, and the seed fluid is obtained, and then the resulting seed liquid is added to 10% inoculation amount. In the medium, 7d was cultured in 28 ± 2 ° C, 200R / min shaker, and the fermentation product was obtained.

[0042] (3) Filting the fermentation product obtained by step (2) is filtered, resulting in fermentation liquid and mycelial body, the fermentation broth extraction, mycerer is extracted with methanol, combined with EtOAc,...

Embodiment 3

[0051] Anti-viral activity assay for Barnocycloalkyl derivative Massarilactone D and MassariLACTONE E:

[0052] (1) Determination of Cell Disease (CPE):

[0053] CPE analysis is used to evaluate the anti-H1N1 influenza virus activity of the helicy cyclic derivative Massarilactone D and Massarilactone E. Collect MDCK cells and count, 20ml 10 by 2% DMEM 5 / ml cells, MDCK cells in 2 × 10 4 The density of a cell / well is inoculated into a 96-well plate and incubated at 37 ° C, and in 5% CO at 37 ° C. 2 Place 24h in. Two-fold continuous diluted spiral lacene derivative Massarilactone D and Massarilactone E and Hydrotective virus (100TCID 50 Mix, incubated at 37 ° C for 30 min, then transferred to a cell in a 96-well plate, and then incubated for 30 min. After incubation, MDCK cells were washed twice with PBS to remove uncomfortable viruses, and then added 1 μg / ml TPCK-trypsin and 0.2% BSA DMEM. After incubation 48 h, an antiviral effect was observed under a microscope. The data was...

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Abstract

The invention belongs to the technical field of microorganisms, and relates to an Amorocoelophoma sp. and application thereof in preparation of a spirolactone derivative. The Amorocoelophoma sp. YE3351 is preserved in China center for type culture Collection on August 7, 2020, the address is Wuhan University, Wuhan City, China, the preservation number is CCTCC NO. M2020407, and the taxonomic name is Amoro-elophoma sp. According to the Amorocoelophoma sp. and the application thereof in preparation of the spirolactone derivative, the Amorocoelophoma sp. YE3351 is separated from the root bark part of Chinese catalpa in Yunnan for the first time, the spirolactone derivative is separated from the metabolite of the Amorocoelophoma sp. YE3351, and the blank of the prior art is filled.

Description

Technical field [0001] The invention belongs to the field of microbial technology, involving a plant endogenic fungi and its application in the preparation of a spiral cyclic derivative. Background technique [0002] EndophyTIC Fungi is separated from plant health tissue, confirmed by surface disinfection experiments that are not surface attachea or pollutifier fungi, generally in higher plants, wood, herb, monocotyla Endomided fungi in the bonus leaf plant. Plant endocarburities have powerful biosynthesis, is an important source of drug pioneering compounds, and their secondary metabolic products have a variety of structural types, and have a variety of biological activity, and the discovery of drug pioneering compounds for microbial derivements has a high research value. . [0003] The spiro compound is a polycyclic compound that co-uses a carbon atom by two monocycles. The spiro compound has a rigid structure with a stable structure, and its chiral ligand has a larger than the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P17/04C12P17/18C12R1/645
CPCC12P17/04C12P17/181
Inventor 吴少华刘思思
Owner YUNNAN UNIV
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