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A tobacco endophytic fungus and its application in the preparation of ergosterol 5,8 peroxide

A technology of ergosterol and peroxide, which is applied in the field of microorganisms, achieves high research value and application value, enhances the ability of the human body to resist diseases, and promotes the effect of tumor cell apoptosis

Active Publication Date: 2021-04-16
TOBACCO RES INST CHIN AGRI SCI ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there is no report about a tobacco endophytic fungus capable of preparing ergosterol 5, 8 peroxide by microbial fermentation and its application

Method used

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  • A tobacco endophytic fungus and its application in the preparation of ergosterol 5,8 peroxide
  • A tobacco endophytic fungus and its application in the preparation of ergosterol 5,8 peroxide
  • A tobacco endophytic fungus and its application in the preparation of ergosterol 5,8 peroxide

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Experimental program
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Embodiment 1

[0027] Isolation and identification of embodiment 1 bacterial strain TE-3

[0028] 1. Isolation of tobacco endophytic fungus TE-3

[0029] The tobacco endophytic fungus TE-3 of the present invention is a bacterial strain obtained from healthy tobacco leaves collected in Enshi Prefecture, Hubei Province.

[0030] Tobacco endophytic fungi were isolated by tissue separation. First, the tobacco leaves are fully rinsed under tap water to remove the soil on the surface; then the rinsed leaf tissue is surface-sterilized according to the three-step method of "75% ethanol (1min)-2.5% sodium hypochlorite (30s)-75% ethanol (1min)" Sterilize, then rinse with sterile water 3 times; cut the leaves into small pieces of 0.5cm×0.5cm in size, insert the cut into PDA medium containing chloramphenicol, and culture in a constant temperature incubator at 28°C for 3 to 7 days ; When mycelium grows from the incision of the tissue in the culture medium, use a sterilized inoculation loop to pick the ...

Embodiment 2

[0035] Preparation and detection of embodiment 2 ergosterol 5,8 peroxide

[0036] 1. Preparation of ergosterol 5,8 peroxide

[0037] Under sterile conditions, pick an appropriate amount of the strain TE-3 that has been cultured on a slant for 3 days; pick an appropriate amount of TE-3 mycelium and transfer it to a 1000 mL Erlenmeyer flask, which is filled with 300 mL of sterilized liquid in advance PDB culture medium; the Erlenmeyer flask was fermented and cultivated at 28°C for 30 days; after the fermentation was completed, the fermentation broth was separated from the mycelium by filtration, and the fermentation broth was continuously extracted 3 times with an equal volume of ethyl acetate, and the 3 extracts were combined. Concentrate under reduced pressure to obtain concentrated solution A; the mycelium was continuously extracted 3 times with 3 times the amount of 80% acetone ultrasonically, combined 3 times with the acetone extract, concentrated under reduced pressure and...

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Abstract

The present invention proposes a method for preparing ergosterol 5,8 peroxide by utilizing tobacco endophytic fungi: under aseptic conditions, the TE-3 mycelium is transferred to the sterilized liquid PDB medium; Leave to ferment, filter and separate the fermentation broth and mycelia; the fermentation broth is extracted 3 times with ethyl acetate, combined and concentrated under reduced pressure to obtain concentrated solution A; the mycelium is ultrasonically extracted 3 times with acetone, concentrated under reduced pressure and then extracted with ethyl acetate 3 times, the concentrated solution B was obtained after merging; A and B were combined to obtain the fermented extract; silica gel was added to mix the sample, petroleum ether: ethyl acetate = 2: 1 for silica gel column chromatography; gel column chromatography was used to separate and obtain The pure product of ergosterol 5, 8 peroxide, the bacterial strain TE‑3 is Arthrinium arundinis TE‑3. The preparation method is suitable for industrial large-scale cultivation, has high yield, and is easy to separate and purify; the fermentation medium has low cost and simple preparation, and can realize industrial-scale production, providing a basis for the research, application and industrial production of canosterol 5,8 peroxide. Drug source.

Description

technical field [0001] The invention relates to a tobacco endophytic fungus and its application in the preparation of ergosterol 5,8 peroxide, belonging to the technical field of microorganisms. Background technique [0002] Ergosterol (ergosterol), also known as ergosterol, exists in large quantities in fungi such as yeasts and molds and certain plants. It is an important phytosterol and has important physiological functions. It can enhance the body's ability to resist diseases. An important source of fat-soluble vitamin D2, which has significant antibacterial and antitumor effects. Ergosterol is also a commonly used chemical raw material in food, feed and pharmaceutical industries, and can be used in the production of drugs such as cortisone and hormone progesterone. Ergosterol 5, 8 peroxide is a derivative of ergosterol C5 and C8 oxidized to form a peroxide bridge. It is a rare type of sterol, which is more common in some lower marine organisms and less common in some fu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P33/20C12R1/645
CPCC12P33/20C12N1/145C12R2001/645
Inventor 张鹏袁晓龙李盼盼张忠锋杜咏梅刘新民
Owner TOBACCO RES INST CHIN AGRI SCI ACAD