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Method for preparing rare ginsenoside by fermentation of ginseng endophyte

A technology of ginsenosides and endophytes, applied in the field of ginseng endophytes, can solve the problems of high cost, low yield, poor specificity, etc., and achieve the effects of short growth cycle, simple medium and rich content

Inactive Publication Date: 2018-11-06
YANBIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chemical methods such as acid hydrolysis, alkali hydrolysis and Smith degradation method, etc., although the operation is simple, but the specificity is poor, it is difficult to obtain a monomeric saponin with a specific structure, and the yield is very low, causing environmental pollution at the same time
In addition, there are also research reports on the synthesis of rare ginsenosides with chemical catalysts, but there are many defects in the above methods of chemically synthesizing ginsenosides, generally requiring multiple protection and deprotection steps, and the yield is low
The biotransformation method uses the degradation of enzymes produced in the body to selectively hydrolyze the side chain sugar groups of saponin molecules to achieve the purpose of structural modification. Compared with chemical methods, it has high specificity, mild reaction conditions, and no Pollution and other characteristics, but ginsenosides obtained from ginseng or Panax notoginseng are used as raw materials, so the cost is high, and it is difficult to obtain a large amount of rare saponins

Method used

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  • Method for preparing rare ginsenoside by fermentation of ginseng endophyte

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Isolation of endophytes:

[0017] First, rinse 6-year-old ginseng roots with running water to remove impurities such as soil on the surface of ginseng, then rinse with 75% ethanol for 1 minute, rinse with sterile water for 4 times, soak in 1% sodium hypochlorite solution for 10 minutes, and rinse with sterile water for 4 times , dry the surface with filter paper, cut off the epidermis and root fibers of ginseng with a sterile knife, and cut them into small pieces of 1mm×5mm×5mm, and then culture them on PDA plate medium for 2 days. thus obtaining a pure culture.

[0018] Bacterial fermentation culture:

[0019] 150mL of LL liquid medium was sterilized at 121°C for 40 minutes using a high-pressure steam sterilizer, placed on a biosafety purification console and cooled to room temperature, inoculated with mixed endophytic bacteria PDA-M, and placed in an air bath shaker After fermentation at 30°C and 150rpm for 4 days, the fermentation product was obtained; a 600 μL sam...

Embodiment 2

[0021] Isolation of endophytes:

[0022] First, rinse 6-year-old ginseng roots with running water to remove impurities such as soil on the surface of ginseng, then rinse with 75% ethanol for 1 minute, rinse with sterile water for 4 times, soak in 1% sodium hypochlorite solution for 10 minutes, and rinse with sterile water for 4 times , dry the surface with filter paper, cut off the ginseng epidermis and root hairs with a sterile knife, and cut them into small pieces of 1mm×5mm×5mm, then place them on the R2A plate culture medium for 2 days, and obtain a single colony after streaking, separation and culture, thus obtaining a pure culture.

[0023] Bacterial fermentation culture:

[0024] Add 0.01mol pyruvate to 150mL of LL liquid medium, use a high-pressure steam sterilizer, sterilize at 121°C for 40 minutes, place it on a biosafety purification bench to cool to room temperature, and inoculate the mixed endophyte R2A-M , placed in an air bath shaker and fermented at 30°C and ...

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Abstract

The invention discloses a method for preparing rare ginsenoside by fermentation of ginseng endophyte. The method comprises the following steps of (1) cleaning the surface of the ginseng, removing bacteria, removing peel or root and whisker, slicing, and obtaining pure culture on a flat plate culture medium; (2) inoculating single or multiple colonies into a liquid fermentation culture medium for culturing for 4 to 14 days; (3) adding 1 to 1.5 volume of n-butanol into the fermentation liquid, fully and uniformly mixing, centrifuging, obtaining a supernatant, and concentrating and drying under the vacuum condition, so as to obtain the rare ginsenoside. The method has the advantages that the rare ginsenoside is produced by fermentation of ginseng endophyte, the growth cycle of the endophyte is short, the growth speed is quick, the culture is easy, the influence by environment factors is avoided, the culture medium is simpler, the content of secondary ginsenoside is high, and the method issuitable for industrialized production; by adding acetone acid, acetyl coenzyme A, sodium acetate and other precursor matters, and lovastatin, eugenol and other enzyme inhibitors, the rare ginsenoside Rg3 can be induced to produce.

Description

technical field [0001] The invention relates to ginseng endophytes and uses thereof, in particular to a method for fermenting ginseng endophytes to prepare rare ginsenosides. Background technique [0002] Ginsenosides are a kind of sterol compounds, which belong to triterpenoid saponins, and are the main pharmacological active ingredients in ginseng medicinal materials. The structure of ginsenoside is divided into two parts, one part is hydrophobic aglycone, and the other part is hydrophilic sugar group. According to the different saponins, ginsenosides can be divided into three categories: oleanolic acid type saponins (such as Ro), panaxadiol type saponins (such as Rb1, Rb2, Rg3, Rd, Rh2, PPD), panaxatriol Type saponins (such as Re, Rg1, Rh1, PPT), among which Rg3, Rh2, etc. exist in wild ginseng and red ginseng with extremely low content, called rare ginsenosides (Rareginsenosides). Rare ginsenosides play an important role in promoting the apoptosis of tumor cells, promo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P33/20
CPCC12P33/20
Inventor 严华玉尹成日金护定
Owner YANBIAN UNIV
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