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Method for preparing ginsenoside Rd through biotransformation of panax notoginseng saponins

A technology of Panax notoginseng saponins and ginsenosides, applied in biochemical equipment and methods, methods based on microorganisms, microorganisms, etc., can solve the problems of high transformation specificity, complex types, low screening success rate, etc., and achieve transformation specificity High, high conversion efficiency

Active Publication Date: 2016-02-17
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The source of strains in the microbial fermentation method is relatively complicated, and it is relatively difficult to screen active or highly active strains, and the success rate of screening is low. Usually, only a few hundred strains are selected to screen a few strains with saponin conversion activity. Generally, the conversion rate is not Very high and the conversion process and conversion product types (including the secondary metabolites of fermenting microorganisms) are relatively complex; extracting microbial crude enzyme method, high conversion specificity, mild conditions, no medium required for microbial fermentation, and clear product targets And it will not produce secondary metabolites of the microorganism itself, but due to the protein properties of the enzyme, it is difficult to extract, isolate and purify the enzyme with saponin conversion activity from the microorganism, and the purified enzyme needs to be dissolved in the buffer to maintain the enzyme activity. At the same time, a series of problems such as difficult enzyme recovery

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1: The method for preparing ginsenoside Rd by transforming Panax notoginseng saponins by this microorganism is carried out as follows:

[0019] (1) Use conventional PDB medium to activate and cultivate endophytic bacteria of the genus Conechaeta Coniochaeta sp.EA-9

[0020] Endophytic bacteria of the genus Conechaeta preserved on a slope at 4°C Coniochaeta The sp.EA-9 bacteria were activated and cultured, and the medium used was PDB medium. The microorganisms were picked from the inclined plane of the preservation test tube and inserted into 250ml Erlenmeyer flasks. Each bottle contained 100ml of sterilized medium. The inoculation process was carried out in a sterile environment Carried out; Sterilization conditions: 121°C, 30min. Placed at 25°C, cultivated on a shaker for 5 days, and set the speed at 150r min -1 ;

[0021] Medium preparation:

[0022] PDB medium: Take 200g of fresh commercially available potatoes, cut them into small pieces of 3-5mm squ...

Embodiment 2

[0026] Embodiment 2: The method for preparing ginsenoside Rd by transforming Panax notoginseng saponins by this microorganism is carried out as follows:

[0027] (1) Use conventional Martin's medium to activate and cultivate endophytic bacteria of the genus Conechaeta Coniochaeta sp.EA-9

[0028] Endophytic bacteria of the genus Conechaeta preserved on a slope at 4°C Coniochaeta The sp.EA-9 bacteria were activated and cultivated. The medium used was Martin's medium. The microorganisms were picked from the inclined plane of the preservation test tube and inserted into 250ml Erlenmeyer flasks. Each bottle contained 100ml of sterilized medium. Environment; Sterilization conditions: 121°C, 25min. Placed at 22°C, cultivated on a shaker for 4 days, and set the speed at 110r min -1 ;

[0029] Medium preparation:

[0030] Martin's Medium: KH 2 PO 4 1.0g, MgSO 4 ·7H 2 O0.5g, peptone 5.0g, glucose 10g, add water to volume 1L.

[0031] (2) Inoculate the activated bacteria in step...

Embodiment 3

[0034] Embodiment 3: The method for preparing ginsenoside Rd by transforming Panax notoginseng saponins by this microorganism is carried out as follows:

[0035] (1) Use conventional Martin's medium to activate and cultivate endophytic bacteria of the genus Conechaeta Coniochaeta sp.EA-9

[0036] Endophytic bacteria of the genus Conechaeta preserved on a slope at 4°C Coniochaeta The sp.EA-9 bacteria were activated and cultivated. The medium used was Martin's medium. The microorganisms were picked from the inclined plane of the preservation test tube and inserted into 250ml Erlenmeyer flasks. Each bottle contained 100ml of sterilized medium. Environment; Sterilization conditions: 121°C, 20min. Placed at 28°C, cultivated on a shaker for 3 days, and set the speed at 180r min -1 ;

[0037] Medium preparation:

[0038] Martin's Medium: KH 2 PO 4 1.0g, MgSO 4 ·7H 2 O0.5g, peptone 5.0g, glucose 10g, add water to volume 1L.

[0039] (2) Inoculate the activated bacteria in st...

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Abstract

The invention discloses a method for preparing ginsenoside Rd through biotransformation of panax notoginseng saponins. According to the method, coniochaetasp.EA-9 or crude enzyme secreted by the coniochaetasp.EA-9 is adopted to conduct biotransformation on panax notoginseng saponins, and the ginsenoside Rd and C-K are obtained. The method is easy to operate, high in conversion specificity, mild in reaction condition, little in microbial secondary metabolite, simple in extraction and fermentation product, high in conversion rate and capable of being used for large-scale preparation of the ginsenoside Rd and achieving industrialization of the ginsenoside Rd.

Description

technical field [0001] The invention relates to a method for obtaining ginsenoside Rd by biotransforming total saponins of Panax notoginseng. The method uses an enzyme substance secreted by microorganisms as a medium to modify the branched chain of glycosidic bonds in the skeleton structure of total saponins to obtain ginsenoside Rd. . Background technique [0002] Ginsenoside Rd is a diol-type ginsenoside, which has a wide range of biological activities and has unique effects on cardiovascular and cerebrovascular, anti-tumor, nervous and immune systems. For example, as a new type of calcium ion antagonist, it can specifically block receptors Dependent calcium ion channel, blocking calcium ion influx leads to neuronal cell death. Because of its complex structure, chemical synthesis is not yet feasible. It needs to be extracted from plants such as Panax notoginseng, ginseng, and Gynostemma pentaphyllum. However, due to the low content in plants and limited plant resources, l...

Claims

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Application Information

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IPC IPC(8): C12P33/20C12R1/645
Inventor 杨晓艳郭从亮崔秀明武双杨野王承潇曲媛
Owner KUNMING UNIV OF SCI & TECH
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