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Method for preparing rare ginsenoside CK by fermenting protopanoxadiol with penicillium adametzii

A technology of protopanaxadiol and adapenicillium, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc.

Inactive Publication Date: 2014-11-12
天津市尖峰天然产物研究开发有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention relates to using aglycon protopanaxadiol as a substrate to carry out directional glycosylation with adapenicillium, and to prepare ginsenoside CK in large quantities [4-5] , there is no literature report at home and abroad

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] (1) Cultivation of microorganisms: inoculate Penicillium adabilis (3.3203) from the solid medium into the pre-prepared liquid potato medium, the liquid potato medium contains 1kg of glucose in 100L; place it in a fully automatic fermenter, The culture temperature is 24 degrees, the aeration ratio is 0.1% (v / v), the stirring rate is 360 rpm / min, and the fermentation culture is 36 hours;

[0019] (2) Dissolve protopanaxadiol in methanol to make 100g / L, take 3L of the substrate drug solution and add it to the fermentation broth prepared in step (1), culture at 24°C, stir at 360rpm / min, and continue to ferment and cultivate 120 hours;

[0020] (3) Separation and purification of transformed products: collect the filtrate after centrifugation and filtration of the fermentation broth, concentrate under reduced pressure to 1 / 10 of the original volume, separate by HP20 macroporous resin column chromatography, and elute with deionized water to 2 times the column retentio...

Embodiment 2

[0025] (1) Cultivation of microorganisms: inoculate Penicillium adabilis (3.3203) from the solid medium into the pre-prepared liquid potato medium, and the liquid potato medium 100L contains 5kg of glucose; put it in a fully automatic fermenter, The culture temperature is 32 degrees, the aeration ratio is 10% (v / v), the stirring rate is 100 rpm / min, and the fermentation culture is 72 hours;

[0026] (2) Dissolve protopanaxadiol in 50% ethanol water solution by volume to prepare 10g / L, take 3L of the substrate drug solution and add it to the fermentation broth prepared in step (1), culture at 32°C, and stir at 100rpm / min, continue to ferment and cultivate for 48 hours;

[0027] (3) Separation and purification of transformed products: collect the filtrate after centrifugation and filtration of the fermentation broth, concentrate under reduced pressure to 1 / 20 of the original volume, separate by D101 macroporous resin column chromatography, and elute with deionized wate...

Embodiment 3

[0029] (1) Culture of microorganisms: inoculate Penicillium adabilis (3.3203) from the solid medium into the pre-prepared liquid potato medium, the liquid potato medium contains 2kg of glucose in 100L; put it in a fully automatic fermenter, The culture temperature is 28 degrees, the aeration ratio is 5% (v / v), the stirring rate is 220rpm / min, and the fermentation culture is 48 hours;

[0030] (2) Dissolve protopanaxadiol in methanol aqueous solution with a concentration of 50% by volume to make 50mg / ml, take 3L of the substrate solution and add it to the fermentation broth prepared in step (1), culture at 28 degrees, and stir at For 220rpm / min, continue to ferment and cultivate for 96 hours;

[0031] (3) Separation and purification of the transformed product: collect the filtrate after centrifugation and filtration of the fermentation broth, concentrate it under reduced pressure to 1 / 15 of the original volume, separate it by AB-8 macroporous resin column chromatogra...

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Abstract

The invention provides a preparation method of ginsenoside CK and relates to a method for preparing 20-O-beta-D-glucopyranosyl-20-protopanoxadiol (ginsenoside CK) by fermenting and converting protopanoxadiol in a full-automatic fermentation tank by means of a bacterial strain. The method comprises the following steps: (1) culture of microorganisms; (2) fermentation and conversion of microorganisms; and (3) chromatographic separation and purification by macroporous resin to finally obtain a compound with the chemical name: 20-O-beta-D-glucopyranosyl-20-protopanoxadiol (ginsenoside CK).

Description

technical field [0001] The present invention relates to the technical field of medicine, in particular to a method for preparing ginsenoside CK (20-O-β-D-glucopyranosyl-20-protopanaxadiol) by fermenting and transforming protopanaxadiol from Penicillium adabilis and A method for preparing ginsenoside CK (20-O-β-D-glucopyranosyl-20-protopanaxadiol) by separating and preparing ginsenoside CK from the subsequent fermentation broth. Background technique [0002] Ginsenosides are the material basis for the efficacy of traditional Chinese medicine ginseng. So far, more than 40 kinds of ginsenoside monomers have been isolated, mainly containing ginsenosides Rb1, Rb2, Rc, Rg1 and Re, accounting for more than 80% of the total ginsenosides [1] , the remaining saponins only account for a small fraction of the total amount of saponins, and are called rare ginsenosides. Although the content of these saponins is very small, they have unique pharmacological activities. Ginsenoside CK is ...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P33/20C12R1/80
Inventor 刘岱琳刘丹孙华庚
Owner 天津市尖峰天然产物研究开发有限公司
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