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Method for preparing rare ginsenoside CK from protopanaxadiol ginsenoside through fermentation of microbacterium oxydans

A technology of oxidative microbacteria and ginsenosides, applied in the field of medicine, can solve the problems of harsh culture conditions and low yield, and achieve the effect of improving the catalytic effect

Active Publication Date: 2016-08-10
NORTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of these intestinal microorganisms are anaerobic bacteria, and the external culture conditions are relatively harsh and the yield is not high. Therefore, a high-efficiency, easy-to-cultivate and ferment and low-cost strain is used to convert and hydrolyze sugar chains to prepare rare ginseng Saponins have important application value

Method used

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  • Method for preparing rare ginsenoside CK from protopanaxadiol ginsenoside through fermentation of microbacterium oxydans
  • Method for preparing rare ginsenoside CK from protopanaxadiol ginsenoside through fermentation of microbacterium oxydans
  • Method for preparing rare ginsenoside CK from protopanaxadiol ginsenoside through fermentation of microbacterium oxydans

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Experimental program
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Effect test

Embodiment 1

[0029] (1) Cultivation of microorganisms: inoculate Microbacterium oxidans (CCTCC AB 205576) from a solid medium into a pre-configured 200L microbacterium fermentation medium containing 10 g / L glucose and 20 g peptone. L, beef extract 20g / L; placed in a fermentation tank, sterilized online (121°C, 20min), microorganism culture temperature 30°C, aeration ratio 5% (v / v), stirring rate 200rpm / min, fermentation time 48h;

[0030] (2) CK transformation preparation: Dissolve ginsenoside Rb1 with a purity of 90% diol group in 50% methanol solution, prepare 6L of 100g / L saponin solution and put it into the above-mentioned fermented microbial culture species, and add 10g heteropoly acid H 3 PW 12 O 40 ·6H 2 O, carry out constant temperature stirring and catalytic reaction. The stirring rate is 200 rpm, the reaction temperature is 30°C, and it is maintained for 96 hours;

[0031] (3) Collection and purification of reaction product CK: Centrifuge the above conversion product, and discard the...

Embodiment 2

[0034] (1) Cultivation of microorganisms: Inoculate Microbacterium oxidans (CCTCC AB 205576) from a solid medium into a pre-configured 150L microbacterium fermentation medium containing 20g / L glucose and 30g / peptone L, beef extract 30g / L; placed in a fermentation tank, sterilized online (121°C, 20min), microorganism cultivation temperature 30°C, aeration ratio 3% (v / v), stirring rate 200rpm / min, fermentation time 120h;

[0035] (2) CK transformation preparation: Dissolve ginsenoside saponins Rd with a purity of 90% diol group in 50% methanol solution, prepare 3L of 100g / L saponin solution and put it into the above-mentioned fermented microbial culture species, and Add 20g heteropoly acid H 4 SiW 12 O 40 ·25H 2 O, carry out constant temperature stirring and catalytic reaction. The stirring rate is 250rpm, the reaction temperature is 30°C, and it is maintained for 96h;

[0036] (3) Collection and purification of reaction product CK: Centrifuge the above conversion product, and disca...

Embodiment 3

[0038] (1) Cultivation of microorganisms: inoculate Microbacterium oxidans (CCTCC AB 205576) from a solid medium into a pre-configured 100L microbacterium fermentation medium containing 30g / L glucose and 30g / L peptone L, beef extract 25g / L; placed in a fermentation tank, sterilized online (121°C, 20min), microorganism culture temperature 35°C, aeration ratio 5% (v / v), stirring rate 200rpm / min, fermentation time 120h;

[0039] (2) CK transformation preparation: Dissolve ginsenoside saponins Rb2 with a purity of 95% diol group in 80% methanol solution, make 2L of 100g / L saponin solution and put it into the above-mentioned fermented microorganism culture species, and Add 40g heteropoly acid H 3 FeW 12 O 40 ·19H 2 O, carry out constant temperature stirring and catalytic reaction. The stirring rate is 250rpm, the reaction temperature is 40℃, and it is maintained for 72h;

[0040] (3) Collection and purification of reaction product CK: Centrifuge the above conversion product, and discar...

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Abstract

The invention relates to a method for preparing rare ginsenoside CK from protopanaxadiol ginsenoside through fermentation of microbacterium oxydans, in particular to a method for preparing 20-O-beta-D-glucopyranose-20-protopanaxadiol ginsenoside from protopanaxadiol ginsenoside Rb1, Rd and the like converted by utilizing fermentation of microbiological strains in a fully-automatic fermentation tank, belongs to the technical field of medicine, and comprises the steps: (1) fermentation cultivation of bacterial strains; conversion of raw materials Rb1, Rd and the like by utilizing fermented microorganisms and taking heteropolyacid HxYW12O40.nH2O with a Keggin structure as a catalyst, wherein Y is selected from P, Si, Fe or Zn, x is 3 or 4, and n is a positive integer ranging from 0 to 30; (3) preparative chromatographic separation and purification of conversion products, so as to finally obtain a compound with the chemical name being 20-O-beta-D-glucopyranose-20-protopanaxadiol ginsenoside.

Description

Technical field [0001] The invention relates to a method for preparing rare ginsenoside CK by fermentation of diol group ginsenosides by Microbacterium oxidans, and specifically relates to a method for Microbacterium oxidans to specifically catalyze glycol type saponins and transform into 20-0-β-D-pyran The method for glucosyl-20-diol group ginsenosides belongs to the technical field of medicine. technical background [0002] In recent years, ginseng has been widely used in the pharmaceutical industry due to its biologically active saponin components. At present, a variety of active saponins have been discovered and have been used in the treatment and prevention of diseases, such as anti-aging, anti-tumor, anti-inflammatory, and myocardial protection. Most saponins are composed of the diol group ginsenosides (Rb1, Rb2, Rc, Rd) and protoginsenosides (Re, Rg1), and the total content accounts for more than 80% of the saponins. It is reported that these saponins account for the main...

Claims

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Application Information

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IPC IPC(8): C12P33/20C12R1/01
CPCC12P33/20
Inventor 范代娣段志广惠俊峰朱晨辉米钰马沛李伟娜马晓轩
Owner NORTHWEST UNIV
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