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Method for preparing rare ginsenoside CK from transformed ginsenoside Rb1 and use thereof

A rare technology of ginsenoside, which is applied in the field of preparation of rare ginsenoside CK, which can solve the problems of low conversion efficiency and failure to meet production requirements, and achieve the effects of high conversion rate, improved absorption and utilization rate, and short conversion cycle

Active Publication Date: 2016-02-03
HUNAN INSTITUTE OF ENGINEERING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although some bifidobacteria have the ability to transform ginsenoside Rb1 to prepare ginsenoside CK, their transformation efficiency is low and cannot meet the production requirements

Method used

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  • Method for preparing rare ginsenoside CK from transformed ginsenoside Rb1 and use thereof
  • Method for preparing rare ginsenoside CK from transformed ginsenoside Rb1 and use thereof
  • Method for preparing rare ginsenoside CK from transformed ginsenoside Rb1 and use thereof

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Experimental program
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Effect test

Embodiment 1

[0028] Bifidobacterium culture and β-glucosidase extraction

[0029] 1. MRS liquid medium is used for anaerobic cultivation of bifidobacteria; the components of MRS medium are: 10.0g casein peptone, 10.0g beef extract, 5.0g yeast powder, 5.0g glucose, 5.0g sodium acetate, 2.0 g diammonium citrate g, Tween 801.0g, dipotassium hydrogen phosphate 2.0g, sodium sulfate heptahydrate 0.2g, manganese sulfate monohydrate 0.05g, calcium carbonate 20.0g, agar 15.0g, distilled water 1.0L, pH6.8. MRS medium was sterilized at 121°C for 20min.

[0030] 2. The specific method of fermentation and cultivation of bifidobacteria is as follows: inoculate bifidobacteria on solid MRS medium, culture anaerobically at 37°C for 24-48 hours, and then pick a single colony of bifidobacteria and inoculate it in MRS liquid medium In the method, under the condition of 37° C., shake anaerobic culture for 24-48 hours to obtain a bifidobacterium fermentation liquid.

[0031] 3. Centrifuge the bifidobacterium ...

Embodiment 2

[0037] Take by weighing an appropriate amount of ginsenoside Rb1 and dissolve it in a small amount of methanol, then mix it with 0.02mol / L, pH4.0-6.0 acetic acid-sodium acetate buffer solution to prepare a 10mg / mL ginsenoside Rb1 solution; The prepared β-glucosidase solution was prepared into 20U / mL enzyme solution; then the ginsenoside Rb1 solution was mixed with 20U / mL β-glucosidase solution according to the volume ratio of 10:1, the pH was 5.0, 45°C Under the condition of reaction for 24 hours, then inactivated in a water bath at 80°C for 20 minutes, centrifuged at 8000 rpm for 5 minutes at 25°C, and the supernatant was dried at 60°C to obtain the rare ginsenoside CK.

Embodiment 3

[0039] Take by weighing an appropriate amount of ginsenoside Rb1 and dissolve it in a small amount of methanol, then mix it with 0.02mol / L, pH4.0-6.0 acetic acid-sodium acetate buffer solution to prepare a 10mg / mL ginsenoside Rb1 solution; The prepared β-glucosidase solution was prepared into 20U / mL enzyme solution; then the ginsenoside Rb1 solution was mixed with 20U / mL β-glucosidase solution according to the volume ratio of 10:1, the pH was 4.5, 55°C Under the condition of reaction for 24 hours, then inactivated in a water bath at 80°C for 20 minutes, centrifuged at 8000 rpm for 5 minutes at 25°C, and the supernatant was dried at 60°C to obtain the rare ginsenoside CK.

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Abstract

The invention discloses a method for preparing rare ginsenoside CK from transformed ginsenoside Rb1 and a use thereof. Ginsenoside Rb1 is transformed into rare ginsenoside CK by beta-glucosidase produced by bifidobacteria and a conversion rate is in a range of 62-68%. The transformed rare ginsenoside CK has very polarity, can be easily absorbed by the human body, and can enter into liver by blood circulation so that metabolism is finished and unique pharmacological effects are obtained. Bifidobacteria are edible probiotics and can produce glycosidase with high catalysis activity, selectivity and stability. The beta-glucosidase produced by bifidobacteria can transform ginsenoside Rb1 into rare ginsenoside CK, and has the advantages of high conversion rate, less side effect, safety, reliability, no pollution and industrial production easiness. The method has a low cost, simple processes and a short conversion period. Beta-glucosidase produced by bifidobacteria has an important application value in preparation of rare ginsenoside CK and the method utilizes the application value.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a method for preparing rare ginsenoside CK (Compound K) by using β-glucosidase produced by bifidobacteria to transform ginsenoside Rb1, and also relates to β-glucosidase produced by the bifidobacteria. Application of glucosidase in preparing rare ginsenoside CK with Rb1 as substrate. Background technique [0002] Ginsenosides are triterpene glycoside compounds and are the main medicinal active ingredients of ginseng. Different types of ginsenosides are derived according to the types and quantities of glycosidic bonds at positions C-3, C-6 and C-20 on the aglycone. According to its content in plants, it is divided into major ginsenosides (Majorginsenosides, such as Rb1, Rb2, Rc, Rd, Re and Rg1, etc.) and rare ginsenosides (Rareginsenosides, such as Rg3, Rh1, Rh2, F1, F2, CK, C-O , CY and C-Mc) etc. Among them, rare ginsenosides refer to saponins whose content in ginseng is ex...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P33/20C12R1/01
Inventor 张儒张变玲李谷才谢涛
Owner HUNAN INSTITUTE OF ENGINEERING
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