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Method for massively producing ginsenoside CK by catalyzing protopanaxadiol ginsenoside through enzyme

A technology for enzymatically catalyzing diols and ginsenosides, applied in the field of biochemical industry, can solve the problems of inability to scale up large-scale production and low catalytic efficiency, and achieve the effects of improved catalytic effect, high yield and low production cost

Inactive Publication Date: 2016-09-07
NORTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, they all catalyze the reaction in the form of a single enzyme. The raw materials for catalysis are generally mixed saponins extracted from Chinese medicinal materials. The catalytic efficiency is low, and large-scale scale-up production cannot be carried out.

Method used

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  • Method for massively producing ginsenoside CK by catalyzing protopanaxadiol ginsenoside through enzyme
  • Method for massively producing ginsenoside CK by catalyzing protopanaxadiol ginsenoside through enzyme
  • Method for massively producing ginsenoside CK by catalyzing protopanaxadiol ginsenoside through enzyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Add 60L volume, pH 5, containing 1Mm Ca 2+ Ionic phosphate buffer, then add 3kg of ginsenoside Rb1 with a purity of 85% (impurities are other types of ginsenosides or polysaccharides), pass N 2 , the ventilation flow rate is 50 L / min, and lasts for 5 minutes, then stops the ventilation and starts the heating-in-line sterilization, and the sterilization condition is 121°C, and maintains for 20 minutes. After cooling down to below room temperature, add pectinase: cellobiase 6:4 compound enzyme 2kg, and 20g heteropoly acid H 3 PW 12 o 40 ·5H 2 O. Keep the temperature of the fermenter at 25° C., the stirring speed at 200 rpm, and catalyze the reaction for 96 hours. After the reaction, put all the liquid in the container and let it stand for 12 hours, discard the supernatant, add phosphate buffer to wash several times, then add water:ethyl acetate to the precipitate for extraction, repeat the extraction three times, and collect The organic phase extract was concentrate...

Embodiment 2

[0029] It is 200L, pH is 7, contains 4mM Ca 2+ Ionic phosphate buffer, then add 30kg of ginsenoside Rd with a purity of 90% (impurities are other types of ginsenosides or polysaccharides), pass N 2 , the ventilation flow rate is 150 L / min, and lasts for 5 minutes, then stops the ventilation and starts to heat in-line sterilization, the sterilization condition is 121°C, and maintains for 20 minutes. After cooling down to below room temperature, add pectinase: 25kg of compound enzyme with cellobiase ratio of 8:2, and 10g of heteropolyacid H 4 SiW 12 o 40 18H 2 O. Keep the temperature of the fermenter at 40°C, the stirring speed at 200rpm, and catalyze the reaction for 120h. After the reaction, put all the liquid in the container and let it stand for 12 hours, discard the supernatant, add phosphate buffer to wash several times, then add water:ethyl acetate to the precipitate for extraction, repeat the extraction three times, and collect The organic phase extract was concent...

Embodiment 3

[0031] It is 300L that volume is added in the full-automatic fermentor of 500L, pH is 9, contains the phosphate buffer solution of 5mM Ca2+ ion, adds the ginsenoside Rb2 (impurity is other types of ginsenosides or polysaccharides), through N 2 , the ventilation flow rate is 200 L / min, and lasts for 5 minutes, then stops the ventilation and starts to heat in-line sterilization, the sterilization condition is 121°C, and maintains for 20 minutes. After cooling down to below room temperature, add pectinase: cellobiase 9:1 compound enzyme 30kg, and 40g heteropolyacid H 3 FeW 12 o 40 28H 2 O. Keep the temperature of the fermenter at 50°C, the stirring speed at 200rpm, and catalyze the reaction for 120h. After the reaction, put all the liquid in the container and let it stand for 12 hours, discard the supernatant, add phosphate buffer to wash several times, then add water:ethyl acetate to the precipitate for extraction, repeat the extraction three times, and collect The organic...

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Abstract

The invention relates to a novel technology for producing CK through enzymatic catalysis and oriented conversion by taking protopanaxadiol ginsenoside as a raw material and separating and purifying the CK. The technology specifically comprises the steps that the protopanaxadiol ginsenoside is dissolved into deionized water, the solution is put into a fermentation tank, and after N2 is introduced into the tank for protection, on-line sterilization is conducted for 20 min at 121 DEG C; a complex enzyme and a catalytic quantity of a heteopoly acid (HxYW12O40.nH2O) catalyst with a Keggin structure are quantitatively added, wherein a mixed enzyme prepared by mixing pectinase with cellobiase in different proportions is adopted as the complex enxyme, Y is selected from P, Si, Fe and Zn, x is 3 or 4, and n is a positive integer ranging from 0 to 30; oriented conversion is conducted under the corresponding conditions to obtain a large quantity of the CK product, and the product is collected and purified to obtain the CK with the purity of 90% or above. The method is high in yield, friendly to the environment and simple in aftertreatment, and a foundation is laid for industrialized production.

Description

technical field [0001] The invention relates to a method for enzymatically catalyzing diol group ginsenosides for large-scale production of ginsenoside CK, which belongs to the field of biochemical industry. Background technique [0002] Ginseng (Ginseng), as a Chinese herbal medicine, has been used in Asia for thousands of years, and its main function is to strengthen the body and prolong life. Ginsenoside is one of the iconic active ingredients in ginseng. Currently, there are more than 150 ginsenosides found in roots, leaves, stems and flowers of ginseng, most of which have biological activities, including anti-tumor, anti-aging, improving memory , anti-inflammatory and liver protection effects. However, a large number of natural saponins (Rb1, Rg1, Re, R1, Rd) contained in ginseng are difficult to be absorbed, and need to be digested by intestinal microorganisms before they can be absorbed by the intestine. Studies have shown that the conversion of saponins into desuga...

Claims

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Application Information

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IPC IPC(8): C12P33/20A61K31/7028
CPCC12P33/20A61K31/7028
Inventor 范代娣段志广惠俊峰米钰马沛朱晨辉李伟娜马晓轩
Owner NORTHWEST UNIV
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