Method for large-scale production of rare ginsenoside from enzyme-catalysis panaxatriol ginsenoside

A technology of ginsenoside and catalytic triol, applied in the direction of fermentation and the like, can solve the problems of low catalytic efficiency, difficulty in large-scale production and the like, and achieve the effects of greatly improved conversion efficiency, high yield and environmental friendliness

Inactive Publication Date: 2017-03-08
NORTHWEST UNIV
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  • Summary
  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

The enzymes reported in the literature that can catalyze the transformation into rare ginsenosides mainly include cellulase, naringinase, pectinase, amylase, glucoamylase, etc. The raw materials for catalysis are generally mixed saponins extracted from Chinese medicinal materials, but the catalytic efficiency Not high, difficult to mass-produce

Method used

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Examples

Experimental program
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Effect test

preparation example Construction

[0014] (2) Preparation of enzyme preparation: take two or more of cellulase, naringinase, pectinase, amylase, and glucoamylase to form an enzyme preparation in a certain ratio, and wait until the temperature in the reactor drops to room temperature Then put it into the reactor, where the total concentration of enzyme is between 50g / L~200g / L.

[0015] (3) The conversion reaction temperature is 25℃~50℃, the stirring speed is 100rpm~360rpm, and the catalytic reaction is 24h~120h under corresponding conditions.

[0016] (4) Separation and purification of conversion products: Because the rare ginsenosides are not easily soluble in water, they are soluble in organic solvents such as ethyl acetate, n-butanol, ethyl butyrate, ethyl lactate, and chloroform, and the conversion rate of the product is relatively high. high. After centrifugal filtration of the conversion solution, the precipitate is collected, the precipitate is washed several times with phosphate buffer, and then a mixture of...

Embodiment 1

[0019] Add 30L of ionic liquid 1-ethyl-3-methyl-imidazole acetate into a reactor with an effective volume of 50L, and then add 1.5kg of triol ginsenoside (detected by ultraviolet method) with a content of 85.7% ( Impurities are mainly polysaccharides), pass N 2 , The ventilation flow is 25L / min, lasting 5min. After cooling to below room temperature, 3 kg of enzyme preparation with cellulase / pectinase ratio of 1:1 was added. Keep the temperature of the reactor at 25°C, the stirring speed at 100rpm, and the catalytic reaction for 120h. After the reaction, put all the liquid out of the container and let it stand for 12 hours, discard the supernatant, add phosphate buffer solution for washing several times, and then add water to the precipitate with a 1:2 solution of ethyl acetate for extraction, and repeat the extraction twice. The organic phase extract was collected and concentrated under reduced pressure to 1 / 20 of the original volume. Use a preparative column for separation a...

Embodiment 2

[0022] Add 60L of ionic liquid 1-ethyl-3-methylimidazole diethyl phosphate into a reactor with an effective volume of 100L, and then add 30kg of triol ginsenoside (detected by ultraviolet method) with a volume of 85.3% ( Impurities are mainly polysaccharides), pass N 2 , The ventilation flow is 200L / min, lasting 1min. After cooling to below temperature, 12 kg of enzyme preparation with cellulase / glucoamylase ratio of 1:1 was added. Keep the temperature of the reactor at 30°C, the stirring speed at 360 rpm, and the catalytic reaction for 72 hours. After the reaction, put all the liquid out of the container and let stand for 12 hours, discard the supernatant, add phosphate buffer solution to wash several times, and then add water to the precipitate with a 2:3 solution of ethyl butyrate for extraction, repeated extraction 3 times , Collect the organic phase extract, concentrate under reduced pressure to 1 / 12 of the original volume. Use a preparative column for separation and pur...

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Abstract

The invention discloses a novel process for producing rare ginsenoside from panaxatriol ginsenoside in ionic liquid through enzyme-catalysis directional transformation and for separating and purifying the rare ginsenoside. Specifically, the method comprises the following steps: dissolving panaxatriol ginsenoside in the ionic liquid and adding the ionic liquid dissolved with the panaxatriol ginsenoside to a reaction kettle, and then adding a complex enzyme preparation in a quantitative mode; conducting the directional transformation under corresponding conditions, so that a great amount of rare ginsenoside is obtained; and collecting a finished product and purifying the product, so that various varieties of rare ginsenoside which are higher than 95% in purity are obtained. The process is high in yield of the rare ginsenoside, environment-friendly, relatively simple in downstream purifying process and conducive to large-scale production.

Description

Technical field [0001] The invention relates to a method for large-scale production of rare ginsenosides by enzyme-catalyzed triol group ginsenosides, which belongs to the field of biochemical industry. Background technique [0002] Ginsenosides are the main medicinal components of plants in the genus Araliaceae (Ginseng, American ginseng, Panax notoginseng, etc.). They have various pharmacological effects such as enhancing immunity, anti-tumor, anti-fatigue, anti-anoxia, anti-aging, and lowering blood sugar. At present, there are more than 150 kinds of ginsenosides isolated from ginseng. They can be roughly divided into two categories: common ginsenosides and rare ginsenosides. Among them, common ginsenosides refer to ginseng that has a high content in the genus Ginseng and is relatively easy to separate and purify. The saponins mainly include protopanaxadiol type ginsenosides Rb1, Rb2, Rb3, Rc, Rd and protopanaxatriol type ginsenosides Re, Rg1, Rg2, etc. Rare ginsenosides are ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P33/20C12P33/00
CPCC12P33/20C12P33/00
Inventor 惠俊峰范代娣马沛段志广米钰朱晨辉马晓轩李伟娜
Owner NORTHWEST UNIV
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