Method for converting astragalosides to prepare cycloastragenol by two-step enzymolysis

A technology of astragaloside IV and cycloastragenol, which is applied in the field of medicine and chemical industry, can solve the problems of three-membered rings that are easy to crack astragalol, and achieve the effects of reduced energy consumption, high purity, and mild preparation conditions

Active Publication Date: 2017-08-18
BEIJING UNIV OF CHEM TECH
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  • Abstract
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Problems solved by technology

[0008] The present invention chooses to adopt a two-step enzymatic hydrolysis method to convert astragaloside IV to prepare cycloastragenol. The purpose of using this mild biological enzymatic method to prepare cycloastragenol is to: 1. Solve the formation of three-membered rings that are easy to crack during the preparation process of traditional chemical methods The problem with the by-product astragalol

Method used

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  • Method for converting astragalosides to prepare cycloastragenol by two-step enzymolysis
  • Method for converting astragalosides to prepare cycloastragenol by two-step enzymolysis
  • Method for converting astragalosides to prepare cycloastragenol by two-step enzymolysis

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Embodiment 1

[0035] Accurately prepare 0.2 mol / L disodium hydrogen phosphate solution and 0.1 mol / L citric acid solution, mix the two solutions at a volume ratio of 1.06:1, and adjust the pH of the buffered saline solution to 5.0 with hydrochloric acid.

[0036] The first step of enzymatic hydrolysis: accurately measure 100mL of enzymatic hydrolysis buffer saline solution and place it in a 250mL beaker, and add 100mg of astragaloside IV with a mass percentage of 10% to it, and heat the system after the substrate astragaloside IV is completely dissolved to 45°C, then add 0.05g of cellulase that has been heated to 45°C into the system, and adjust the pH of the system to 4.6 with hydrochloric acid, and finally place the reaction system on a magnetic heating and stirring instrument at a stirring speed of 200r / min , The temperature is 45 ℃ and fully reacted for 60h. Afterwards, 1 mL of the sample was passed through a 0.22 μm filter membrane and detected by high-performance liquid chromatography...

Embodiment 2

[0041] Accurately prepare 0.2 mol / L disodium hydrogen phosphate solution and 0.1 mol / L citric acid solution, mix the two solutions at a volume ratio of 1.06:1, and adjust the pH of the buffered saline solution to 5.0 with hydrochloric acid.

[0042] The first step of enzymatic hydrolysis: accurately measure 100mL of enzymatic hydrolysis buffer saline solution and place it in a 250mL beaker, and add 100mg of astragaloside IV with a mass percentage of 10% to it, and heat the system after the substrate astragaloside IV is completely dissolved to 50°C, then add 0.05g of cellulase that has been heated to 50°C into the system, and adjust the pH of the system to 5.0 with hydrochloric acid, and finally place the reaction system on a magnetic heating stirring instrument at a stirring speed of 200r / min , The temperature is 50 ℃ and fully reacted for 48h. Afterwards, 1 mL of the sample was passed through a 0.22 μm filter membrane and detected by high-performance liquid chromatography. Th...

Embodiment 3

[0047] Accurately prepare 0.2 mol / L disodium hydrogen phosphate solution and 0.1 mol / L citric acid solution, mix the two solutions at a volume ratio of 1.06:1, and adjust the pH of the buffered saline solution to 5.0 with hydrochloric acid.

[0048] The first step of enzymatic hydrolysis: accurately measure 100mL of enzymatic hydrolysis buffer saline solution and place it in a 250mL beaker, and add 100mg of astragaloside IV with a mass percentage of 10% to it, and heat the system after the substrate astragaloside IV is completely dissolved to 55°C, then add 0.05g of cellulase that has been heated to 55°C into the system, and adjust the pH of the system to 5.5 with hydrochloric acid, and finally place the reaction system on a magnetic heating and stirring instrument at a stirring speed of 200r / min , The temperature is 55 ℃ and fully reacted for 72h. Afterwards, 1 mL of the sample was passed through a 0.22 μm filter membrane and detected by high-performance liquid chromatography...

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Abstract

The invention discloses a method for converting astragalosides to prepare cycloastragenol by two-step enzymolysis; the method comprises the specific steps of using astragalosides as a substrate, using two different hydrolases to hydrolyze and break beta-xyloside bond at site C3 of the astragalosides and beta-glucoside bond at the site C6 respectively to obtain aglycone cycloastragenol of the astragalosides, extracting, performing silica-gel column chromatography, recrystallizing with ethanol, and purifying to obtain the finished cycloastragenol up to 95% and higher in purity. The problem that damage of three-membered ring structure of astragalosides during the preparation of cycloastragenol by a chemical process causes massive byproducts is solved, the defects of traditional cycloastragenol preparation methods, such as low substrate conversion rate, step complexity and environmental pollution, are overcome. The method has the advantages that substrate specificity is high, the substrate astragalosides is completely converted, the steps are simple, the conditions are mild, the cost is low, and the method is a mild biological preparation method, has no environmental pollution and is suitable for industrial production.

Description

technical field [0001] The invention belongs to the field of medicine and chemical industry, and in particular relates to a method for preparing cycloastragenol by converting astragaloside IV through a two-step enzymatic hydrolysis method. Background technique [0002] Astragaloside IV is the main active ingredient of the traditional Chinese medicine Astragalus, which has various effects such as improving immunity, scavenging free radicals, lowering blood pressure, protecting liver and diuresis, and antibacterial. Cycloastragenol is the aglycone of astragaloside. Studies have shown that it has anti-oxidation, immunity-enhancing and anti-aging effects, and it is easier to absorb in the intestine and permeate the membrane than astragaloside. Another study shows that astragaloside exerts its drug effect in the body mainly by being partially decomposed into cycloastragenol. The most important thing is that cycloastragenol is currently the only active molecule in the world that ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P33/20C07J53/00
CPCC07J53/004C12P33/20
Inventor 袁其朋程磊雨
Owner BEIJING UNIV OF CHEM TECH
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