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Method for preparing icariin by using biphasic enzymatic reaction

A technology of icariin and enzymatic reaction, applied in the direction of fermentation, etc., can solve the problems of unsteady, sustainable and effective reaction process, waste of enzyme resources, etc., and achieve the effect of saving enzyme cost, high yield, and reducing the amount of enzyme

Pending Publication Date: 2021-06-15
XIAN GIANT BIOGENE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the enzyme reaction generally needs to be carried out in a complete aqueous solution environment, the products and by-products produced during the reaction may inhibit the efficiency of the enzyme reaction during the reaction, making the reaction process unable to be carried out stably, continuously and effectively, which indirectly wastes a lot of Enzyme resource

Method used

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  • Method for preparing icariin by using biphasic enzymatic reaction
  • Method for preparing icariin by using biphasic enzymatic reaction
  • Method for preparing icariin by using biphasic enzymatic reaction

Examples

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Embodiment 1

[0059] The enzyme preparation product that adopts in this patent is to form through liquid submerged fermentation purification and refinement by fungal strain, purchased from sunson Xiasheng Enzyme Preparation Co., Ltd. α-L-rhamnosidase (α-L-rhamnosidase, EC 3.2.1.40) is a hydrolase, which mainly acts on α-1,2, α-1,3, α-1,4, α- The 1,6 and α1-linked glycosidic bonds can hydrolyze the non-reducing α-L-rhamnosidic bond bound to the terminal of the substrate, releasing rhamnose and the corresponding ligand at the same time. The enzyme complies with GB1886. 174-2016 "National Food Safety Standard Enzyme Preparations for Food Industry". The enzyme of this product is a light yellow-brown powder preparation, without the enzyme activity of other components. Enzyme activity of this product: see product inspection report. The optimum temperature for enzyme products is 45°C-60°C, and the effective temperature is 40°C-65°C. PH value: the optimum pH value is 4.0-5.0, and the effective p...

Embodiment 2

[0071] Compared with Example 1, the present embodiment is only different in that the protective agent "starch" is removed.

[0072] (1) Preparation of Epimedium extract: Get crushed Epimedium and add 70% ethanol for extraction, the ratio of solid to liquid is 1:30, concentrate and dry under reduced pressure to obtain Epimedium extract;

[0073] (2) Two-phase enzymatic hydrolysis reaction: first add an appropriate amount of buffer to the enzyme coated with starch, stir in a water bath at 40°C for 30 minutes, then centrifuge at 5000 rpm for 10 minutes to remove the starch in the enzyme preparation, and perform the following reaction. Add the enzyme / Epimedium extract in 1:5 (g / g) to the chloroform / buffer (1:10) reaction system, and the solid-liquid ratio is 1:10. After shaking and mixing, carry out enzymatic hydrolysis reaction. Sampling and detection according to the reaction time, when the conversion rate of Epimedin C reaches more than 95% (the content of Epimedin C in the ra...

Embodiment 3

[0084] Compared with Example 1, this example differs only in the use of different organic phases.

[0085] (1) Preparation of Epimedium extract: Get crushed Epimedium and add 70% ethanol for extraction, the ratio of solid to liquid is 1:30, concentrate and dry under reduced pressure to obtain Epimedium extract;

[0086] (2) Two-phase enzymatic hydrolysis reaction: add starch-coated enzyme / Epimedium extract at 1:5 (g / g) to n-butanol / buffer (1:10) reaction system, solid-liquid The ratio is 1:10. After shaking and mixing, the enzymatic hydrolysis reaction was carried out. Sampling and detection according to the reaction time, when the conversion rate of Epimedin C reaches more than 95% (the content of Epimedin C in the raw material itself is relatively high, so for the accuracy of the data, the conversion rate of Epimedin C is used as the judgment of the reaction end point), Finish the reaction, cool to room temperature, separate the n-butanol phase and the buffer phase through...

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Abstract

The present application relates to a method for preparing icariin using a biphasic enzymatic reaction. The reaction uses an enzyme to convert epimedin A / B / C in an epimedium extract into an enzymatic hydrolysate icariin. The method comprises the following steps: preparing an epimedium extract; carrying out the biphasic enzymatic reaction, wherein a first phase is a buffer phase and the first phase has a good solubility with respect to epimedin A / B / C in the epimedium extract, and a second phase is an organic phase and the second phase has a good solubility with respect to icariin as the enzymatic hydrolysate; and separating and purifying the product icariin, wherein an enzyme used for the biphasic enzymatic reaction is coated with non-water-soluble starch, so that the enzyme is located at the phase interface of the first phase and the second phase when the biphasic enzymatic reaction is carried out.

Description

technical field [0001] The application belongs to the field of biochemical industry, and in particular relates to a method for preparing icariin from an epimedium extract by using a biphasic enzymatic reaction. Background technique [0002] Epimedium is a traditional Chinese medicine in my country. It has the functions of nourishing kidney and strengthening yang, strengthening muscles and bones, and enhancing immunity. It is often used to treat osteoporosis, promote regeneration and differentiation of myocardial cells, and delay aging. The main active ingredients in Epimedium are flavonoids, which mainly exist in the form of glycosides in Epimedium, among which icariin is a flavonoid with the above-mentioned effects that scholars have studied more in recent years one of the compounds. However, due to the difference in epimedium growth environment and picking period, the content of icariin varies greatly, and its basic content is usually below 2%. Relatively speaking, the c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/60
CPCC12P19/60
Inventor 范代娣范翠英段志广董杨芳黄蓉严建亚
Owner XIAN GIANT BIOGENE TECH CO LTD
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