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Process for preparing low sugar icariin or aglycone by enzymic hydrolysis of icariin glycosylate

A technology of icariin and huoside sugar, applied in the direction of fermentation, etc., can solve the problems of poor purpose, destructive glucoside, pollution, etc., achieve the effect of strong purpose, overcome destructiveness, and increase enzyme production

Inactive Publication Date: 2004-02-11
金凤燮
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The activity of polysaccharide-based icariin is low, while the activity of low-glycosyl icariin or icariin is higher. In order to obtain icariin or low-glycosyl icariin, acid-base hydrolysis has been reported (Sun Pengyue etc., Chinese Journal of Medicinal Chemistry, 1998, 30 (4), 281-284), but described method is destructive to glucoside, serious pollution, purpose poor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] a. Take the high-temperature aerobic bacteria Clostridium thermocopriea (document 1) in a medium containing 3% corn flour, 1% enzyme-producing inducer-Epimedium extract (dry matter), shake at a temperature of 60°C Cultivate for 30-60 hours, sterilize by centrifugation; add 65% saturated ammonium sulfate to precipitate the enzyme protein in the supernatant (containing the enzyme mixture), dialyze with 0.02M, pH5 acetate buffer, centrifuge to remove the slag, freeze-dry, Get glucosidase.

[0042] b. Mix 0.2 gram of the above-mentioned glucosidase, 3 grams of the mixture of icariin 1, 2, and 3 or the monomer, and 150 ml of acetate buffer (0.2M, pH 5.0) to make icariin 1, 2 The mixture of , 3 accounts for 0.1-10% of the total reaction volume, and is stirred and reacted at a temperature of 75° C. for 1 hour.

[0043] c. After the reaction, 300 ml of ethanol was added to precipitate the enzyme protein, filtered to remove the enzyme protein precipitate, and the filtrate was e...

Embodiment 2

[0046] a. Take the high-temperature aerobic bacteria Clostridium thermocopriea (document 1) in a medium containing 4% corn flour, 0.2% enzyme-producing inducer-Epimedium extract (dry matter), and shake at a temperature of 60°C Cultivate for 30-60 hours, sterilize by centrifugation, add ethanol to the supernatant (containing the mixed solution of enzyme) to precipitate the enzyme protein, and freeze-dry to obtain the glucosidase.

[0047] b. Mix 0.3 gram of the above-mentioned glucosidase, 2 grams of the mixture of icariin 1, 2, and 3, and 120 milliliters of acetic acid buffer (0.2M, pH5.0) to make the icariin 1, 2, and 3 The mixture accounts for 0.1-10% of the total reaction volume, and the reaction is stirred and reacted at a temperature of 5° C. for 36 hours.

[0048] c. After the reaction, 300 ml of ethanol was added to precipitate the enzyme protein, filtered to remove the enzyme protein precipitate, and the filtrate was evaporated to dryness under reduced pressure to obta...

Embodiment 3

[0051]a. Take the high-temperature aerobic bacteria Clostridium thermocopriea (document 1) in a medium containing 3% corn flour, 1% enzyme-producing inducer-ginseng extract (dry matter), and shake culture at a temperature of 60°C for 30 -60 hours, centrifuge to sterilize, add ethanol to the supernatant (containing the mixed solution of enzyme) to precipitate the enzyme protein, collect the protein, and freeze-dry to obtain the glucosidase.

[0052] b. Mix 0.4 gram of above-mentioned glucosidase, 2 gram of icariin 4, 5, 6 mixture, 120 milliliters of phosphate buffer (0.2M, pH5.0) to make the icariin 4, 5, 6 The mixture accounts for 0.1-10% of the total reaction volume, and the reaction is stirred for 18 hours at a temperature of 60°C.

[0053] c. After the reaction, 300 ml of ethanol was added to precipitate the enzyme protein, the protein precipitate was removed by filtration, and the filtrate was evaporated to dryness under reduced pressure to obtain 1.5 g of crude product of...

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PUM

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Abstract

The enzymic hydrolysis process of icariin glycosylate to prepare low sugar icariin or aglycone includes the following steps: taking enzyme capable of hydrolyzing icariin glycosylate; mixing the enzyme capable of hydrolyzing icariin glycosylate, icariin and buffering liquid for reaction for 1-36 hr at temperature 4-75 deg.c and pH 3-9; and extracting low sugar icariin or aglycone. The said process has the advantages of no pollution, high low sugar icariin or aglycone converting rate and less damage to effective component.

Description

Technical field: [0001] The invention relates to a method for preparing low-sugar icariin or aglycon, in particular to a method for preparing low-sugar icariin or aglycon by hydrolyzing icariin glycosyl with enzyme. Background technique: [0002] Epimedium is a commonly used traditional Chinese medicine, which is a plant of the genus Epimedium (Epimedium genus) in the family Berberridaceae. The Pharmacopoeia of the People's Republic of China (2000 Edition) recorded the following five Epimedium medicinal materials, which are the dry aerial parts. That is, Epimedium brevicornum Maxim, Epimedium sagittatum, Epimedium pubescens, Epimedium wushanenes, Epimedium koreanum Nakai, etc. Epimedium mainly contains isopentenyl-substituted flavone glycosides (icariin) and acetyl-substituted compounds, but its acetyl group is easily removed during the processing of Epimedium and extraction of icariin, so The main glycosides in Epimedium are flavonoid glycosides, namely icariin. Its main...

Claims

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Application Information

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IPC IPC(8): C12P19/44
Inventor 鱼红闪金凤燮姚新生
Owner 金凤燮
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