Method for preparing isoquercetin and quercetin by enzymatic method and hydrolyzing rutin

A technology for isoquercetin and quercetin, which is applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., can solve the problems of less isoquercetin, complicated processes, and inability to produce quercetin, and achieves high productivity. The effect of large output, simple and safe operation

Inactive Publication Date: 2006-06-28
SUNFLOWER PHARM GRP (TIANJIN) DRUG RES INST CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is not only complex in process, but also part of the solid support will participate in enzymolysis and produce undesirable products, especially the dissolution of radiloside is slight, and the produced isoquercetin (isoquercetin) is less, let alone produce quercetin white

Method used

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  • Method for preparing isoquercetin and quercetin by enzymatic method and hydrolyzing rutin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] a. With the thermoaerobic bacteria Bacillus sp.JF (document 1), in 0.01mol MgSO containing 3% corn flour, 1% enzyme-producing inducer - Sophora japonica extract (dry matter) 4 culture medium at 60°C for 30-40 hours, centrifuge and sterilize to obtain an enzyme-containing mixture, add 70% ethanol to precipitate the enzyme protein, collect the protein, and freeze-dry to obtain rutin-hydrolyzing α-rhamna Glycosyl enzymes.

[0020] b. Dissolve 4 grams of the above enzyme in 150 milliliters of 0.02 M phosphate buffer of pH 6; mix 9 grams of rutin monomer with 103 milliliters of 0.02 M phosphate buffer of pH 7 and 47 milliliters of 95% ethanol. The above two solutions are mixed, the concentration of the reactant rutin is 0.01-10%, the concentration of ethanol is 0-35%, and the reaction is stirred and reacted at a temperature of 60° C. for 10 hours.

[0021] c. After the reaction, add 24 grams of NaOH to the solution, filter to remove the protein precipitate, add 300ml 2N HCl...

Embodiment 2

[0024] a. with Aspergillus oryzae in the culture medium containing 4% corn flour, 1% enzyme-producing inducer-Sophora japonica extract (dry matter), cultured under ventilation at 30°C for 30-40 hours, and then sterilized, The culture medium was precipitated with 75% saturated ammonium sulfate, and the precipitate was collected, diluted to 1 / 5 of the volume of the culture medium with 0.02M sodium acetate buffer solution of pH 5.8, dialyzed, and centrifuged to remove slag, which was the enzyme solution.

[0025] b. Take 50 milliliters of the above-mentioned enzyme solution, 10 grams of rutin monomer, 5 liters of pH6 0.02M phosphate buffer and 50 milliliters of 95% ethanol and mix evenly, so that the concentration of the reactant rutin is 0.01-10%, and the concentration of ethanol 0-35%, stirred and reacted at a temperature of 70°C for 2 hours.

[0026] c. After the reaction, add 24 grams of NaOH to the solution, filter to remove the protein precipitate, add 300ml 2N HCl to the f...

Embodiment 3

[0029] a. Use Aspergillus niger (Aspergillus niger) in the medium containing 4% malt extract and 0.5% ginseng extract, ventilate and cultivate at a temperature of 30°C for 30-40 hours, sterilize after cultivation, and use 80% saturated sulfuric acid for the culture solution Precipitate the enzyme protein with ammonium, collect the precipitate, dissolve it with 0.02M sodium acetate buffer solution with pH 5.8, dialyze to remove ammonium sulfate, centrifuge to remove residue, and freeze-dry to obtain the enzyme.

[0030] b. Get 7 grams of the above-mentioned enzyme and dissolve in 200 milliliters of pH5.8 0.02M sodium acetate buffer; get 12 grams of commercially available rutin monomer and 115 milliliters of 0.02M sodium acetate buffer of pH5.8 and 85 milliliters 95% ethanol is mixed evenly, the concentration of the reactant rutin is 0.01-10%, and the concentration of ethanol is 0-35%, and the reaction is stirred and reacted at a temperature of 15° C. for 24 hours.

[0031] c. A...

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Abstract

The method for preparing isoquercetin and quercetin by adopting enzyme method to hydrolyze rutin glycosyl includes the following steps: using the enzyme capable of hydrolyzing rutin glycosyl, mixing said enzyme, rutin, buffer solution and ethyl alcohol and making them produce reaction for 2-24 hr. and its reaction temp. is 15-70 deg.C, pH value is 4-8, rutin concentration is 0.1-10% and ethyl alcohol concentration is 0-35%, so that the isoquercetin and quercetin can be extracted, and the purity of said invented products is greater than 90%.

Description

Technical field: [0001] The invention relates to a method for preparing isoquercetin and quercetin, in particular to a method for preparing isoquercetin and quercetin by enzymatically hydrolyzing sugar groups on rutin. Background technique: [0002] Rutin is a flavonoid compound, which is widely distributed in plants in nature, and exists in the roots, stems, leaves, flowers, fruits, and seeds of many plants. Some commonly used traditional Chinese medicines, such as Huai Mi, rue, buckwheat, seabuckthorn, ginkgo, wolfberry, motherwort, Bupleurum, Prunella vulgaris, aloe, eucalyptus leaves and tobacco leaves contain rutin, among which Huai Mi, buckwheat, eucalyptus The leaf content is the highest, and the higher one is Sophora japonica L. (the flower bud of the leguminous plant Sophora japonica L.), the content can reach more than 20%, and it is the main raw material for extracting rutin in my country's pharmaceutical industry. [0003] From the chemical structure point of vie...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/06C07D311/38C12R1/01C12R1/645
Inventor 鱼红闪金凤燮
Owner SUNFLOWER PHARM GRP (TIANJIN) DRUG RES INST CO LTD
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