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Method for preparing aglycon of soybean isoflavone glycoside base by enzymatic method hydrolyzing soybean

A technology of isoflavone glycosides and isoflavone aglycones, which is applied in the field of preparing soybean isoflavone glycosides by enzymatic hydrolysis, and can solve the problems of serious pollution and large damage of soybean isoflavone aglycones

Inactive Publication Date: 2004-03-24
沈阳天乐保化品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to obtain soybean isoflavone aglycone with high activity, the acid-base hydrolysis method has been reported (Edited by Need Life, Natural Chemistry, Science Press, 1996, p603-604), but the damage to soybean isoflavone aglycone is large and the pollution also more serious

Method used

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  • Method for preparing aglycon of soybean isoflavone glycoside base by enzymatic method hydrolyzing soybean
  • Method for preparing aglycon of soybean isoflavone glycoside base by enzymatic method hydrolyzing soybean
  • Method for preparing aglycon of soybean isoflavone glycoside base by enzymatic method hydrolyzing soybean

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] a. Take the high-temperature aerobic bacteria Clostridium thermocopriea (document 1) in a medium containing 3% corn flour, 1% enzyme-producing inducer-soybean extract (dry matter), and shake culture at a temperature of 60°C 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 acetic acid buffer solution, centrifuge to remove residue, and freeze-dry to obtain soybean isoflavone glycosides enzyme.

[0027] b. Mix 2 grams of soybean isoflavone glucosidase, 30 grams of genistin (Genistin), 1500 milliliters of acetate buffer (0.2M, pH5.0) and 150 milliliters of ethanol, so that genistin accounts for the total reaction volume 0.1-10% of the mixture, stirred and reacted for 1 hour at a temperature of 75°C.

[0028] c. add 6000 milliliters of ethanol after the reaction, filter and remove the protein precipitate, and the filtrate is evaporat...

Embodiment 2

[0032] a. Aspergillus niger (Aspegillus niger) was cultured with agitation and ventilation at a temperature of 28-30°C for 50-100 hours in a medium containing 4% malt extract, 1% enzyme-producing inducer-soybean extract, Centrifuge, use 60-75% saturated ammonium sulfate to precipitate enzyme protein in supernatant (containing enzyme mixture), collect protein, dissolve in phosphate buffer (0.02M, pH5. 0), the ammonium sulfate is removed by dialysis, and the residue is removed by centrifugation, which is the enzyme solution.

[0033] b. Mix 4 grams of soybean genistin (Genistin), 100 milliliters of phosphate buffer (0.02M, pH5.0) and 50 milliliters of the above-mentioned enzyme solution, so that genistin accounts for 0.1-10% of the total reaction volume, React at a temperature of 4°C for 40 hours.

[0034] c. adding 1 / 3 of the total reaction volume of n-butanol for extraction three times, and evaporating to dryness under reduced pressure to obtain crude aglycone. A silica gel ...

Embodiment 3

[0039] a. Using Aspergillus oryzae in a culture medium containing 5% wheat bran extract, 0.2% enzyme-producing inducer—soybean isoflavone or rutin, or 1% extract of its flavonoid-derived plant, Stirring and ventilating at a temperature of 28-30°C for 50-100 hours, centrifuging to obtain an enzyme-containing mixture, adding ethanol to 70% to precipitate the enzyme protein, collecting the protein, and freeze-drying to obtain the enzyme.

[0040] b. Treat three kinds of soybean isoflavone glycosides or mixed glycosides with the same method as in Example 1 and Example 2, and convert more than 80% of them into aglycones.

[0041] The above enzymes were separated and purified by the DEAE-Cellulose ion exchange resin column method and BioRed protein preparation chromatography (document 3), and the molecular weight was determined by SDS electrophoresis method (document 4). The molecular weight of the enzyme was 53,000.

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Abstract

The method for preparing soybean isoflavone aglycone by using enzyme method to hydrolyze soybean isoflavone glycoside glycosyl includes the following steps: using the enzyme capable of hydrolyzing soybean isoflavone glycoside glycosyl, mixing said enzyme, soybean isoflavone glycoside and buffer solution to make reaction for 1-40 hr., its reaction temp. is 4-75 deg.C and pH value is 3-9, so as to extract soybean isoflavone aglycone. Said invention not only can use enzyme to hydrolyze monoglycoside-soybean glycoside and genistein glycoside and convert them into correspondent aglycone, but also can use enzyme to hydrolyze soybean glycoside and genistein glycoside mixed glycoside and convent them into the mixed aglycone. Said product can be used as raw material for making medicine and health-care product.

Description

Technical field: [0001] The invention relates to a method for preparing isoflavone aglycone, in particular to a method for preparing its aglycone by enzymatically hydrolyzing soybean isoflavone glycosides. Background technique: [0002] Soybean isoflavones are high in soybeans and soybean cotyledons, mainly containing three aglycones (daidzein, soybean isoflavone aglycone and daidzein), collectively referred to as soybean isoflavone aglycone; (daidzein, genistein and daidzein), collectively referred to as soybean isoflavone glycosides. Its chemical structure is as follows: [0003] Daidzein Soybean Isoflavones (Genistein) Glycitein [0004] [0005] Daidzin Genistin [0006] [0007] Glycitin [0008] [0009] Soy isoflavones are phytoestrogens, which can promote the secretion of sex hormones and growth factors, and have the activities of improving women's menopausal syndrome, preventing osteoporosis and cancer. The physiological activity of soybean isoflavone ...

Claims

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Application Information

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IPC IPC(8): C12P17/06
Inventor 金凤燮鱼红闪芦明春张春枝谢明杰
Owner 沈阳天乐保化品有限公司
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