Method for enzyme hydrolysis of soybean isoflavone for producing genistein and daidzin aglycon

A technology of soybean isoflavones and genistein, applied in the direction of fermentation, etc., can solve the problems of unsafe conditions, difficult strong acid conditions, and many by-products of strong acid hydrolysis, and achieve the effects of product safety, easy operation, and reduced production costs

Inactive Publication Date: 2006-12-27
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Claims
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Problems solved by technology

More than 95% of the soybean isoflavones prepared in this way are still in the bound form, and the content of free forms of genistein and daidzein is very small, which is difficult to regulate the physiological functions of the human body
Some people in my country use acid

Method used

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  • Method for enzyme hydrolysis of soybean isoflavone for producing genistein and daidzin aglycon
  • Method for enzyme hydrolysis of soybean isoflavone for producing genistein and daidzin aglycon
  • Method for enzyme hydrolysis of soybean isoflavone for producing genistein and daidzin aglycon

Examples

Experimental program
Comparison scheme
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Example Embodiment

[0036] Example 1 Preparation of soybean β-glucosidase and hydrolysis of soybean isoflavones

[0037] Soybeans (Dongnong No.42, provided by Soybean Research Institute of Northeast Agricultural University), 500.0g of whole beans, soaked in water at 20°C for 20 hours, rinsed with cold water. Add 0.1mol·L to the soaked soybeans -1 , 5000mL of pH 6.60 phosphate buffer solution, homogenized with an electric homogenizer. Soy milk was centrifuged at 4000 rpm for 3 min. 0.1mol·L for supernatant -1 The HCl was acidified to pH 5.0, and then centrifuged at 8000r / min at 4°C for 10min in an automatic refrigerated centrifuge. The supernatant is the crude enzyme. Add ammonium sulfate to the crude enzyme to make the saturation reach 75%, and let it stand for 20 hours. 8000r / min, 4℃, refrigerated centrifugation for 10min, collect the precipitated protein, and dissolve it in 0.05mol·L -1 , PH5.0 disodium hydrogen phosphate-citric acid buffer solution. Pour the enzyme solution into a dialysis bag, di...

Example Embodiment

[0039] Example 2 T. reesei β-glucosidase (commercial enzyme is fermented with T. reesei to obtain cellulase, and β-glucosidase is extracted and purified from cellulase) to hydrolyze soybean isoflavone powder to prepare genistein and Daidzein

[0040] Take 100g of 20% soybean isoflavone powder (provided by Heilongjiang Province Grain Research Institute), add disodium hydrogen phosphate-citric acid buffer solution to adjust the pH to 4.5, add β-glucosidase (β) according to the enzyme-substrate concentration ratio of 5% The specific activity of monoglucosidase is 230,000 U / g), hydrolyzed with shaking in a water bath at 50°C for 90 min. Then it was separated by centrifugal filtration, and the separated soy isoflavone aglycon precipitate was washed with water (water volume 4 / 5 of the original liquid) at 70° C. for 30 min and left overnight. After centrifugal filtration, the obtained precipitate was washed with water (4 / 5 of the original liquid) at 70°C for 15 minutes, centrifuged and s...

Example Embodiment

[0041] Example 3 Method for preparing high-content genistein and daidzein

[0042] If it is necessary to prepare high-content genistein and daidzein, high-purity soybean isoflavone powder can be used as a substrate for hydrolysis. Before hydrolysis, the soybean isoflavone powder is further purified and then hydrolyzed. The specific method is as follows:

[0043] Accurately weigh the soybean isoflavone powder, add the extractant acetone according to 1:20, heat and stir reflux for 3 hours at 60°C, centrifuge the extract at 4000r / min for 20min, and take the upper liquid. The precipitation was extracted again with 1:10 acetone at 60°C under magnetic heating and stirring at reflux for 1 h, centrifuged, and the two extracts were combined. The acetone was removed by rotary evaporation under reduced pressure. The purified soy isoflavones are used for enzymatic hydrolysis of the substrate. After hydrolyzing according to the above-mentioned optimal hydrolysis conditions, the hydrolyzed solu...

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Abstract

This invention provides the method of producing genistein and daidzein by enzymatic hydrolysis of soybean isoflovone. It uses the soybean isoflovone with different purity as raw material, and hydrolyse it with Trichoderma beta- glucosidase or bean beta- glucosidase. Then solvent extraction, ultrafiltration and freeze centrifugal isolation and purification are performed to get genistein and daidzein. This invention can use hydrolysis material with different purity. The enzyme used can be purchased directly or prepared by yourself. The buffer solution can recycle. It is easy to operate and with relatively high transformation rate (>85%). The total content of soybean isoflovone daidzein is high. By using the extracted bean beta- glucosidase from soybean came from northeast region of china, the problem of poor edible security with enzyme generated by microbiological fermenting method can be resolved. This invention is suitable for the industralization production because the low cost with commercial enzyme.

Description

(1) Technical field [0001] The invention relates to a method for producing genistein and daidzein from soybean isoflavone powder with different concentrations. More specifically, the present invention uses self-extracted and purified soybean β-glucosidase from Northeast soybeans and commercialized Trichoderma reesei β-glucosidase to hydrolyze soybean isoflavones, and uses solvent extraction, ultrafiltration and freezing Separation and purification methods such as centrifugation, and a method for producing genistein and daidzein from soybean isoflavone powder. (2) Background technology [0002] Soy isoflavones are a class of secondary metabolites formed during soybean growth. There are 12 kinds of soy isoflavones naturally present in soybeans, which can be divided into 3 categories, namely daidzin, genistin and glycitin. There are four forms of glucoside, acetyl glucoside and malonyl glucoside (Kudou et al., 1991a; Kudou et al., 1991b). The chemical structural formulas of ...

Claims

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Application Information

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IPC IPC(8): C12P17/06
Inventor 张永忠孙艳梅徐晶井乐刚李冬梅
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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