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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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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 hydrolysis to obtain free form soybean isoflavones, but it is not easy to industrialize under strong acid conditions
Since there are many by-products of strong acid hydrolysis, and the hydrolyzate is mainly used in food or medicine, there are also potential unsafe factors

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
Effect test

Embodiment 1

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

[0037] 500.0 g of whole soybeans (No. Dongnong No. 42, provided by the Soybean Research Institute of Northeast Agricultural University) were soaked in water at 20° C. for 20 hours, and rinsed with cold water. Add 0.1mol·L to soaked soybeans -1 , pH 6.60 phosphate buffer solution 5000mL, electric homogenizer homogenization. Soymilk was centrifuged at 4000 rpm for 3 min. Supernatant with 0.1mol·L -1 acidified with HCl to pH 5.0, and centrifuged for 10 min at 4°C in an automatic refrigerated centrifuge at 8000 r / min. The supernatant obtained is the crude enzyme. Add ammonium sulfate to the crude enzyme to make the saturation reach 75%, and let stand for 20 hours. 8000r / min, 4°C, refrigerated and centrifuged for 10min, collected precipitated protein, dissolved in 0.05mol·L -1 , pH 5.0 in disodium hydrogen phosphate-citric acid buffer solution. Pour the enzyme solution into ...

Embodiment 2

[0039] Example 2 Reesei Trigerase β-glucosidase (commercial enzyme fermented by Reesei Trigerase to obtain cellulase, extracted and purified from cellulase to obtain β-glucosidase) hydrolyzes soybean isoflavone powder to prepare genistein and Daidzein

[0040] Get 100g 20% ​​soybean isoflavone powder (provided by Heilongjiang Grain Research Institute), add disodium hydrogen phosphate-citric acid buffer solution to adjust pH to 4.5, add β-glucosidase (β-glucosidase (β The specific activity of a glucosidase is: 230,000 U / g), and it is hydrolyzed by shaking in a water bath at 50°C for 90 minutes. Then centrifuge and filter to separate, and the separated soy isoflavone aglycone precipitate is washed with water (the water volume is 4 / 5 of the original liquid) at 70° C. for 30 minutes, and left overnight. Centrifugal filtration and separation, the obtained precipitate was washed with water (the volume of water was 4 / 5 of the original liquid) at 70°C for 15 minutes, centrifugal filt...

Embodiment 3

[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 soybean isoflavone powder, add leaching agent acetone at a ratio of 1:20, heat, stir and reflux for extraction at 60°C for 3h, centrifuge the extract at 4000r / min for 20min, and take the upper layer. The precipitate was extracted again with 1:10 acetone at 60° C. under magnetic heating, stirring and reflux for 1 h, centrifuged, and the two extracts were combined. Acetone was removed by rotary evaporation under reduced pressure. The purified soy isoflavones are used for enzymatic hydrolysis of the substrate, and after hydrolysis according to the above-mentioned optimal enzymatic hydrolysis condition...

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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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