Method for hydrolyzing soybean isoflavone by enzyme

A technology of soybean isoflavone and enzymatic hydrolysis, applied in the direction of fermentation, etc., can solve the problems of unrecyclable enzymes, difficult product separation, high cost and high cost, and achieve the effect of good industrial enlargement prospect and simple equipment and process.

Active Publication Date: 2007-12-12
TSINGHUA UNIV
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Problems solved by technology

The stability of the products obtained by acid and alkaline hydrolysis has been questioned (Zhang Chen, Yang Xiaoquan, Kong Huiqing. The research status of the stability of soybean isoflavone components under different processing conditions. Soybean Science, 2006, 25(1): 73- 76), and due to a large amount of acid a

Method used

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  • Method for hydrolyzing soybean isoflavone by enzyme
  • Method for hydrolyzing soybean isoflavone by enzyme
  • Method for hydrolyzing soybean isoflavone by enzyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1, the preparation of immobilized β-glucosidase

[0056] 1. Preparation of immobilized β-glucosidase

[0057] Prepare chitosan pellets with a diameter of about 2.5mm and very uniform according to the method described in the literature (Peng Zhiying, Yue Zhenfeng, Zhang Xuebing. Preparation of microsphere-shaped immobilized α-glucosidase. Journal of South China University of Technology (Natural Science Edition), 2001 , 29(6): 56-59 and Nitin W. Fadnavis, Gurrala Sheelu, Bezavada Mani Kumar. Gelatin Blends with Alginate: Gels for Lipase Immobilization and Purification, Biotechnol. Prog. 2003, 19: 557-564). The specific method is as follows: use 1% acetic acid by volume to prepare a 25g / 100ml chitosan solution, let it stand for 30 minutes, and use a suction tip to cooperate with a small funnel to make a dripping device as shown in Figure 3. The suction tip is Jill Mori 1ml and 200μl tips, the funnel is a 10ml small funnel.

[0058] Just below the distance of 2...

Embodiment 2

[0075] Embodiment 2, immobilized enzyme two-phase hydrolysis soybean isoflavones

[0076] 14.8 mg of 30% strength soybean isoflavones and 1.48 U of immobilized β-glucosidase were added to 5 ml of 0.1 M pH=5 citric acid-sodium citrate buffer and 5 ml of ethyl acetate. The reactions were all carried out in a constant temperature shaker at 40° C. for three hours, and the contents of daidzin, genistin, daidzein and genistein were determined according to the above-mentioned HPLC method.

[0077] The results showed that the amount of daidzein obtained by the immobilized enzyme hydrolysis was 0.97mg, the amount of daidzein was 0.25mg at the initial stage of the reaction, and the amount of daidzein that should be obtained by the complete hydrolysis of daidzin was 0.96mg; The amount of genistein is 1.50 mg, the amount of genistein at the beginning of the reaction is 0.22 mg, and the amount of genistein that should be obtained by complete hydrolysis of genistein is 1.64 mg. According t...

Embodiment 3

[0078] Embodiment 3, application comparison of immobilized β-glucosidase in single-phase system and biphasic system

[0079] Two treatments were set up in the experiment, one of which was a single-phase system, and 14.8 mg of 30% soybean isoflavones and 1.48 U of immobilized β-glucose were added to 10 ml of 0.1M, pH=5 citric acid-sodium citrate buffer glycosidase; another treatment is a biphasic system, 14.8 mg of 30% specification soybean isoflavones and 1.48 U of immobilized β - Glucosidase. The reactions were all carried out in a constant temperature shaker at 40°C, samples were taken every hour, and the contents of daidzin, genistin, daidzein and genistein were determined according to the above-mentioned HPLC method, and the reaction was performed for three hours. The results are shown in Fig. 9 and Fig. 10, not only the product cannot be obtained in the single-phase system, but also the hydrolysis rate is slightly lower than that in the two-phase system.

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Abstract

The invention discloses a method for hydrolyzing soybean isoflavone with biphase biological method. It is to hydrolyze glycoside soybean isoflavone by using immobilized soybean isoflavone glycoside hydrolase in biphase system that is composed of organic solvent and buffering solution, and get aglycone soybean isoflavone. The solubility of said organic solvent in said soybean isoflavone is better than that in water. The invention is characterized by effective hydrolysis of soybean isoflavone, no speical smell of product, and simple device and process.

Description

technical field [0001] The invention relates to a method for enzymatically hydrolyzing soybean isoflavones. Background technique [0002] Enzyme immobilization is to immobilize the biocatalyst enzyme on a certain carrier. With the development of biotechnology, enzymes as catalysts have the advantages of good specificity, high catalytic efficiency, and mild reaction conditions, and are increasingly replacing traditional catalysts in industrial applications. However, the main disadvantage of the industrial application of enzymes as catalysts is that they are expensive and cannot be reused. In 1916, Nelson and Griffin adsorbed sucrase on activated carbon and alumina for the first time, realizing the immobilization of enzyme for the first time (R.F.Taylor, Protein immobilization, Marcel Dekker, Inc: New York, 1991, v). Since then, the enzyme immobilization technology has developed rapidly, and so far there have been very mature immobilization methods: adsorption, covalent cros...

Claims

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Application Information

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IPC IPC(8): C12P17/06
Inventor 林章凛张涛黄哲
Owner TSINGHUA UNIV
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