A method for fermenting and transforming soybean isoflavone glycosides into aglycones from Grifola frondosa

A technology for isoflavone glycosides and Grifola frondosa, which is applied in the field of Grifola frondosa fermenting and transforming soybean isoflavone glycosides to generate aglycones, can solve the problems that the conversion efficiency of β-glucosidase needs to be improved, and achieves improved activity and absorption rate, no The effect of organic solvent residue and cost reduction

Inactive Publication Date: 2019-01-04
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, Aspergillus is generally considered to be an excellent strain for producing β-glucosidase, especially Aspergillus niger has the highest efficiency, but it is limited to the laboratory development stage, and the conversion efficiency of Aspergillus niger β-glucosidase still needs to be improved

Method used

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  • A method for fermenting and transforming soybean isoflavone glycosides into aglycones from Grifola frondosa
  • A method for fermenting and transforming soybean isoflavone glycosides into aglycones from Grifola frondosa
  • A method for fermenting and transforming soybean isoflavone glycosides into aglycones from Grifola frondosa

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1 Grifola frondosa liquid fermentation produces β-glucosidase

[0027] 1. Method for measuring the activity of β-glucosidase

[0028] (1) Preparation of standard curve

[0029] Accurately weigh 13.90 mg of pNP (p-nitrophenol), dissolve it in distilled water and dilute to 100 mL. Take six 10mL volumetric flasks, draw 100, 200, 300, 400, 500, and 600 μL of pNP solution to bottles 1-6 respectively, and use 1mol / L Na 2 CO 3 The solution was made to volume and mixed evenly. The pNP concentrations corresponding to bottles 1-6 were 0.01, 0.02, 0.03, 0.04, 0.05, and 0.06 mmol / L. Measure the absorbance at 400 nm with distilled water as a control and draw a standard curve.

[0030] (2) Determination of β-glucosidase activity in the sample

[0031] Take 1 mL of the homogenized fermentation broth in a centrifuge tube, and centrifuge at 10,000 rpm for 10 min at 4°C. Mix 100 μL supernatant with 200 μL 5mM pNPG, incubate at 50°C for 30 min, add 2 mL 1mol / L Na 2 CO 3 ...

Embodiment 2

[0043] Example 2 β-glucosidase biotransformation of soybean isoflavone glycosides

[0044] 1. β-glucosidase biotransformation of soybean isoflavone glycosides to produce aglycones

[0045] Take 3-5 g of soybean isoflavone extract as a substrate, mix it with 100 mL of homogenized fermentation broth, and carry out a stirring reaction (transformation) at 40° C. to 60° C. for 12 to 20 hours to obtain soybean isoflavone aglycone. (optimization of the above parameters, as described in part 3 of this example)

[0046] 2. HPLC detection of soybean isoflavones

[0047] Liquid chromatography was used for detection, and the chromatographic column was C18, 4.6 mm×250 mm, with a particle size of 5 μm; the mobile phases A and B were acetonitrile and pH=3.0 phosphoric acid aqueous solution respectively, and the gradient elution conditions were shown in Table 3. The flow rate was 1.0 mL / min, the detection wavelength was 260 nm, the injection volume was 10 μL, and the column temperature was ...

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Abstract

The invention discloses a method for generating aglycon by converting grifola frondosa into soybeam isoflavone glycoside by fermenting. The method comprises the following steps: inoculating grifola frondosa to a culture medium to carry out fermentative culture, so that grifola frondosa mycelium fermentation liquor is obtained; carrying out homogenate treatment on the fermentation liquor, so that a grifola frondosa mycelium is broken, and beta-glucosidase is released to the fermentation liquor, thereby obtaining crude enzyme; and mixing the crude enzyme with soybeam isoflavone glycoside, and reacting the obtained mixture, so that soybean isoflavone glycoside is obtained. According to the method disclosed by the invention, through the deep fermentation of grifola frondosa, a large amount of beta-glucosidase is obtained, and then aglycon is prepared by converting the beta-glucosidase into soybeam isoflavone glycoside, the activity of the obtained beta-glucosidase is high, the product conversion rate is high, and the activity and absorption rate of soy isoflavone are greatly improved; and the converted product is also rich in biological active substances such as grifola frondosa ectoenzyme and mycelia polysaccharides, thereby laying a foundation for the development and industrial production of novel function materials.

Description

technical field [0001] The invention relates to the technical field of biotransformation, in particular to a method for fermenting and transforming soybean isoflavone glycosides into aglycones from Grifola frondosa. Background technique [0002] Soy isoflavones are a type of secondary metabolites formed during the growth of soybeans. There are a total of 12 types of soybean isoflavones naturally occurring in soybeans, which are divided into two types: free aglycones (Aglycon) and bound glycosides (Glycosides). , free aglycones account for 2%-3% of the total, including genistein, daidzein and glycitein; glycosides account for 97%-98% of the total, mainly It exists in the form of glucoside, acetyl glucoside and malonyl glucoside. [0003] Soy isoflavones have 2 or 3 hydroxyl groups and aromatic rings, which are very similar in structure to estrogen secreted by the human body and have weak estrogenic activity. Therefore, soy isoflavones are called natural phytoestrogens. In a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/06C12R1/645
Inventor 何国庆杨浣漪崔美林
Owner ZHEJIANG UNIV
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