Process for preparing soybean isoflavone aglycon by microorganism enzyme method
A technology of isoflavone aglycone and soybean isoflavone, which is applied in the field of preparation of soybean isoflavone aglycone, can solve the problems of strict requirements for filtration equipment and technical conditions, many raw materials for β-glucosidase, and increased filtration and refining costs, etc. Achieve the effects of improving bioavailability and biological activity, filling the gap in the preparation process, and improving bioavailability
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Embodiment 1
[0039] Raw material: 10% soybean ketone flavone coarse powder (product of Tianjin Jianfeng Natural Products Co., Ltd.)
[0040] Species: Aspergillus oryzae 3042 Shanghai Brewing Research Institute
[0041] Preparation method of culture slant subculture medium: PDA medium preparation
[0042] 1. Preparation of β-glucosidase by solid fermentation method and determination of enzyme activity
[0043] (1) the preparation of seed medium and its bacterial culture:
[0044]Precisely weigh 10.0g of soluble starch, 5.0g of glucose, 0.5g of potassium dihydrogen phosphate, 0.5g of magnesium sulfate, 2.5g of yeast extract, and 1.0g of potassium nitrate, dissolve and dilute to 500ml with distilled water, pH 6.5, and dispense into 500ml In a conical flask, 80ml per bottle, sterilize at 121°C for 15min. Aseptic operation, pick an appropriate amount of fresh spores of rice koji 3042 on the slant, transfer it into the seed medium, and culture at 27°C, 160r / min shaking for 42h. That is, the ...
Embodiment 2
[0064] 1. Preparation of soybean isoflavone aglycone by hydrolysis of 5% soybean isoflavone powder
[0065] The enzyme liquid used for hydrolysis is the same as 10% soybean isoflavone powder hydrolysis experiment: the raw material used is 5% soybean isoflavones (10% soybean isoflavone powder produced by Huabei Pharmaceutical Co., Ltd. is added with 1 times the amount of extracted soybean isoflavones, and then dried. The content of daidzein before transformation is 3.37%, the content of genistein is 1.12%, the content of daidzin is 1.21%, the daidzein is 0.25%, the genistein is 0.02%, and the glycitein 0.04%.
[0066] (1) Weigh 5% soybean isoflavones 100g, add 192ml of crude enzyme solution according to the ratio of substrate and enzyme 2300U / g, add distilled water to 1000 (substrate concentration is 0.1g / ml), pH is natural, at temperature 45 Under the condition of stirring speed of 180 r / min, the transformation was carried out for 8 hours to terminate the reaction. The enzyme ...
Embodiment 3
[0072] 1. Hydrolysis of 20% soybean isoflavone powder to prepare soybean isoflavone aglycone
[0073] The enzyme liquid used in hydrolysis is the same as the hydrolysis experiment of 10% soybean isoflavone powder: the raw material used is 20% soybean isoflavone powder (Tianjin Jianfeng Natural Products Co., Ltd.), the content of daidzein is 7.98%, the content of genistein is 9.51%, soybean 2.62% of glycosides, 0.72% of daidzein, 0.61% of genistein, and 0.19% of glycidin.
[0074] (1) Weigh 100g of 20% soybean isoflavones, add 250ml of crude enzyme solution according to the ratio of substrate and enzyme 3000U / g, add distilled water to 1000 (substrate concentration is 0.1g / ml), pH is natural, at temperature 45 Under the condition of stirring speed of 180 r / min, the transformation was carried out for 8 hours to terminate the reaction. The enzyme transformation solution was centrifuged at 10000 r / min at 4°C, and the precipitate was collected at 70°C. -Drying under reduced pressur...
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