Process for preparing soybean isoflavone aglycon by microorganism enzyme method

Abstract

The invention discloses a method for producing soybean isoflavone aglycone through microbe's enzyme method, and is culturing Aspergillus oryzae with soybean isoflavone powder as raw materials for fermentation to produce beta-glucosidase, and converting soybean isoflavone to soybean isoflavone aglycone. The method is characterized in comprising the following preparation steps: the said raw materials are 2-90% soybean isoflavone powder, the said strain is Aspergillus oryzae 3042, preparing seed culture liquid with the strain, preparing solid fermentation medium, fermenting to obtain fermenting product containing beta-glucosidase, adding soybean isoflavone powder for preparing enzyme converting solution, centrifuging for separation to obtain deposit, drying at low temperature to obtain soybean isoflavone aglycone. The invention avoids the restriction of the raw materials purity, converting rata can reach to over 90%. The invention has the advantages of short procedure, low cost, high bioavailability.

Description

technical field

[0001] The invention relates to a method for preparing soybean isoflavone aglycone with soybean isoflavone, in particular to a preparation method for converting soybean isoflavone aglycone into soybean isoflavone aglycone by microbial enzymatic method. Background technique

[0002] Soybean isoflavone (sorbean isoflovone) refers to a class of compounds with 3-phenylchromone as the core, which is a soybean nutritional ingredient with various biological activities. At present, scientists have discovered a total of 15 isomers of soybean isoflavones, which are divided into two categories: free aglycone and bound glycoside. Among them, glucosides account for about 95-98% (w / w) of the total amount of soybean isoflavones, and aglycones account for 2-5% (w / w) of the total amount of soybean isoflavones. The latest research from the Rakult Microbiology Research Center in Tokyo, Japan shows that free aglycones are smaller and more hydrophobic than bound glycosides, so t...

Examples

Embodiment 1

[0039] Raw materials: 10% daidzein and flavone coarse powder (product of Tianjin Jianfeng Natural Products Co., Ltd.)

[0040] Strain: Aspergillus oryzae 3042 Shanghai Brewing Research Institute

[0041] The preparation method of the slant subculture medium of the strain: PDA medium preparation

[0042] 1. Preparation of β-glucosidase by solid fermentation method and determination of enzyme activity

[0043] (1) Preparation of seed culture medium and strain cultivation thereof:

[0044]Accurately 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, 1.0g of potassium nitrate, dissolve and dilute to 500ml with distilled water, pH6.5, and dispense to 500ml In Erlenmeyer flasks, 80ml per bottle, sterilized at 121°C for 15min. Aseptically operate, pick an appropriate amount of rice koji 3042 spores from the fresh slant, transfer them into the seed medium, and cultivate them at 27°C and 160r / min ...

Embodiment 2

[0064] 1. Prepare 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 test: raw material 5% soybean isoflavones (the 10% soybean isoflavone powder that North China Pharmaceutical Co., Ltd. Prepared from defatted soybean flour) Before conversion, the content of daidzein is 3.37%, the content of genistin is 1.12%, the content of daidzein is 1.21%, the content of daidzein is 0.25%, the content of genistein is 0.02%, and the content of glycitein is 0.04%.

[0066] (1) Weigh 100g of 5% soy isoflavones, add 192ml of crude enzyme solution according to the ratio of enzyme to substrate 2300U / g, add distilled water to 1000 (substrate concentration is 0.1g / ml), pH is natural, at a temperature of 45 Under the condition of stirring speed of 180 rpm at ℃, the conversion was carried out for 8 hours, and the reaction was terminated. The enzyme conversion liquid was cent...

Embodiment 3

[0072] 1. Prepare soybean isoflavone aglycone by hydrolysis of 20% soybean isoflavone powder

[0073] The enzyme liquid used for hydrolysis is the same as 10% soybean isoflavone powder hydrolysis test: the raw materials used are 7.98% daidzin content in 20% soybean isoflavone powder (Tianjin Jianfeng Natural Products Co., Ltd.), 9.51% genistin content, soybean Glycoside 2.62%, daidzein 0.72%, genistein 0.61%, glycitein 0.19%.

[0074] (1) Weigh 100g of 20% soy isoflavones, add 250ml of crude enzyme solution according to the ratio of enzyme to substrate 3000U / g, add distilled water to 1000 (substrate concentration is 0.1g / ml), pH is natural, at a temperature of 45 Under the condition of stirring speed of 180 rpm at ℃, the conversion was carried out for 8 hours, and the reaction was terminated. The enzyme conversion liquid was centrifuged at 10000 r / min at 4 °C, and the precipitate was collected at 70 °C. Drying under reduced pressure under the condition of -0.1Mpa, and pulveri...

technical field

[0001] The invention relates to a method for preparing soybean isoflavone aglycone with soybean isoflavone, in particular to a preparation method for converting soybean isoflavone aglycone into soybean isoflavone aglycone by microbial enzymatic method. Background technique

[0002] Soybean isoflavone (sorbean isoflovone) refers to a class of compounds with 3-phenylchromone as the core, which is a soybean nutritional ingredient with various biological activities. At present, scientists have discovered a total of 15 isomers of soybean isoflavones, which are divided into two categories: free aglycone and bound glycoside. Among them, glucosides account for about 95-98% (w / w) of the total amount of soybean isoflavones, and aglycones account for 2-5% (w / w) of the total amount of soybean isoflavones. The latest research from the Rakult Microbiology Research Center in Tokyo, Japan shows that free aglycones are smaller and more hydrophobic than bound glycosides, so t...

Examples

Embodiment 1

[0039] Raw materials: 10% daidzein and flavone coarse powder (product of Tianjin Jianfeng Natural Products Co., Ltd.)

[0040] Strain: Aspergillus oryzae 3042 Shanghai Brewing Research Institute

[0041] The preparation method of the slant subculture medium of the strain: PDA medium preparation

[0042] 1. Preparation of β-glucosidase by solid fermentation method and determination of enzyme activity

[0043] (1) Preparation of seed culture medium and strain cultivation thereof:

[0044]Accurately 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, 1.0g of potassium nitrate, dissolve and dilute to 500ml with distilled water, pH6.5, and dispense to 500ml In Erlenmeyer flasks, 80ml per bottle, sterilized at 121°C for 15min. Aseptically operate, pick an appropriate amount of rice koji 3042 spores from the fresh slant, transfer them into the seed medium, and cultivate them at 27°C and 160r / min ...

Embodiment 2

[0064] 1. Prepare 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 test: raw material 5% soybean isoflavones (the 10% soybean isoflavone powder that North China Pharmaceutical Co., Ltd. Prepared from defatted soybean flour) Before conversion, the content of daidzein is 3.37%, the content of genistin is 1.12%, the content of daidzein is 1.21%, the content of daidzein is 0.25%, the content of genistein is 0.02%, and the content of glycitein is 0.04%.

[0066] (1) Weigh 100g of 5% soy isoflavones, add 192ml of crude enzyme solution according to the ratio of enzyme to substrate 2300U / g, add distilled water to 1000 (substrate concentration is 0.1g / ml), pH is natural, at a temperature of 45 Under the condition of stirring speed of 180 rpm at ℃, the conversion was carried out for 8 hours, and the reaction was terminated. The enzyme conversion liquid was cent...

Embodiment 3

[0072] 1. Prepare soybean isoflavone aglycone by hydrolysis of 20% soybean isoflavone powder

[0073] The enzyme liquid used for hydrolysis is the same as 10% soybean isoflavone powder hydrolysis test: the raw materials used are 7.98% daidzin content in 20% soybean isoflavone powder (Tianjin Jianfeng Natural Products Co., Ltd.), 9.51% genistin content, soybean Glycoside 2.62%, daidzein 0.72%, genistein 0.61%, glycitein 0.19%.

[0074] (1) Weigh 100g of 20% soy isoflavones, add 250ml of crude enzyme solution according to the ratio of enzyme to substrate 3000U / g, add distilled water to 1000 (substrate concentration is 0.1g / ml), pH is natural, at a temperature of 45 Under the condition of stirring speed of 180 rpm at ℃, the conversion was carried out for 8 hours, and the reaction was terminated. The enzyme conversion liquid was centrifuged at 10000 r / min at 4 °C, and the precipitate was collected at 70 °C. Drying under reduced pressure under the condition of -0.1Mpa, and pulveri...

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