JPH0714927B2 - Method for producing isoflavone compound - Google Patents

Description

TECHNICAL FIELD The present invention relates to a process for producing an isoflavone compound from soybean, particularly an isoflavone compound containing a large amount of aglycones thereof.

<Conventional Technology and Problems> Soybean contains isoflavone compounds such as daidzin, glycitin, genistin, daidzein, and genistein. Its physiological activity is estrogen, antioxidant,
Anti-hemolytic action, anti-bacterial action, anti-lipidemia, anti-cholesterol action are known, and recently, anti-cancer effects such as cancer cell differentiation inducing action and oncogene inhibitory action have been confirmed, and their usefulness has been noted. There is.

Among these isoflavone compounds, aglycones such as daidzein and genistein are mainly used for the medicinal effects such as carcinostatic action, not glycosides.

As a method for obtaining an isoflavone compound from an extract of soybean, for example, JP-A-62-126186 is mentioned. However, since 95% or more of soybean exists as a glycoside, it can be obtained by this method. The isoflavone compound is mainly composed of glycosides, and it is possible to obtain an extremely small amount of aglycone.

<Means for Solving Problems> The inventors of the present invention investigated a method for inexpensively obtaining a large amount of highly useful aglycone among isoflavone compounds. As a result, soy isoflavone is an action of β-glucosidase in soybean. The sugar bond was easily cleaved to convert it to its aglycone. The conversion was 50 ℃, pH6.3.
We obtained the knowledge that

The present invention was made based on such findings,
At the time of producing an isoflavone compound from a soybean extract or a soybean ground product, any one of dipping step, grinding step or enzymatic reaction step after grinding, or two or more so that β-glucosidase activity in soybean is maximized In the step of 1, the soybean or ground soybean product is heated to 45 to 55 ° C, which is a method for producing an isoflavone compound.

Hereinafter, the present invention will be specifically described.

Any soybean may be used as a raw material as long as the enzyme is not deactivated, and for example, low-denaturation defatted soybean powder, crushed soybean, dehulled soybean, whole soybean, and the like can be used.

In order to produce an isoflavone compound from these soybean extracts, soybeans are soaked in 5 to 10 times warm water at 45 to 55 ° C., and the soaked water is used as a refined raw material for soaking. Since the ratio is about 10% higher than in soybeans,
It can be a suitable refined raw material.

When producing an isoflavone compound from a ground soybean product, the soybean soaked as described above is ground with soaking water, or soaked in room temperature water, and the soybean soup is 45-
It may be ground at 55 ° C, or after grinding, it may be heated to 45 to 55 ° C to cause an enzymatic reaction, but preferably the immersion is carried out at 45 to 55 ° C and this is carried out at 45 to 55 ° C together with immersion water. Grind and hold the resulting grind at 45-55 ° C for several hours. This greatly increases the percentage of aglycone in soybeans.

The soybean extract or soybean ground product thus obtained is used as a purification raw material, and there are two methods for purification of the isoflavone compound.

One is purification with a solvent, which is a raw material for refining, that is, soybean soaked water or ground soybeans, is pulverized by hot air drying, freeze drying, etc., degreased with n-hexane or petroleum ether, and the residue is dried with ethyl ether. It is a method of extracting only isoflavone aglycone.

On the other hand, the isoflavone compound is reflux-extracted with a hydrous alcohol from the powdered raw material for purification, and the extract is concentrated and dried to dryness by a conventional method, and then dissolved in a small amount of hydrous alcohol. , YMC-GEL ODS-A type 60-01 (manufactured by Yamamura Chemical Laboratory Co., Ltd.) and Diaion
After adsorbing on HP-20 (manufactured by Mitsubishi Kasei Co., Ltd.) etc.,
It is a method of thoroughly washing with water, eluting phenolic acid with about 20% hydrous alcohol, and then obtaining an isoflavone compound category rich in aglycone with 80% hydrous alcohol.
Also, after washing with water, if the glycoside fraction is flown out with about 40% hydrous alcohol, it is possible to obtain only the aglycone category by elution of 80% hydrous alcohol.

Incidentally, the reverse phase resin used here is easy to wash and regenerate with an organic solvent such as alcohol or acetone.
Can be used repeatedly.

It is also possible to directly adsorb the purification raw material onto the resin without pulverizing it for purification, and in the case of immersion water, as it is, or in the case of ground material, filtrate filtered by a conventional method or centrifuged. The supernatant is brought into contact with the resin in a reverse phase system and then purified in the same manner as above.

The effects of the present invention will be described below with reference to experimental examples.

Experimental Example 1 Hulled soybeans are immersed in 5 times the amount of water at 20 to 80 ° C. for 6 hours, and the soaked water and the soybeans are immediately cooled and frozen. It was dried with a freeze drier, pulverized, a certain amount of the mixture was refluxed and extracted with 80% methanol, and a fixed amount was analyzed by high performance liquid chromatography (Model 209D manufactured by Waters).

The results are shown in Table 1.

Experimental Example 2 Dehulled soybeans were immersed in 5 times the amount of warm water at 50 ° C. adjusted to pH 5.5 to 11 for 2 hours, and the immersion water was acidified immediately and immediately thereafter.
After filtration with a 0.45 μm filter, analysis was performed by high performance liquid chromatography in the same manner as in Experimental Example 1.

The results are shown in Table 2.

As is clear from the results of Experimental Examples 1 and 2, the immersion temperature is 45 to 55 ° C.
At pH 6.3, the exchange of isoflavone aglycone is maximized.

<Examples> Examples are shown below.

Example 1 5 kg of dehulled soybeans were dipped in warm water 25 at 50 ° C. and soaked for 2 hours while keeping the temperature at 50 ° C.

Then, the immersion water was concentrated with an evaporator and dried to obtain 450 g of a purified raw material. This was degreased with n-hexane using a Soxhlet extractor. Then, the residue was sufficiently dried and then extracted with ethyl ether to obtain 0.5 g of isoflavone aglycone.

Example 2 Dehulled soybeans dipped in the same manner as in Example 1 together with immersion water at 50 ° C.
Was ground for 1 hour, dried at a freeze dryer and powdered to obtain 4.1 kg of purified raw material. In the same manner as in Example 1, degreasing with n-hexane and extraction of aglycone with ethyl ether were carried out.
7.2 g of isoflavone aglycone was obtained. The aglycone ratio was 60% or more after milling at 50 ° C and holding at that temperature for 1 hour.

Example 3 50 kg of water at 50 ° C. was added to 10 kg of low-denaturation defatted soybean (Nisshin Soya Flower) and stirred for 1 hour. This was dried with hot air by spray drying to obtain a purified raw material. 5 for refined raw materials
Isoflavones were extracted with a double amount of 80% hot methanol and dried under reduced pressure to obtain 103 g of a crude isoflavone fraction. This was redissolved in a small amount of methanol and used as a packing material for ODS-A.
Φ70 packed with Type 60-01 (made by Yamamura Chemical Laboratory Co., Ltd.)
It was adsorbed by passing through a column of mm × 100 cm.

Next, the phenolic acid and isoflavone glycoside fractions were flown out with 40% methanol, removed, and then eluted with 80% methanol, which was evaporated to dryness under reduced pressure, and aglycone 9.
Got 5g.

Example 4 1 kg of the synthetic adsorbent Diaion HP-20 (manufactured by Mitsubishi Kasei Co., Ltd.) was added to immersion water 25 obtained by dipping dehulled soybeans in the same manner as in Example 1 and stirred for 1 hour. While adsorbing the isoflavone compound. Then, the resin was filtered off, washed with 20% ethanol to remove phenolic acid, and then eluted with 80% ethanol to obtain an isoflavone compound. The weight of this after drying under reduced pressure was 1.1 g. The isoflavone compound contained about 40% aglycone.

Claims (1)

[Claims] 1. When producing an isoflavone compound from a soybean extract or a soybean ground product, any one of a dipping step, a grinding step and an enzyme reaction step after grinding so that the β-glucosidase activity in soybean is maximized. Or, in two or more steps, soybeans or ground soybeans at 45 to 55 ° C
A method for producing an isoflavone compound, which comprises heating to