KR20150079234A - Isoflavone-biotransforming activity and processes for preparing isoflavone aglycone using the same - Google Patents

Abstract

The present invention relates to a strain having an excellent isoflavone transformation activity and a preparation method of isoflavone aglycone by using the same. More particularly, the preparation method of a high-purity isoflavone aglycone comprises the steps of: (a) extracting soybeans or germinated soybeans with water and recovering and filtering an extract thereof to obtain a filtrate; (b) culturing lactic acid bacteria by inoculating thereof to the filtrate filtered in the step (a); (c) recovering a precipitate by inducing an immersion of a culture medium cultured in the step (b) for 6 to 24 hours at 20°C and; (d) adsorbing the recovered precipitate by using a methacrylate type resin; and (e) eluting the adsorbed material in the step (d) by using a mixed solvent of alcohol and water, and separating a high-purity isoflavone aglycone. In addition, the present invention uses water and an ethanol without using hazardous solvents which are conventionally used. Accordingly, the preparation process is relatively simple, economical and has a high yield compared with a conventional method of separating an isoflavone aglycone.

Description

Isoflavone biotransforming activity and processes for preparing isoflavone aglycone using same The present invention relates to a method for producing isoflavone aglycone,

More particularly, the present invention relates to a method for producing isoflavone aglycone, which is superior in isoflavone conversion efficiency, and more particularly, to a method for producing isoflavone aglycone using soybean or germinated soybean by extracting with water, recovering the extract, step; (b) inoculating and culturing the filtered filtrate in step (a) with lactic acid bacteria; (c) recovering the precipitate by inducing precipitation of the cultured medium cultured in step (b) at 20 DEG C for 6 to 24 hours; (d) adsorbing the recovered precipitate using a methacrylate type resin; And (e) eluting the adsorbed material of step (d) with a mixed solvent of water and water to separate the high purity isoflavone aglycon.

Soy has been used as an important food resource of mankind and it is known that it is rich in protein, essential fatty acid, phospholipid, vitamin E and calcium, and shows various physiological activities when it is ingested by humans. The physiological activity of soybeans is known to be effective for prevention of various diseases such as antihypertensive effect, thrombolytic ability and blood cholesterol lowering effect, and also good for brain activity and skin beauty as well as anticancer, anti-mutation and anti-aging. In addition, soybean is a versatile food that can be transformed into various forms such as soybean oil, tofu, and soybeans, including kochujang, soybean paste, soybean yogurt, and so on.

Recently, interest has been focused on isoflavones among functional ingredients of soybean, and isoflavones have been reported to have preventive and therapeutic effects on cancer, heart disease, osteoporosis and kidney disease. The content of isoflavones varies greatly depending on the variety and cultivation conditions of the soybean, which is usually about 0.6 to 3.1 mg%, and a loss of about 30 to 50% of the isoflavone in the production of tofu and soybean protein.

Isoflavone is a natural compound that exists in plant species such as soybean, plant, and 칡. It has a similar structure to estrogen, a kind of female hormone, and has a physiological function such as phytoestrogen. Isoflavone is a group of compounds with the molecular formula of 12 kinds, including genistein, daidzein, glycitein and its glucose glycosides, further acetylated and malonyl, It is known as an isomer present in the state of a glycoside, and is a substance mainly present in the form of a glycoside. In addition, they have been considered only as simple pigments in the past, but recent studies have shown that anti-cancer effects (Barnes, J., Nutr., 125, 777s, (1995); Akiyama et al., J. Biol. Chem., 262 (1987) Molteni et al., J. Nurt., 125, 751s (1995), Prevention of osteoporosis (Shino et al., Life Sci., 42 (11), 1123 (Naim et al., J. Agric. Food Chem., 24 (6), 1174 (1976)), aldehyde (1993)) and prostate cancer prevention (Peterson and Barnes, The Prostate, 22, 335 (1993)). It is still being revealed.

Most of isoflavones exist in the form of glycosides in their natural state. These isoflavone glycosides are not absorbed as they are not decomposed by gastric acid in the body. They are absorbed after being decomposed by enzymes secreted from microorganisms in the colon, (U.S. Patent No. 5,506,211). On the other hand, isoflavone aglycon has the advantage of being absorbed directly from the stomach and small intestine without any additional transformation in the body. Accordingly, a method of producing isoflavone aglycon having a high water absorption rate is strongly desired.

As a method for producing such an isoflavone aglycon from soybeans, there is a method disclosed in U.S. Patent No. 5,679,806. That is, soy bean is ground, and then ethyl alcohol or methyl alcohol solvent is added thereto so that the ratio of soybean to solvent is 1: 3 to 1:10, followed by heating at 40 to 70 ° C for extraction. Diluting it with water to obtain crude extract, and then filtering it to obtain a crude extract of isoflavone from which insoluble components have been removed; Purifying the isoflavone using an adsorption resin made of polystyrene as a substrate or an adsorbent resin made of a polyacrylic substrate in order to remove impurities such as water-soluble phenol, protein or saccharide remaining in the crude extract; And then eluting it with ethyl alcohol or methyl alcohol, decomposing it with an acid to produce an isoflavone aglycon, followed by filtration, drying and crystallization to obtain an isoflavone aglycon.

However, the above-mentioned method has an advantage that each component such as isoflavone aglycones, such as genistein, daidzein and glycitein, can be obtained in high purity. However, as described above, when a large amount of solvent is used during extraction, The time and cost of ethanol recovery can be increased. In order to obtain the aglycon of isoflavone, the structure of isoflavone can be expected to be changed by treating with acid, and in order to increase the purity, There is a need for this.

Korean Patent Publication No. 90-13878 discloses a method for producing a food material containing a glycoside substance such as isoflavonoid by ultrafiltration of a tofu waste solution in the production of food materials and health drinks from tofu waste liquids , Which requires a further improvement process to produce high purity isoflavone, i.e., isoflavone aglycon, by simple isoflavone production.

In addition, Korean Patent Publication Nos. 96-704927, 96-704928, 96-704929, 98-24261 and 98-32766 disclose a method for converting an isoflavone to an aglycon using an enzyme However, the above methods are not economical since expensive enzymes must be used in sufficient amounts.

Recently, a new manufacturing method for improving the above problems has been disclosed. Korean Patent No. 1,198,426 discloses a method for adsorbing isoflavone on a column packed with adsorbent resin, comprising the steps of adsorbing isoflavone glycoside on a column packed with adsorbent resin, Circulating an enzyme solution in an upflow and bubble infusion system to convert the adsorbed glycoside isoflavone to an unglycosylated isoflavone and recovering the converted unglycosylated isoflavone, Lt; / RTI >

However, this technique requires a large amount of resin and developing solvent (ethanol or fermentation alcohol) used in column chromatography, and it requires a complicated manufacturing process and high cost.

Therefore, there is a need to develop a method for producing isoflavone aglycon that is simple, economical, and yieldable without using an organic solvent.

Patent Document 1: U.S. Patent No. 5,506,211 Patent Document 2: U.S. Patent No. 5,679,806 Patent Document 3: Korean Patent Publication No. 90-13878 Patent Document 4: Korean Unexamined Patent Publication No. 96-704927 Patent Document 5: Korean Patent Publication No. 96-704928 Patent Document 6: Korean Patent Publication No. 96-704929 Patent Document 7: Korean Patent Publication No. 98-24261 Patent Document 8: Korean Patent Publication No. 98-32766 Patent Document 9: Korean Patent No. 1,198,426

Non-Patent Document 1: Barnes, J. et al. Nutr., 125, 777s, 1995 Non-Patent Document 2: Akiyama et al., J. Biol. Chem., 262 (2), 5592, 1987 Non-Patent Document 3: Molteni et al. Nur., 125, 751s, 1995 Non-Patent Document 4: Shino et al., Life Sci., 42 (11), 1123, 1988 Non-Patent Document 5: Fotsis et al., Proc. Natl. Acad. Sci. USA, 90, 2690, 1993 Non-Patent Document 6: Naim et al. Agric. Food Chem., 24 (6), 1174, 1976 Non-Patent Document 7: Keung and Vallee, Proc. Natl. Acad. Sci. USA, 90, 1247, 1993 Non-Patent Document 8: Peterson and Barnes, The Prostate, 22, 335, 1993

Therefore, the inventors of the present invention have found that an isoflavone aglycon production method which is relatively simple, economical, and yieldable compared to the conventional separation method using water and ethanol without using a harmful organic solvent that has been conventionally used Thereby completing the present invention.

It is therefore an object of the present invention

(a) extracting soybeans or germinated soybeans with water to recover an extract, and then filtering to obtain a filtrate;

(b) inoculating and culturing the filtered filtrate in step (a) with lactic acid bacteria;

(c) recovering the precipitate by inducing precipitation of the cultured medium cultured in step (b) at 20 DEG C for 6 to 24 hours;

(d) adsorbing the recovered precipitate using a methacrylate type resin; And

(e) eluting the adsorbed substance of step (d) with a mixed solvent of water and water to separate high purity isoflavone aglycon.

It is another object of the present invention to provide a high purity isoflavone aglycon produced by the process of the present invention.

In order to achieve the above object,

(a) extracting soybeans or germinated soybeans with water to recover an extract, and then filtering to obtain a filtrate;

(b) inoculating and culturing the filtered filtrate in step (a) with lactic acid bacteria;

(c) recovering the precipitate by inducing precipitation of the cultured medium cultured in step (b) at 20 DEG C for 6 to 24 hours;

(d) adsorbing the recovered precipitate using a methacrylate type resin; And

(e) eluting the adsorbed material of step (d) with a mixed solvent of water and water to separate high purity isoflavone aglycon.

In order to accomplish still another object of the present invention, there is provided a high purity isoflavone aglycon produced by the process of the present invention.

Hereinafter, the present invention will be described in detail.

(a) extracting soybeans or germinated soybeans with water to recover an extract, and then filtering to obtain a filtrate;

(b) inoculating and culturing the filtered filtrate in step (a) with lactic acid bacteria;

(c) recovering the precipitate by inducing precipitation of the cultured medium cultured in step (b) at 20 DEG C for 6 to 24 hours;

(d) adsorbing the recovered precipitate using a methacrylate type resin; And

(e) eluting the adsorbed material of step (d) with a mixed solvent of water and water to separate high purity isoflavone aglycon.

The soybean or germinated soybean is one of soybean and plant. Examples of the physiologically active substance of soybean include isoflavone, dietary fiber, soy oligosaccharide, lecithin, saponin, fic acid and trypsin inhibitor. In the present invention, isoflavone is extracted and used .

The isoflavone aglycon refers to an unglycosylated form of isoflavone. In the present invention, the glyphosate-type isoflavone is fermented and converted into a non-glycoside form. The aglycon form of isoflavones increases the absorption of isoflavones into the body, and the isoflavone aglycon includes both genistein, daidzein and glycitein.

The lactic acid bacterium is a microorganism having excellent isoflavone biotransformation ability and is a means used for fermentation. The type of the lactic acid bacterium is not particularly limited as long as it is a lactic acid bacterium. Preferably, lactobacillus plantarum PMO08 ( Lactobacillus plantarum PMO08), Bifidobacterium longum and Lactobacillus casei can be used, and more preferably Lactobacillus plantarum PMO08 ( Lactobacillus casei ) plantarum PMO08) may be used. The above Lactobacillus plantarum PMO08 ( Lactobacillus plantarum PMO08) is a strain developed by Pulmuone Holdings Co., Ltd., which is described in Korean Patent No. 10-0264361. In addition, the lactic acid bacteria used in the present invention are novel strains having excellent isoflavone biotransformation ability as compared with general lactic acid bacteria.

The methacrylate type resin is used as a chromatographic resin column for adsorbing and separating isoflavones. The type of the methacrylate type resin is not particularly limited, (E.g., Diaion HP2MG and Diaion HP1MG from Mitsubishi Chemical) and Amberlite XAD 7HP from Rohm & Haas.

≪ Formula 1 >

Hereinafter, the method for producing high purity isoflavone aglycon of the present invention will be described step by step.

(a) Soybean or Germination soybeans  Extracting with water to recover the extract, and then filtering to obtain a filtrate

In step (a), soybean or germinated soybean is extracted with water, the extract is recovered, and then filtered to obtain a filtrate.

Although the recovery of the extract is not limited to a specific method, it may be performed by adding hot water at 50 to 100 DEG C to soybeans or germinated soybeans. When the temperature of water is lower than 50 ° C, the extraction efficiency of isoflavone is low. When it exceeds 100 ° C, it is preferable to extract at a temperature of 50 to 100 ° C because water boils, more preferably 80 to 90 ° C . The amount of water added is preferably 4 to 10 parts by weight based on 1 part by weight of the plant, and the reaction time is preferably 30 minutes to 90 minutes.

The filtration is not limited to a specific method, but any method may be used as long as it is capable of removing the filtration foil from the extract. Preferably, a filter network or centrifugal separation may be used, and when using a filter network, it is preferable to use a filter network of 150 to 200 mesh.

(b) incubating and culturing the filtered filtrate in step (a)

In step (b), the filtrate obtained in step (a) is inoculated with lactic acid bacteria.

On the other hand, lactic acid bacteria of step (b) of the present invention Lactobacillus bacteria Planta column PMO08 (Lactobacillus plantarum PMO08), Bifidobacterium ronggeom (Bifidobatacterium longum) and Lactobacillus casei (Lactobacillus casei ) can be selected.

The lactic acid bacterium is a microorganism having excellent isoflavone biotransformation ability and is used for fermentation, and as described above.

Preferably, the culture conditions are such that the tablets are added so that the concentration of the isoflavone of the filtrate filtered in the step (a) is 5 to 30%, sterilized at 85 ° C for 30 minutes and cooled to 30 ° C, To 5% to 10% of the culture medium, and the mixture is preferably cultured at 30 DEG C for 2 days to 5 days with stirring.

(c) recovering the precipitate by inducing precipitation of the cultured medium cultured in step (b) at 20 DEG C for 6 to 24 hours

In step (c), the culture solution cultured in step (b) is induced to precipitate at 20 DEG C for 6 to 24 hours to recover the precipitate.

After the step of recovering the precipitate, centrifuging the further recovered precipitate may be included. The impurity (excess supernatant) is further removed through centrifugation to produce isoflavone of high purity. At this time, in the centrifugal separation method, it is preferable that the collected precipitate is centrifuged at 5,000 to 7,000 rpm for 10 to 20 minutes.

In addition, it may include neutralizing the precipitate obtained. Neutralization refers to the method used to adjust the pH of a precipitate to a suitable pH as a food additive by controlling the pH to near neutral. Preferably, the alkali solution may be added so that the pH of the precipitate is 6 to 8, and the alkali solution is preferably sodium hydroxide or potassium hydroxide.

(d) recovering the recovered precipitate Methacrylate system  Suzy( 메록acrylate type  resin adsorption step

In step (d), the recovered precipitate is adsorbed using a methacrylate type resin.

Meanwhile, the methacrylate resin of step (d) of the present invention may be a resin having a structure represented by the following formula (1).

≪ Formula 1 >

The methacrylate type resin refers to a means used as a chromatography resin column for adsorbing and separating isoflavones, and has been described above.

The methacrylate type resin used in the above and the precipitate dissolved in the aqueous solution are preferably used in a weight ratio of 10 to 30: 1.

(e) eluting the adsorbed substance of step (d) with a mixed solvent of water and water to separate high purity isoflavone aglycon

In step (e), the adsorbed material of step (d) is eluted with a mixed solvent of water and water to separate high purity isoflavone aglycon.

The step (e) may include the steps of: i) eluting with water to remove a water-soluble substance; And

Ii) eluting with a mixed solvent of water and water to separate the high purity isoflavone aglycon.

(I) of step (e) refers to a method used for removing a water-soluble substance from water when eluting isoflavone aglycon to increase the recovery rate, and it is preferable that water is used at a rate of 0.5-2 SV As shown in Fig.

SV is the abbreviation of space velocity, which is a value obtained by dividing the flow rate per hour in the column by the volume of the adsorbent resin, and BV is the volume of the bed volume, which means the volume occupied by the filler in the column.

In the step (ii) of the step (e), the mixing ratio of the alcohol and water may be 1: 1 to 9.

(V / w) ethanol solution to a 50% (v / w) ethanol solution as an eluting solvent. The ethanol concentration of the isoflavone aglycon in the step (e) And gradually elongating while gradually increasing. At this time, the velocity of the supernatant and developing solvent may preferably be SV 1 to 3, and the volume may preferably be BV 5 to 10, more preferably 3 to 5 times.

Thus, the present invention provides high purity isoflavone aglycon produced by the process of the present invention. At this time, the high purity may be a high purity isoflavone aglycon having a purity of 50 wt% or more. The high purity isoflavone aglycon of the present invention has a very high purity as compared with that prepared by the conventional production method.

According to the production method of the present invention, a high purity isoflavone aglycon can be obtained in an amount smaller than that of the conventional alcohol.

These effects are well illustrated in the comparative example of the present invention.

In Comparative Example of the present invention, the adsorption rate of isoflavone and the amount of alcohol used according to the adsorbent resin were compared with each other according to the method disclosed in Korean Patent No. 1,198,426 (Comparative Example 1, Comparative Example 2 and Table 1). As a result, when methacrylate type resin synthetic resin was used, as shown in Table 1, when a certain amount of soybean extract (BV 40) passed through the conventional method, the adsorption rate of the glycoside isoflavone rapidly However, as shown in Table 1, when the washing and adsorption were repeated according to the method of the present invention, it was confirmed that the adsorption rate of the glycoside isoflavone was maintained at 90% or more even when it was flowed more than 100 times of the column.

As a result of comparing the amount of alcohol required for recovery of the glycoside isoflavone adsorbed on the synthetic resin of the comparative example and the synthetic resin of the present invention, 2,000 ml of 80% alcohol was used in the comparative example. However, the methacrylate synthetic resin Was able to recover all of the adsorbed glycoside isoflavones even when 1,500 ml of 50% alcohol was used.

The effect of the method of the present invention is to improve the yield of high purity isoflavone aglycon, which is harmless to the human body as compared with the conventional method, by using methacrylate synthetic resin.

The method of producing isoflavone aglycone according to the present invention is characterized by comprising: (a) extracting soybean or germinated soybean with water, recovering the extract, and then filtering to obtain a filtrate; (b) inoculating and culturing the filtered filtrate in step (a) with lactic acid bacteria; (c) recovering the precipitate by inducing precipitation of the cultured medium cultured in step (b) at 20 DEG C for 6 to 24 hours; (d) adsorbing the recovered precipitate using a methacrylate type resin; And separating the high purity isoflavone aglycon by eluting the adsorbed material of step (d) with a mixed solvent of a water and a water, and separating the high purity isoflavone aglycon from water and ethanol without using a harmful organic solvent which is usually used, , The production process is relatively simple, economical, and the yield is relatively higher than the conventional separation method of isoflavone aglycon.

FIG. 1 is a diagram showing the process for producing a high purity isoflavone aglycon of the present invention.
2 is an HPLC chromatogram of soybean extract.
Figure 3 is an HPLC chromatogram of isoflavone aglycon prepared according to the method of the present invention.

Hereinafter, the present invention will be described in detail.

However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

< Example  1>

Preparation of high purity isoflavone aglycon

<1-1> Recovery of soybean extract and obtaining filtrate

The soybean or germinated soybean was subjected to hot water extraction by adding water at 50 to 100 캜. When the temperature of water is less than 50 캜, the extraction efficiency of isoflavone is low. When the temperature exceeds 100 캜, water boils. Therefore, 4 to 10 parts by weight of water is added to 1 part of the plant at a temperature of 50 to 100 캜 And extracted by stirring for 30 minutes to 90 minutes.

After extracting the soybean extract extracted in the above step, the filtrate was obtained by filtration using a filter mesh of 150 to 200 mesh.

<1-2> Inoculation and culture of microorganisms

Tablets were added to the filtrate of Example <1-1> so that the concentration of isoflavone was 5 to 30%, sterilized at 85 DEG C for 30 minutes, cooled to 30 DEG C and then added with fresh microorganism having isoflavone biotransformation Lactobacillus plantarum PMO08 ( Lactobacillus plantarum PMO08) KFCC 11028 was inoculated at 5 to 10% of the liquid amount and cultured at 30 占 폚 for less than 2 days, more than 5 days with stirring.

&Lt; 1-3 > Precipitation and recovery of precipitate

The culture broth cultured in Example <1-2> was induced to precipitate at 20 ° C for 6 to 24 hours, separated into supernatant and precipitate, and the precipitate was recovered. The recovered precipitate was centrifuged at 5,000 to 7,000 rpm for 10 to 20 minutes to obtain a precipitate from which impurities (surplus supernatant) were removed.

To 1 part by weight of the obtained precipitate, 1.0 to 1.4 parts by weight of distilled water was added, and then 25 to 35% (w / v) of sodium hydroxide or potassium hydroxide was added and stirred for 20 to 40 minutes to obtain a pH of 6 to 8 Neutralized.

<1-4> Separation and purification of precipitate

The precipitate obtained in Example <1-3> was dissolved in water to prepare a water-soluble solution, which was then separated and purified using a methacrylate type resin column, which was a separation column for separation adsorption chromatography. The methacrylate type resin and the precipitate dissolved in the aqueous solution were used at a weight ratio of 10 to 30: 1.

In addition, when performing the separation and purification process, the first eluate is passed through BV 5 to 10 at a speed of SV 0.5 to 2 using water to remove the water-soluble substance, and then 10% (v / w) The ethanol solution was eluted sequentially with increasing ethanol concentration to 50% (v / w) ethanol solution.

Fractions were prepared by concentrating only the isoflavone fraction in the fractions. At this time, when the content of isoflavone aglycon in the fraction was less than 10%, the separated and purified process was repeated after collecting and concentrating under reduced pressure. The fractions having an isoflavone aglycone content of 10% or more were also collected, and the filtrate was concentrated under reduced pressure and the separation and purification process was repeated. The filtrate was concentrated under reduced pressure and then dried to obtain isoflavone aglycon &Lt; / RTI &gt;

< Comparative Example  1>

On adsorbent resin  Of isoflavones Adsorption rate  compare

As a comparative example, the adsorption rates of isoflavones were compared as disclosed in Korean Patent No. 1,198,426. The content of glyoxal isoflavone contained in the soybean extract was analyzed by High Performance Liquid Chromatography (HPLC). The soybean extract solution was dissolved in 100% methanol through ultrasonic treatment, and the liquid component obtained by filtration was subjected to HPLC to determine glycosyl isoflavone content.

Organization: Shimadzu Prominence LC-20AD

Column: Symmetry C18 4.6 (i, d) x 250 mm

Mobile phase A: acetonitrile (containing 1% acetic acid): distilled water = 15: 85

Mobile phase B: acetonitrile (containing 1% acetic acid): distilled water = 65: 35

Flow rate: 1.0 ml / min

Detector: UV detector 254 nm

The test method was the same as that of the comparative example except that only the type of adsorbent resin was different. The content of glyoxal isoflavone contained in the soybean extract used in the adsorption rate comparative test was 7 mg / g.

In the case of the comparative example, soybean extract was prepared and 12,000 ml of soybean extract was flowed in a column filled with 1,000 ml of Diaion HP-20 synthetic resin for 6 hours at a time, 12,000 ml of purified water (BV 12) Lt; / RTI &gt; and slowly washed through the column. This procedure was repeated three times for 3 days to adsorb 36,000 ml (BV 36) of soybean extract to the column. Every two hours, the purified product passed through the column was recovered to remove the adsorbed glycoside isoflavone The content was analyzed by HPLC.

As a control, methacrylate type resin synthetic resin used in the present invention was packed in a column of 1,000 ml, and the soybean extract solution was continuously loaded and adsorbed on the column as in the comparative example. 12,000 ml of soybean extract was adsorbed onto the column, and 12,000 ml of purified water (BV 12) was slowly passed through the column for 12 hours to wash the column. This procedure was repeated three times for 3 days to adsorb 36,000 ml (BV 36) of soybean extract to the column. Every two hours, the purified product passed through the column was recovered to remove the adsorbed glycoside isoflavone The content was analyzed by HPLC.

As a result, according to the conventional method, as shown in Table 1, when a certain amount of the soybean extract (BV 40) passes through, the adsorption rate of the glycoside isoflavone sharply drops, but washing and adsorption are repeated according to the method of the present invention As shown in Table 1, it was confirmed that the adsorption rate of the glycoside isoflavone was maintained at 90% or more even when it was flowed more than 100 times of the column.

Comparison of the maximum adsorption amount of adsorbed resin when loading soybean extract BV25 BV40 BV55 BV70 BV85 BV100 BV115 BV130 Comparative Example 1 98.5 95.3 77.8 69.3 54.1 11.1 0 0 Invention 99.4 98.6 97.7 96.4 95.7 95.1 88.4 76.4

< Comparative Example  2>

On adsorbent resin  Comparison of alcohol consumption for recovery of isoflavones

The amount of alcohol required for recovery of the glycoside isoflavone adsorbed on the synthetic resin of the comparative example and the present invention was compared. As a result, 2,000 ml of 80% alcohol was used in the comparative example, but the methacrylate synthetic resin proposed in the present invention was able to recover all of the adsorbed glycoside isoflavones even when 1,500 ml of 50% alcohol was used.

As described above, the strain having excellent isoflavone conversion ability of the present invention and the method for producing isoflavone aglycon using the same, comprises the steps of: (a) extracting soybean or germinated soybean with water, recovering the extract, and then filtering to obtain a filtrate; (b) inoculating and culturing the filtered filtrate in step (a) with lactic acid bacteria; (c) recovering the precipitate by inducing precipitation of the cultured medium cultured in step (b) at 20 DEG C for 6 to 24 hours; (d) adsorbing the recovered precipitate using a methacrylate type resin; And separating the high purity isoflavone aglycon by eluting the adsorbed material of step (d) with a mixed solvent of a water and a water, and separating the high purity isoflavone aglycon from water and ethanol without using a harmful organic solvent which is usually used, , It is more likely to be used industrially because the production process is relatively simple, economical, and the yield is higher than the conventional separation method of isoflavone aglycon.

Claims (7)

(a) extracting soybeans or germinated soybeans with water to recover an extract, and then filtering to obtain a filtrate;
(b) inoculating and culturing the filtered filtrate in step (a) with lactic acid bacteria;
(c) recovering the precipitate by inducing precipitation of the cultured medium cultured in step (b) at 20 DEG C for 6 to 24 hours;
(d) adsorbing the recovered precipitate using a methacrylate type resin; And
(e) eluting the adsorbed material of step (d) with a mixed solvent of water and water to separate the high purity isoflavone aglycon.
The method according to claim 1, wherein the lactic acid bacterium of step (b) is selected from the group consisting of Lactobacillus plantarum PMO08, Bifidobatacterium longum , and Lactobacillus casei . The method of claim 1, wherein the isoflavone aglycon is selected from the group consisting of Lactobacillus casei , Lactobacillus casei , and Bifidobacterium longum .
The method for producing high purity isoflavone aglycone according to claim 1, wherein the methacrylate resin of step (d) is a resin having a structure of the following formula (1).
&Lt; Formula 1 >

2. The method of claim 1, wherein step (e)
I) eluting with water to remove the water-soluble material; And
Ii) eluting with a mixed solvent of alcohol and water to separate high purity isoflavone aglycon.
[Claim 5] The method according to claim 4, wherein the mixing ratio of the alcohol in the step (ii) is 1: 1 to 9: 1.
A high purity isoflavone aglycon produced by the method of claim 1.
The high purity isoflavone aglycon according to claim 6, wherein the high purity is a purity of at least 50% by weight.

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