KR102212800B1 - Manufacturing method of fermented liquid of liquorice extract with increased amount of bioactive phytoestrogen and enriched calcium and the fermented liquid produced therefrom - Google Patents

Abstract

The present invention extracts phytoestrogen from licorice, which has a high content of phytoestrogen of a plant female hormone, has been scientifically reported in Korea and abroad, has proven safety, and increases the concentration of the active form through fermentation of lactic acid bacteria. Through technology, the concentration of active ingredients is higher than that of simple extracts, and minerals derived from calcium and deep ocean water are added to help relieve various symptoms caused by a lack of female hormones and prevent osteoporosis, and can be consumed without side effects. It relates to a method for preparing a fermented broth for enhancing active phytoestrogen and fortified calcium of licorice extract, which can be prepared in various forms such as liquid, granules, and pills, and a fermented broth prepared therefrom.

Description

Manufacturing method of fermented liquid of liquorice extract with increased amount of bioactive phytoestrogen and enriched calcium and the fermented liquid produced therefrom}

The present invention extracts phytoestrogen from licorice, which has a high content of phytoestrogen of a plant female hormone, and a mechanism of action has been scientifically reported in Korea and abroad, and its safety is proven and highly recognized,

Through technology that increases the concentration of active form by fermentation of lactic acid bacteria, the concentration of active ingredients is higher than that of simple extracts,

Calcium and minerals derived from deep ocean water are added to help relieve various symptoms caused by a lack of female hormones and prevent osteoporosis.

It relates to an active phytoestrogen increase and calcium-enhanced fermentation broth manufacturing method of licorice extract and a fermentation broth prepared therefrom, which can be prepared in various forms such as liquids, granules, pills, etc. that can be consumed without side effects.

Licorice is a perennial herbaceous plant whose origin is Inner Mongolia and belongs to the legume family. The root is used as a medicinal material. It has a sweet taste, no poison, and a warm energy, so it is often used as a component of herbal medicine.

Recently, research on the physiologically active ingredients of licorice is active, and it is widely used not only as a component of herbal medicine, but also as a processed food ingredient. In particular, the vegetable estrogen component of licorice is attracting attention in foreign countries. The physiologically active substances of licorice include glycyrrhizin, a triterpenoid-based saponin known as a sweetener, and lliquiritin, liquiritigenin, and isoliquiritin, which are flavonoid-based phytoestrogen components.

The present invention extracts the plant female hormone phytoestrogen from the licorice and increases the concentration of the active form (non-glycosyl form) by fermentation of lactic acid bacteria, and the concentration of the active ingredient is higher than that of the simple extract, and it is derived from calcium and deep sea water. This invention relates to a fermentation broth that is added to one mineral to help relieve various symptoms caused by a lack of female hormones and prevent osteoporosis.

In relation to the present invention, in the'composition using licorice extract' of Korean Patent Registration No. 10-0177832 (registration date 1998.11.19), a composition for a functional food using a licorice extract and a technology for a functional food containing the functional food composition Is disclosed,

In Korean Patent Registration No. 10-0586813 (Registration Date 2006.05.29)'Mixed herbal extracts and health foods for osteoporosis prevention or treatment containing them', herbal medicines containing licorice are extracted from alcohol or alcoholic aqueous solution to prevent or treat osteoporosis Technology for the extract is disclosed,

In Korean Patent Registration No. 10-1544532 (registration date 2015.08.07)'a composition for preventing or treating female menopausal diseases containing baekshou extract', for a food composition for preventing or improving female menopausal diseases containing licorice extract The technology is disclosed.

However, the conventionally disclosed techniques have been scientifically reported in Korea and abroad to present through the present invention, extract pitestrogen from licorice with proven safety and high recognition, and increase the concentration of the active form through fermentation of lactic acid bacteria. Through technology, the concentration of active ingredients is higher than that of simple extracts, and minerals derived from calcium and deep sea water are added, so there is no disclosure on a fermentation broth manufacturing technology that helps to relieve various symptoms caused by a lack of female hormones and prevent osteoporosis. Also, it is difficult to derive a fermentation broth manufacturing technology from the disclosed technologies.

Korean Patent Registration 10-0177832 (Registration date 1998.11.19) Korean Patent Registration 10-0586813 (Registration Date 2006.05.29) Korean Patent Registration 10-1544532 (Registration date 2015.08.07)

The present invention extracts phytoestrogen from licorice, which has a high content of phytoestrogen of a plant female hormone, and a mechanism of action has been scientifically reported in Korea and abroad, and its safety is proven and highly recognized,

Through technology that increases the concentration of active form by fermentation of lactic acid bacteria, the concentration of active ingredients is higher than that of simple extracts,

Calcium and minerals derived from deep ocean water are added to help relieve various symptoms caused by a lack of female hormones and prevent osteoporosis.

It is an object of the present invention to provide a method for producing an active phytoestrogen and calcium-enhanced fermentation broth of licorice extract and a fermentation broth prepared therefrom that can be prepared in various forms such as liquids, granules, and pills that can be consumed without side effects. .

To achieve the above object,

The present invention is deep sea water or purified water; And mixing and extracting herbs to prepare a herbal extract (S10),

Adjusting the concentration of the herbal extract (S20) and,

A step (S30) of forming a mixture by mixing the herbal extract and calcium whose concentration is adjusted through the step (S20),

Sterilizing the mixture (S40),

Step (S50) of inoculating lactic acid bacteria into the mixture sterilized through the step (S40),

It provides a method for producing a fermentation broth for increasing active phytoestrogen and calcium enrichment of licorice extract, comprising the step (S60) of fermenting a mixture inoculated with lactic acid bacteria through the step (S50) to prepare a fermentation broth.

In addition, as prepared through the above-described manufacturing method, it provides an active phytoestrogen increase and calcium-enhanced fermentation broth of licorice extract, forming any one of a liquid, pill, or granule. The fermentation broth can be used as a vegetable female hormone supplement for relieving menopausal symptoms.

The licorice extract fermentation broth according to the present invention has the following effects.

first. By extracting phytoestrogen, a vegetable female hormone, from licorice and increasing the concentration of the active form (non-glycoside type) through fermentation of lactic acid bacteria, the concentration of the active ingredient is higher than that of the simple extract.

second. Calcium and minerals derived from deep sea water are added to help alleviate various symptoms caused by a lack of female hormones and prevent osteoporosis.

third. There is no worries about side effects, and it has the advantage of being able to provide vegetable supplements in various forms such as liquids, granules, and pills.

1 is a manufacturing process diagram according to the method for producing an active phytoestrogen increase and calcium-enhanced fermentation broth of the licorice extract of the present invention.
Figure 2 is a graph showing fermentation characteristics according to the concentration of licorice extract before and after inoculation of Lactobacillus plantarum DK119 (119) isolated from kimchi.
Figure 3 is a graph showing fermentation characteristics according to the concentration of licorice extract before and after inoculation of Lactobacillus casei DK128(128) isolated from kimchi.
Figure 4 is a graph showing fermentation characteristics according to the concentration of licorice extract before and after inoculation of commercial lactic acid bacteria (Chr. Hansen. Hoersholm Danmark) Bifidobacterium lactis BB-12 (BB).
5 is a graph showing fermentation characteristics according to the inoculation concentration of lactic acid bacteria Lactobacillus plantarum DK119 (119) isolated from kimchi.
Figure 6 is a graph showing fermentation characteristics according to the inoculation concentration of lactic acid bacteria Lactobacillus casei DK128 (128) isolated from kimchi.
7 is a graph showing fermentation characteristics according to the inoculation concentration of commercial lactic acid bacteria (Chr. Hansen. Hoersholm Danmark) Bifidobacterium lactis BB-12 (BB).
Figure 8 is a graph showing the fermentation characteristics of licorice extracted with Deep Sea Water-DSW and inoculated with the lactic acid bacteria Lactobacillus plantarum DK119 (119) isolated from kimchi.
Figure 9 is a graph showing the fermentation characteristics of licorice inoculated with Lactobacillus casei DK128 (128) extracted with Deep Sea Water-DSW and isolated from kimchi according to the present invention.
Figure 10 is a graph showing the fermentation characteristics of licorice inoculated with Bifidobacterium lactis BB-12 (BB) extracted with Deep Sea Water-DSW and commercial lactic acid bacteria (Chr. Hansen. Hoersholm Danmark) according to the present invention.
11 is a graph showing the change in reducing sugar content of the licorice fermentation broth extracted with deep sea water (DSW) according to the present invention.
12 is a graph showing the change in the content of phytoestrogen using Liquiritin as an active ingredient in the licorice fermentation broth extracted with deep ocean water (DSW) according to the present invention.
13 is a graph showing the change in the content of phytoestrogen using Liquiritigenin as an active ingredient in the licorice fermentation broth extracted with deep ocean water (DSW) according to the present invention.
14 is a graph showing the change in the content of phytoestrogen using Isoliquiritin as an active ingredient in the licorice fermentation broth extracted with deep ocean water (DSW) according to the present invention.
15 is a graph showing the change in the content of Phytoestrogen using Isoliquiritigenin as an active ingredient in the licorice fermentation broth extracted with deep ocean water (DSW) according to the present invention.
16 is a graph showing the effect of inoculation of lactic acid bacteria Lactobacillus plantarum DK119 (119) isolated from kimchi according to the present invention, and the effect of calcium addition on the fermentation of licorice extract.
17 is a graph showing the effect of inoculation of lactic acid bacteria Lactobacillus casei DK128 (128) isolated from kimchi according to the present invention, and the effect of calcium addition on fermentation of licorice extract.
18 is a graph showing the effect of inoculation of Bifidobacterium lactis BB-12 (BB), calcium addition on the fermentation of licorice extract, according to the present invention, commercial lactic acid bacteria (Chr. Hansen. Hoersholm Danmark).
19 is a graph showing the effect of calcium addition on the change of reducing sugar content of fermented licorice broth according to the present invention.
20 is a graph showing the effect of calcium addition on the content of phytoestrogen using Liquiritin as an active ingredient in a licorice fermentation broth according to the present invention.
21 is a graph showing the effect of calcium addition on the content of phytoestrogen using Liquiritigenin as an active ingredient in a licorice fermentation broth according to the present invention.
22 is a graph showing the effect of calcium addition on the content of phytoestrogen using Isoliquiritin as an active ingredient in a licorice fermentation broth according to the present invention.
23 is a graph showing the effect of calcium addition on the content of Phytoestrogen using Isoliquiritigenin as an active ingredient in a licorice fermentation broth according to the present invention.

Hereinafter, the technical configuration according to the present invention presented above will be described in detail.

As mentioned above,

As shown in Figure 1, the active phytoestrogen increase and calcium-enhanced fermentation broth production method of licorice extract according to the present invention includes: deep sea water or purified water; And mixing and extracting herbs to prepare a herbal extract (S10),

Adjusting the concentration of the herbal extract (S20) and,

A step (S30) of forming a mixture by mixing the herbal extract and calcium whose concentration is adjusted through the step (S20),

Sterilizing the mixture (S40),

Step (S50) of inoculating lactic acid bacteria into the mixture sterilized through the step (S40),

It comprises a step (S60) of fermenting the mixture inoculated with lactic acid bacteria through the step (S50) to prepare a fermentation broth.

The fermentation broth according to the present invention extracts phytoestrogen, a vegetable female hormone from licorice, and increases the concentration of the active form (non-glycosyl form) through fermentation of lactic acid bacteria, so that the concentration of the active ingredient is higher than that of the simple extract, and also in calcium and deep sea water. Derived minerals are added to help relieve various symptoms caused by a lack of female hormones and prevent osteoporosis.

The manufacturing process of the fermentation broth having such characteristics will be described in each step.

[Medicinal herb extract manufacturing step (S10)]

This step (S10) includes deep sea water or purified water; It is a step of mixing and extracting herbs to prepare a herbal extract.

The herbal extract is prepared by mixing 1.0 to 20.0 wt% of licorice chopped into slices and 80.0 to 99.0 wt% of deep sea water, and then extracting it at 100 to 120°C for 8 to 15 hours using an herbal extractor.

If the amount of licorice is less than 1.0 wt%, there is a problem that the phytoestrogen content is low, and if it exceeds 20.0 wt%, there is a problem that extraction is not smooth, so the amount of licorice is 1.0 ~ It is preferable to limit it within the range of 20.0 wt%.

The amount of the deep sea water or purified water is determined according to the amount of licorice, and is preferably limited to a range of 80.0 to 99.0 wt% according to the amount of licorice.

[Herbal extract concentration control step (S20)]

This step (S20) is a step of adjusting the concentration of the herbal extract extracted through the step (S10), by measuring the dry solid concentration of the herbal extract is adjusted to a concentration of 1 ~ 5%. Standardization of the production process can be achieved through such concentration control.

[Step of configuring a mixture of herbal extracts and calcium (S30)]

This step (S30) is a step of forming a mixture by mixing the herbal extract and calcium whose concentration is adjusted from the step (S20).

The mixture is composed by mixing 0.05 to 0.2 parts by weight of calcium based on 100.0 parts by weight of the herbal extract.

If the amount of calcium is less than 0.05 parts by weight, calcium fortification is insignificant and there is a problem in highlighting calcium fortification in the product, and if it exceeds 0.2 parts by weight, there is a problem in solubility and taste, so calcium mixed with the herbal extract The amount of used is preferably limited to within the range of 0.05 to 0.2 parts by weight.

The calcium is one or two or more selected from fermented calcium, calcium citrate, or calcium lactate.

[Mixture sterilization step (S40)]

This step (S40) is a step of sterilizing the mixture prepared through the step (30).

The sterilization is performed by heating at 80 to 100 °C for 20 to 40 minutes. More specifically, the mixture is heated at 90° C. for 30 minutes.

[Step of inoculating lactic acid bacteria in the mixture (S50)]

This step (S50) is a step of inoculating the lactic acid bacteria in the sterilized mixture through the step (S40).

The lactic acid bacteria are Lactobacillus plantarum DK119 (119), Lactobacillus casei DK128 (128) isolated from kimchi; Or commercial lactic acid bacteria (Chr. Hansen. Hoersholm Danmark) Bifidobacterium lactis BB-12 (BB) as any one or two or more selected from, and inoculated with 0.01 to 0.2 parts by weight based on 100 parts by weight of licorice extract.

If the inoculation is less than 0.01 parts by weight, there is a problem that fermentation does not occur, and if it exceeds 0.2 parts by weight, there may be destruction of physiologically active ingredients due to a rapid increase in acidity, so the inoculation is performed in 100 parts by weight of licorice extract. It is preferable to make it within the range of 0.01 to 0.2 parts by weight.

[Mixture fermentation step (S60)]

This step (S60) is a step of fermenting a mixture inoculated with lactic acid bacteria to prepare a fermentation broth, which is prepared by fermenting at 30 to 38°C for 3 to 5 days.

The fermentation broth prepared as described above may be prepared in various forms, such as a pill or granule form, as needed, in addition to a liquid form.

Hereinafter, the production process of the active phytoestrogen and calcium-enhanced fermentation broth of the licorice extract according to the present invention will be described through test examples.

1) Sample extraction and calcium addition

Licorice slices (cut herbal medicine) are mixed with deep sea water (QBM Co., Ltd.) or purified water so that 1.0 ~ 20.0 wt% is obtained, and extracted for 12 hours using an herbal extractor.

After adjusting the concentration of the extract, 0.25 g (calcium content 52.5 mg) per 100 mL of the licorice extract is added to calcium (100% Gulgak, KMF Co., Ltd., water-soluble fermented calcium S).

2) Lactobacillus inoculation and fermentation

Licorice extract is sterilized by heating at 80 ~ 100 ℃ for 30 minutes, and fermented using 3 kinds of lactic acid bacteria in freeze-dried form.

Lactobacillus plantarum DK119(119), Lactobacillus casei DK128(128) and Chr. Bifidobacterium lactis BB-12 (BB) of Hansen (Hoersholm Danmark) was inoculated with 0.01 ~ 0.2% (w/v) and fermented at 37° C. for 4 days.

3) pH and titratable acidity measurement

For all samples except for the measurement of the number of lactic acid bacteria, the fermentation broth is centrifuged at 3,000 rpm for 10 minutes using a centrifuge (Union 32R, Hanil Scientific Inc. Kimpo, Korea) to precipitate the lactic acid bacteria, and the supernatant is used as a sample.

pH is measured using a pH meter (Orion 3 star, Thermo Fisher Scientific Inc., Beverly, MA, USA).

For titration acidity, dilute the sample with distilled water to take 25 mL, titrate with 0.1 N NaOH using 0.1% phenolphthalein as an indicator, and convert the 0.1 N NaOH volume consumed to neutralize the dilution into %-lactic acid content. Indicate.

4) Analysis of reducing sugar

Quantitative analysis is performed according to the DNS (3,5-dinitrosalicylic acid) method.

5) Analysis of major phytoestrogen components of licorice (indicator)

The main phytoestrogen indicators of licorice are the glycoside forms of liquiritin and isoliquiritin, and their non-glycoside forms of liquirigenin and isoliquiritigenin by HPLC Analyze by method.

After fermentation, the sample is centrifuged at 3,000 rpm for 10 minutes, the supernatant is diluted with 50%-EtOH, filtered through a 0.45 µm filter, and used for analysis.

All solvents and mobile phases used for HPLC analysis and sample preparation are HPLC grade, and J.T. The product of Baker (Phillipsburg, NJ, USA) was used, and the standard material was purchased and used from Sigma (St. Louis, MO, USA).

The HPLC system used for the analysis was Dionex Summit HPLC system (Germany), P680 HPLC pump, ASI-100 automated sample injector. It consists of a thermostated column compartment TCC-100, UVD 340U UV/VIS detector, and processes data analyzed through the Dionex Chromelon chromatography data system.

The analysis conditions are shown in Table 1 below.

Parameters Condition HPLC model
Column
Detector
Flow rate
Column oven temperature
Injection volume Dionex 4000i
YMC Hydrosphere C18 (S-5μm,12 nm, 250 x 4.6 mmI.D.)
UV 250 nm
1.0 mL/min (gradient)
30℃
20 μL

The mobile phase is 100%-Acetonitril (Sol. A) and 0.1%-Acetic acid (Sol. B), and the gradient as in Table 2 below.

Time(min) 100%-Acetonitril (Sol. A) 0.1%-Acetic acid (Sol. B) 10
15
20
30 10%
15%
50%
10% 90%
75%
50%
90%

[ result ]

1) Fermentation characteristics of licorice with different extract concentration and inoculum concentration

2 to 4,

10%-Licorice extract (w/w) was inoculated with 0.1% of three types of lactic acid bacteria (w/v), and the pH was measured before fermentation (D0) and from 1 day (D1) to 4 days (D4) of fermentation. The licorice extract fermented with reached the pH of 4 in D1 and showed the fastest fermentation rate. 119 and BB were also lower than pH 4 after D2.

The titratable acidity change of licorice liquor fermented with 10%-licorice extract (w/w) was inoculated with 0.1%, and the fermented licorice liquor rose the fastest, followed by an increase in titratable acidity of 119 and BB.

In addition, FIGS. 2 to 4 show a result of inoculating (w/v) three strains with 0.1% of 20%-licorice water extract and measuring the pH change. It can be seen that the pH decreases rapidly to pH 3.8 on the first day of fermentation (D1), and it can be seen that the pH decreases rapidly in the fermentation broth 119 and BB.

The titratable acidity change of the licorice extract inoculated with 20%-licorice extract (w/w) 0.1%-fermented licorice solution increased the fastest, followed by the increase in titratable acidity of 119 and BB.

In conclusion, as can be seen from the drawings shown in Figs. 2 to 4, the fermentation of 10%-licorice extract and 20%-licorice extract is 0.1% of 119, 128 and BB inoculation fermentation depending on the lactic acid bacteria pH and titratable acidity There are slight differences, but it was confirmed that fermentation proceeds rapidly by all three strains.

5 to 7 are inoculated with 20%-licorice extract with 119, 128 and BB with 0.2%-and measured pH and titratable acidity to analyze fermentation characteristics and compared with 0.1%-inoculation results.

As shown in Figs. 5 to 7, the fermentation of 20%-licorice extract showed similar fermentation rates when 119, 128 and BB were inoculated with 0.1% and 0.2%- and fermented. That is, fermentation took place efficiently in both 10% and 20%-licorice extract with 0.1%-inoculation.

2) Fermentation Characteristics of Licorice Licorice Extracted from Deep Ocean Water

8 to 10, the fermentation characteristics of the water extract and the sea water extract were very similar, so that the pH of the fermentation day per day was sharply lowered and the titratable acidity increased. It was found that the number of lactic acid bacteria decreased after 1 day of fermentation (D1). These fermentation characteristics were similar in fermentation with 119, 128 and BB strains.

As shown in Figure 11, looking at the change in reducing sugar during the fermentation process of 20%-licorice extracted with deep seawater, it begins to decrease from day 1 of fermentation in all 119, 128 and BB bacteria, and in particular, 0.1%-128 inoculation groups The reduction in reducing sugar was found to be the greatest.

These results are consistent with changes in pH and titratable acidity. In addition, it is estimated that deep sea water extract shows a greater reduction in reducing sugar than water extract, and deep sea water promotes the fermentation of lactic acid bacteria.

3) Effect of deep sea water on the change of phytoestrogen content in licorice fermentation broth

119, 128 and 0.1% BB are inoculated and fermented with 20% licorice extract extracted from deep ocean water, and vegetable estrogen liquiritin, isoliquiritin, isoliquiritin, which are representative bioactive substances of licorice by fermentation of lactic acid bacteria The purpose of this study is to analyze the degree of conversion to the aglycone forms, liquiritigenin and isoliquiritigenin, and then investigate the effect of deep ocean water on biotransformation.

Figure 12 shows the change of the content of liquorice by fermentation, and the liquoitin content (Water group) of the liquorice fermentation broth extracted with water is about 15 to 20 in the fermentation broths 119 and 128 after 4 days of fermentation (D4) compared to before fermentation. It shows a decrease of about %, and about 30% in BB fermentation broth.

The change in the content of liquorite in the extract extracted with deep ocean water (DSW group) is similar to the result of the extract extracted with water. After 4 days of fermentation (D4), the 119 and 128 fermentation broths showed a decrease of 15 to 20%. It can be seen that it is reduced by about 30%.

13 shows the change in the content of requeritygenin by fermentation, and the content of requeritygenin (Water group) of the extract extracted with water is about 4 days after fermentation (D4) by fermentation of 119 and 128 compared to before fermentation. It increased 2.3 times and increased by about 4 times by BB fermentation.

The change in the content of liquorite in the extract extracted with deep ocean water (DSW group) is similar to the result of the extract extracted with water, and increased by about 2.3 times by 119 and 128 fermentation 4 days after fermentation (D4), and about 2.3 times by BB fermentation. It can be seen that it increases four times.

Table 3 and FIG. 14 below show the change in the content of isoliquaritin by fermentation, and the isoliquaritin content (Water group) of the licorice extract extracted with water was compared with before fermentation 4 days after fermentation (D4) 119 and 128 It decreased by about 44 ~ 55% in the fermented broth and by 84% in the BB fermented broth.

The change in the content of isoliquaritin in the extract extracted with deep sea water (DSW group) is similar to the result of the extract extracted with water, and decreased by about 45 to 55% by 119 and 128 fermentation 4 days after fermentation (D4), and BB fermentation liquid It can be seen that about 85% decrease in

Change of isoliquaritin content in fermented licorice extract extracted from sea water (ug/mL) 119-0.1% vaccination 128-0.1% vaccination BB-0.1% vaccination Day DSW Water DSW Water DSW Water D0 86.97 85.34 86.97 85.34 86.97 85.34 D4 46.88 47.66 38.23 38.23 12.11 13.24

Table 4 and FIG. 15 below show the change in the content of isoliqueritygenin by fermentation, and the isoliqueritygenin content (Water group) of the licorice extract extracted with water was compared with before fermentation 4 days after fermentation (D4). The contents were 119 and 128, which increased about 2.7 times in the fermented broth and about 4 times in the BB fermented broth.

Even in the extract extracted with deep ocean water (DSW group), the content of D4 after 4 days of fermentation was 119 and 128, which was increased by about 2.7 times in the fermented broth, and about 4 times in the BB fermented broth.

Changes in isoliquerigenin content in fermented licorice extract extracted from sea water (ug/mL) 119-0.1% vaccination 128-0.1% vaccination BB-0.1% vaccination Day DSW Water DSW Water DSW Water D0 4.94 4.82 4.94 4.82 4.94 4.82 D4 13.26 13.21 13.43 13.33 20.93 19.63

In conclusion, it can be seen that the content of liqueritin decreased by fermentation of 119, 128 and BB lactic acid bacteria, whereas the amount of aglycon, liquoritgenin, increased, resulting in efficient bioconversion.

In addition, isoliquaritin decreased and aglycon-type isoliqueritigenin increased, showing that bioconversion of the physiologically active substance in the form of phytoestrogen, the glycoside of licorice, to a non-glycoside form occurs by fermentation.

4) Fermentation Characteristics of Calcium-Added Licorice Extract

Figure 16 is a comparison of the pH, titratable acidity, and number of lactic acid bacteria of the calcium-added licorice fermentation broth (DSW-Ca group) fermented by adding calcium to the licorice extract and inoculating 119-0.1% with the group without adding calcium (DSW group) One result.

The pH decreased less in the group to which calcium was added, but titratable acidity increased more significantly. It was found that the number of lactic acid bacteria decreased less from the 2nd day of fermentation (D2). That is, the addition of calcium was found to change the fermentation properties of licorice extract.

17 is a result of comparing the pH, titratable acidity, and number of lactic acid bacteria of the calcium-added licorice fermentation broth (DSW-Ca group) obtained by inoculating 128-0.1% of the inoculated fermentation with the group without calcium addition (DSW group).

Similar to the result of fermentation at 119, the decrease in pH was small in the group to which calcium was added, and titratable acidity increased more significantly. The number of lactic acid bacteria decreased from the second day of fermentation (D2), and the addition of calcium in the fermentation broth by 128 was found to have an effect by changing the fermentation characteristics of the licorice extract.

18 is a result of comparing the pH, titratable acidity, and number of lactic acid bacteria of the calcium-added licorice fermentation broth (DSW-Ca group) obtained by inoculating BB-0.1% with the group without calcium addition (DSW group).

Similar to the results of fermentation at 119 and 128, the decrease in pH was less in the group to which calcium was added, and titratable acidity increased more significantly. The number of lactic acid bacteria decreased from the 2nd day of fermentation (D2), and the addition of calcium in the fermented broth by BB changed the fermentation characteristics of the licorice extract and had an effect.

As shown in FIG. 19, as a result of analyzing the effect of calcium addition on the change of reducing sugar content in licorice fermentation broth, it was found that the reducing sugar reduction of the fermentation broth with calcium addition was large in fermentation by 119, 128 and BB inoculation. Seems to be promoted.

5) Effect of calcium addition on phytoestrogen content change (bioconversion) in licorice fermentation broth

As shown in Fig. 20, the liquoitin content of the extract obtained by fermenting the licorice extract (DSW) to which calcium was not added at 119, 128 and BB was compared to before fermentation (D0) 4 days after fermentation (D4), respectively. 15%, 19%, and 30% decreased, whereas the content of liquoritin in the calcium-added extract (DSW+Ca) decreased 18%, 25% and 40% after 4 days of fermentation (D4). There was a tendency that the decrease of

As shown in Figure 21, the liquorice genin content of the extract obtained by fermenting the licorice extract (DSW) without adding calcium at 119, 128 and BB was 4 days after fermentation (D4) compared to before fermentation (D0). While the requerity genin content of the extract (DSW+Ca) added with calcium increased by 2.31 times, 2.9 times and 4 times, respectively, after 4 days of fermentation (D4), 2.9 times, 3.4 times and 4.5 times increased. There was a tendency to have a slight effect on the increase of xenin.

As shown in FIG. 22, the isoliquaritin content of the extract obtained by fermenting the licorice extract (DSW) without adding calcium at 119, 128 and BB was 4 days after fermentation (D4) compared to before fermentation (D0). The content of isoliquaritin in the extract (DSW+Ca) added with calcium decreased by 43%, 56%, and 87%, respectively, while the content of isoliquaritin decreased by 51%, 63% and 93% after 4 days of fermentation (D4), resulting in a decrease in calcium. There was a tendency to influence the reduction of tin.

As shown in FIG. 23, the extract obtained by fermenting the licorice extract (DSW) without adding calcium at 119 and 128 was increased by about 2.7 times after fermentation 4 days (D4) compared to before fermentation (D0), and fermented by BB. One extract increased about 4 times, whereas the isoliquaritin content of the calcium added extract (DSW+Ca) increased about 3.1 times by 119 and 128 4 days after fermentation (D4) and about 4.5 times by BB fermentation. .

In conclusion, the addition of calcium showed a tendency to promote the conversion of liquoritin to liquoritgenin and from isoliquaritin to isoliquerigenin.

The active phytoestrogen increase and calcium-fortified fermentation broth manufacturing method and fermentation broth prepared from licorice extract according to the present invention extract phytoestrogen, a vegetable female hormone, from licorice By increasing it, the concentration of active ingredients is higher than that of simple extracts, and minerals derived from calcium and deep sea water are added to help alleviate various symptoms caused by a lack of female hormones and prevent osteoporosis. .

Claims (10)

Step (S10) of preparing a licorice extract by mixing 1.0 to 20.0 wt% of finely chopped licorice and 80.0 to 99.0 wt% of deep sea water or purified water, and then extracting it at 100 to 120°C for 8 to 15 hours using an herbal extractor. ,
Measuring the dry solid concentration of the licorice extract and adjusting the concentration to 1 ~ 5% (S20),
Comprising a mixture by mixing 0.05 to 0.2 parts by weight of any one or two or more types of calcium selected from fermented calcium, calcium citrate or calcium lactate with respect to 100.0 parts by weight of the licorice extract whose concentration is adjusted through the step (S20) (S30) Wow,
Sterilizing the mixture by heating at 80 to 100° C. for 20 to 40 minutes (S40),
Step (S50) of inoculating any one or two selected from Lactobacillus plantarum DK119 (119) or Lactobacillus casei DK128 (128), which are lactic acid bacteria isolated from kimchi, to the sterilized mixture through the step (S40);
The mixture inoculated with the lactic acid bacteria through the step (S50) is fermented at 30 to 38° C. for 3 to 5 days to form a fermentation broth,
A method for producing an active phytoestrogen increase and calcium-enhanced fermentation broth of licorice extract comprising the step of preparing the fermentation broth in any one form selected from liquid, pill, or granule (S60).














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