KR20170061959A - Preparation method of fermented ginseng and fermented black-red ginseng with active ginsenoside heightening rate absorption - Google Patents

Abstract

A fermented ginseng or fermented black ginseng containing active ginsenoside having enhanced absorption rate in the body and a processed product of black ginseng are provided. A step of preparing nano-powdered ginseng or black red gins through a pulverizing process of ginseng or black ginseng, adding water to the ginseng powder or ginseng powder and sterilizing it, A second step of inoculating the red ginseng with the lactic acid bacteria, a third step of fermenting the nano-powdered ginseng or red ginseng by culturing the lactic acid bacterium, the fourth step of aging the culture solution obtained after the third step, And a fifth step of sterilizing the sample. Accordingly, it is possible to provide a fermented ginseng and fermented black ginseng production method containing the active forminsenoside (Rg2, Rh1, Rg3, and 20-ginsenoside Rg3) with enhanced body water absorption rate. Furthermore, by low-temperature aging of the culture broth obtained by fermenting ginseng or red ginseng using lactic acid bacteria, the content of ginsenoside Rg3 and 20-ginsenoside Rg3, which are excellent in anti-fatigue, suppression of cancer metastasis and excellent anticancer effect, can be increased. In addition, fermented ginseng and fermented black ginseng biologically converted into active formsinoside can be produced by a relatively simple process without going through the neutralization process and extraction and purification, so that they can be effectively used as food, cosmetics or pharmaceuticals.

Description

[0001] The present invention relates to a method for preparing fermented ginseng or fermented black ginseng extracts containing active-type ginsenosides,

The present invention relates to fermented ginseng or fermented black ginseng extract, and more particularly, to a fermented ginseng or fermented black ginseng extract containing active ginsenosides having enhanced body water absorption.

Ginseng is a perennial herbaceous perennial belonging to the genus Oga and Ginseng. It is known about 11 species on the earth. Panax ginseng CA Meyer, Panaxquinquefolium L., Panaxnotoginseng FH Chen, ( Panaxjaponicus CA Meyer). Korean ginseng ( Panax ginseng CA Meyer) is native to the Far East region (lat. 33 ~ 48, Korea, northern Manchuria, part of Russia) and has excellent efficacy. Panaxquinquefolium L. is naturally grown and cultivated in the United States and Canada, and Panaxnotoginseng FH Chen is cultivated wild or in the southwestern part of Guangxi province from the southeastern part of Yunnan province in China. Panaxjaponicus CA Meyer is also distributed in Japan, Southwest China, and Nepal.

As the research on the major ingredients and pharmacological effects of ginseng has been announced, it has been widely recognized as a natural health food. Its demand is gradually expanding. It is accelerated by economic growth and improvement of living standard and widely used as a popular natural health food . In addition, ginseng has been used in various forms such as psychiatric, nervous system diseases and diabetes in the form of herbal medicines in Asian countries such as Korea. Saponin, which is a main ingredient of ginseng, has been widely used as an antioxidant, Activity and so on.

The ingredients of ginsenosides contained in ginseng and their pharmacological efficacy are shown in Table 1 below.

Types of Gin Senocides Effectiveness Ginsenoside Rb1 The present invention relates to the use of a compound of formula I in the manufacture of a medicament for the treatment or prevention of central venous thrombosis, Anti-inflammatory, promote cholesterol metabolism Ginsenoside Rb2 The present invention relates to a method for inhibiting the production of lipid peroxidation, comprising the steps of: accelerating sugar and fat metabolism; antidiabetic action; promotion of protein and lipid synthesis; reduction of high cholesterol and atherosclerosis; improvement of stressful food decline; inhibition of tumor angiogenesis; Promoting proliferation and DNA synthesis Gin Senocide Rc Promotion of serum cholesterol and RNA synthesis, promotion of protein and lipid synthesis, analgesic action, promotion of corticosterone secretion, promotion of prostacyclin biosynthesis, inhibition of renal glomerular hypertrophy Gin Senocide Rd Promotion of adrenocortical stimulating hormone and corticosterone secretion, suppression of hypertrophied kidney glomeruli Gin Senocide Re Promotion of adrenocorticotropic hormone and corticosterone secretion, analgesia, vasodilation, hyperthermia stress, promotion of bone marrow cell DNA, RNA, protein and lipid synthesis, liver injury protection, promotion of cholesterol metabolism Ginsenoside Rg1 It has been shown that the antitumor effect of the immunostimulating, platelet aggregation inhibition, enhancement of memory and learning function, anti-fatigue, antisstress, central excitement, vasodilation, anti-inflammatory and neo-blood- Promotes secretion, promotes cholesterol metabolism Ginsenoside Rg2 Anti-stress, immune enhancement, antitumor, improvement of memory decline, inhibition of platelet aggregation, induction of acetylcholine, inhibition of catecholamine secretion and inhibition of intracellular calcium influx, antithrombin, Ginsenoside Rg3 Anti-stress, immune enhancement, anti-tumor and anti-cancer effect, inhibition of cancer cell metastasis, inhibition of platelet aggregation and thrombosis, inhibition of liver injury, inhibition of anticancer drug resistance Ginsenoside Rh1 Inhibition of hepatotoxicity, promotion of differentiation of tumor cells, inhibition of platelet aggregation, Ginsenoside Rh2 Inhibiting cancer cell proliferation and invasion, promoting cancer cell division induction, inhibiting tumor growth, anti-inflammation, anti-allergy, promoting insulin secretion Compound K Inhibition of cancer cell metastasis, inhibition of carcinogenicity and resistance to cisplatin, strong anti-cancer effect on cells

Generally, ginseng is used in the form of red ginseng prepared by washing white ginseng or ginseng dried at room temperature with purified water and treating ginseng with steam at about 98 ° C to 100 ° C. Among them, red ginseng is known to have far better pharmacological efficacy than white ginseng. In recent years, a method has been used in which the active ingredient of ginseng is easily absorbed into the human body, and a method of fermenting ginseng using intestinal microorganism or lactic acid bacteria and treating the enzyme, as a method for strengthening components present in trace amounts in ginseng.

However, when a ginseng extract having a high content of ginsenoside Rg3 and 20-ginsenoside Rg3 is prepared, generation of carcinogens such as HMF (hydroxylmethylfurfural) produced during the heat treatment can not be avoided and heat and / Since the neutralization process must necessarily be carried out according to the acid treatment process and the extraction and purification process must be separately performed, there is a problem that it is not easy to use as a direct food product.

In addition, most of the microorganisms used to convert the saponins creatures in Peddie OKO Gus non-edible (Pediococcussp.), Lou Kono in Stock (Leuconostocsp.), Seusok Aspergillus (Aspergillus sp . ), Eubacterium sp. , Fusobacterium sp. sp . ) Or the genus Bacteroides sp . ) And the like, the metabolites produced therefrom must be subjected to ultrafiltration or separate purification (extraction and purification) steps separately, which makes it difficult to use them directly as food, and thus it is difficult to use them in real life.

Korean Patent No. 10-228510 Korean Patent No. 10-479803 Korean Patent No. 10-856790 Korean Patent No. 10-866504

Hasegawa H., J. Pharmacol. Sci., 2004, 95 (2): 153-157. Tawab M. A. et al., Drug Metab. Dispos., 2003, 31 (8): 1065-1071.

The problem to be solved by the present invention is to use saponin contained in ginseng, red ginseng or black red ginseng, that is, ginsenoside, as an active form ginsenoside (Rg2, Rh1, Rg3 and 20 - ginsenoside Rg3), and a fermented black ginseng fermented red ginseng.

Another problem to be solved by the present invention is to prepare fermented ginseng and fermented black red ginseng biologically converted into active forminsenoside by a relatively simple process without going through the neutralization process and extraction and purification.

According to one aspect of the present invention, there is provided a method for preparing fermented ginseng or fermented black ginseng extract. The method for preparing fermented ginseng or fermented black ginseng comprises the steps of preparing ginseng or black ginseng by pulverizing ginseng or black ginseng, adding water to the ginseng or black ginseng powder A third step of fermenting the nano-powdered ginseng or the red ginseng by culturing the lactic acid bacterium, a third step of fermenting the nano-powdered ginseng or the red ginseng after sterilization, A fourth step of aging the culture solution, and a fifth step of sterilizing the aged solution obtained after the fourth step.

The first step may include a step of firstly grinding the ginseng or the red ginseng to 2 to 5 mm and a step of secondly grinding the ginseng or the red ginseng after the first grinding to 0.3 to 15 탆.

The fourth step of aging the culture solution obtained after the third step may be performed at 35 캜 to 45 캜 for 12 to 72 hours.

The fermented ginseng or fermented black ginseng was prepared by mixing the compound K (20-O-? -D-glucopyranosyl-20 (S) -protonacaxadiol), ginsenoside Rh1, ginsenoside Rh2, ginsenoside Rg2 and 20 - < / RTI > ginsenoside Rg3.

The lactic acid bacterium may be one that is at least one selected from Enterococcus (Enterococcus) genus Streptomyces Cocos (Streptococcus) or in the group consisting of Bifidobacterium genus (Bifidobacterium).

The fifth step of sterilizing the aging solution obtained after the fourth step may be performed at 80 캜 to 95 캜 for 12 hours to 48 hours.

The lactic acid bacteria may be inoculated at a concentration of 1% to 5%.

The lactic acid bacteria may be cultured for 1 to 5 days.

The fifth step may further include a step of aging the supernatant obtained by centrifuging after the sterilization treatment, and then concentrating or lyophilizing the supernatant.

According to the present invention, there can be provided a fermented ginseng and a fermented black ginseng production method containing active-type ginsenosides (Rg2, Rh1, Rg3 and 20-ginsenoside Rg3) with enhanced in-vivo absorption rate. Furthermore, by low-temperature aging of the culture broth obtained by fermenting ginseng or red ginseng using lactic acid bacteria, the content of ginsenoside Rg3 and 20-ginsenoside Rg3, which are excellent in anti-fatigue, suppression of cancer metastasis and excellent anticancer effect, can be increased.

Further, according to the present invention, fermented ginseng and fermented black ginseng biologically converted into active type ginsenoside are produced through a relatively simple process without passing through a neutralization process and extraction and purification, and thus can be usefully used as foods, cosmetics or pharmaceuticals have.

The technical effects of the present invention are not limited to those mentioned above, and other technical effects not mentioned can be clearly understood by those skilled in the art from the following description.

FIG. 1 is a flowchart illustrating a method of preparing fermented ginseng and fermented black ginseng extract according to an embodiment of the present invention.
FIG. 2 is a graph of HPLC (High performance liquid chromatography) analyzing saponins of ginseng according to an embodiment of the present invention and ginseng according to a comparative example.
FIG. 3 is a graph showing changes in serum saponin concentration of active ginsenosides Rg2, Rh1, Rg3 or 20-ginsenoside Rg3, which are respectively observed when oral administration of the fermented ginseng extract according to an embodiment of the present invention and the ginseng extract according to the comparative example, Fig.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings.

While the invention is susceptible to various modifications and alternative forms, specific embodiments thereof are shown by way of example in the drawings and will herein be described in detail. Rather, the intention is not to limit the invention to the particular forms disclosed, but rather, the invention includes all modifications, equivalents and substitutions that are consistent with the spirit of the invention as defined by the claims.

It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

FIG. 1 is a flowchart illustrating a method of preparing fermented ginseng and fermented black ginseng extract according to an embodiment of the present invention.

1, the method for preparing fermented ginseng and fermented black ginseng according to the present invention comprises a first step (S100) of preparing nano-powdered ginseng or red ginseng by pulverizing ginseng or black ginseng (S100) (S200) in which water is added to sterilized ginseng or black ginseng and sterilized, and the lactic acid bacteria are inoculated into the nano-powdered ginseng or red ginseng (S200). The lactic acid bacterium is cultured to prepare the nanoparticulated ginseng or black ginseng (S400) for fermenting the culture solution obtained after the third step, and a fifth step (S500) for sterilizing the fermentation solution obtained after the fourth step (S300).

More specifically, the ginseng may be any of ginseng itself and ginseng processed materials, and is not particularly limited. For example, the ginseng may include ginseng, red ginseng, red ginseng, black ginseng, black ginseng, black ginseng, white ginseng, ginseng, ginseng fruit, ginseng leaf, ginseng extract or ginseng powder. For example, it can include Korean ginseng, Korean ginseng, American ginseng, Panax ginseng, Himalayan ginseng or Vietnamese ginseng. In addition, the ginseng may be sterilized by a conventional method if necessary.

The black ginseng can be formed through the black and red ginseng process using the ginseng as the raw ginseng. First, the raw material ginseng can be dried to have a moisture content of 10% to 14% in ginseng to be in the form of dried ginseng. Next, the dried ginseng is placed in a sealed container together with water, and heated to a certain temperature under a certain pressure to form black ginseng. For example, the pressure of the sealed container may be 2.0 × 10 ⁵ Pa to 1.0 × 10 ⁶ Pa. The heating can be carried out for 8 to 12 hours while maintaining 85 to 180 ° C.

Then, the ginseng or the red ginseng can be pulverized at least twice. For example, the ginseng or black ginseng can be firstly pulverized to 2 to 5 mm. Then, the ginseng or the red ginseng that has been first pulverized may be secondly pulverized to 0.3 to 15 탆 to prepare ultrafine powdered ginseng or red ginseng. At this time, the secondary pulverization may use an ACM pulverizer, but is not limited thereto.

When the fermentation process is performed using ultrafine ginseng or black ginseng powder which has undergone the nano-powdering process, a bioconversion yield that converts ginsenoside, an active ingredient of ginseng or black ginseng, to active ginsenoside The effect is increased. In addition, while conventional techniques have fermented ginseng extract or concentrate to increase the yield of active ginsenoside bioconversion, the present invention can produce active ginsenoside without extraction, Or black ginseng is not subjected to the extraction and / or concentration process, which is advantageous in manufacturing cost and environmentally friendly.

In addition, since the present invention keeps the whole-body type ginseng or black red ginseng, it is possible to naturally contain all the components known in the academic world as various active ingredients of ginseng such as various minerals and acidic polysaccharides in addition to the active ingredient ginsenoside, Of fermented black ginseng or fermented black ginseng extract.

Next, water may be added to the nano-powdered ginseng or black ginseng and sterilized, and the lactic acid bacteria may be inoculated into the nano-powdered ginseng or black ginseng (S200).

The lactic acid bacteria may be lactic acid bacteria having saponin biotransformation ability. The lactic acid bacterium having the saponin biotransformation ability can be used as long as it is a microorganism commonly used in the field of ginseng fermentation.

With lactic acid bacteria having the ability of saponin biotransformation may include genus Lactobacillus (Lactobacillus) genus Streptomyces Cocos (Streptococcus) or Enterococcus genus (Enterococcus). For example, the Lactobacillus (Lactobacillus) in lactic acid bacteria from Lactobacillus rhamnose ATCC 7469 ju (Lactobacillus rhamnose ATCC 7469), Lactobacillus biasing Li DSM 20243 ju (Lactobacillus gasseri DSM 20243), Lactobacillus mali (Lactobacillus gmali ATCC 27304) , Lactobacillus plantarum ATCC 14947, Lactobacillus plantatum ATCC 10241, or Lactobacillus lactis . In addition, the Lactobacillus plantarum ATCC 14477 can be used as an active ingredient . In addition, it may include Streptomyces Cocos write a brush loose (Streptococcus thermophilus) from Lactobacillus genus Streptomyces Cocos (Streptococcus).

In addition, the saponin biotransformation lactic acid bacteria having the ability is used alone or be used as a mixed culture, preferably Enterococcus Fish Titanium (Enterococcus faecium), Enterococcus faecalis bend (Enterococcus faecalis , Lactobacillus casei , Lactobacillus acidophillus , Bifidobacterium ( Bifidobacterium longum ) and Bifidobacterium infantis .

Preferably with the lactic acid bacteria used in fermented ginseng and fermented black ginseng extract production process of the invention Advantageously, Enterococcus (Enterococcus) genus, one from the group consisting of genus Streptomyces Cocos (Streptococcus) in or Bifidobacterium (Bifidobacterium) Or two or more of them may be selected.

For example, Enterococcus sp. faecium , Enterococcus faecalis ) can be used.

The lactic acid bacteria may be inoculated at a concentration of 1% to 5%.

Then, the lactic acid bacteria may be cultured to ferment the nano-powdered ginseng or the red ginseng (S300). More specifically, the inoculated lactic acid bacteria can be cultured for 1 to 5 days. Thus, the nano-powdered ginseng or black ginseng can be fermented. In addition, the content of ginsenosides of fermented ginseng or black ginseng can be increased.

Then, the culture solution obtained after the fermentation of the nano-powdered ginseng or the red ginseng is aged (S400). Fermented ginseng or black ginseng culture can be aged at a specific temperature for a certain period of time to utilize the fermentation enzyme present in the lactic acid bacteria. The fermented ginseng or fermented black ginseng was prepared by mixing the compound K (20-O-? -D-glucopyranosyl-20 (S) -protonacaxadiol), ginsenoside Rh1, ginsenoside Rh2, ginsenoside Rg2 and 20 - < / RTI > ginsenoside < RTI ID = 0.0 > Rg3. ≪ / RTI > In particular, Rg2, Rg3, Rh1, and 20-Rg3, which are known to have excellent anti-fatigue, suppression of cancer metastasis and anti-diabetic effect, can be increased compared to other ginsenosides.

For example, the aging treatment may be performed at 35 캜 to 45 캜 for 12 hours to 72 hours.

Then, the aged solution obtained after the aging treatment can be sterilized (S500). The sterilization treatment may be performed intermittently at 80 ° C to 95 ° C for 12 hours to 48 hours. At this time, the fermented black ginseng extract can be prepared by concentrating or lyophilizing the supernatant obtained by centrifuging the culture solution obtained after the sterilization treatment.

The fermented ginseng and fermented black ginseng extract according to an embodiment of the present invention is a relatively simple process including a low temperature aging treatment and is a process for producing ginsenoside, an active ingredient of ginseng or red ginseng, as an active ginsenoside Converted fermented ginseng and fermented red ginseng can be produced. Accordingly, it is possible to exclude the generation of carcinogens such as HMF (Hydroxylmethylfurfural), which is generated during the heat treatment process in the conventional fermented ginseng production process, and it is also possible to exclude the neutralization process that necessarily goes through the heat and / The present invention can provide a fermented ginseng and a fermented black ginseng extract preparation method that can be easily used in real life without being subjected to purification.

In addition, the microorganisms of the genus Lactobacillus sp. And Bifidobaterium sp., Which are lactic acid bacteria that specifically grow on black ginseng, are cultured to convert the metabolites such as ginsenosides Rg1 and Rb1 into ginsenosides Rg3, Compound K, and Ginsenoside F1 to produce a lactic acid fermentation broth, a fermented powder, a concentrate, and a fermented probiotic preparation containing a ginsenoside component having a high bioavailability and improving the antibacterial activity of the ginseng extract, A composition can be prepared.

Furthermore, since the other medium components for culturing the lactic acid bacteria are not contained except for the black ginseng extract, the process can be simplified and the loss of the product content by the additives can be reduced.

≪ Example 1 >

Fermented red ginseng  Method of manufacturing extract

Ginseng over 4 years old is selected as raw ginseng and cleaned.

The raw material ginseng is dried so that the content of water in ginseng is 14% or less.

The dried dried ginseng is put into a sealed container with water and maintained at a high pressure of 1.0 × 10 ⁶ Pa. At this time, water is injected so that the dried ginseng is not immersed. Then, it is heated for 10 hours while maintaining the temperature at 125 캜, and black red ginseng is formed.

Then, the thus-prepared black ginseng was put into a hammer mill, firstly pulverized into particles having a size of 46 to 74 mu, and the first pulverized product was put into an ACM pulverizer to be secondly pulverized into particles having a size of 13 to 23 um, The pulverized tea is put into a urethane coated dry or wet ball mill and is ground to a size of 500 nm into nanoparticles to prepare a black ginseng powder for fermented black ginseng production.

Then, 250 ml of water is added to 50 g of the prepared red ginseng powder, dissolved oxygen of the fermentation broth is removed using nitrogen gas, and sterilized at 85 캜. Next, Enterococcus faecium was inoculated at a concentration of about 5% and then incubated at 37 DEG C for 2 days with stirring.

Subsequently, the incubator was depressurized, and the inside air was replaced with a nitrogen gas for culturing the microorganism, followed by aging at a low temperature of about 25 캜 for 24 hours.

After the aging treatment, the resulting culture was concentrated in vacuo at 85 캜 to produce fermented black ginseng extract (i.e. fermented black ginseng).

≪ Example 2 >

Fermentation American red ginseng  Method of manufacturing extract

US red ginseng was prepared in the same manner as in Example 1 except that American ginseng (fire ginseng) was used instead of Korean ginseng, and then Enterococcus faecalis ) instead of Enterococcus ficulis ( Enterococcus faecalis) to the fermentation American black ginseng was prepared by fermentation and is the American black ginseng in the same manner as in Example 1 except for using.

≪ Example 3 >

Fermentation Blackhead red ginseng  Method of manufacturing extract

In the same manner as in Example 1 except that Korean black ginseng was used instead of Korean ginseng, black red ginseng was prepared and then Enterococcus faecalis) was prepared in place of the fermentation jukjeol black ginseng in the same manner as in Example 1, except for using Enterococcus lactis (Enterococcus lactis).

<Example 4>

Fermentation Himalayan black red ginseng  Method of manufacturing extract

Fermented Himalayan black ginseng was prepared in the same manner as in Example 1 except that Himalayan ginseng was used instead of Korean ginseng.

&Lt; Example 5 >

Fermentation Vietnamese red ginseng  Method of manufacturing extract

Fermented ginseng was prepared in the same manner as in Example 1, except that Vietnamese black ginseng was used instead of Korean ginseng as in Example 1.

FIGS. 2A and 2B are graphs of HPLC (high performance liquid chromatography) analysis of saponin as an active ingredient of fermented black ginseng according to Comparative Example 1 and Example 1 of the present invention, respectively.

&Lt; Comparative Example 1 &

Heat treatment and Organic solvent  Method of manufacturing ginseng extract by extraction process

100 g of fresh ginseng was placed in a sealed container and heated at 130 ° C for 2 hours.

The processed ginseng was extracted with 200 ml of methanol to obtain a methanol extract, which was then removed by evaporation of methanol. The remaining residue was suspended in 100 ml of water and extracted three times with 100 ml of ether. The remaining aqueous layer was extracted three times with 100 ml portions of saturated butanol, To obtain a butanol extract solution. (See Example 1 of Korean Patent No. 228,510)

Korean Patent No. 228,510 The column chromatography step for obtaining the Rg3 fraction in Example 1 was omitted for direct comparison with the extract.

&Lt; Comparative Example 2 &

Production method of ginseng extract using lactic acid fermentation

500 g of 6 - year - old white ginseng purchased from Kyungdong market was cut and extracted 5 times with 5 L of MeOH and concentrated under reduced pressure to obtain 25 g. After dissolving in 30 ml of water, it was extracted 4 times with 1500 ml of BuOH and concentrated to obtain 10 g of ginseng BuOH fraction. 10 g (wet weight) of Bifidobacterium KK-1 (Accession No .: KCCM 10364) and Bifidobacterium KK-2 (Accession No .: KCCM 10365) were added and incubated at 37 ° C for 72 hours. Extraction with BuOH And concentrated to obtain 4 g of processed ginseng (refer to Korean Patent No. 479,803, Example 1)

&Lt; Comparative Example 3 &

Lactobacillus sp.  Method for manufacturing fermented ginseng extract using lactic acid fermentation

The ginseng concentrate was diluted 10 times with distilled water and sterilized at 121 ° C under 1.5 atm for 15 minutes. Then, the sterilized ginseng concentrate was added with Lactobacillus alimentarius M-2 or Leuconostoc mensenteroides M-3 strain was inoculated. The inoculum amount was 1% (v / v) (corresponding to 10 ⁷ -10 ⁸ cfu) with respect to the total volume of the ginseng concentrate. After inoculation, the cells were cultured at 30 DEG C for 120 hours. (Refer to Korean Patent No. 856,790, Example 4)

&Lt; Comparative Example 4 &

Lactobacillus sp.  Using lactic acid bacteria fermentation Fermented red ginseng  Method of manufacturing extract

50 g of a phosphate buffer solution (50 mM, pH 5.0) was added to 5 g of red ginseng powder purchased from Geumsan, and the suspension was sterilized by autoclaving. The isolated strains Lactobacillus plantanum P2 and Lactobacillus brevis M2, which were precultured in a MRS medium, were inoculated to a pressurized sterilized red ginseng suspension at 2% of the total culture medium and incubated for 5 days with gentle stirring at 37 캜 Fermented red ginseng was prepared. The red ginseng fermented by inoculation with Lactobacillus plantanum P2 was designated as 'P2 fermented red ginseng' and the red ginseng fermented by inoculation with Lactobacillus brevis M2 as 'M2 fermented red ginseng'.

In addition, red ginseng extract (60 brix) was diluted to 15 brix, pH was adjusted to 6.0, and pressure sterilization was performed. The isolates P2 and M2, preincubated with MRS, were inoculated at 2% level of the total culture medium and incubated at 37 ℃ for 5 days with gentle agitation at 37 ℃ to prepare red ginseng fermented product. (Refer to Korean Patent No. 866,504, Example 3)

&Lt; Test Example 1 >

Measurement of compositional change of ginseng extract

The compositional changes of the ginseng extracts obtained in Examples 1, 2 and 3 and Comparative Examples 1, 2, 3 and 4 of the present invention were measured.

5 g of each of the obtained ginseng extract is precisely weighed and taken in a 100 ml concentrated flask. Then, 50 ml of saturated water-saturated butanol was added to the reaction mixture, and the mixture was refluxed with a reflux condenser. The mixture was heated at 70 ° C to 80 ° C for about 1 hour in a water bath, cooled, filtered and the same operation was repeated twice. The filter paper was washed with water saturated butanol at 10 ° C, and the filtrate and the washing solution were combined and placed in a 250-ml separatory funnel, shaken well with 20 ml of water and washed with water. The saturated water-saturated butanol extract solution was transferred to a concentrated flask which was previously diluted with water and concentrated under reduced pressure in a water bath to remove butanol. 50 ml of ether was added to the residue, and the mixture was degassed by heating in a water bath for 30 minutes at 36 ° C in a water bath. Respectively. The obtained residue was completely dissolved by adding 10 times methanol, filtered using a filter (0.45 mu m), and quantitatively analyzed for ginsenoside components by high performance liquid chromatography (HPLC). The results are shown in Table 2 below. The HPLC conditions were as follows.

Organization: Shimadzu Prominence

Column: Gromsil ODS 4.6 (i, d) x 250 mm

Mobile phase A: acetonitrile: distilled water = 15: 85

Mobile phase B: acetonitrile: distilled water = 80: 20

Flow rate: 1.0 / min

Detector: ELSD (Alltech)

division Ginsenoside (mg / g) Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Ginsenoside Rb1 0.6 0.5 0.5 2.7 2.8 2.8 2.8 Ginsenoside Rb2 0.3 0.3 0.3 1.5 2.1 2.1 2.1 Gin Senocide Rc 0.2 0.2 0.2 1.6 2.0 2.0 2.0 Gin Senocide Rd 0.1 0.2 0.2 0.8 1.1 1.1 1.1 Ginsenoside Rg2 1.4 1.6 1.9 0.7 0.9 0.3 0.3 Ginsenoside Rg3 7.8 8.2 8.3 6.7 7.8 7.2 6.9 ²⁰ - Ginsenoside Rg3 10.2 10.4 11.2 5.8 6.2 6.1 6.7 Ginsenoside Rh1 1.9 1.7 1.9 0.5 0.7 0.8 0.9 Ginsenoside Rh2 1.1 0.8 0.9 0.1 0.2 0.1 0.1 Compound K 0.7 0.6 0.6 0 0 0 0

As shown in Table 2, in the case of the ginseng extract of Comparative Example 1 prepared through heat treatment and organic solvent extraction without fermentation with lactic acid bacteria or intestinal bacteria, the content of ginsenoside Rg3 was relatively high, but satisfactory The ginsenoside extract of Comparative Example 2 prepared through acid treatment and lactic acid fermentation showed an increase in the content of ginsenoside Rg3, but the content of Rg2, Rh1 and 20-ginsenoside Rg3 was still lower than that of Comparative Example 1 . In addition, the fermented ginseng extracts of Comparative Examples 3 and 4, which had undergone lactobacillus fermentation in Lactobacillus sp . , Were comparable to those of Comparative Examples 1 and 2.

In contrast, the fermented ginseng and the fermented red ginseng extract (examples) prepared according to the present invention had a relatively high content of active ginsenosides Rg2, Rh1, Rg3 and 20-Rg3 as compared with the comparative examples.

FIG. 3 is a graph showing the activity of the active ginsenosides Rg2, Rh1, Rg3 or 20-ginsenoside Rg3 in the serum of the fermented black ginseng extract according to Example 1 of the present invention and the ginseng extract according to Comparative Example 1, And a graph showing changes in concentration over time.

&Lt; Test Example 2 &

Measurement of bioavailability of active ginsenosides

40 g of the ginseng extract prepared in Example 1 was orally administered to 60 to 70 kg of a healthy adult (male, n = 3) once, followed by 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, Blood samples (5 ml) were taken at 8 hours, 12 hours and 24 hours to analyze saponin and saponin metabolites present in the blood.

5 ml of blood was collected for each hour. The analytical conditions were as follows. The ginsenosides were extracted and purified using a solid phase extraction method. Then, methanol was added to elute ginsenosides adsorbed to the solid phase extractor adsorbent, ) And quantitatively analyzed for ginsenoside components by high performance liquid chromatography (HPLC). The results of measuring the changes with time in the serum saponin derivatives are shown in FIG.

It is known that saponin known as pharmacological component of ginseng has a glycoside structure, so that saponin is not absorbed by itself but is bioconverted to saponin metabolites (compound K, ginsenoside F1, etc.) by intestinal microorganisms and then absorbed into the body (Hasegawa H., J. Pharmacol . Sci., 2004, 95 (2): 153-157; Tawab MA et al., Drug Metab. Dispos., 2003, 31 (8): 1065-1071.).

As can be seen in Fig. 3, the active absorption of ginsenosides Rg2, Rh1, Rg3 and 20 ginsenoside Rg3 components showed maximum absorption in 30 minutes to 2 hours, respectively. Comparing the examples of the present invention and the comparative example, it can be seen that the absorption rate of fermented black ginseng extract according to the present invention is increased 20 times or more.

It should be noted that the embodiments of the present invention disclosed in the present specification and drawings are only illustrative of specific examples for the purpose of understanding and are not intended to limit the scope of the present invention. It will be apparent to those skilled in the art that other modifications based on the technical idea of the present invention are possible in addition to the embodiments disclosed herein.

Claims (9)

A first step of preparing ginseng or black ginseng which has been nano-powdered by pulverizing ginseng or black ginseng;
A second step of adding water to the nano-powdered ginseng or black ginseng and sterilizing the ginseng or the ginseng powder, and then inoculating the nano-powdered ginseng or black ginseng into the lactic acid bacteria;
Culturing the lactic acid bacteria to ferment the nano-powdered ginseng or red ginseng;
A fourth step of aging the culture solution obtained after the third step; And
And a fifth step of sterilizing the obtained aging solution after the fourth step. The method according to claim 1,
In the first step,
Firstly crushing the ginseng or black ginseng to 2 to 5 mm; And
And secondly pulverizing the ginseng or black ginseng after the first pulverization step to 0.3 탆 to 15 탆, wherein the fermented ginseng or the fermented black ginseng extract is fermented. The method according to claim 1,
And the fourth step of aging the culture solution obtained after the third step is carried out at 35 ° C to 45 ° C for 12 hours to 72 hours. The method according to claim 1,
The fermented ginseng or fermented black ginseng was prepared by mixing the compound K (20-O-? -D-glucopyranosyl-20 (S) -protonacaxadiol), ginsenoside Rh1, ginsenoside Rh2, ginsenoside Rg2 and 20 - &lt; / RTI &gt; ginsenoside Rg3. &Lt; RTI ID = 0.0 &gt; 21. &lt; / RTI &gt; The method according to claim 1,
The lactic acid bacteria is Enterococcus (Enterococcus) genus, method of producing Streptomyces Cocos (Streptococcus) or genus Bifidobacterium in fermented ginseng or red ginseng extract fermented black will be at least one selected from the group consisting of genus (Bifidobacterium). The method according to claim 1,
Wherein the fifth step of sterilizing the aged solution obtained after the fourth step is performed at 80 캜 to 95 캜 for 12 hours to 48 hours. The method according to claim 1,
Wherein the lactic acid bacterium is inoculated at a concentration of 1% to 5%. The method according to claim 1,
Wherein the lactic acid bacterium is cultured for 1 to 5 days. The method according to claim 1,
Wherein the fifth step further comprises a step of aging the supernatant obtained by centrifuging after the sterilization treatment and then concentrating or lyophilizing the fermented ginseng or fermented black ginseng extract.

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