JP2003238424A - Biologically converted ginseng and method of producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide biologically converted ginseng composition in which the components having antiallergic action, antioxidant action or anticancer action are largely enhanced. <P>SOLUTION: The biologically converted ginseng composition has a ratio of 0.1 of [20-O-β-D-glucopyranosyl-20-(S)-protopanaxadiol + ginsenoside F2]/[ginsenoside Rc + ginsenoside Rd + ginsenoside Rb1 + ginsenoside Rb2]. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a ginseng composition and a method for producing the same, and more particularly to a biotransformed ginseng composition having an enhanced anti-cancer effect through a process for converting ginseng components by lactic acid bacteria and enterobacteria and a method thereof. It relates to a manufacturing method.

[0002]

2. Description of the Related Art The present application is filed in Korean Patent Application No. 20.
No. 02-5369 (filing date: 2002.01.30), which is a priority claim application based on "Method for producing biotransformed ginseng composition".

About 11 kinds of ginseng, which is a perennial herbaceous root belonging to the genus Ginseng genus Ginseng in plant taxonomy, are known on the earth, and there are the following four kinds as typical species. 1) Ginseng (scientific name: Panax ginseng C.
A. Meyer) -Variously grown or cultivated in the Far East Asia region (latitudes 33 to 48 north, South Korea, northeastern China, part of Russia). It has an extremely excellent medicinal effect among the ginseng species. 2) American ginseng (scientific name: Panax quinquefol
ium L. ) -Grown or cultivated in the USA and Canada. 3) Nana Ginseng (scientific name: Panax notoginsen
g (Burk) F.I. H. Chen) -Grown or cultivated in the southeastern part of Yunnan Province, China to the southwestern part of Guangxi Province. 4) Takebushi (Scientific name: Panax japonicus
C. A. Meyer) -Distributes throughout Japan, Southwestern China, and Nepal.

The ginseng is mainly used in Asian countries such as Korea, China and Japan as a crude drug against various diseases such as psychiatric, nervous system diseases and diabetes. Furthermore, saponin, which is a main component of ginseng, has been proved to have effects such as tonicity, tonicity, sedation, hematopoiesis, and antihypertension.

Further, in recent years, IH-901, IH-902, IH-9, which are metabolites of the saponins by microorganisms,
03 or IH-904, in addition to the immunopotentiating action,
It has been proved that it has an extremely strong tumor angiogenic action or cancer cell metastasis inhibiting action, and development of an anti-cancer ginseng composition using these is expected.

The above IH-901, IH-902, IH-
903 and IH-904 are products of final metabolism by microorganisms such as ginsenoside-Rb1, Rb2, Rc, and Rd of the protopanaxadiol system, and have a secretion inhibitory action of type IV collagenase, anti-angiogenic (cancer,
A powerful cancer cell metastasis inhibitory effect (anti-met) through the inhibitory effect on diabetes, rheumatism) activity and platelet aggregation
astatic activities).

In particular, IH-901 has little toxicity and side effects, inhibits normal differentiation of leukemia cells (HL-60 cells), and thus extremely strongly suppresses abnormal cell growth, and IH-901. It has been revealed through pharmacological experiments and stability experiments that the apoptosis-inducing effect of is as strong as that of cisplatin which is widely used as an anticancer therapeutic agent.

On the other hand, Korean Patent Application No. 1980-
No. 4291 "Method for producing lactic acid bacterium ginseng beverage" states that'organic nitrogen content is 0.2-0.8% and the production content of glucose is obtained by enzymatically decomposing the ginseng residue, which is a component of ginseng's characteristic scent. When the concentration of lactic acid bacteria is 3% or more, the pH is adjusted to 3.8 to 4.8 with organic acid, insoluble high molecular proteins and fibers are removed, and lactic acid bacteria are cultivated in the eluate. A method of adding a scent component specific to ginseng 'is disclosed.

Korean Patent Application No. 1988-125
No. 02 “Method for producing activated lactic acid bacteria drinking water using carrot”
'Is a raw juice or extract of ginseng, or after cultivating lactic acid bacteria in a medium in which the enzyme has been made to act on the ginseng boiled or boiled catabolite, skim milk, skim milk powder, carbonated water,
A method for producing carbonated drinking water of ginseng lactic acid bacteria and frozen desserts having high nutritional value by adding and mixing vitamins is disclosed.

Korean Patent Application No. 1996-237
No. 50 “Fermented milk composition containing ginseng or live ginseng and method for producing the same” is 0.1 to 0.6 cm after grinding.
0.001 to ginseng or live ginseng
2. Ginseng or live ginseng, including the step of adding 2.39% by mass and the step of further adding one or more components selected from the group consisting of water, vitamins, sugars, organic acids, fruits, grains and vegetables A method for producing a fermented milk composition containing the same is disclosed.

According to these methods, a composition containing a ginseng scent component or a ginseng component in an existing fermented milk or a lactic acid bacterium beverage is obtained, but the ginseng-containing composition and the like are simply added with a ginseng component. It is not possible to obtain a ginseng composition containing a high concentration of an active ingredient such as IH-901 which is a biometabolite of ginseng saponin.

[0012]

The object of the present invention to solve such problems is to provide a biotransformed ginseng composition in which an anticancer component derived from ginseng, such as IH-901, is significantly enhanced, and to prepare the same. To provide a method.

Another object of the present invention is to provide a biotransformed ginseng composition capable of maximizing the efficacy of the active ingredient contained in the ginseng raw material and a method for producing the same.

Still another object of the present invention is to provide a biotransformed ginseng composition having beneficial effects on the human body, such as antiallergic action, aging prevention and colorectal cancer / liver damage prevention, and a method for producing the same. Especially.

[0015]

The biotransformed ginseng composition of the present invention for solving the above-mentioned problems is a biotransformed ginseng composition comprising (20-O-β-D-glucopyranosyl-2
0 (S) -protopanaxadiol + ginsenoside F2) / (ginsenoside Rc + ginsenoside Rd
+ Ginsenoside Rb1 + Ginsenoside Rb2)
Is a bio-converted ginseng composition having a ratio of 0.1 or more.

In the biotransformed ginseng composition of the present invention, the biotransformed ginseng composition is a biotransformed ginseng composition derived from lactic acid bacteria or intestinal bacteria, and the lactic acid bacterium is Bifidobacterium KK. -1, lactic acid bacteria selected from the group consisting of Bifidobacterium KK-2, Bifidobacterium K-103, Bifidobacterium K-506, wherein the intestinal bacterium is Prevotella The composition may be one or more intestinal bacteria selected from the group consisting of Oris and Fusobacterium K-60.

Part of lactic acid bacteria or intestinal bacteria biotransforms compounds in foods and drinks and drugs to produce biotransformants having higher physiological activity than the original compounds.

As shown in FIG. 2, ginsenoside-Rb, which is a compound contained in carrot which is a raw material of the present invention.
1, ginsenoside-Rb2, ginsenoside-Rc, etc. are lactic acid bacteria (Lactic acid bacteria).
a) and intestinal bacteria
a), which are metabolized by the primary intermediate metabolites ginsenoside-Rd, compound-O, ginsenoside-Mc
Ginsenoside-F2, I, a primary and secondary intermediate metabolite
H-902 (Compound-Y), IH-903 (Compound-Mc), and the final metabolite IH-90
1 (Compound-K) is produced.

According to the above constitution, since the ratio is 0.1 or more, the dose of the biotransformed ginseng composition and 20-O
Provided is a biotransformation ginseng composition which exerts a sufficient effect in balance with the contents of β-D-glucopyranosyl-20 (S) -protopanaxadiol (IH-901) and its precursor ginsenoside F2 it can. Also,
It is needless to say that the higher this ratio is, the more preferable, but since the IH-901 or its precursor is a biotransformation product derived from microbial cells such as lactic acid bacteria and enterobacteria, this ratio is substantially It ranges from 0.1 to 10.0,
When it is 0.5 or more and 10.0 or less, a biotransformed ginseng composition in which the effect and the dose are more surely balanced is provided.

Further, the biotransformed ginseng composition of the present invention can be configured such that the biotransformed ginseng composition is an anti-allergic substance that suppresses the release of histamine.

Further, the biotransformed ginseng composition of the present invention may be configured such that the biotransformed ginseng composition is an antioxidant having an antioxidant effect by radical scavenging.

Further, the biotransformed ginseng composition of the present invention can be configured such that the biotransformed ginseng composition is an anticancer agent.

Further, the biotransformed ginseng composition of the present invention can be configured such that the biotransformed ginseng composition is a substance for preventing colon cancer or liver damage, which inhibits the production of β-glucuronidase. .

[0024] According to the above constitution, an antiallergic agent, an antioxidant and an anticancer agent, in which the biotransformation ginseng component is greatly enhanced, are provided.

Further, the biotransformed ginseng composition of the present invention can be configured such that the biotransformed ginseng composition is a beverage containing a biotransformed ginseng component.

[0026] According to this constitution, there is provided a beverage containing a bio-converted ginseng component having antiallergic action, antioxidant action and anticancer action.

The method for producing a biotransformed ginseng composition of the present invention for solving the above-mentioned problems is as follows. After the raw material of ginseng is suspended in water, lactic acid bacteria or intestinal bacteria are added and cultured to obtain a biotransformed ginseng liquid. And a concentration step of concentrating, freezing, or drying the biotransformed ginseng solution that has undergone the biotransformation step, or collecting the supernatant after centrifugation to obtain a biotransformed ginseng concentrate. It is a method for producing a carrot composition.

According to this constitution, it is possible to produce a biotransformation composition derived from cells such as lactic acid bacteria and enterobacteria, from which impurities and precipitates have been removed.

Further, the method for producing a biotransformed ginseng composition of the present invention further comprises, as the ginseng raw material, a group consisting of ginseng, red ginseng, white ginseng, ginseng, ginseng leaves, ginseng extract and ginseng powder. One or more selected substances can be used.

The method for producing a biotransformed ginseng composition of the present invention further comprises, as the lactic acid bacteria, Bifidobacterium KK-1 and Bifidobacterium KK-2.
One or more lactic acid bacteria selected from the group consisting of can be used.

The method for producing a biotransformed ginseng composition of the present invention further comprises, as the lactic acid bacteria, Bifidobacterium K-103 and Bifidobacterium K-5.
One or more lactic acid bacteria selected from the group consisting of 06 can be used.

The method for producing a biotransformed ginseng composition of the present invention further comprises, as the lactic acid bacterium, Bifidobacterium KK-1, Bifidobacterium KK-2, Bifidobacterium K-103. And one or more lactic acid bacteria selected from the group consisting of Bifidobacterium K-506 can be used.

Further, in the method for producing a biotransformed ginseng composition of the present invention, one or more enterobacteria selected from the group consisting of Prevotella orris and Fusobacterium K-60 are used as the enterobacteria. can do.

In the method for producing a biotransformed ginseng composition of the present invention, further, after the concentration step, an extraction step of administering a preset solvent to the biotransformed ginseng concentrate to obtain a biotransformed ginseng extract Can be provided.

Further, in the method for producing a biotransformed ginseng composition of the present invention, after the extraction step, the biotransformed ginseng extract is added to a beverage stock solution prepared by adding purified water to a sweetener and stirred. A stirring step for obtaining a biotransformed ginseng mixture,
A dilution step of adding purified water to the biotransformed ginseng mixed solution to obtain a biotransformed ginseng beverage solution can be adopted.

Further, the method for producing the biotransformed ginseng composition of the present invention further comprises one or more selected from the group consisting of citric acid, sodium citrate, a herbal medicine extract, and taurine, in the beverage stock solution in the stirring step. Can be added.

Further, the method for producing a biotransformed ginseng composition of the present invention further comprises a drying step after the extraction step to freeze-dry the biotransformed ginseng extract to obtain a biotransformed ginseng powder. can do.

According to the above constitution, the saponin component contained in the ginseng raw material is biotransformed to generate a metabolite having a high physiological activity, and thus a biotransformation having an antiallergic action, an antioxidant action and an anticancer action. A biotransformed ginseng composition with significantly enhanced ginseng components can be produced, and ginseng leaves, which were not suitable for use as raw materials for ginseng production, can also be used as raw materials because of their low content of active ingredients such as saponin. Cost can be reduced.

[0039]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described in detail below with reference to the drawings.

FIG. 1 is a flow chart showing a method for producing a bio-converted ginseng beverage solution, which is an example of the bio-converted ginseng composition of the present invention. As shown in FIG. 1, the method for producing a biotransformed ginseng composition of the present invention comprises a biotransformation step (S10) and a concentration step (S10).
20), including an extraction step (S30), a drying step (S40),
The method may further include a stirring step (S50) and a diluting step (S60).

The biotransformed ginseng composition was prepared as follows.

[Bioconversion step] After the ginseng raw material is suspended in water, lactic acid bacteria or intestinal bacteria are added to the suspension for a certain period of time, preferably 24 hours to 72 hours, more preferably 48 hours.
The culture was performed for 72 hours to 72 hours to obtain a biotransformed ginseng solution.

Here, as the ginseng raw material, one or more natural ginsengs selected from the group consisting of raw ginseng, red ginseng, white ginseng, ginseng and ginseng leaves, the ginseng extract of the natural ginseng, or the natural ginseng. Ginseng powder and the like can be used.

The lactic acid bacterium is not particularly limited as long as it can metabolize the ginseng saponin component and produce IH-901 which is a biotransformant.
103, Bifidobacterium K-506, Bifidobacterium KK-1 and Bifidobacterium
One or more lactic acid bacteria selected from the group consisting of KK-2 can be preferably used.

Of these, the Bifidobacterium K
K-1 is the deposit number KCCM-10364 (Deposit date: 2
002.03.22), Bifidobacterium KK
-2 is the deposit number KCCM-10365 (Deposit date: 20
02.03.22) at the Korean Culture Center of Korea.
Microorganisms).

The intestinal bacterium is not particularly limited as long as it can produce IH-901 which is a biotransformant.
Preferably Prevotella Oris (Prevotella
oris) and Fusobacterium K-60 can be used.

[Concentration Step] The biotransformed ginseng liquid was directly concentrated, frozen, dried, or centrifuged, and the supernatant was collected to obtain a biotransformed ginseng concentrated liquid.

[Extraction Step] The biotransformed ginseng extract was obtained by administering a preset solvent to the biotransformed ginseng concentrate.

The above-mentioned solvent is preferably a solvent capable of dissolving and extracting the active ingredient contained in the biotransformed ginseng concentrate, and specifically, water, a lower alcohol such as methanol or ethanol, a supercritical fluid, or these. Can be used.

[Drying Step] The biotransformed ginseng extract was freeze-dried to obtain biotransformed ginseng powder.

[Stirring Step] The above-mentioned biotransformed ginseng extract or biotransformed ginseng powder was added to a beverage stock solution prepared by adding purified water to a sweetener and stirred to obtain a biotransformed ginseng mixture.

Here, one or more of citric acid, sodium citrate, a herbal medicine extract, and taurine can be added to the beverage stock solution. As the herbal medicine extract, omimomi (gomishi), jujuba (jujube), cinnamon (keihi), 杸 杞 (kuko), seisei (osei), and Kibanaogi can be used.

[Diluting Step] A preset amount of purified water was added to the biotransformed ginseng mixed solution to obtain a biotransformed ginseng beverage solution.

The biotransformed ginseng composition of the present invention produced by the above process is (20-O-β-D-glucopyranosyl-20 (S) -protopanaxadiol + ginsenoside F2) / (ginsenoside Rc + ginsenoside
Rd + ginsenoside Rb1 + ginsenoside R
The ratio of b2) is maintained in the range of 0.1 or more.

Below, based on Examples and Comparative Examples,
The content of the present invention will be described in more detail. Needless to say, the present invention is not limited to the examples below.

Example 1 100 g of ginseng powder obtained by extracting ginseng with water and drying it was suspended in water, and then lactic acid bacteria Bifidobacterium K-103 and Bifidobacterium
The K-506 strain was added and cultured for 72 hours, and this was centrifuged to collect the supernatant, which was then concentrated to obtain a biotransformed ginseng concentrate.

(Example 2) Ginseng was finely chopped and sterilized, suspended in water, and then lactic acid bacterium Bifidobacterium K-
103 and Bifidobacterium K-506 strain were added and cultivated for 48 hours, and the mixture was centrifuged to collect the supernatant, which was concentrated and freeze-dried to obtain a biotransformed ginseng powder.

(Example 3) After extracting ginseng with water and drying it, 100 mg of the ginseng powder obtained was suspended with water, and then lactic acid bacteria Bifidobacterium KK-1 and Bifidobacterium -Add the KK-2 strain and culture for 72 hours, centrifuge this, collect the supernatant, then concentrate,
A bio-converted carrot concentrate was obtained.

(Example 4) A saponin fraction obtained by extracting 1 kg of white ginseng with methanol by cooling was further extracted with butanol, and the saponin fraction was suspended in water, followed by lactic acid bacterium Bifidobacterium KK-1. And Bifidobacterium KK-2 strain were added, and the mixture was cultured for 72 hours, centrifuged to collect the supernatant, and then concentrated to obtain a biotransformed ginseng concentrate.

[Experiment 1: Content analysis 1] White ginseng powder (2 g)
After adding water (500 ml) to Bifidobacterium KK-1 (depositee: Korea Microbial Conservation Center, deposit number: KCCM-10364, deposit date: 2002.03.
22) and Bifidobacterium KK-2 (depositee: Korea Microbial Conservation Center, deposit number: KCCM-1036)
5, deposit date: 2002.03.22) was added 1 g each, and the mixture was cultured at 37 ° C. for 72 hours and concentrated under reduced pressure to obtain a concentrated biotransformed ginseng solution of Example 3.

The biotransformed ginseng concentrate of Example 3 (2
g) and white ginseng (2 g) respectively in methanol (10
The extract was extracted 3 times with 0 ml), concentrated, suspended in water, and extracted 3 times with ether (100 ml). Furthermore, after extraction with butanol (100 ml) three times, the butanol fraction was concentrated, the concentrate was dissolved in methanol, and thin layer chromatography (TLC; Shimadzu TLC s) was performed.
From the analysis by canner CS-9301PC),
The content (%) of each component with respect to the total amount of saponin was determined.
The results are shown in Table 1 below.

In the TLC analysis, trichloromethane / methanol / water (mass ratio 65: 35: 1) was used as a developing solvent.
0) was used as a color reagent in a 5% sulfuric acid / methanol solution.

[0063]

[Table 1]

[Experiment 2: Content Analysis 2] Water (100 ml) was added to the white ginseng saponin fraction (1 g), and then Bifidobacterium KK-1 and Bifidobacterium KK-2 were added.
1 g of each of the above strains was added and cultured at 37 ° C. for 48 hours, and this was concentrated under reduced pressure to obtain a concentrated biotransformed ginseng solution of Example 4. Biotransformed ginseng concentrate of Example 4 (1 g)
And the white ginseng saponin fraction (1 g) were dissolved in methanol and analyzed by TLC in the same manner as in Experiment 1 to determine the content (%) of each component with respect to the total amount of saponin. The results are shown in Table 2 below.

[0065]

[Table 2]

[Experiment 3: Content analysis 3] Ginseng powder (2 g) obtained by extracting ginseng with water and drying it was suspended in water (500 ml), and then Bifidobacterium K-103 and Bifido 1 g of each strain of bacterium K-506 was added, and the mixture was cultured at 37 ° C. for 72 hours and then concentrated to obtain a biotransformed ginseng concentrate of Example 1. Also, dried ginseng (2 g) sterilized by slicing live ginseng was suspended in water (500 ml), and 1 g each of Bifidobacterium K-103 and Bifidobacterium K-506 strains were suspended. In addition, the mixture was cultured at 37 ° C. for 48 hours, concentrated, and then treated in Example 2.
To obtain a biotransformed carrot concentrate.

The biotransformed ginseng concentrate (2 g each) and white ginseng (2 g) were extracted with methanol (100 ml) three times and concentrated, and then suspended in water and ether (10 g).
It was extracted 3 times with 0 ml). In addition, butanol (100
(ml) three times, the butanol fraction was concentrated, the concentrate was dissolved in methanol, and TL was added as in Experiment 1.
It analyzed by C and calculated | required the content rate (%) of each component with respect to the total amount of saponins. The results are shown in Table 3 below.

[0068]

[Table 3]

In order to examine the anticancer action of the biotransformed ginseng composition of the present invention, the following Experiment 4 was carried out to examine the cancer cell growth inhibitory effect.

[Experiment 4: Anticancer effect analysis] Human liver cancer-derived cell line (HepG2), human lung cancer-derived cell line (A-549), mouse bone marrow-derived cell line (P-38)
8), the mouse myeloid leukemia cell line (L-1210)
0% fetal bovine serum (FBS), 1% antibiotics-antimycotics
s) and 2.2 g / L sodium bicarbonate supplemented RPMI 1640 medium.

HepG2, A-549 is 0.2
After treating with 5% trypsin, the cells were taken out from the culture flask and placed in a 96-well culture plate.
180 μl was spread according to the number of cells so that each cell would be ³ × 10 ⁴ cells / well, and the cells were allowed to adhere in a CO ₂ incubator (37 ° C.) saturated with 5% carbon dioxide (CO ₂ ) for 24 hours. Also, P-388 and L-1210
180 μl of cells were spread according to the number of cells so that each cell would be ⁴ × 10 ⁴ cells / well, and 5 hours for 5 hours.
Stabilized in a CO ₂ incubator saturated with% CO ₂ .

The biotransformed ginseng concentrate of Example 4 or the extract obtained by soaking white ginseng in methanol with cold was further extracted with butanol, and the saponin fraction was adjusted to a concentration of 10 mg / ml. After sterilizing with high pressure steam, each culture plate had a final concentration of 1 per well.
It was added so that the concentration would be mg / ml, and the cells were cultured for 48 hours in a CO ₂ incubator (37 ° C.) saturated with 5% CO ₂ . Thereafter, 20 mg / ml of tetrazolium (MTT) reagent was added to each well in an amount of 50 μl, and the mixture was further reacted in a CO ₂ incubator for 4 hours. After removing the medium, 100 μl of dimethyl sulfoxide (DMSO) was added to the precipitate. Using an absorbance measuring device (ELISA reader) for enzyme-labeled immunoassay (ELISA), the absorbance (5
The respective ED ₅₀ values were derived by measuring (at 80 nm). The results are shown in Table 4 below. The standard and blank for measurement were prepared by the corresponding known method.

[0073]

[Table 4]

From Table 4, it was found that the biotransformed ginseng composition of the present invention exerts an excellent anticancer effect.

In order to examine the antiallergic effect of the biotransformed ginseng composition of the present invention, the following Experiment 5 was conducted to examine the inhibitory effect on the antigen-antibody reaction.

[Experiment 5: Antiallergic effect analysis 1] Rat mast cells (RBL-2H3 cells), 10% fetal bovine serum, and L-
Culture medium containing glutamine (Dulbeccos' Mod
if the Eagle's Medium: DMEM)
The cells were placed in a culture dish and cultured in a 5% CO ₂ incubator (37 ° C., wet type), and then cells fixed to the culture dish were suspended using a trypsin-EDTA solution, which was separated and collected.

The collected RBL-2H3 cells were added to 24
5 × 10 ⁵ each on a 24-well culture plate
After cell / well splitting, 0.5 μg / mouse of mouse immunoglobulin E (IgE) monoclonal antibody
Each of them was added in an amount of ml, and cultured in a 5% CO ₂ incubator (37 ° C.) for 12 hours to sensitize the antibody.

The sensitized cells were treated with a Siraganian buffer (119 mM sodium chloride, 5 mM potassium chloride, 0.4 mM magnesium chloride, 25 mM piperazine-N, N'-).
Bis (2-ethanesulfonic acid): PIPES), 40m
0.5 ml of M sodium hydroxide, pH 7.2) was added for washing and then 5.6 mM glucose, 1 mM calcium chloride. , 0.16 ml of Silaghanian buffer solution to which 0.1% bovine serum albumin (BSA) was further added, and the mixture was incubated in a 5% CO ₂ incubator (37 ° C.) for 10 minutes.

After culturing for 10 minutes, the biotransformed ginseng concentrate (0.04 ml) of Example 4 and the extract obtained by chilling white ginseng with methanol were further extracted with butanol to obtain a saponin fraction (0 .04 ml), or a known antiallergic agent, disodium cromoglycate (Disodi
um cromolycate), and after 20 minutes,
Antigen (dinitrophenol-bovine serum albumin 1 μg
/ 2 ml) was added, and the cells were activated by culturing at 37 ° C. for 10 minutes and then centrifuged at 2000 rpm for 10 minutes, and the collected supernatant (0.025 ml) was cultured in 96 wells (96 well). Transferred to plate. 1m here
After adding 0.025 ml of M p-NAG culture solution (0.1 M citrate buffer containing p-nitrophenyl-N-acetyl-β-D-glucosamide, pH 4.5), 3
After culturing at 7 ° C for 60 minutes, the reaction was stopped by adding 0.2 ml of 0.1 M sodium carbonate or sodium hydrogen carbonate aqueous solution, and enzyme-labeled immunoassay (ELISA) was performed.
The respective IC ₅₀ values were derived by measuring the absorbance (at 405 nm) using a light absorption measuring device (ELISA reader) for use. The results are shown in Table 5 below. The standard and blank for measurement were prepared by the corresponding known method.

[0080]

[Table 5]

From Table 5, it was found that the biotransformed ginseng composition of the present invention exhibited an excellent antiallergic action.

[Experiment 6: Antiallergic Effect Analysis 2-Histamine Release Inhibition] According to the biotransformed ginseng composition of the present invention,
The effect of inhibiting histamine release was examined.

Physiological solution
solution: 137 mM sodium chloride, 2.
7 mM calcium chloride, 1 mM magnesium chloride hexahydrate, 5.6 mM glucose, 1 unit / ml heparin, 5 mM phosphate buffer, pH 7.2)
The rat (male Wistar rat, 200
20 ml was administered intraperitoneally (± 20 g). Furthermore, the abdomen was lightly massaged for 2 minutes, and then the peritoneal drainage liquid was extracted from the inside of the peritoneum with a syringe. 300 peritoneal fluid drained
It was centrifuged for 5 minutes at 4.times.g and 4.degree. C., and washed several times with the physiological solution.

Saponin fraction obtained by further extracting the peritoneal effluent (2.5 ml) after washing, the biotransformed ginseng concentrate of Example 4 and the extract obtained by chilling white ginseng with methanol and further extracting with butanol. (Ginseng extract: non-biotransformed ginseng concentrate), or a known anti-allergic agent, Disodium cromoglyca (Disodium cromoglyca)
te) suspended at various concentrations in the physiological solution (0.5 m
1) and were mixed and incubated at 37 ° C. for 5 minutes.

To the reaction solution (3.0 ml) after culturing, 0.5 parts of a histamine-releasing substance (compound 48/80) were added.
After mixing with ml and culturing at 37 ° C for 10 minutes, 2500 ×
g, centrifuged at 4 ° C. for 10 minutes to collect the supernatant, and the supernatant (0.6 ml) was added with distilled water (1.4 ml), 1N sodium hydroxide (0.4 ml) and 1% o-phthalaldehyde. -Methanol solution (0.1 ml) was added and allowed to react at room temperature for 4 minutes, then 3N hydrochloric acid (O.2 ml) was added to stop the reaction, and the fluorescence spectrophotometer (Jasco FP
-750, excitation wavelength 353nm, fluorescence wavelength 438n
The amount of histamine was measured using m), each IC ₅₀ of
Derived the value. The results are shown in Table 6 below. The standard and blank for fluorescence spectroscopic measurement were prepared by the corresponding known method.

Since the peritoneal effluent contains many mast cells, it can be considered that the measured histamine is almost derived from mast cells.

[0087]

[Table 6]

From Table 6, the histamine release inhibitory effect of the biotransformed ginseng composition of the present invention was shown to be lower than that of the ordinary ginseng extract (IC ₅₀ ), and the known anti-allergic agent, Disodium cromoglycol. Kate (Dis
It was found that the effect can be exhibited at a concentration almost as low as that of the odium cromolycate). Therefore, the biotransformed ginseng composition of the present invention is
It is considered that it can be used as an anti-allergic substance by suppressing the release of histamine.

[Experiment 7: Antiallergic Effect Analysis 3] The following method of Inagaki et al. (Int. Arch. Allerg)
y Appl. Immunol., 87, 254-25
9, 1988), the antiallergic effect of the biotransformed ginseng composition of the present invention was examined.

Dinitrophenol-Bovine Serum Albumin (Dinitrophenol-bovine ser)
um albumin: DNP-BSA)
E serum (10 μg) was diluted with physiological saline and anesthetized with ether (male ICR mice 20-2).
5 g) was injected into the back and then manually sensitized.

As a control example A, 48 hours after the manual sensitization, DNP-BSA (0.2 mg) and the fluorescent dye (eva) were used.
A physiological saline solution containing ns blue (1.6 mg) (0.
2 ml) into the tail vein and 30 minutes later, cervical dislocation
(Cervical dislocation).

As a trial example, DNP-BSA (0.2 m
g) and fluorescent dye (evans blue, 1.6 mg)
Inject physiological saline containing water (0.2 ml) into the tail vein 1
The saponin fraction obtained by extracting the biotransformed ginseng concentrate of Example 4 and white ginseng cold-soaked with methanol further before extraction with butanol (general ginseng extract: non-biotransformed ginseng concentrate) ), Or a known anti-allergic agent, Disodium cromoglycate (Disodium cro
50 mg / kg, 5 each
Physiological saline dissolved or suspended at 0 mg / kg or 100 mg / kg was orally administered or intraperitoneally injected.

A certain portion was cut from the back of the mouse of the control example A or the trial example, put into a test tube, and 1N potassium hydroxide (0.7 ml) was added thereto, followed by culturing at 37 ° C. overnight.

After culturing, 4 ml of a mixed solution of 0.6N phosphoric acid and acetone (mass ratio 5:13) was added to the tester, and the mixture was shaken and filtered to extract the fluorescent dye by colorimetry (620
Then, the inhibition rate of allergic reaction was derived according to the following formula (1). The results are shown in Table 7 below.

Here, dinitrophenol-bovine serum albumin (Dinitrophenol-bovine)
Serum albumin (DNP-BSA) was prepared by the same procedure as Control Example A except that IgE serum was not sensitized, and colorimetric determination was performed.

[0096]

[Equation 1]

[0097]

[Table 7]

From Table 7, it was found that the biotransformed ginseng composition of the present invention exhibited an excellent antiallergic action.

[Experiment 8: Antioxidant effect-prevention of aging associated with radical scavenging] The biotransformation ginseng composition of the present invention
The 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging effect was investigated.

60 μM DPPH / ethanol solution (5
00 μl) and a sample group (each 500 μl) of the biotransformed ginseng concentrate of Example 4, and the saponin fraction obtained by further extracting the extract obtained by cold soaking white ginseng with methanol (general ginseng). Extract: non-biotransformed ginseng concentrate) or caffeic acid known to have a very strong radical scavenging effect was mixed, and this mixed sample solution was reacted at room temperature for 30 minutes. Absorbance of this mixed solution (520n
The concentration of the sample at which 50% of DPPH radicals were removed (IC ₅₀ ) was determined by measuring (in m). The results are shown in Table 8 below. In the blank, ethanol was used instead of the DPPH / ethanol solution in the mixed sample solution, and in the standard, water was used instead of the sample in the mixed sample solution.

Furthermore, the active oxygen (Superoxide Anion R) produced by the biotransformation ginseng composition of the present invention is used.
Adical) elimination effect was investigated.

0.05M aqueous sodium carbonate solution (pH
10.2, 900 μl), 3 mM xanthine (Xant
hine: Sigma), 3 mM disodium ethylenediaminetetraacetate (EDTA: Sigma), 1.
5 μg / ml BSA (Sigma), 0.75 mM
Nitro blue tetrazolium (NBT: Sigma)
50 μl of each of the above sample groups (50 μl
1) and 0.1 μg / ml xanthine oxidase (50 μl) were added and reacted for 30 minutes, and then 6 mM copper chloride was added to stop the reaction. After measuring the absorbance (at 560 nm) after stopping the reaction,
The concentration of the sample in which 50% of DPPH radicals are removed (IC
₅₀ ) asked. The results are shown in Table 8 below. The standard and blank for measurement were prepared by the corresponding known method.

[0103]

[Table 8]

From Table 8, it was found that the 50% radical scavenging concentration of the biotransformed ginseng composition of the present invention was lower than that of ordinary ginseng extract, although not as low as caffeic acid. From this, it is considered that the biotransformed ginseng composition of the present invention has an excellent antioxidant effect and can be used as a substance for preventing aging of a living body.

[Experiment 9: β-glucuronidase inhibitory effect on colon cancer / liver injury-induced] β-glucuronidase, which is an enzyme produced by intestinal bacteria, causes colon cancer and liver damage. Therefore, a substance that inhibits this enzyme is colon. It is known to have an effect of preventing cancer and liver damage (Reference: D.
H. Kim, I.K. S. Jang, J .; B. Par
k, S. W. Lee, Protective roll
es of musics in experiment
ntal colon rcinogenesis. A
rch. Pharm. Res. 18, 79-83,
1995; D. H. Kim, S.K. B. Shim,
N. J. Kim, I.D. S. Jang, β-glucu
ronidase-inhibitory activ
ity and hepatoprotective e
effects of Ganoderma lucidu
m. Biol. Pharm. Bull. 22, 1
62-164, 1999).

The biotransformed ginseng composition of the present invention provides β-
The glucuronidase inhibitory effect was evaluated by the method of Kim et al.
Kim, S.K. B. Shim, N.M. J. Kim,
I. S. Jang, -Glucuronidase-
inhibitory activity and he
patoprotective effects ofG
anodema lucidum. Biol. Ph
arm. Bull. 22, 162-164, 199
9).

First, Escherichia coli (Es isolated in human intestine
chelicia coli HGU-3) strain was cultured, and the β-glucuronidase enzyme solution was purified according to the method of Kim et al. 50 mM ginseng buffer (0.38 ml)
And 20 mM p-nitrophenyl-β-D-glucuronide (Sigma, USA) (0.02 ml),
The saponin fraction (general ginseng extract: non-extract) obtained by further extracting the enzyme solution (0.05 ml), the biotransformed ginseng concentrate of Example 4 and the white ginseng cold-soaked extract with butanol Biotransformed ginseng concentrate), or sugar acid-1,4-lactone (saccharicacid) known to have a very strong β-glucuronidase activity inhibitory action
1,4-lactone) was mixed and reacted for 30 minutes, and then the reaction was stopped by adding 0.2N sodium hydroxide (0.5 ml).

After the reaction was stopped, the absorbance (at 405 nm) was measured to determine the concentration (IC ₅₀ ) of the sample that inhibits the β-glucuronidase activity by 50%. The results are shown in Table 8 below. The standard and blank for measurement were prepared by the corresponding known method.

[0109]

[Table 9]

From Table 9, it was found that the β-glucuronidase activity inhibitory concentration of the biotransformed ginseng composition of the present invention was lower than that of common ginseng extract, although not as low as sugar acid. From this, the biotransformed ginseng composition of the present invention is excellent in β-glucuronidase inhibitory effect,
It is considered that it can be used as a substance for preventing colorectal cancer and liver damage.

(Other Matters) The ginseng raw material of the biotransformed ginseng composition is not limited to the raw materials shown in the above examples, but selected from the group consisting of ginseng, red ginseng, white ginseng, ginseng and ginseng leaves. Even if one or more natural ginseng, the ginseng extract of the natural ginseng, or the ginseng powder of the natural ginseng is used, it is confirmed that a biotransformed ginseng composition exhibiting the excellent effect of the present invention is provided. There is.

[0112] Further, the microbial cells that biotransform the ginseng raw material are not limited to the microbial cells shown in the above-mentioned examples, but Bifidobacterium KK-1 and Bifidobacterium KK can be used.
-2, one or more lactic acid bacteria selected from the group consisting of Bifidobacterium K-103 and Bifidobacterium K-506, or selected from the group consisting of Prevotella oris and Fusobacterium K-60 It has been confirmed that a bio-converted ginseng composition exhibiting the excellent effect of the present invention can be provided by using one or more enterobacteria. It is needless to say that known lactic acid bacteria and enterobacteria can be used as well as the above-mentioned bacterial cells.

The carrot beverage, which is an example of the biotransformed carrot composition of the present invention, can be provided as follows, for example. First, add purified water to fructose, glucose and sucrose to add 95
After heating to 0 ° C to dissolve, the mixture is gradually cooled to 70 ° C, and citric acid, sodium citrate and sodium benzoate are added thereto and dissolved with stirring. At this time, while stirring the Gokakin extract and taurine,
It may be further dissolved. The biotransformed ginseng powder derived from the biotransformed ginseng concentrate was added to this solution, and the mixture was thoroughly stirred, and then appropriate purified water was added to a total volume of 100 ml to contain the biotransformed ginseng extract (250 mg). Beverage (100 ml) can be provided.

[0114]

INDUSTRIAL APPLICABILITY As described above, according to the biotransformation ginseng composition and the method for producing the same of the present invention, biotransformation in which components having antiallergic action, antioxidant action and anticancer action are greatly enhanced. A carrot composition can be provided.

[0115] Further, according to the biotransformed ginseng composition and the method for producing the same according to the embodiment of the present invention, the saponin component contained in the ginseng raw material is biotransformed to generate a metabolite having high physiological activity. Ginseng leaves, etc., which had a low content of active ingredients such as saponin and were unsuitable for use as a raw material for man-made products, can also be used as raw materials.

[Brief description of drawings]

1 is a flow chart showing a method for producing a biotransformed ginseng composition according to a preferred embodiment of the present invention.

FIG. 2 is a schematic diagram showing the biotransformation process of ginseng saponin.

[Explanation of symbols]

S10: Biotransformation process S20: Concentration process S30: Extraction process S40: Drying process S50: Stirring process S60: Dilution process

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 17/16 A61P 35/00 35/00 37/08 37/08 39/06 39/06 43/00 111 43/00 111 113 113 A61K 31/704 // A61K 31/704 A23L 2/00 F (72) Inventor Sung Zhong Hwan South Korea Republic of Korea Seoul Special City Gangnamgu Apgujeongdong 369-1 Hyundai Apartment 31-304 (72) Inventor Beunga Seoul Special City Sonpagu Shin Chungdong Jangmi Apartment 3 Cha 2-705 (72) Inventor Hanmyung South Korea Seoul Special City Gangnamgu Apgujeongdong 369-1 Hyundai Apartment 31-304 (72) Inventor Chungmin Kyung Seoul Special City Seoul Seochog Bambe 4 Dong 858-33 (72) Inventor Bak Eun Gyung Seoul Special City Gangnam Gyuluon 1 Dong Doshige Val apartment 109-1012 F-term (reference) 4B017 LC03 LG15 LK21 LP01 LP02 LP05 4B018 LB08 LE05 MD64 MD85 MD87 ME06 ME10 MF01 MF06 MF13 4C086 AA01 AA02 AA04 EA11 GA17 MA01 MA04 MA16 MA52 NA14 ZA75C20C41 ZC13 Z41 Z13 Z13 AA02 AA03 BC32 BC59 CA10 CA37 MA16 MA52 NA14 ZA75 ZA89 ZB13 ZB26 ZC13 ZC20 ZC41 4C088 AB18 AC05 AC11 AD22 BA08 BA13 BA31 CA25 MA17 MA52 NA14 ZA75 ZA89 ZB13 ZB26 ZC13 ZC20 ZC41

Claims (18)

[Claims] 1. A biotransformed ginseng composition comprising: (20-O-β-D-glucopyranosyl-20 (S)-
Protopanaxadiol + ginsenoside F2) /
The ratio of (ginsenoside Rc + ginsenoside Rd + ginsenoside Rb1 + ginsenoside Rb2) is
A bioconverted ginseng composition, which is 0.1 or more. 2. The biotransformed ginseng composition according to claim 1, wherein the biotransformed ginseng composition is derived from lactic acid bacteria or enterobacteria. 3. The lactic acid bacterium is Bifidobacterium
KK-1, Bifidobacterium KK-2, Bifidobacterium K-103, Bifidobacterium K
At least one lactic acid bacterium selected from the group consisting of -506, wherein the intestinal bacterium is at least one intestinal bacterium selected from the group consisting of Prevotella orris and Fusobacterium K-60. The biotransformed ginseng composition according to claim 2. 4. The biotransformed ginseng composition according to claim 1, wherein the biotransformed ginseng composition is an antiallergic substance that suppresses the release of histamine. 5. The biotransformed ginseng composition according to claim 1, wherein the biotransformed ginseng composition is an antioxidant having an antioxidant effect by radical scavenging. 6. The biotransformed ginseng composition according to claim 1, wherein the biotransformed ginseng composition is an anticancer agent. 7. The biotransformed ginseng composition according to claim 1, wherein the biotransformed ginseng composition is a preventive agent for colon cancer or liver damage, which inhibits the production of β-glucuronidase. 8. The biotransformed ginseng composition according to claim 1, wherein the biotransformed ginseng composition is a beverage containing a biotransformed ginseng component. 9. A method for producing a biotransformed ginseng composition, comprising a step of suspending a ginseng raw material in water and then culturing by adding lactic acid bacteria or enterobacteria to obtain a biotransformed ginseng liquid. Concentration, freezing of the biotransformed ginseng liquid that has undergone the biotransformation process,
Alternatively, a method of producing a biotransformed ginseng composition, comprising a concentration step of obtaining a biotransformed ginseng concentrate by drying or centrifuging the supernatant. 10. One or more substances selected from the group consisting of raw ginseng, red ginseng, white ginseng, ginseng, ginseng leaf, ginseng extract and ginseng powder are used as the ginseng raw material. The method for producing the biotransformed ginseng composition according to claim 9. 11. The Bifidobacterium KK-1 and Bifidobacterium KK-2 are used as the lactic acid bacteria.
The method for producing a biotransformed ginseng composition according to claim 9, wherein one or more lactic acid bacteria selected from the group consisting of: 12. The lactic acid bacteria are Bifidobacterium K-103 and Bifidobacterium K-5.
The method for producing a biotransformed ginseng composition according to claim 9, wherein one or more lactic acid bacteria selected from the group consisting of 06 is used. 13. The lactic acid bacterium is selected from the group consisting of Bifidobacterium KK-1, Bifidobacterium KK-2, Bifidobacterium K-103 and Bifidobacterium K-506. The method for producing a biotransformed ginseng composition according to claim 9, wherein one or more lactic acid bacteria are used. 14. The biotransformation according to claim 9, wherein one or more enterobacteria selected from the group consisting of Prevotella orris and Fusobacterium K-60 is used as the enterobacteria. A method for producing a carrot composition. 15. The method according to claim 9, further comprising, after the concentration step, an extraction step of administering a preset solvent to the biotransformed ginseng concentrate to obtain a biotransformed ginseng extract. A method for producing a bio-converted ginseng composition. 16. After the extraction step, a stirring step of mixing a beverage stock solution prepared by adding purified water to a sweetener and the biotransformed ginseng extract solution to obtain a biotransformed ginseng mixed solution, and the biotransformed ginseng. The method for producing a biotransformed ginseng composition according to claim 15, further comprising: a dilution step of adding purified water to the mixed solution to obtain a biotransformed ginseng beverage solution. 17. The undiluted beverage solution in the stirring step further comprises:
The method for producing a biotransformed ginseng composition according to claim 16, which comprises one or more substances selected from the group consisting of citric acid, sodium citrate, a crude drug extract, and taurine. 18. The biotransformed ginseng composition according to claim 15, further comprising a drying step of freeze-drying the biotransformed ginseng extract to obtain a biotransformed ginseng powder after the extraction step. Method of manufacturing things.