KR102106018B1 - Method For Producing Steaming Ginseng Berry Fermented Extracts Having Preventive and Improving Effects of Liver Damage - Google Patents

Abstract

The present invention relates to a method for producing a steamed and fermented ginseng berry extract. The method comprises steps of: steaming and drying ginseng berries to prepare primarily steamed ginseng berries; steaming and drying the primarily steamed ginseng berries to prepare secondarily steamed ginseng berries; steaming and drying the secondarily steamed ginseng berries to prepare tertiarily steamed ginseng berries; carrying out an extraction for the tertiarily steamed ginseng berries with an extraction solvent to obtain a steamed ginseng berry extract; and inoculating Lactobacillus casei GFC 1 strain (accession number: KCTC18333P) into the steamed ginseng berry extract. The steamed ginseng berry extract prepared from the present invention shows a remarkable increase in a content of ginsenoside contained in ginseng, thereby having an excellent effect of preventing or ameliorating liver damage.

Description

Method for Producing Steaming Ginseng Berry Fermented Extracts Having Preventive and Improving Effects of Liver Damage

The present invention relates to a method for preparing fermented ginseng berry fruit with prevention or improvement activity of liver damage.

Ginseng ( Panax ginseng CA Meyer ) is a plant belonging to the genus Agapiaceae and has been used in Korea, China, and Japan for more than 2,000 years, and has been used for the purpose of preventing disease and extending life. The efficacy of ginseng known to date is the action on the central nervous system, anti-carcinogenic action, anti-cancer activity, immune function control action, anti-diabetic action, liver function anti-efficacy effect, cardiovascular disorder improvement, anti-atherosclerosis action, blood pressure control action, improvement of climacteric disorders , Effects on osteoporosis, anti-stress action, anti-fatigue action, antioxidant activity, anti-aging, etc.

Ginseng is a non-saponin-based bioactive substance, including more than 40 ginsenosides, which are glycosides, and contains various useful ingredients for the human body, such as Polyacetylene, Phenolic compounds, Acid polysaccharides, Peptides, and Alkaloids.

Ginsenoside, the representative bioactive component of ginseng, means saponin contained in ginseng, and ginseng contains various types of ginsenoside components. Ginsenosides are evenly distributed on the ground and the basement of ginseng. In particular, it is known that ginsenosides have different composition as well as ginsenoside content depending on the part of ginseng root (root), ginseng leaf, and ginseng fruit.

Ginseng ginsenosides account for about 5% of ginseng dryness, and glucose, glucose, arabinose, xylose, and ramnose are on the trimarpenoid-based Dammarane skeleton. (Rhamnose) is a neutral glycoside. Protopanaxadiol (PPD) -based saponins for 2 individuals (positions 3 and 12) depending on the number of -OH attached to the non-sugar moiety, and protopanax triols for 3 individuals (positions 3, 6 and 12) It is called (Protopanaxatriol; PPT) series saponins. Glucose, arabinose, xylose, rhamnose, and the like are combined with -OH at the R1, R2, and R3 positions of the PPD and PPT. More than 30 types of ginsenosides have been reported separately from ginseng, but the significant amount is detected when extracting and analyzing actual ginseng. Six types of Rb1, Rb2, Rc, Rd, Re and Rg1 occupy more than 90% of the total saponin, and the rest Saponin ingredients are low in content.

Ginsenosides are not absorbed as they are in the human body, but are broken down by microorganisms such as Bacteriodes, Lactobacillus, Fusobacterium, Eubacterium, and Provetella species present in the human intestine, and their metabolites are absorbed. In the case of ginsenoside Rb1, it is sequentially converted to ginsenoside Rd, compound K (hereinafter abbreviated as CK) and protopanaxadiol by intestinal microorganisms. PPD-based ginsenosides Rb1, Rb2, Rc, and Rd are metabolized and absorbed in the intestine as 20-O-β-D-glucopyranosyl-20 (S) -protopanaxadiol (CK). PPT-based ginsenosides Re, Rf and Rg1 are metabolized to Rh1, F1 and Protopanaxatriol in the intestine. PPD-based ginsenosides Rb1, Rb2, Rc, and Rd are converted into 20 (R)-/ 20 (S) -ginsenoside Rg3 by intestinal microorganisms, and Rg3 is converted to Rh2 by intestinal microorganisms.

In recent years, such a medicinal ingredient of ginseng is easily absorbed into the human body, and a method of fermenting ginseng using microorganisms or lactic acid bacteria in the intestine or processing enzymes is used as a method of strengthening components present in a very small amount in ginseng. Korean Registered Patent Publication No. 10-1017378 relates to `` a method for manufacturing fermented ginseng, fermented red ginseng, and Bacillus subtilis new strains derived from Cheonggukjang suitable for it, '' using ginseng using microorganisms from Bacillus isolated from Cheonggukjang And fermented ginseng and fermented red ginseng, which are characterized by enhancing the content of ginsenoside Rd by fermenting red ginseng, and technology related to Bacillus substilis new strains derived from Cheonggukjang.

In addition, Korean Registered Patent Publication No. 10-1240192 relates to `` Ginseng fermentation extract for improving insulin secretion using Lactobacillus acidophilus Ｋ-59 strain, Ｋ-59 strain and a manufacturing method thereof, '' and newly identified and identified Ginsenosides Rg1, Rb1, Rg1 or Re, which are components with low bioavailability of ginseng, are converted into ginsenosides Rh1, Rg3, Rh2 or CK using the Lactobacillus acidophilus K-59 (KFCC11467P) strain. A method of conversion is disclosed.

Thus, the present inventors prepared ginseng fruit by steaming and drying them step by step using ginseng fruit, and extracting and fermenting it to produce fermented ginseng fruit, ginsenoside contained in ginseng. ) Content was significantly improved to confirm that the alcoholic liver damage can be prevented or improved, and the present invention has been completed.

Republic of Korea Registered Patent Publication No. 10-1017378 Republic of Korea Registered Patent Publication No. 10-1240192

The main object of the present invention is to provide a steamed ginseng fruit fermentation product capable of enhancing the content of ginsenosides contained in ginseng fruit and a method for manufacturing the same.

Another object of the present invention is to provide a health functional food for preventing or improving alcoholic liver damage comprising the steamed ginseng fruit fermentation product as an active ingredient.

In order to achieve the above object, the present invention comprises the steps of steaming and drying the ginseng fruit to produce a primary steamed ginseng fruit; Preparing a second steamed ginseng fruit by steaming and drying the first steamed ginseng fruit; Preparing a third steamed ginseng fruit by steaming and drying the second steamed ginseng fruit; Extracting the third steamed ginseng fruit with an extraction solvent to obtain steamed ginseng fruit extract; It provides a method for producing fermented ginseng fruit ginseng, characterized in that it comprises the step of inoculating and fermenting Lactobacillus casei GFC 1 (Accession No .: KCTC18333P) strain to the steamed ginseng fruit extract.

In the present invention, the primary steamed ginseng fruit is prepared by steaming at 80 to 90 ° C for 3 to 5 hours, and then drying at 50 to 60 ° C for 15 to 30 hours, and the secondary steamed ginseng fruit is It is prepared by steaming at 80 to 90 ° C for 2 to 5 hours, then dried at 50 to 60 ° C for 15 to 30 hours, and the third steamed ginseng fruit is steamed at 80 to 90 ° C for 2 to 5 hours, and then , Dried at 50 to 60 ° C. for 15 to 30 hours.

In the present invention, the fermentation is prepared by adding 0.5 to 1.0 g of lactose to steamed ginseng fruit extract, and then producing Lactobacillus casei GFC 1 strain (Accession No .: KCTC18333P) in the medium. Characterized in that the culture by inoculation.

In the present invention, the steamed ginseng fruit fermentation product is ginsenoside Rg2 (ginsenoside Rg2), ginsenoside Rg3 (ginsenoside Rg3), ginsenoside Rh2 (ginsenoside Rh2), ginsenoside Rh3 (ginsenoside Rh3), ginsenoside Ginsenoside Rh23 (ginsenoside Rh23), ginsenoside Rk1 (ginsenoside Rk1), ginsenoside Rg5 (ginsenoside Rg5), and ginsenoside F4 (ginsenoside F4) comprising at least one active ingredient selected from the group consisting of do.

In the present invention, the fermented ginseng fruit fermentation product is ginsenoside Rg2 (ginsenoside Rg2) 1.0 to 1.5 mg / g, ginsenoside Rg3 (ginsenoside Rg3) 0.5 to 1.2 mg / g, ginsenoside Rh2 (ginsenoside Rh2) 0.3 to 0.7 mg / g, ginsenoside Rh3 (ginsenoside Rh3) 0.6 to 1.5 mg / g, ginsenoside Rh23 (ginsenoside Rh23) 0.1 to 0.5 mg / g, ginsenoside Rk1 (ginsenoside Rk1) 1.0 to 3.0 mg / g, ginsenoside Rg5 (ginsenoside Rg5) 1.0 to 3.0 mg / g and ginsenoside F4 (ginsenoside F4) 1.0 to 3.0 mg / g.

In addition, the present invention is ginsenoside Rg2 (ginsenoside Rg2) 1.0 ~ 1.5mg / g, ginsenoside Rg3 (ginsenoside Rg3) 0.5 ~ 1.2 mg / g, ginsenoside Rh2 (ginsenoside Rh2) 0.3 ~ 0.7 mg / g, Ginsenoside Rh3 (ginsenoside Rh3) 0.6 to 1.5 mg / g, ginsenoside Rh23 (ginsenoside Rh23) 0.1 to 0.5 mg / g, ginsenoside Rk1 (ginsenoside Rk1) 1.0 to 3.0 mg / g, ginsenoside Rg5 ( ginsenoside Rg5) 1.0 to 4.0 mg / g and ginsenoside F4 (ginsenoside F4) 1.0 to 3.0 mg / g to provide a fermented ginseng fruit characterized in that it contains an active ingredient.

In the present invention, the fermented ginseng fruit is characterized in that it has an effect on inhibiting liver damage and anti-inflammatory.

In addition, the present invention provides a health functional food for preventing or improving liver damage comprising fermented ginseng fruit.

The fermentation product of steamed ginseng fruit prepared from the manufacturing method of the present invention is excellent in preventing or improving liver damage because the content of ginsenoside contained in ginseng is significantly enhanced. Accordingly, it is possible to have a low production cost and high productivity, and thus economic efficiency is high.

1 is a graph showing the results of the test for preventing liver cell damage efficacy (GOT / AST) of steamed ginseng fruit fermentation obtained from Example 1 of the present invention.
Figure 2 is a graph showing the results of the liver cell damage prevention efficacy test (GPT / ALT) of steamed ginseng fruit fermentation obtained from Example 1 of the present invention.
3 is a graph showing the expression level of SOD (Superoxide dismutase), an enzyme related to antioxidant activity of steamed ginseng fruit fermentation obtained from Example 1 of the present invention.

In the description of the present invention, when it is determined that detailed descriptions of related known technologies or configurations may unnecessarily obscure the subject matter of the present invention, detailed descriptions thereof will be omitted.

In addition, terms to be described later are terms defined in consideration of functions in the present invention, which may vary according to a user's or operator's intention or practice, and thus the definition should be made based on the contents of the present specification describing the present invention.

Thus, the present inventors prepared ginseng fruit by steaming and drying them step by step using ginseng fruit, and extracting and fermenting it to produce fermented ginseng fruit, ginsenoside contained in ginseng. It was predicted that the content of) could be significantly enhanced to prevent or improve alcoholic liver damage.

In this example, steamed ginseng fruit is prepared by steaming and drying step by step using the fruit of ginseng, and fermented steamed ginseng fruit is produced through extraction and fermentation to confirm the composition and content of specific ginsenosides.

Therefore, the present invention, in one aspect, steaming and drying the ginseng fruit to produce a primary steamed ginseng fruit; Preparing a second steamed ginseng fruit by steaming and drying the first steamed ginseng fruit; Preparing a third steamed ginseng fruit by steaming and drying the second steamed ginseng fruit; Extracting the third steamed ginseng fruit with an extraction solvent to obtain steamed ginseng fruit extract; It relates to a method for producing fermented ginseng fruit ginseng, characterized in that it comprises the step of inoculating and fermenting Lactobacillus casei GFC 1 strain (Accession No .: KCTC18333P) to the steamed ginseng fruit extract.

A more detailed method of preparing steamed ginseng fruit fermentation is as follows.

In the present invention, "steamed ginseng fruit fermentation product" refers to a product obtained by steaming and drying the ginseng fruit step by step to produce steamed ginseng fruit, and extracting and fermenting it.

According to a preferred embodiment of the present invention, the primary steamed ginseng fruit is prepared by steaming at 80 to 90 ° C for 3 to 5 hours, and then drying at 50 to 60 ° C for 15 to 30 hours to make saponin as an active ingredient. It is preferred in terms of conversion and increasing polysaccharide content.

If the primary steamed ginseng fruit is steamed at less than 80 ° C, heat may not be effectively transferred to the tissues within the ginseng fruit, and a problem that effective saponins contained in the ginseng fruit are not effectively converted may occur. In addition, when steaming ginseng fruit that is primarily steamed above 90 ° C, saponin components are destroyed and pharmacological activity is reduced, which is not preferable.

In addition, if the primary steamed ginseng fruit is steamed for less than 3 hours, saponin is not efficiently converted, and thus, an active ingredient is not sufficiently generated. When steamed for more than 5 hours, the saponin component is destroyed and pharmacological activity is reduced. Therefore, it is not preferable.

In addition, when the primary steamed ginseng fruit is dried, it is not preferable because it may cause a problem of destroying active ingredients while being exposed to heat for a long time when it is out of 15 to 30 hours at 50 to 60 ° C.

According to a preferred embodiment of the present invention, the secondary steamed ginseng fruit is prepared by steaming at 80 to 90 ° C for 2 to 5 hours, and then drying at 50 to 60 ° C for 15 to 30 hours to make saponin as an active ingredient. It is preferred in terms of conversion and increasing polysaccharide content.

If the second steamed ginseng fruit is steamed at less than 80 ° C, effective saponin contained in the ginseng fruit is not effectively converted, which may cause a problem that the content of the active ingredient is significantly reduced. In addition, when steaming the second steamed ginseng fruit in excess of 90 ° C, it is not preferable because the high temperature can easily break the tissue and cause the problem of destroying the active ingredients.

In addition, when the second steamed ginseng fruit is steamed for less than 2 hours, saponin is not efficiently converted, so that an active ingredient is not sufficiently generated, and when steamed for more than 5 hours, the saponin component is destroyed and pharmacological activity is reduced. Therefore, it is not preferable.

In addition, when the second steamed ginseng fruit is dried, it is not preferable because when it is out of the range of 15 to 30 hours at 50 to 60 ° C., it may cause a problem of destroying active ingredients while being exposed to heat for a long time.

According to a preferred embodiment of the present invention, the third steamed ginseng fruit is prepared by steaming at 80 to 90 ° C for 2 to 5 hours and then drying at 50 to 60 ° C for 15 to 30 hours to make saponin as an active ingredient. It is preferred in terms of conversion and increasing polysaccharide content.

If the third steamed ginseng fruit is steamed at less than 80 ° C, the effective saponin contained in the ginseng fruit is not effectively converted, which may cause a problem that the content of the active ingredient is significantly reduced. In addition, when steaming ginseng fruit that has been steamed to a temperature exceeding 90 ° C., it is not preferable because tissues are easily broken and burned due to high temperature, which may cause problems of destroying active ingredients.

In addition, when the 3rd steamed ginseng fruit is steamed for less than 2 hours, saponin is not efficiently converted, so that an active ingredient is not sufficiently generated. When it is steamed for more than 5 hours, the saponin component is destroyed and the pharmacological activity is reduced. Therefore, it is not preferable.

In addition, when the third steamed ginseng fruit is dried, it is not preferable because when it is out of the range of 15 to 30 hours at 50 to 60 ° C., it may cause a problem of destroying active ingredients while being exposed to heat for a long time.

If the fermentation product is prepared only with the steaming ginseng fruit of step 1 or 2 without going through the steaming process of step 3, the minor ginsenosides generated by repeating the steaming and drying process are not sufficiently generated, and the content of the polysaccharide is reduced to effect efficacy This is reduced, and it is not preferable because a problem in which pharmacological components such as ginsenosides contained in the fermented ginseng ginseng fruit finally produced may be significantly reduced may occur.

According to a preferred embodiment of the present invention, the fermentation is prepared by adding 0.5 to 1.0 g of lactose to steamed ginseng fruit extract, followed by Lactobacillus casei GFC 1 strain (accession number: KCTC18333P) is preferably inoculated and cultured.

At this time, it is very important to add lactose in an optimal content to make the growth of the strain more smooth and increase the activity of the enzyme. If lactose is added to the extract of steamed ginseng fruit in an amount of less than 0.5 g, it is not preferable because it does not produce enzyme activity smoothly, and if it is added in excess of 1.0 g, the growth of the strain is hindered and the enzyme secretion is reduced. This is not preferred.

According to a preferred embodiment of the present invention, it is possible to prepare the steamed ginseng fruit extract by extracting the third steamed ginseng fruit with an extraction solvent.

The extraction is preferably added to the extraction solvent in a content of 1 to 50 times the weight of solid content of steamed ginseng fruit. For example, 100 parts by weight of steamed ginseng fruit may be extracted by adding an extraction solvent at 100 to 5000 parts by weight.

Specifically, the content of the extraction solvent may be 1.5 to 30 times, more specifically 5 to 20 times or 5 to 15 times the weight of steamed ginseng fruit.

The extraction solvent may be water, an alcoholic organic solvent, or a mixture thereof. Specifically, the water may be alkaline water. Specifically, the organic solvent may include one or more of anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms, for example, ethanol, propanol, methanol, n-butanol, and mixtures thereof. The extraction solvent may be used alone or in combination of two or more.

According to a preferred embodiment of the present invention, the final steamed ginseng fruit fermentation product can be prepared by inoculating and fermenting the steamed ginseng fruit extract with Lactobacillus casei GFC 1 strain (Accession No .: KCTC18333P).

At this time, fermentation using Lactobacillus casei GFC 1 ( Lactobacillus casei GFC 1) strain (Accession No .: KCTC18333P) is preferred in terms of inducing the conversion and production of saponins.

The steamed ginseng fruit fermentation product may be obtained by centrifuging at 8000 rpm for 20 minutes to remove the cells, followed by filtration and sterilization.

In addition, the fermentation product of steamed ginseng fruit may have a sugar content of 5.0 to 50.0 brix.

The steamed ginseng fruit fermentation product may be processed and used in various forms such as dried powder and concentrated filtrate. The extract prepared from the fermented ginseng fruit is not particularly limited, but may be used, for example, in the form of tincture, concentrate, extract, and flow extract containing hydrous alcohol as a leaching solvent.

The fermented ginseng fruit obtained as described above is ginsenoside Rg2 (ginsenoside Rg2), ginsenoside Rg3 (ginsenoside Rg3), ginsenoside Rh2 (ginsenoside Rh2), ginsenoside Rh3 (ginsenoside Rh3), ginsenoside Rh23 (ginsenoside Rh23), ginsenoside Rk1 (ginsenoside Rk1), ginsenoside Rg5 (ginsenoside Rg5), and may include one or more active ingredients selected from the group consisting of ginsenoside F4 (ginsenoside F4).

At this time, the fermented ginseng fruit fermentation product is ginsenoside Rg2 (ginsenoside Rg2) 1.0 to 1.5 mg / g, ginsenoside Rg3 (ginsenoside Rg3) 0.5 to 1.2 mg / g, ginsenoside Rh2 (ginsenoside Rh2) 0.3 to 0.7 mg / g, ginsenoside Rh3 (ginsenoside Rh3) 0.6 to 1.5 mg / g, ginsenoside Rh23 (ginsenoside Rh23) 0.1 to 0.5 mg / g, ginsenoside Rk1 (ginsenoside Rk1) 1.0 to 3.0 mg / g, gins Ginsenoside Rg5 (ginsenoside Rg5) 1.0 to 4.0 mg / g and ginsenoside F4 (ginsenoside F4) 1.0 to 3.0 mg / g may include active ingredients.

More specifically, the steamed ginseng fruit fermentation product contains ginsenoside Rg3 (ginsenoside Rg3) 0.5 to 1.2 mg / g, which is more than ginseng fruit extract containing about 0.2 mg / g ginsenoside Rh3 (ginsenoside Rh3). It may be increased by ~ 6 times or more.

In addition, the steamed ginseng fruit fermentation product contains ginsenoside Rh3 (ginsenoside Rh3) 0.6-1.5 mg / g, which is 3-7 than ginseng fruit extract containing ginsenoside Rh3 (ginsenoside Rh3) of about 0.2mg / g. It may be more than doubled.

In addition, steamed ginseng fruit fermentation products include 0.1 to 0.5 mg / g of Rh23 (ginsenoside Rh23), which may be significantly increased than ginseng fruit extract containing about 0.0mg / g of Rh23 (ginsenoside Rh23).

In addition, the steamed ginseng fruit fermentation product includes ginsenoside Rk1 (ginsenoside Rk1) 1.0 to 3.0 mg / g, which is 10-30 than ginseng fruit extract containing ginsenoside Rk1 (ginsenoside Rk1) about 0.1 mg / g. It may be more than doubled.

In addition, fermented ginseng fruit fermentation products include ginsenoside Rg5 (ginsenoside Rg5) 1.0-4.0 mg / g, which is 5-15 compared to ginseng fruit extract containing ginsenoside Rg5 (ginsenoside Rg5) about 0.2mg / g. It may be more than doubled.

In addition, the steamed ginseng fruit fermentation product contains ginsenoside F4 (ginsenoside F4) 1.0-3.0 mg / g, which is 2-8 compared to ginseng fruit extract containing about 0.4 mg / g ginsenoside F4 (ginsenoside F4). It may be more than doubled.

As described above, steamed ginseng fruit fermentation products have more excellent pharmacological components, such as ginsenosides, than the generally prepared ginseng fruit extract, and thus have better efficacy.

As described above, steamed ginseng fruit is prepared by steaming and drying it step by step using ginseng fruit, and when extracting and fermenting it to produce fermented ginseng fruit, ginsenosides contained in ginseng (Ginsenoside) ) Content can be significantly improved to impart an effect of preventing or improving alcoholic liver damage.

On the other hand, the present invention is a ginsenoside Rg2 (ginsenoside Rg2) 1.0 ~ 1.5mg / g, ginsenoside Rg3 (ginsenoside Rg3) 0.5 ~ 1.2 mg / g, ginsenoside Rh2 (ginsenoside Rh2) prepared by the above-mentioned manufacturing method ) 0.3 to 0.7 mg / g, ginsenoside Rh3 (ginsenoside Rh3) 0.6 to 1.5 mg / g, ginsenoside Rh23 (ginsenoside Rh23) 0.1 to 0.5 mg / g, ginsenoside Rk1 (ginsenoside Rk1) 1.0 to 3.0 mg / g, ginsenoside Rg5 (ginsenoside Rg5) 1.0 to 4.0 mg / g and ginsenoside F4 (ginsenoside F4) 1.0 to 3.0 mg / g relates to fermented ginseng fruit characterized in that it contains an active ingredient. .

According to a preferred embodiment of the present invention, the fermented ginseng berry fruit may be effective in inhibiting liver damage and anti-inflammatory.

On the other hand, the present invention relates to a health functional food for preventing or improving liver damage comprising the fermented ginseng fruit.

According to a preferred embodiment of the present invention, the health functional food for preventing or improving liver damage may include 1.0 to 5.0mg, preferably 2.0 to 4.0mg, more preferably 2.5 to 3.0mg of fermented ginseng fruit. have.

The health functional food may be provided as a liquid beverage formulation, and as an example of the health functional food, steamed ginseng fruit fermentation product is powdered using a method such as spray drying, and these powders are purified, pills, powders, tablets , It can be provided in any one or more solid dosage form selected from capsulants.

Hereinafter, the present invention will be described in more detail through examples. These examples are only intended to explain the present invention in more detail, and according to the gist of the present invention, the scope of the present invention is not limited by these examples to those skilled in the art to which the present invention pertains. It will be self-evident.

<Production Example 1> Preparation of steamed ginseng fruit

(1) After putting ginseng fruit into the washing machine and completely removing foreign substances such as soil, the ginseng fruit was put into a steamer and primary steamed at 90 ° C for 5 hours.

(2) The primary steamed ginseng fruit was dried at 60 ° C for 30 hours.

(3) The primary steamed and dried steamed ginseng fruits were second steamed at 85 ° C for 3 hours.

(4) The second steamed ginseng fruit was dried at 60 ° C for 30 hours.

(5) Second steaming and dried steamed ginseng fruit were steamed at 85 ° C for 3 hours.

(6) The third steamed ginseng fruit was dried at 60 ° C for 30 hours.

<Production Example 2> Preparation of primary steamed ginseng fruit extract

1 kg of the primary steamed ginseng fruit was added to 10 kg of 30% ethanol and repeatedly extracted three times at 80 ° C. for 3 hours, the filtrate was mixed, and concentrated under reduced pressure to 60 brix to prepare a primary steamed ginseng fruit extract.

<Production Example 3> Preparation of secondary steamed ginseng fruit extract

1 kg of the second steamed ginseng fruit was added to 10 kg of 30% ethanol and repeatedly extracted three times at 80 ° C. for 3 hours, and the filtrate was mixed and concentrated under reduced pressure to 60 brix to prepare a second steamed ginseng fruit extract.

<Production Example 4> Preparation of tertiary steamed ginseng fruit extract

1 kg of the third steamed ginseng fruit was added to 10 kg of 30% ethanol and repeatedly extracted three times at 80 ° C. for 3 hours, and the filtrate was mixed and concentrated under reduced pressure to 60 brix to prepare a third steamed ginseng fruit extract.

<Example 1> Preparation of steamed ginseng fruit fermentation

1 to 2 g of ginseng fruit is added to 100 g of MRS medium, and the medium is inoculated with Lactobacillus casei GFC 1 strain (Accession No .: KCTC18333P), and cultured with stirring for 3 days at 37 ° C. Cultured. After mixing 3 wt% ginseng fruit extract 10 wt% and 90 wt% water containing 1.0 g lactose in Preparation Example 4 to form a fermentation medium, sterilizing it at 80 ° C. for 1 hour and cooling it to room temperature to ferment The medium for the preparation was prepared.

Lactobacillus casei GFC 1 ( Lactobacillus casei GFC 1) strain (Accession No .: KCTC18333P) previously pre-cultured was inoculated with 3 parts by weight based on 100 parts by weight of the prepared medium, and then incubated for 7 days at 37 ° C, followed by centrifugation. To obtain a culture supernatant. The fermentation product was centrifuged at 8000 rpm for 20 minutes to remove the cells, followed by filtration and sterilization. The fermentation product was concentrated under reduced pressure to 60 brix to prepare a final steamed ginseng fruit fermentation product.

<Comparative Example 1> Preparation of ginseng fruit extract

Ginseng fruit extract was prepared by adding 1 kg of unsteamed ginseng fruit to 10 kg of 30% ethanol and repeatedly extracting it three times at 80 ° C. for 3 hours, mixing the obtained filtrate, and then concentrating under reduced pressure to 60 brix.

<Comparative Example 2> Preparation of tertiary steamed ginseng fruit extract

1 kg of the third steamed ginseng fruit prepared in Preparation Example 4 was added to 10 kg of ethanol and repeatedly extracted three times at 80 ° C. for 3 hours, and the filtrate was mixed and concentrated under reduced pressure to 60 brix to extract the third steamed ginseng fruit extract. It was prepared.

<Comparative Example 3> Preparation of fermented ginseng fruit

After mixing 10% by weight of ginseng fruit extract of Comparative Example 1 and 90% by weight of water containing 1.0 g of lactose to form a fermentation medium, sterilization treatment at 80 ° C. for 1 hour and cooling to room temperature results in a medium for fermentation Was prepared.

Next, Lactobacillus casei GFC 1 ( Lactobacillus casei GFC 1) strain (Accession No .: KCTC18333P) pre-cultured in the same manner as in Example 1 was inoculated at 3 parts by weight with respect to 100 parts by weight of the prepared medium, and at 37 ° C. After suspension culture for 7 days, the culture supernatant was obtained by centrifugation. The fermentation product was centrifuged at 8000 rpm for 20 minutes to remove the cells, followed by filtration and sterilization. The fermentation product was concentrated under reduced pressure to 60 brix to prepare a final ginseng fruit fermentation product.

<Comparative Example 4> Preparation of primary steamed ginseng fruit

After preparing the fermentation medium by mixing 90% by weight of water containing 10% by weight of the first steamed ginseng fruit extract of Preparation Example 2 and 1.0g of lactose, and then sterilizing it at 80 ° C for 1 hour, cooling it to room temperature to ferment The medium for the preparation was prepared.

Next, Lactobacillus casei GFC 1 ( Lactobacillus casei GFC 1) strain (Accession No .: KCTC18333P) pre-cultured in the same manner as in Example 1 was inoculated at 3 parts by weight with respect to 100 parts by weight of the prepared medium, and at 37 ° C. After suspension culture for 7 days, the culture supernatant was obtained by centrifugation. The fermentation product was centrifuged at 8000 rpm for 20 minutes to remove the cells, followed by filtration and sterilization. The fermentation product was concentrated under reduced pressure to 60 brix to prepare a final ginseng fruit fermentation product.

<Comparative Example 5> Secondary steamed ginseng fruit fermentation

After preparing the fermentation medium by mixing 90% by weight of water containing 10% by weight of the second steamed ginseng fruit extract of Preparation Example 3 and 1.0g of lactose, sterilizing it at 80 ° C for 1 hour and cooling it to room temperature to ferment The medium for the preparation was prepared.

Next, Lactobacillus casei GFC 1 ( Lactobacillus casei GFC 1) strain (Accession No .: KCTC18333P) pre-cultured in the same manner as in Example 1 was inoculated at 3 parts by weight with respect to 100 parts by weight of the prepared medium, and at 37 ° C. After suspension culture for 7 days, the culture supernatant was obtained by centrifugation. The fermentation product was centrifuged at 8000 rpm for 20 minutes to remove the cells, followed by filtration and sterilization. The fermentation product was concentrated under reduced pressure to 60 brix to prepare a final ginseng fruit fermentation product.

. <Test Example 1> Ginsenoside content analysis (HPLC; High Performance Liquid Chromatography)

Supernatant obtained by adding 5 g of water-saturated butanol to 1 g of steamed ginseng fruit fermented product and ginseng fruit extract prepared from Example 1 and Comparative Examples 1 to 5, and then taking the supernatant of the butanol mixture solution and repeating this process twice Was concentrated under reduced pressure. The obtained concentrate was dissolved in methanol (HPLC grade), filtered through a 0.2 μm membrane filter, and used as a sample for HPLC analysis. In the HPLC analysis, a C18 column (3.0x50 mmm, ID 5 μm) was used to detect UV 203 nm. The mobile phase was given gradient using Acetonitrile (solvent A) and Water (solvent B) and separated at a flow rate of 1.6 ml / min, and the gradient conditions of the mobile phase are shown in Table 1.

Time (min) Mobile phase Acetonitrile (solvent A) Water (solvent B) 0.00 17 83 10.0 17 83 18.0 23 77 32.0 31 69 45.0 48 52 60.0 72 28 68.0 90 10 78.0 90 10 80.0 17 83 90.0 17 83

Table 1 shows the results of the changes in the ginsenoside components of Example 1 and Comparative Examples 1 to 5. At this time, the content unit is mg / g.

Re Rd Rg2 Rg3 Rh2 Rh3 Rh23 Rk1 Rg5 F4 Example 1 2.34 0.32 1.48 1.15 0.68 1.42 0.44 2.88 3.05 2.90 Comparative Example 1 11.51 9.08 1.62 0.18 0.23 0.21 0.00 0.10 0.11 0.39 Comparative Example 2 4.32 3.54 2.04 0.67 0.40 0.38 0.02 2.39 2.54 0.81 Comparative Example 3 9.54 8.52   3.58 0.38 0.33 0.29 0.15 0.09 0.12 0.42 Comparative Example 4 6.21 3.33 2.52 4.86 0.54 0.51 0.20 0.84 1.67 0.99 Comparative Example 5 5.36 1.86 1.47 3.34 0.59 0.89 0.29 1.56 2.08 2.22

As a result, as shown in Table 2, as a result of analyzing the ginsenoside components and contents of Example 1 and Comparative Examples 1 to 5, ginsenosides Rh2, Rh3, Rh23, Rk1, Rg5 of Example 1 And it was confirmed that the content of F4 is remarkably high. The content of ginsenoside Re and Rd is reduced through the process of steaming and fermenting ginseng fruit

This was confirmed that the effective ginsenoside contained in the ginseng fruit was effectively converted by fermenting the ginseng fruit in steps 1 to 3 by steaming and drying step by step, and then fermenting into a strain.

<Test Example 2> Test for preventing damage to liver cells by alcohol

(1) Experiment of induction of liver toxicity by ethanol administration

The liver disease inhibitory activity of the fermented ginseng fruit and ginseng fruit extract prepared from Example 1 and Comparative Examples 1 to 5 was confirmed.

First, alcoholic liver disease was induced by oral administration of 25% ethanol to a 5 week-old male Balb / c mouse at a dose of 5 g / kg once a day for a total of 7 days. Mice that were orally administered 10 times a day with the amount of 2.0mg / mouse and 3.0mg / g in the amount of steamed ginseng fruit or ginseng fruit extract prepared in Example 1 and Comparative Examples 1 to 3 from 3 days before ethanol administration GOT / AST, GPT / ALT and LDH were quantified in the serum of the sample to confirm liver disease activity of the sample. At this time, the serum GPT / ALT, GOT / AST and LDH in the blood were quantified using serum samples on the first day after the ethanol administration was finished, and the results are shown in Table 3 and FIGS. 1 to 2 below.

GPT / ALT (U / L) GOT / AST (U / L) LDH (U / L) Control 30 ± 0.55 52 ± 1.31 330 ± 1.10 Experiment group (EtOH only) 52 ± 0.92 91 ± 1.27 480 ± 1.98 Example 1 32 ± 0.74 49 ± 0.18 360 ± 1.13 Comparative Example 1 49 ± 0.90 78 ± 0.16 420 ± 2.15 Comparative Example 2 44 ± 0.84 69 ± 0.14 405 ± 1.27 Comparative Example 3 40 ± 0.89 71 ± 0.28 404 ± 2.24 Comparative Example 4 40 ± 0.75 62 ± 0.15 390 ± 2.22 Comparative Example 5 36 ± 0.95 58 ± 0.15 360 ± 2.09

As a result, as shown in Table 3, AST / ALT in the blood elevated by administration of ethanol was compared to the liver disease inhibitory effect shown in 2 mg / mouse dose of ginseng fruit extract and steamed ginseng fruit extract of Comparative Examples 1 to 5, Example 1 It was confirmed that the effect of inhibiting liver disease at 2mg / mouse dose of fermented ginseng fruit was significantly improved.

In addition, Figures 1 to 2 are graphs by specifying the blood concentrations of GOT / AST and GPT / ALT, which are hepatotoxicity indices, for liver toxicity induction results by administration of fermented ginseng fruit of Example 1. As shown in FIGS. 1 and 2, it was confirmed that the blood concentrations of GOT / AST and GPT / ALT were all increased in the experimental group in which 25% ethanol was administered at 5 g / kg than the control group not administered with 25% ethanol. Accordingly, it can be confirmed that liver cells are damaged and toxicity is expressed by administering 25% ethanol. Quantitative determination of GOT / AST and GPT / ALT in serum of mice that were orally administered 10 times a day with the amount of 1,2, 3 and 4mg / mouse fermented ginseng fruit fermented in Example 1 from 3 days before ethanol administration By doing so, the activity of inhibiting liver disease in the sample was confirmed. As a result, the AST / ALT increased by ethanol administration showed liver disease suppressing effect at the 2 mg / mouse dose of the steamed ginseng fruit fermentation of Example 1.

(2) Analysis of antioxidant activity in liver tissue

In order to investigate how the fermented ginseng fruit fermented products or ginseng fruit extracts prepared in Examples 1 to 5 affect the expression of SOD (Superoxide dismutase), an enzyme related to the antioxidant activity present in the liver, in the liver tissue, The expression level was quantified. In the same manner as in the above experimental conditions, liver disease caused by alcohol was induced, and the measurement of SOD was analyzed by quantifying SOD in liver tissue by dissolving liver tissue on day 1 after ethanol administration was finished.

SOD (U / mL) Control 1.82 ± 0.12 Experiment group (EtOH only) 1.14 ± 0.09 Example 1 2.24 ± 0.12 Comparative Example 1 1.62 ± 0.15 Comparative Example 2 1.59 ± 0.18 Comparative Example 3 1.81 ± 0.19 Comparative Example 4 1.98 ± 0.13 Comparative Example 5 2.01 ± 0.13

As a result, as shown in Table 4 above, the ginseng fruit extract of Comparative Examples 1 to 5 and steamed ginseng fruit fermentation 2mg / mouse 2mg / mouse dose increased compared to the expression level of SOD prepared in Example 1 steamed ginseng fruit fermentation 2mg It was confirmed that the increase in the expression level of SOD in the / mouse dose was remarkably high.

In addition, as shown in Figure 3, the fermented ginseng ginseng fruit prepared in Example 1 increased from the 1.0 mg / mouse dose to the expression level of SOD, and a significant increase was observed from the 3 mg / mouse dose, which was effective in antioxidant activity. I could confirm that there is.

(3) liver and spleen weight measurement

In order to confirm the stability of the fermented ginseng fruit prepared in Example 1, changes in the spleen and liver weight of the experimental animals according to the administration were measured.

Liver weight (mg) Spleen weight (mg) Control 1482.82 ± 22.81 96.25 ± 1.22 Experiment group (EtOH only) 1263.93 ± 31.69 89.13 ± 1.01 Example 1 1283.55 ± 26.72 90.08 ± 1.39

As a result, as shown in Table 5, it was found that the long-term weight reduction by administration of the fermented ginseng ginseng prepared in Example 1, as well as the long-term weight in the treatment of ethanol did not have a significant effect. In addition, no significant change was observed in the weight gain rate during the test period.

<Test Example 3> Inflammatory factor suppression test

To confirm the anti-inflammatory activity effects of steamed ginseng fruit fermented products prepared from Examples 1 and 5 and Comparative Examples 1 to 5, steamed ginseng fruit fermented products prepared from Examples 1 and Comparative Examples 1 to 5, and The degree of inhibition of Nitric Oxide (NO) was measured by diluting the ginseng fruit extract to a concentration of 50 (µg / ml), respectively.

RAW 264.7 cell, a type of macrophage, was counted and dispensed equally to 1.0 x 10 ⁴ cells / well in a ²⁴ well plate using DMEM medium containing 1% penicillin / scraptomycin and 10% fetal bovineserum (FBS). , Cultured at 37 ° C. and 5% carbon dioxide for 24 hours.

After mixing the fermented ginseng fruit and ginseng fruit extracts prepared from Example 1 and Comparative Examples 1 to 5 with the medium for each concentration in a cell cultured for 24 hours, 1 ml was added to each well and added at 37 ° C. and 5% CO ₂ conditions. It was reacted in an incubator for 24 hours. At this time, LPS (lipo poly saccharide), an inflammatory factor that expresses NO, was treated together at a concentration of 1 μg / ml and reacted at 37 ° C. and 5% carbon dioxide for 24 hours. Then, after taking only the supernatant of each well separately, take 100 ml of the culture solution into a 96 well plate using a Nitric Oxide detection kit for each well, 50 μl of Griess reagent A (N-1-Naphthylethylenediamine (NEDHC)) and Griess reagent 50 μl of B (Sulfanilamide) was added and then reacted for 10 minutes, respectively. The absorbance was measured at 540 nm using an ELISA plate reader, and the results are shown in Table 6 below.

division NO production concentration (μM) Positive control 13.3 ± 0.16 Eumseong Control 51.8 ± 0.22 Example 1 11.1 ± 0.20 Comparative Example 1 39.0 ± 0.17 Comparative Example 2 38.2 ± 0.14 Comparative Example 3 19.9 ± 0.24 Comparative Example 4 14.6 ± 0.29 Comparative Example 5 15.5 ± 0.18

As a result, as shown in Table 6, it was confirmed that Example 1 has an effect of suppressing the production of Nitric oxide (NO) in a concentration-dependent manner than Comparative Examples 1 to 5.

Since the specific parts of the present invention have been described in detail above, it will be apparent to those skilled in the art that this specific technique is only a preferred embodiment, and the scope of the present invention is not limited thereby. will be. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (8)

Ginseng fruit is steamed at 80 to 90 ° C for 3 to 5 hours, and then dried at 50 to 60 ° C for 15 to 30 hours to prepare a primary steamed ginseng fruit;
Preparing the second steamed ginseng fruit by steaming the first steamed ginseng fruit at 80 to 90 ° C for 2 to 5 hours, and then drying at 50 to 60 ° C for 15 to 30 hours;
Preparing a third steamed ginseng fruit by drying the second steamed ginseng fruit at 80 to 90 ° C for 2 to 5 hours, and then drying at 50 to 60 ° C for 15 to 30 hours;
Extracting the third steamed ginseng fruit as an extraction solvent to obtain steamed ginseng fruit extract;
Fermentation of steamed ginseng fruit having liver damage prevention or improvement efficacy, comprising inoculating and fermenting Lactobacillus casei GFC 1 strain (Accession number: KCTC18333P) to the steamed ginseng fruit extract. Method of making water.


delete The method of claim 1, wherein the fermentation is prepared by adding 0.5 to 1.0 g of lactose to steamed ginseng fruit extract, and then to the medium, Lactobacillus casei GFC 1 strain (Accession No .: KCTC18333P). Method of producing a fermented ginseng fruit ginseng having the effect of preventing or improving liver damage, characterized by inoculating and culturing.
According to claim 1, wherein the fermented ginseng fruit fermentation product is ginsenoside Rg2 (ginsenoside Rg2), ginsenoside Rg3 (ginsenoside Rg3), ginsenoside Rh2 (ginsenoside Rh2), ginsenoside Rh3 (ginsenoside Rh3), ginseng It contains at least one active ingredient selected from the group consisting of ginsenoside Rh23, ginsenoside Rk1, ginsenoside Rg5, and ginsenoside F4. Method for producing fermented ginseng fruit with steam that prevents or improves liver damage.
According to claim 1, wherein the fermented ginseng fruit fermentation product is ginsenoside Rg2 (ginsenoside Rg2) 1.0 ~ 1.5mg / g, ginsenoside Rg3 (ginsenoside Rg3) 0.5 ~ 1.2 mg / g, ginsenoside Rh2 (ginsenoside Rh2) ) 0.3 to 0.7 mg / g, ginsenoside Rh3 (ginsenoside Rh3) 0.6 to 1.5 mg / g, ginsenoside Rh23 (ginsenoside Rh23) 0.1 to 0.5 mg / g, ginsenoside Rk1 (ginsenoside Rk1) 1.0 to 3.0 mg / g, ginsenoside Rg5 (ginsenoside Rg5) 1.0 to 4.0 mg / g, and ginsenoside F4 (ginsenoside F4) 1.0 to 3.0 mg / g. Method of manufacturing fermented ginseng fruit.
Ginsenoside Rg2 (ginsenoside Rg2) 1.0 to 1.5 mg / g, Ginsenoside Rg3 (ginsenoside Rg3) 0.5 to 1.2 mg / g, Ginsenoside Rh2 (ginsenoside Rh2) 0.3 to 0.7 mg / g, Ginsenoside Rh3 ( ginsenoside Rh3) 0.6 to 1.5 mg / g, ginsenoside Rh23 (ginsenoside Rh23) 0.1 to 0.5 mg / g, ginsenoside Rk1 (ginsenoside Rk1) 1.0 to 3.0 mg / g, ginsenoside Rg5 (ginsenoside Rg5) 1.0 to Steamed ginseng fruit fermentation product comprising 4.0 mg / g and ginsenoside F4 (1.0 to 3.0 mg / g) of active ingredient.
The fermentation product of steamed ginseng fruit according to claim 6, wherein the fermented product of steamed ginseng fruit has an effect on inhibiting liver damage and anti-inflammatory.
Health functional food for preventing or improving liver damage, including fermented ginseng fruit of claim 6.

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