KR20150143253A - Ginseng preparations containing abundant ginsenoside using cladosporium sp. and leuconostoc citreum, and method thereof - Google Patents

Abstract

The present invention relates to a producing method of a red ginseng formulation with an increased compound K content, and a cosmetic composition for skin whitening, comprising a red ginseng formulation produced by a producing method of the present invention as an active ingredient. The present invention is to co-culture Cladosporium sp. and Leuconostoc citreum strains by inoculating both Cladosporium sp. and Leuconostoc citreum strains. The red ginseng extract with an increased ginsenoside content than an existing ginsenoside is produced by using the producing method of the present invention and, specifically, the extract has effects of enhancing anti-cancer, treating cancer, reducing cancer drug toxicity, preventing cancer and a thrombotic disease, and whitening, due to increasing the compound K in ginsenosides when using the method of the present invention.

Description

TECHNICAL FIELD [0001] The present invention relates to a red ginseng preparation having increased ginsenoside content and a method for producing the red ginseng preparation using the clodosporium and leuconosporidiocyte strains. AND LEUCONOSTOC CITREUM, AND METHOD THEREOF}

The present invention relates to a method for preparing a red ginseng preparation having enhanced ginsenoside components and a cosmetic composition for skin whitening comprising the red ginseng preparation as an active ingredient.

Ginseng is a botanical plant belonging to Panax ginseng (Araliaceae). It is a medicinal herb that has long been known to have efficacy and efficacy in the Orient.

Recently, according to the scientific progress of the research on the pharmacological effect of ginseng, the specific ginsenoside component is separated from the ginseng, and its chemical structure is revealed. Meanwhile, as the efficacy and the pharmacological effect of ginseng are known, Techniques related to processing and manufacturing methods of ginseng for efficiently and economically manufacturing a functional product reinforced with a specific component of saponin by developing different and different effects have been developed and these technologies are attracting attention.

There is a known method of producing black ginseng by Gujung Gupo with a classical method in relation to the technique of strengthening saponin of a specific ingredient contained in ginseng. There is a method of heat treatment by treatment of red ginseng extract and extract at high temperature and high pressure, There is a disadvantage in that the time required for the manufacturing process is too long. In the high-temperature and high-pressure treatment method, the time required for the manufacturing process can be shortened, but the cost due to the energy consumption due to high temperature and high pressure increases and the process stability is low .

The method using microorganisms (lactic acid bacteria), the method of hydrolysis with acids and enzymes, and the like, have advantages in that they can selectively enhance specific saponins, but it is difficult to select and cultivate lactic acid bacteria , The process is complicated and difficult to manage. In the hydrolysis method, the selectivity and conversion efficiency of the specific saponin differ depending on the kind of the acid and the kind of the enzyme. Generally, the selectivity and conversion efficiency But it has a merit that the process is simple and can remarkably lower the production cost. Especially, if the optimum condition of the selection of the acid or the enzyme suitable for the specific saponin conversion and the process is developed, the industrial competitiveness is the greatest.

Korean Patent Laid-Open Publication No. 10-2010-0098208 discloses a method for enhancing the saponin of a specific component contained in ginseng, wherein the content of Rg3 and Rh2 is increased. However, this method has a complicated process There are disadvantages.

In order to overcome the problems of the prior art, the inventors of the present invention have found that a red ginseng preparation capable of increasing Compound K content, which is a minor amount of ginsenoside, by using Cladosporium sp. Strain and Leuconostocetium sp. Strain The inventors of the present invention have completed the present invention by developing a technique for producing a red ginseng preparation and confirming that the whitening effect of the red ginseng preparation is excellent.

Accordingly, it is an object of the present invention to provide a method for preparing a red ginseng preparation using a clodospholium strain and a leuconosporotium citrate strain.

It is also an object of the present invention to provide a cosmetic composition for whitening skin, which contains, as an active ingredient, a red ginseng preparation having an increased ginsenoside content.

In order to achieve the above object, the present invention provides a method for preparing a red ginseng preparation having an increased ginsenoside content, comprising the steps of:

a) adding red ginseng extract to a nutrient medium and sterilizing the same;

b) Inoculating the medium with Cladosporium sp. and liquid culture to obtain a primary culture;

c) Second inoculation of leuconostoc citreum into the primary culture and liquid culture to obtain a secondary culture; And

d) adding butanol (BuOH) to the secondary culture and sonication to isolate the butanol fraction.

In another embodiment of the present invention, the ginsenoside is a compound K.

In another embodiment of the present invention, the red ginseng extract is added in a concentration of 0.01 to 5% by volume.

In still another embodiment of the present invention, the sterilization is performed at 100 to 150 ° C in the step a).

In another embodiment of the present invention, the sterilization is performed for 10 to 30 minutes in the step a).

In another embodiment of the present invention, the cladosporium is added in a concentration of 10 to 30% (v / v).

In another embodiment of the present invention, the leucono-Stokelium is added in a concentration of 10 to 30% (v / v).

In another embodiment of the present invention, the liquid culture in the step b) is performed for 1 to 5 days.

In another embodiment of the present invention, the liquid culture in the step c) is performed for 1 to 3 days.

In yet another embodiment of the present invention, the butanol is added in a ratio of 3: 7 to 7: 3 relative to the secondary culture.

In another embodiment of the present invention, the ultrasonic treatment is performed for 10 minutes to 120 minutes.

The present invention also provides a cosmetic composition for skin whitening comprising an active ingredient of a red ginseng preparation having an increased ginsenoside content prepared by the above production method.

In one embodiment of the present invention, the ginsenoside is a compound K.

The present invention can produce a red ginseng preparation containing a high content of trace ginsenosides of red ginseng extract by inoculating cladosporium, followed by second inoculation with leuconosporidiocyte and co-culture. This can solve the disadvantage that the production yield is lowered as the red ginseng goes through the separation and purification process, and accordingly the unit price is increased. Despite the relatively low unit cost of the red ginseng preparations, the red ginseng preparations thus prepared contain a high content of compound k, and are excellent in anti-wrinkle, antioxidant and whitening effects, resulting in a variety of effects In the functional cosmetics market, which has a high growth rate by improving the disadvantages of existing cosmetic raw materials that have problems, it has strengthened the compound k which is excellent in raw material and anti-cancer, cancer treatment, reduction of anticancer drug toxicity, cancer prevention, immunity enhancement and prevention of thrombosis It can be a raw material or a product of food having functionality.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic view showing a step of preparing a red ginseng formulation according to the present invention.
FIG. 2 is a graph showing the results of a comparison between the red ginseng extract inoculated with neither cladosporium nor leuconosporidiocytoma, the red ginseng extract only inoculated with cladosporium, and the red ginseng extract inoculated with both clodosporium and leucono stock ≪ tb >< TABLE >
FIG. 3 is a graph showing the results of a comparison between the red ginseng extract inoculated with neither clodospholium nor leuconosporous citrate, the red ginseng extract only inoculated with cladosporium, and the red ginseng extract inoculated with both clodospholium and leucono stock Lt; RTI ID = 0.0 > HPLC-ELSD < / RTI >
FIG. 4 is a graph showing the effect of the red ginseng extract, which is not inoculated with any of the cadosporium and leuconoistocardium bacteria, the red ginseng extract inoculated only with cladosporium or leuconosporidiocytes, and the cladosporium and leucono stock Of the present invention is a graph showing the inhibition of tyrosinase activity of the red ginseng extract inoculated.

The present inventors prepared a red ginseng preparation by inoculating Cladosporium sp. And leuconostoc citreum strains to produce a red ginseng preparation having a significantly increased ginsenoside content, The present inventors have completed the present invention by developing a cosmetic composition for skin whitening comprising an active ingredient of a red ginseng preparation produced by the method of the present invention.

That is, the object of the present invention is to provide a method for preparing a red ginseng preparation containing a large amount of ginsenoside compound K, which comprises the following steps:

a) adding red ginseng extract to a nutrient medium and sterilizing the same;

b) Inoculating the medium with Cladosporium sp. and liquid culture to obtain a primary culture;

c) Second inoculation of leuconostoc citreum into the primary culture and liquid culture to obtain a secondary culture; And

d) adding butanol (BuOH) to the secondary culture and sonication to isolate the butanol fraction.

The red ginseng preparations according to the present invention are characterized in that the ginsenoside component is remarkably increased, and the content of the compound k in the ginsenoside components is remarkably increased.

The present inventors compared the red ginseng extract inoculated with the strain used in the present invention with the red ginseng extract alone inoculated with the cladosporium alone and the red ginseng extract inoculated with the clodospholium and leuconostocetium inoculum As a result, Compound K of the red ginseng extract prepared by the method of the present invention was remarkably increased (see Example 2 and Example 3), and it was confirmed that the whitening effect was remarkably increased (see Example 4).

Therefore, by using the method of the present invention, a functional product in which the content of ginsenoside is remarkably increased can be provided.

The steps of the invention are described in more detail below.

In the step a), the red ginseng extract is added to the nutrient medium and sterilized. The red ginseng extract may be any substance as long as it is extracted from red ginseng as a material.

In one embodiment of the present invention, the red ginseng extract may be added to the nutrient medium at a concentration of 0.01 to 5% by volume, preferably 0.1 to 3% by volume, more preferably, At 1 vol% concentration, it can have a good ginsenoside increase effect.

In another embodiment of the present invention, the sterilization may be performed at 100 to 150 ° C for 10 minutes to 30 minutes, more preferably 110 to 130 ° C, And the time is not limited to the above range.

The step b) is a step of firstly inoculating cladosporium in a medium and culturing the medium to obtain a primary culture. The clodosporium culture broth is added by homogenization and can be inoculated with 10-30% (v / v), preferably 15-25% (v / v), and most preferably Can be inoculated 20% (v / v). If the cadosporium is used in an amount less than the above range, the effect of increasing ginsenoside may not be obtained. If cadosporium is used in the above range, impurities may be generated in the subsequent concentration process.

In another embodiment of the present invention, the liquid culture can be performed for 1 to 5 days, and most preferably for 3 days, showing excellent ginsenoside-increasing effect.

The step c) is a step of inoculating the culture medium with leuconosporidiocyte twice and liquid culture to obtain a secondary culture solution. The leucomoniostositium culture medium is added by homogenization, and can be inoculated at 10 to 30% (v / v), preferably at 15 to 25% (v / v) Can be inoculated with 20% (v / v). Use of less than the above range of Ryukono Stock Potassium may not attain the effect of increasing ginsenoside, and if more Ryukonosutoritium than the above range is used, there is a possibility that impurities are generated in the subsequent concentration process.

In another embodiment of the present invention, the liquid culture can be performed for 1 to 3 days, and most preferably for 2 days, showing excellent ginsenoside increase effect.

In step d) of the present invention, butanol is added to the secondary culture and the butanol fraction is separated by ultrasonication. The amount of the butanol may be 3: 7 to 7: 3 relative to the culture medium, May be added at a ratio of 5: 5, but it is not limited thereto, so long as the amount of the culture can be concentrated.

The ultrasonic treatment may be performed for 10 minutes to 120 minutes, preferably for 1 hour, followed by concentration under reduced pressure.

In addition, the cosmetic composition of the present invention contains an active ingredient of a red ginseng preparation having increased ginsenoside content prepared by the method of the present invention as an active ingredient and can be applied to various creams such as skin, lotion, essence, nutritional cream, massage cream and cleansing cream , Basic cosmetics such as cleansing foams, soaps, ultraviolet screening agents and packs, and color cosmetics including powders, foundations and makeup bases. In the cosmetic composition of each formulation, the ingredients other than the red ginseng preparations according to the present invention can be mixed and selected without difficulty by those skilled in the art depending on the formulations of the other cosmetic preparations or the purpose of use. To facilitate the distribution of the composition there may be included various excipients which are acceptable for the cosmetic composition, acting as a diluent, dispersant or carrier for the active ingredient in the composition. Examples of excipients other than water or excipients containing water include emollients, solvents, wetting agents, preservatives, emollients, surfactants, organic and inorganic pigments, ultraviolet absorbers, preservatives, bactericides, antioxidants, pH adjusters , Fragrances, thickeners or powders, and the like. However, the present invention is not limited thereto.

In the cosmetic composition of the present invention, the red ginseng preparation having an increased ginsenoside content prepared by the preparation method is preferably contained in an amount of 0.01 to 5% by weight based on the total weight of the cosmetic composition, but is not limited thereto.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the following examples.

[Example]

Example 1. Preparation of red ginseng extract

The dried red ginseng extract (KRG) was added to nutrient broth at a concentration of 1% and sterilized at 121 ° C for 20 minutes.

Cladosporium sp. The culture broth was homogenized and inoculated 20% (v / v) in KRG supplemented nutrient medium and cultured at 24 ℃ for 3 days. After this, Leuconostoc citreum was inoculated with 20% (v / v) and co-cultured at 30 ° C for 2 days.

BuOH (butanol) was added 1: 1 to the incubated flask and sonicated for 1 hour. The BuOH layer was separated and concentrated under reduced pressure to prepare red ginseng extract.

Example 2. Confirmation of ginsenoside content

To confirm the ginsenoside content of the red ginseng extract of the present invention, the concentrated BuOH fraction was dissolved in MeOH, and then subjected to TLC analysis by 0.2 μm syringe filter.

The TLC (TLC silica gel 60 F254) used for the analysis of red ginseng extract was used, and ginsenoside TLC analysis was performed by dissolving butanol fractions on a silica gel TLC plate and washing with chloroform: ethanol (8: 1 ) For 40 minutes, followed by spraying with 10% H ₂ SO ₄ solution and heating at 100 ° C for 5 minutes. The ginsenosides patterns in the samples were compared and analyzed.

As a result, as shown in FIG. 2, the compound K content was remarkably increased in the red ginseng extract obtained by inoculation with the clodospholium and the leuconosporous citrate.

Example 3. Analysis of ginsenoside content of fermented red ginseng inoculated with strain

The ginsenoside content of the red ginseng extract co-cultured inoculated with the cladosporium and leuconosporidiocyte of the present invention was compared with the red ginseng extract inoculated with neither cladosporium nor leuconospora citrate Analysis. In this example, HPLC (agilent) as the analyzer for the analysis and ELSD (agilent) as the vaporization light scattering detector were used for HPLC analysis.

In this example, the mobile phase was analyzed at room temperature using acetonitrile and water at a flow rate of 1 ml / min and a sample injection volume of 10 μl.

As a result, as shown in Fig. 3, compared to the red ginseng extract in which none of the cadosporium and leuconosporidiocyte were inoculated, the present invention in which the cadosporium and the leuconosporidiocyte were inoculated and co- The compound K content of the red ginseng extract was significantly increased.

Example 4. Comparison of whitening efficacy with single strain fermented red ginseng

Experiments were conducted to compare the whitening efficacy of KRG obtained by inoculating KRG, cladosporium, and leuconosporidiocyte only inoculated with the cladosporium of the present invention.

110 units of tyrosinase and 1.5 mM of DOPA (dihydroxyphenylalanine) were added to 50 mM sodium phosphate buffer and reacted at 37 ° C for 10 minutes and then measured at UV 450 nm. As a positive control, albutin was used to inhibit the activity of tyrosinase, a melanin pigment synthetase.

As a result, as shown in Fig. 4, the KRG obtained by inoculation with the clodospholium and leuconostocetium in the symbiotic culture showed the tyrosinase activity to be 25 [mu] g compared with the KRG obtained by single inoculation with clodospholium or leuconosporidiocyte alone % Inhibition and similar efficacy to albutin, a positive control.

From the above results, it is expected that KRG obtained by inoculating cladosporium and leuconosporidiocytoma and co-culturing it can significantly increase whitening efficacy.

It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive.

Claims (13)

A method for preparing a red ginseng preparation having increased ginsenoside content comprising the steps of:
a) adding red ginseng extract to a nutrient medium and sterilizing the same;
b) Inoculating the medium with Cladosporium sp. and liquid culture to obtain a primary culture;
c) Second inoculation of leuconostoc citreum into the primary culture and liquid culture to obtain a secondary culture; And
d) adding butanol (BuOH) to the secondary culture and sonication to isolate the butanol fraction.
The method according to claim 1,
Wherein the ginsenosides are compound K. < RTI ID = 0.0 > 15. < / RTI >
The method according to claim 1,
Wherein the red ginseng extract is added in a concentration of 0.01 to 5% by volume.
The method according to claim 1,
Wherein the sterilization in step a) is carried out at 100 to 150 < 0 > C.
The method according to claim 1,
Wherein the sterilization is performed for 10 to 30 minutes in the step a).
The method according to claim 1,
Characterized in that the cladosporium is added in a concentration of 10 to 30% (v / v).
The method according to claim 1,
Characterized in that the leucono-Stokelium is added in a concentration of 10 to 30% (v / v).
The method according to claim 1,
Wherein the liquid culture in step b) is performed for 1 to 5 days.
The method according to claim 1,
Wherein the liquid culture in step c) is performed for 1 to 3 days.
The method according to claim 1,
Characterized in that the butanol is added in a ratio of 3: 7 to 7: 3 relative to the secondary culture.
The method according to claim 1,
Wherein the ultrasonic treatment is performed for 10 to 120 minutes.
12. A cosmetic composition for skin whitening comprising an active ingredient of a red ginseng preparation having an increased ginsenoside content produced by the production method according to any one of claims 1 to 11.
13. The method of claim 12,
Wherein the ginsenoside is a compound K. < RTI ID = 0.0 > 18. < / RTI >

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