KR100954851B1 - Preparation method of fermented ginseng concentrate or powder having high concentration of IH-901 - Google Patents

Abstract

The present invention relates to a method for preparing fermented ginseng concentrate or fermented ginseng powder containing high concentration of IH-901 (20-O-β-D-glucopyranosyl-20 (S) -protopanaxadiol). Specifically, the present invention relates to a method for preparing fermented ginseng concentrate or fermented ginseng powder containing a large amount of IH-901 by fermenting ginseng extract by adding pectinase and beta galactosidase to ginseng extract or ginseng solution. Fermented ginseng concentrate or fermented ginseng powder production method of the present invention is a method that can easily prepare a fermented ginseng concentrate or fermented ginseng powder containing a high concentration of IH-901, and also the fermented ginseng concentrate prepared by the present invention or Fermented ginseng powder contains high concentration of IH-901 and can be effectively used as a functional food composition.

IH-901, Fermented Ginseng Concentrate, Fermented Ginseng Powder

Description

Preparation method of fermented ginseng concentrate or fermented ginseng powder containing high concentration of IH-901 {Preparation method of fermented ginseng concentrate or powder having high concentration of IH-901}

The present invention relates to a method for preparing fermented ginseng concentrate or fermented ginseng powder containing high concentration of IH-901 (20-O-β-D-glucopyranosyl-20 (S) -protopanaxadiol). Specifically, the present invention relates to a method for preparing fermented ginseng concentrate or fermented ginseng powder containing a large amount of IH-901 by fermenting ginseng extract by adding pectinase and beta galactosidase to ginseng extract or ginseng solution.

Ginseng (the root of Panax ginseng CA Meyer.Araliaceae) is one of the traditional medicines used in the treatment of various diseases in Asian countries such as China, Korea and Japan. In particular, ginsenoside (ginseng saponin, saponin), the main active ingredient of ginseng, has anti-aging, anti-inflammatory, antioxidant activity in the central nervous system and cardiovascular and immune system (Wu JY, et al., J. Immunol. , 148: 1519 -25, 1992;.. Lee FC , Facts about ginseng, the elixir of life Hollyn International . New Jersey, 1992; Huang KC., The pharmacology of Chinese herbs . CRC Press . Florida, 1999), antidiabetic activity (Chang HM., Pharmacology and application of Chinese material medica. Vol 1. World Scientific . Singapore, 1986) and anti-tumor activity (Sato K, et al., Biol. Pharm. Bull . 17: 635-9, 1994; Mochizuki M, et al., Biol. Pharm. Bull. 18: 1197-1202, 1995).

On the other hand, the ginsenosides are metabolized by human intestinal microorganisms after ingestion and their metabolites are also known to have various physiological activities (Karikura M, et al., Chem. Pharm. Bull , 39: 2357-61, 1991 Kanaoda M, et al., J. Tradit.Med . 11: 241-5, 1994; Akao T, et al., Biol. Pharm. Bull. 21: 245-9, 1998). For example, Rb1, Rb2, and Rc, the protoparnaxadiol-based ginsenosides, are identified by IH-901 (20-O-β-D-glucopyranosyl-20 (S) -prototype by human intestinal microorganisms. Panaxanadiol) (Hasegawa H, et al., Planta Medica 63: 463-40, 1997; Tawab MA, et al., Drug Metab. Dispos . 31: 1065-71, 2003) All ginsenosides Re and Rg1 are metabolized to ginsenoside Rh1 or ginsenoside F1 (Hasegawa H, et al., Planta Medica 62: 453-7, 1996; Tawab MA, et al., Drug Metab. . 31: 1065-71, 2003), the metabolite, IH-901, Rh1, and F1 is to exhibit the various physiological activities.

In particular, the IH-901 is mainly anti-diabetic (Yun Suk Choi et.al., J Ginseng Res . 31 (2): 79-85, 2007) and are known to have an immunopotentiating effect and also to induce anti-metastatic or anti-cancer effects by preventing tumor invasion or preventing chromosomal alterations and tumor formation (Wakabayashi C, et al., Oncol. Res. 9: 411-7, 1998; Lee SJ, et al., Cancer Lett. 144: 39-43, 1999).

For this reason, efforts have been extensively attempted to convert the ginsenosides to the metabolites. Some researchers have attempted to produce genuine prosapogenin or sapogenin by chemical synthesis, weak acid hydrolysis, alkali decomposition (Han BH, et al., Planta Medica 44: 146-9, 1982; Chen Y, et al., Chem. Pharm. Bull. 35: 1653-5, 1987; Elyakov GB, et al., Synthesis of the ginseng glycosides and their analogs.Proc . 6th Int. Ginseng Symp. Seoul 74-83, 1993). However, since these methods cause various side reactions such as epimerization, hydration, hydroxylation, etc., recently, enzymes (Ko SR, et al., Biosci. Biotechnol. Biochem. 64: 2739) -43, 2000; Ko SR, et al., Planta Med. 69: 285-6, 2003) and enterobacteria (Hasegawa H, et al., Planta Medica 63: 463-40, 1997; Bae EA, et al. , J. Microbial , Biotechnol. 13: 9-14, 2003), and various methods for converting ginsenosides under mild conditions have been studied.

In particular, methods have been studied using enzymes to prepare the IH-901 from the ginsenoside. Specifically, a method of preparing IH-901 by hydrolyzing a diol-based ginsenoside with naringinase enzyme or the die A method for preparing IH-901 has been developed by orally administering all-type ginsenosides to mice and then digesting them in the large intestine. However, the production method was not only extremely low production yield of IH-901 but also various secondary metabolites were produced to produce high purity IH-901 (Karikura et al., Chem. Pharm. Bull. 38: 2859 , 1990).

Accordingly, a method of preparing IH-901 by reacting beta galactosidase enzyme with ginsenosides Rb1, Rb2, Rc, Rd or a mixture thereof (Patent Application No. 2002-32039) was developed. A method for preparing IH-901 by reacting an agent and pectinase with purified ginsenosides (Patent Application No. 2001-67964) has been developed, and the enzyme ginsenosides with cerulases or beta galactosidase enzymes have been developed. A method of preparing IH-901 by reacting with Rb1, Rb2, Rc, Rd or mixtures thereof (Patent Application No. 2000-43170) has been developed.

The manufacturing method is used to separate each ginsenoside rather than the ginseng concentrate as a main raw material is very complicated, the method is not suitable for the production of fermented ginseng concentrate containing a large amount of IH-901. Therefore, the fermented product prepared by the above method is difficult to use directly as a food, and there is a problem that the production amount of IH-901 is small and it is difficult to expect its efficacy.

The present inventors overcome the above problems and while studying the method for producing a fermented ginseng concentrate or fermented ginseng powder containing a high concentration of IH-901, mixing the pectinase and beta galactosidase under specific conditions The present invention was completed by confirming that fermented ginseng concentrate or fermented ginseng powder containing high concentration of IH-901 by fermentation under specific conditions was added to ginseng extract or ginseng solution.

Accordingly, an object of the present invention is to provide a method for preparing fermented ginseng concentrate or fermented ginseng powder containing high concentration of IH-901.

In order to solve the above problems, the present invention is prepared by adding water or an organic solvent to the ginseng to prepare a ginseng extract, mixed with the pectinase and beta galactosidase to the ginseng extract, fermented the ginseng extract Thereafter, it provides a method for producing a fermented ginseng concentrate, comprising the step of concentrating.

In addition, the present invention is prepared by adding water to the ginseng to prepare a ginseng solution, mixed with the pectinase and beta galactosidase to the ginseng solution and then fermented, then extracted the fermented ginseng solution using water or an organic solvent Thereafter, it provides a method for producing a fermented ginseng concentrate, comprising the step of concentrating.

The present invention also provides a method for producing a fermented ginseng powder, characterized in that the dried fermented ginseng concentrate is dried under reduced pressure or freeze-dried.

In another aspect, the present invention provides a functional food composition comprising the fermented ginseng concentrate or fermented ginseng powder prepared as an active ingredient.

Hereinafter, the present invention will be described in detail.

In the method of preparing the fermented ginseng concentrate of the present invention, the ginseng extract prepared by adding water or an organic solvent to ginseng is prepared, and the pectinase and beta galactosidase are prepared in the ginseng extract prepared in step (a). After mixing and adding (b) and fermenting the ginseng extract after the step (b), characterized in that it comprises a step (c) of concentration.

Hereinafter, the method of preparing the fermented ginseng concentrate of the present invention will be described in detail step by step.

In the step (a) ginseng is not limited to this, but preferably selected from the group consisting of the ginseng, white ginseng, ginseng, dried ginseng, red ginseng, Taegeuk ginseng, extracts thereof, powders thereof, and combinations thereof. In one embodiment of the ginseng, white ginseng, ginseng and ginseng powder was used (see <Example 1>).

In the step (a), the organic solvent is not limited thereto, but preferably refers to one selected from the group consisting of acetonitrile, dioxane, dimethyl sulfoxide, methanol, ethanol, 1-propanol, 2-propanol, and combinations thereof. In one embodiment of the present invention, alcohol (ethanol) was used (see <Example 1>).

Preparing ginseng extract in the step (a) refers to the preparation of the ginseng extract by the method known to those skilled in the art by adding water or the organic solvent to the ginseng, wherein the water and the organic solvent is used by appropriately mixing It is preferable. The mixing and use of the organic solvent is because the solubility of the intermediate reactants produced by the enzymatic reaction can be increased to finally improve the content of IH-901 in the fermented ginseng concentrate or the fermented ginseng powder. Therefore, in one embodiment of the present invention, ginseng extract was prepared by mixing water and ethanol at a ratio of 1: 1 and adding 500% to the total substrate (see <Example 1>).

Adding the pectinase and beta galactosidase to the ginseng extract prepared by the step (a) in the step (b) is a mixture of pectinase and beta galactosidase in a ratio of 1: 1 to 100, and This refers to adding 1 to 50% by weight relative to the total weight of the ginseng extract prepared in step (a). In particular, when the pectinase or beta galactosidase is added alone, the content of IH-901 in the fermented ginseng concentrate is relatively lower than the case where the pectinase and beta galactosidase are mixed and added as described above. It confirmed (see <Experimental example 1>). Therefore, it is one of the features of the present invention to add the pectinase and beta galactosidase by mixing as described above.

In particular, the present inventors can increase the IH-901 content of the fermented ginseng concentrate or fermented ginseng powder by adding the pectinase and the beta galactosidase to the ginseng extract in the above amount through the numerous experiments. Revealed.

Fermenting the ginseng extract from the step (c) in the step (b) is added to the ginseng extract by mixing the pectinase and beta galactosidase as described above, at pH 3 ~ 8 and 10 ~ 70 ℃ It means fermentation for 1 ~ 72 hours. The pH is not limited as long as the pectinase and beta galactosidase enzymes maintain activity, but may be preferably pH 3-8, and most preferably pH 4-6. The pH is not limited so long as it does not lower the activity of the enzyme, but may be adjusted by adding phosphate, citric acid, sodium citrate buffer solution to the ginseng extract. In addition, the reaction temperature is not limited as long as the pectinase and beta galactosidase enzyme maintains activity, but preferably 10 to 70 ° C, and most preferably 50 ° C. In addition, the reaction time is not limited as long as the enzyme maintains activity, but may be preferably 1 to 72 hours, and most preferably 48 hours. In one embodiment of the present invention, after adding the citric acid to the ginseng extract prepared in step (a) to adjust the pH to 3 ~ 4, the pectinase and beta galactosidase is added to the mixture, at 50 ℃ Fermentation was carried out for 48 or 72 hours (see <Example 1>).

Concentrating the ginseng extract fermented in the step (c) is not limited to this, but the concentrated fermented ginseng extract as it is or can be concentrated by taking only the supernatant by centrifugation, it can be carried out using a vacuum decompression concentrator. . In one embodiment of the present invention, the ginseng extract fermented in step (c) was concentrated under reduced pressure at 80 ℃ (see <Example 1>).

In addition, the method for preparing a fermented ginseng concentrate of the present invention is prepared by adding water to ginseng to prepare a ginseng solution, and adding fermented ginseng solution to pectinase and beta galactosidase, and then fermenting the ginseng solution to water or organic matter. After extraction using a solvent, characterized in that it comprises the step of concentrating. More specifically, ginseng solution was prepared by adding water to ginseng according to a conventional method, followed by fermentation by adding pectinase and beta galactosidase to the prepared ginseng solution under the same conditions as described above. Ginseng solution is extracted under the same conditions as described above using water or an organic solvent, characterized in that it comprises the step of concentrating.

The method for preparing fermented ginseng powder of the present invention is characterized in that the fermented ginseng concentrate prepared as described above is dried under reduced pressure or lyophilized. It may be carried out using a hot air dryer or a freeze dryer which is commonly used for drying under reduced pressure or freeze drying.

Functional food composition of the present invention is characterized by containing the fermented ginseng concentrate or fermented ginseng powder according to the present invention as an active ingredient. The fermented ginseng concentrate or fermented ginseng powder according to the present invention contains a large amount of IH-901 known to have excellent immunity enhancement, antidiabetic and anticancer effects, and the functional food composition of the present invention containing it as an active ingredient also enhances immunity, It is obvious that the antidiabetic and anticancer effects are excellent.

The functional food composition of the present invention can be prepared by adding fermented ginseng concentrate or fermented ginseng powder or a mixture thereof according to the present invention as it is or by processing according to methods known to those skilled in the art, and additionally adding other food ingredients. The added amount of the fermented ginseng concentrate or fermented ginseng powder according to the present invention can be appropriately determined by those skilled in the art according to the purpose of use of the functional food composition. In general, the fermented ginseng concentrate or fermented ginseng powder of the present invention is preferably contained in 0.1 to 100% by weight based on the total weight of the functional food composition. However, in the case of long-term intake for the purpose of health control, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

The food ingredients that may be added to the functional food composition of the present invention may vary depending on the type of functional food, and are not limited thereto as long as they do not cause side effects when mixed with fermented ginseng concentrate or fermented ginseng powder according to the present invention.

The functional food composition of the present invention can be used in the form of soft or hard capsules, tablets, granules, drops and tea bags filled by dry powdering according to a conventional method, and foods for immune enhancement, antidiabetic and anticancer effects. It can be used in various ways. The food to which the composition can be added is not limited thereto, and may be, for example, various foods, beverages, gums, candy, confectionery, tea, vitamin complexes, health functional foods, and the like.

As described above, the method for preparing fermented ginseng concentrate or fermented ginseng powder according to the present invention is added directly to the ginseng extract or ginseng solution by fermentation under specific conditions by adding fermented pectinase and beta galactosidase under specific conditions. It is a method to easily prepare fermented ginseng concentrate or fermented ginseng powder containing IH-901, and the fermented ginseng concentrate or fermented ginseng powder prepared by the present invention contains high concentration of IH-901 and is a functional food. It can be effectively used as a composition.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

<Example 1>

Preparation of Fermented Ginseng Concentrate according to the Present Invention

<1-1> Preparation of Fermented Ginseng Concentrate Using Rice and White Ginseng

Add 100 ml of water to 7 g of ginseng extract extracted 4 times each at 80 ℃ for 3 hours with 5 g of rice and white ginseng mixed with water and ethanol (1: 1 mixing ratio, 500% of raw materials), and then add pectinase and Beta galactosidase was added by 0.3 and 1g, respectively, and reacted for 48 hours at 50 ° C., pH 4-5, and then filtered and concentrated at 80 ° C. to prepare 6.3 g of the fermented ginseng concentrate according to the present invention.

<1-2> Preparation of Fermented Ginseng Concentrate Using Rice Ginseng

After extracting 10 g of ginseng with 100 ml of water, 7 g of the extract was adjusted to pH 3-4 with citric acid, followed by 0.3 and 1 g of pectinase and beta galactosidase, respectively, and reacted at 50 ° C. for 48 hours. Let This was filtered and concentrated to prepare 6.5 g of the fermented ginseng concentrate (I) according to the present invention.

In addition, after adjusting the extract 7.2g obtained by extracting 10g of ginseng with the same mixture of water and ethanol as in <Example 1-1> to pH 3-4 with citric acid, the reaction, filtration and concentration in the same manner as described above 6.8 g of fermented ginseng concentrate (II) according to the present invention was prepared.

<1-3> Preparation of Fermented Ginseng Concentrate Using White Ginseng

After concentrating 7 g of 540 g of the extract obtained by extracting 1 kg of white ginseng with 5 L of ethanol and suspending it again with water, 0.3 mg and 1 g of pectinase and beta galactosidase were added thereto, respectively. In the same manner as in the reaction, filtration and concentration was prepared 5.2 g of fermented ginseng concentrate according to the present invention.

<1-4> Preparation of Fermented Ginseng Concentrate Using Fresh Ginseng

Grind fine ginseng 6g and suspend the sterilized with water and adjust to pH 3 ~ 4.5 with citric acid, 0.3, 1g of pectinase and beta galactosidase, respectively, reacted at 50 ℃ for 72 hours Extracted 4 times at 80 ° C. for 4 hours with the same mixture of water and ethanol as in <Example 1-1>, filtered and concentrated to prepare 3.5 g of the fermented ginseng concentrate according to the present invention.

<1-5> Preparation of Fermented Ginseng Concentrate Using Ginseng Powder

7 g of ginseng powder was suspended in water, and then adjusted to pH 3 to 4.5 with citric acid. The reaction and extraction were carried out in the same manner as in <Example 1-4>, followed by filtration and concentration of 3.7 g of the fermented ginseng concentrate according to the present invention. Was prepared.

Comparative Example 1

Preparation of Fermented Ginseng Concentrate Using Pectinase Only

100 g of water was added to 7 g of ginseng extract prepared in the same manner as in <Example 1-1>, and 1.3 g of pectinase was added thereto. each Put the reaction and concentration under reduced pressure in the same manner as in <Example 1-1> to prepare a fermented ginseng concentrate 5.6g.

Comparative Example 2

Preparation of Fermented Ginseng Concentrate Using Only Beta-galactosidase

100 g of water was added to 7 g of the ginseng extract prepared in the same manner as in <Example 1-1>, and then 1.3 g of beta galactosidase was added thereto. each Put the reaction and concentration under reduced pressure in the same manner as in <Example 1-1> to prepare a fermented ginseng concentrate 5.4g.

Experimental Example 1

IH-901 content of fermented ginseng concentrate according to the present invention

Analyzing IH-901 content (20-O-β-D-glucopyranosyl-20 (S) -protopanaxadiol) of the fermented ginseng concentrate prepared according to the present invention prepared in Example 1-1. 1 g of the fermented ginseng concentrate was extracted three times with 60 ml of saturated butanol, and the concentrated butanol fraction was dissolved in 120 mg of methanol, and the content of IH-901 was analyzed by high pressure liquid chromatography. The result is shown in FIG.

In addition, IH-901 content of the fermented ginseng concentrate prepared in <Comparative Example 1> or <Comparative Example 2> was analyzed in the same manner as above.

Meanwhile, the content of IH-901 was also analyzed in the same manner as above for the ginseng extract prepared in <Example 1-1>.

All the above analysis results were compared in Table 1 below.

IH-901 content of fermented ginseng concentrate according to the present invention  ingredient Content by Ingredient Ginseng Extract of Example 1-1 Fermented Ginseng Concentrate of <Example 1-1> Fermented Ginseng Concentrate of <Comparative Example 1> Fermented Ginseng Concentrate of <Comparative Example 2> Ginsenoside (Rb1) 17.6mg / g 1.2mg / g 6.3 mg / g 3.2mg / g Ginsenoside (Rb2) 14.6mg / g 1.1mg / g 7.1mg / g 5.7 mg / g Ginsenoside (Rc) 18.9 mg / g 0.1mg / g 9.3 mg / g 6.3 mg / g Ginsenoside (Rd) 7.9 mg / g 1.4 mg / g 2.2 mg / g 4.2mg / g IH-901 0mg / g 36mg / g 24mg / g 10mg / g

As shown in Table 1, the fermented ginseng concentrate according to the present invention not only contains a large amount of IH-901 which is not present in the ginseng extract, but also contains 3.6 times as much as the beta galactosidase. , It was found that it contained 1.5 times as much as the administration of only pectinase.

<Example 2>

Preparation of Fermented Ginseng Powder According to the Present Invention

<2-1> Preparation of Fermented Ginseng Powder Using Rice and White Ginseng

100 ml of water was added to 7 g of the ginseng extract prepared in the same manner as in <Example 1-1>, followed by 0.3 and 1 g of pectinase and beta galactosidase, respectively, at 50 ° C. and pH 4-5. After reacting for an hour and drying under reduced pressure at 80 ° C., 4.23 g of fermented ginseng powder according to the present invention was prepared.

<2-2> Preparation of Fermented Ginseng Powder Using Ginseng Powder

7 g of ginseng powder was suspended in water, and then adjusted to pH 3 to 4.5 with citric acid, and 0.3 and 1 g of pectinase and beta galactosidase, respectively, were reacted at 50 ° C. for 24 hours, and the <Example 1-1 After extracting 4 times at 80 ° C. for 4 hours with a mixture of water and ethanol, the extract was dried under reduced pressure to prepare 2.6 g of the fermented ginseng powder according to the present invention.

<2-3> Preparation of Fermented Ginseng Powder Using Fermented Ginseng Concentrate

150 mg of the fermented ginseng concentrate prepared in <Example 1-1> was heated to 60 ° C. and left for 1 hour, and further stirred by adding 50 mg of arrowwood extract, followed by drying under reduced pressure. 100 mg of fermented ginseng powder containing the extract was prepared.

1 is a graph showing the results of performing high pressure liquid chromatography on the fermented ginseng concentrate according to the present invention.

Claims (9)

(a) preparing ginseng extract by adding water or an organic solvent to ginseng; (b) adding pectinase and beta galactosidase to the ginseng extract prepared in step (a); (C) a method of producing a fermented ginseng concentrate comprising the step of concentrating the ginseng extract after the step (b), and then concentrated. (a) adding ginseng to water to prepare ginseng solution; (b) adding a mixture of pectinase and beta galactosidase to the ginseng solution prepared in step (a), followed by fermentation; (c) extracting the fermented ginseng solution prepared in step (b) by adding water or an organic solvent, and then concentrating the fermented ginseng concentrate. According to claim 1 or 2, wherein in the step (a) ginseng is selected from the group consisting of the ginseng, white ginseng, ginseng, dried ginseng, red ginseng, Taegeuk ginseng, extracts thereof, powders thereof and combinations thereof. Method for preparing fermented ginseng concentrate. According to claim 1 or 2, wherein the ratio of the pectinase and beta galactosidase in step (b) is 1: 1 ~ 100, the ginseng extract or ginseng solution prepared in step (a) Method for producing a fermented ginseng concentrate, characterized in that 1 to 50% by weight of the mixture of the pectinase and beta galactosidase by weight. The method for preparing fermented ginseng concentrate according to claim 1 or 2, wherein the fermentation in step (b) is performed at pH 3-8 and 10-70 ° C for 1-72 hours. A method for producing a fermented ginseng powder, characterized by drying under reduced pressure or freeze drying the fermented ginseng concentrate prepared in claim 1 or 2. A functional food composition comprising the fermented ginseng concentrate prepared by the method of claim 1 or 2 as an active ingredient. The functional food composition of claim 7, wherein the food composition has any one or more effects selected from immuno-enhancing, anti-diabetic and anti-cancer effects. A functional food composition comprising the fermented ginseng powder prepared by the method of claim 6 as an active ingredient.

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