KR101328179B1 - Preparation method of fermented red ginseng having high ginsenoside contents using surface culture - Google Patents

Abstract

The present invention relates to a method for producing red ginseng having a high ginsenoside content using ginseng fermented with Lactobacillus brevis M2 (Accession No. KCTC11390BP).

Description

Preparation method of fermented red ginseng with high content of ginsenosides using surface culture {PREPARATION METHOD OF FERMENTED RED GINSENG HAVING HIGH GINSENOSIDE CONTENTS USING SURFACE CULTURE}

The present invention relates to a method of fermenting the surface of ginseng and manufacturing fermented red ginseng using the surface fermented ginseng.

Red ginseng is prepared by washing, increasing steaming, steaming, primary drying (water content of 35-40%), storage aging, secondary drying (about 16% water content), and shaping, and are pale yellow or light brown. . When heat-treatment of red ginseng during the manufacturing process of red ginseng causes chemical changes of ingredients, the content of existing bioactive substances increases with the generation of new bioactive substances that do not exist in fresh ginseng or white ginseng. The major components that change during the manufacturing process of red ginseng are saponin-based and non-saponin-based. According to the saponin-based components, 22 kinds of white ginseng and 22 kinds of red ginseng have been reported separately. 12 kinds of saponins, four kinds of saponins that exist only in white ginseng have been identified.

Saponins of ginseng have physiological effects such as anticancer activity, antioxidant activity, arteriosclerosis and hypertension prevention, liver function improvement, antifatigue, antistress action, anti-aging, brain activity promotion, anti-inflammatory activity, allergic disease treatment and protein synthesis ability. Korean red ginseng has excellent physiological activities such as antioxidant, lowering blood pressure, improving alcoholic hyperlipidemia and lowering blood sugar.

In spite of the diverse and excellent pharmacological activity of red ginseng, the difference in the distribution and activation of individual intestinal microorganisms may actually indicate the difference in the efficacy of red ginseng. In order to overcome this problem, research and development on fermented red ginseng that convert or increase red ginseng into the components of the final metabolite after fermentation (fermentation) using intestinal microorganisms have been conducted.

For example, Korean Patent Laid-Open No. 10-2010-0079874 provides fermented red ginseng fermented by culturing lactic acid bacteria in red ginseng, and Korean Laid-open Patent No. 10-2010-0074382 provides a ginseng fermented product inoculated with fermented lactic acid bacteria in ginseng powder. Korean Patent No. 10-0866504 describes fermented red ginseng fermented by inoculating Lactobacillus brevis into red ginseng powder.

However, the techniques were fermented by deep culture in strains ginseng or red ginseng, not by surface culture (solid culture). The present inventors continue the study on the method of manufacturing fermented red ginseng, by culturing the surface of ginseng with Lactobacillus brevis M2 (Accession No. KCTC11390BP) to prepare a surface fermented ginseng, using the same to prepare red ginseng In this case, it was found that the ginsenoside content of the fermented red ginseng significantly increased and completed the present invention.

An object of the present invention is to provide a method for producing red ginseng with a high ginsenoside content.

In order to achieve the above object, the present invention provides a method of fermenting the surface of ginseng and producing fermented red ginseng using the surface fermented ginseng.

Fermented red ginseng prepared by the method of the present invention is characterized by a high content of ginsenosides compared to general red ginseng.

1 is red ginseng main ginseng (left) and misam (right) fermented surface.
Figure 2 shows the component changes during the surface fermentation of root main ginseng.
Figure 3 shows the changes of the components during the surface fermentation of root rice ginseng.

According to the present invention,

Preparing ginseng;

Inoculating the ginseng Lactobacillus brevis M2 ( Lactobacillus brevis ) M2 (Accession No. KCTC11390BP), and incubating on the surface of the ginseng; And

It relates to a method for producing fermented red ginseng comprising the step of producing red ginseng by increasing the surface cultured ginseng.

Further, according to the present invention,

Preparing ginseng; and

Inoculating the ginseng Lactobacillus brevis M2 ( Lactobacillus brevis ) M2 (Accession No. KCTC11390BP), and the method for producing a fermented ginseng comprising the step of culturing on the surface of the ginseng.

The present invention also relates to fermented red ginseng or fermented ginseng prepared by the above method.

The present invention also relates to a food composition comprising the fermented red ginseng or fermented ginseng of the present invention as an active ingredient.

Hereinafter, the present invention will be described in detail.

The ginseng of the present invention may be ginseng generally used in the production of red ginseng, and the kind thereof is not particularly limited. For example, the ginseng of the present invention may be ginseng, white ginseng, camphor ginseng, and the like, but is not limited thereto.

The ginseng of the present invention may use any part of the root of ginseng. For example, the ginseng of the present invention may be main ginseng or misam, or may be used together. In addition, in the practice of the present invention, the main and ginseng may be separated and fermented, or may be fermented as it is. You can also use a mixture of red ginseng fermented with red ginseng and fermented red ginseng.

The culture of the present invention may be any period in which ginsenoside content can be sufficiently enhanced, and the culture period is not particularly limited. However, in consideration of the fermentation efficiency, the culture of the present invention is preferably carried out for 2 to 30 days. This is because the strain for fermentation takes more than two days to fully grow, and because the significant change in the content of ginsenosides in the culture for more than 30 days is less efficient over time. However, this is in consideration of the fermentation efficiency and economical, even if the fermentation for a period of 2 to 30 days is not outside the scope of the present invention.

Surface culture of this invention is also called solid culture.

The surface culture of the present invention, unlike the conventional deep culture method of grinding and liquefying ginseng or red ginseng, and then fermentation, by culturing microorganisms on the surface of the ginseng in the form of maintaining the shape without grinding, the inside of the ginseng Rather, it induces the surface to ferment.

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Fermented red ginseng of the present invention has an increased ginsenoside content as compared to red ginseng that has not undergone fermentation. The fermented red ginseng of the present invention has a higher content of Rh2, Rg3, and compound K than red ginseng without fermented red ginseng.

The food of the present invention is not limited to, but not limited to, health supplements, health functional foods, functional foods, and the like, and includes adding fermented red ginseng or fermented ginseng of the present invention to natural foods, processed foods, and general food materials.

The food composition comprising the fermented red ginseng or fermented ginseng of the present invention, the fermented red ginseng or fermented ginseng may be added as it is, or may be used with other food or food compositions, and may be suitably used according to a conventional method. The mixed amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, improvement or therapeutic treatment). Generally, fermented red ginseng or fermented ginseng of the present invention may be added in an amount of 0.1 to 90 parts by weight, preferably 1 to 60 parts by weight, based on 100 parts by weight of the raw material of the food or beverage.

 There is no particular limitation on the kind of the food. The food composition comprising the fermented red ginseng or fermented ginseng may be used in the form of oral preparations, such as tablets, hard or soft capsules, solutions, suspensions, etc., these preparations are acceptable conventional carriers, such as oral In the case of the preparation for administration, an excipient, a binder, a disintegrant, a lubricating agent, a solubilizer, a suspending agent, a preservative, an extender, etc. can be prepared.

Examples of the food to which the fermented red ginseng or fermented ginseng may be added include meat, sausage, bread, chocolate, candy, snacks, confectionary, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, Teas, drinks, alcoholic beverages and vitamin complexes, and other nutritional supplements, but are not limited to these types of foods.

Hereinafter, the present invention will be described in more detail with reference to the following examples and experimental examples. However, the following examples and experimental examples are intended to illustrate the contents of the present invention, but the scope of the invention is not limited by the examples and the experimental examples.

≪ Materials and methods >

Used strain

The strain used in the present invention was used in experiments while passaged at 37 ° C. for 20-30 days with Lactobacillus brevis M2 (Microorganism Accession No. KCTC11390BP).

Fresh ginseng

Fresh ginseng used as a natural medium in the present invention was purchased through the 6 year old ginseng harvested in 2007 in the Jeungpyeong area of Chungbuk through the Jeungpyeong farming ginseng research group was used while stored in a cold room at -5 ℃ to prevent decay and quality degradation.

Preculture of Strains

MRS medium (Difco Laboratories, dextrose 20.0 g / L; meat peptone 10.0 g / L; beef extract 10.0 g / L; yeast extract 5.0 g / L; sodium acetate 5.0 g / L; disodium phosphate 2.0 g / L; ammonium citrate 2.0 g / L; tween 80 1.0 g / L; magnesium sulfate 0.1 g / L, manganese sulfate 0.05 g / L), and the strain was preincubated at 37 ° C. for 48 hours.

Sample extraction and yield measurement

To obtain an aqueous extract, 10 times the amount of tertiary distilled water was added to 100 g of the dried sample, and the mixture was extracted three times with stirring for 3 hours in a 40 ° C. constant temperature water bath, and 70% ethanol and 70% methanol solvent were extracted with water-soluble extract. Extracted in the same way. Then, the extracts were collected and filtered through a filter paper (Whatman No. 2, Whatman International Ltd., England), and the filtrate was concentrated under reduced pressure with a rotatory evaporator (Rotavapor R215, BUCHI, Switzeland) to remove each solvent, and then freeze dryer (three-way cold heat). Extraction yield was calculated by lyophilization with SF-DSF 12, Busan, Korea).

Analysis of Hydrolyzed Ginseng and Red Ginseng

Polyphenol content was determined according to the Folin-Denis method (Dewanto, V., Wu, X., & Liu, RH (2002b) .Processed sweet corn has higher antioxidant activity.Journal of Agricultural and Food Chemistry, 50, 49594964.) The total and acid sugar contents were measured by phenol-sulfuric acid method and Blumenkrantz and Asboe-Hansen method (Blumenkrantz, N. and Asboe-Hansen, G. (1973) Anal. Biochem., 54, 484).

Analysis of ginsenosides was carried out according to the following method using HPLC. 1 g of each concentrate was weighed, and 50 mL of 70% ethanol was added thereto, extracted twice in an 80 ° C shaking incubator, and filtered using Whatman # 1 paper. The filtrate was dried under reduced pressure using a vacuum concentrator and dissolved by adding 50 mL of distilled water. 2 mL of diethyl ether was added and mixed with 1 mL of the solution, followed by centrifugation at 1500 rpm for 10 minutes to remove diethyl ether, followed by mixing with 1.5 mL of water-saturated butanol, and then recovering the butanol layer. 1.5 mL of distilled water was added to the butanol layer, which was recovered by repetition three times, mixed and centrifuged to remove the water layer. The procedure was repeated twice to wash water-soluble impurities. The butanol layer thus obtained was dried by injecting N 2 gas at 40 ° C., and 0.5 mL of methanol was added thereto, followed by filtration through a 0.45 μm membrane filter to be used as a sample for HPLC analysis. HPLC analysis was performed by ELSD using HPLC equipped with Prevail Carbohydrate ES 5μ column (Alltech, USA) and performed under the conditions as indicated in Table 1 and Table 2 (Gradient Table) below. On the other hand, ginsenoside analysis includes 14 standard materials (compound K, Rh2, Rh1, Rg5, Rk1, Rg2, Rg3, Rg1, Rf, Re, Rd, Rb2, Rc, A standard curve was prepared using Rb2) and the content was calculated from each peak area ratio.

A: Acetonitrile: Water: IPA = 80: 5: 15

B: Acetonitrile: Water: IPA = 67: 21: 12

<Example 1 and Example 2>

10% of Lactobacillus brevis M2 (KCTC11390BP) incubated in MRS basal medium was inoculated into main ginseng (Example 1) and rice ginseng (Example 2) of 6-year-old ginseng, respectively, and surface culture (solid culture) at 37 ° C for 20-30 days. The fresh ginseng surface fermentation product (solid fermentation product) was prepared. And red ginseng was prepared using this (FIG. 1).

&Lt; Comparative Example 1 &

Six-year-old ginseng was used as Comparative Example 1.

Comparative Example 2

Red ginseng was prepared using 6 years old ginseng. Specifically, the fresh ginseng was steamed at 85 to 95 ° C. for 4 to 5 hours, followed by primary hot air drying. Then, natural drying was performed to finally prepare red ginseng so that the moisture content was less than 14%.

<Comparative Example 3 and Comparative Example 4>

The general red ginseng prepared in Comparative Example 2 was separated into main ginseng and rice ginseng, and then lyophilized and ground. 50 mL of phosphate buffer solution (50 mM, pH 5.0) was added to each of 5 g of the main ginseng powder and the ginseng powder, and then suspended by autoclaving. The autoclaved red ginseng suspension was inoculated with 2% of Lactobacillus brevis M2 (KCTC11390BP) pre-cultured in MRS and incubated for 5 days with gentle stirring at 37 ° C. In this case, the fermented product of the red ginseng main ginseng powder was Comparative Example 3, and the fermented product of the red ginseng misam powder was used as Comparative Example 4.

&Lt; Comparative Example 5 &

10% of Lactobacillus brevis KCTC 3102 cultured in MRS basal medium was inoculated into main ginseng of 6-year-old ginseng, and surface culture (solid culture) at 37 ° C. for 20-30 days to prepare a fresh ginseng surface fermentation product (solid fermentation product). And red ginseng was prepared using this.

<Experiment 1> Ginsenoside content change according to the culture presence and method

As a result of measuring the ginsenoside content change according to the presence or absence of the cultivation method, it was confirmed that the ginsenoside content was increased in red ginseng regardless of the culture.

However, in the case of the red ginseng after the strain fermentation (Example 1, Example 2, Comparative Examples 3 to 5), the ginsenoside content increase was more pronounced than that of the red ginseng prepared by the general method (Comparative Example 2). In particular, the amount of increase of Example 1 and Example 2 prepared by red ginseng after surface fermentation of ginseng with Lactobacillus brevis M2 (KCTC11390BP) was significantly higher, and these were absorbed as well as ginsenosides such as Rh2, Rg3 and compound K. The content of ginsenoside metabolite, a prone form, increased significantly overall (Table 3).

Therefore, when preparing fermented red ginseng, it was confirmed that it is advantageous to prepare red ginseng having high ginsenoside content by surface fermenting ginseng and increasing the surface fermented ginseng rather than fermenting it.

Panaxdiol: Rb1 + Rc + Rb2 + Rd + Rg3,

Panaxtriol: Rg1 + Re + Rf + Rh1 + Rg2,

Metabolite = CK + Rh1 + Rg5 + Rk1 + Rg3 + Rg2 + Rh2 + F2.

.

<Experimental Example 2>

Example 1 and Example 2 confirmed to have a high ginsenoside content in Experimental Example 1 were mixed at 7: 3, and the ginsenoside content was measured. At this time, a mixture of Comparative Example 3 and Comparative Example 4 7: 3 and Comparative Example 4 was used as a control.

As a result, the mixture of Comparative Examples 3 and 4 had a total ginsenoside content of 143.1 mg / g, which was somewhat lower than the surface fermentation mixture (the mixture of Examples 1 and 2), and was easily absorbed. Phosphorus metabolite content was about 11.4 mg / g, showing a similar content.

Mixture of Example 1 and Example 2 7: 7 Mixture of Example 1 and Example 2

Mixture of Comparative Example 3 and Comparative Example 4: 7: 3 Mixture of Comparative Example 3 and Comparative Example 4

.

Experimental Example 3 Examination of Extraction Yield

Hydrothermal extraction of Example 1, Example 2, and Comparative Example 2 was carried out using the decoction method, and the yield was measured. Specifically, 500 parts by weight of water was added to 100 parts by weight of fermented red ginseng and red ginseng, which was heated for 12 hours to prepare a hot water extract, and the yield was calculated based on the content of the solid content (Equation 1).

<Formula 1>

Yield = (solid weight of red ginseng extract / weight of red ginseng) ⅹ100

As a result, the yields of the main ginseng fermented product (Example 1), micro ginseng fermented product (Example 2) and general home ginseng (Comparative Example 2) did not show a big difference, respectively, at 45.7%, 45.5 and 32.3%. However, red ginseng fermentation products (Examples 1 and 2) were found to have a higher extraction yield than 32.3% of general red ginseng (Comparative Example 2). It was estimated that the red ginseng's tissues or polymer components were disintegrated or decomposed through the fermentation process of mycelium, resulting in high extraction rate under the same hydrothermal extraction conditions.

Experimental Example 4 Changes in Components during Surface Fermentation of Fresh Ginseng

<4-1> Changes in Components of Surface Ginseng during Fermentation

10% of Lactobacillus brevis M2 (KCTC11390BP) incubated in MRS basal medium was inoculated into the main ginseng of 6-year-old ginseng, and surface changes (solid culture) were performed at 37 ° C for 20 to 30 days, and the changes of the components according to the fermentation time were measured. .

As a result, the total sugar content tended to decrease with the fermentation time and the acidic sugar uronic acid showed the highest content in 6-9 days. On the other hand, the content of polyphenols increased slightly during the fermentation process, but the change of the tendency with fermentation time was not significant. The number of bacteria involved in fermentation during surface fermentation tended to increase as the fermentation time increased, and after 6 days of fermentation, the number of bacteria increased slowly (Fig. 2).

<4-2> Changes in Components during Surface Fermentation of Rice Ginseng

10% of Lactobacillus brevis M2 (KCTC11390BP) incubated in MRS basal medium was inoculated into the ginseng of 6-year-old ginseng, and surface changes (solid culture) were performed at 37 ° C for 20 to 30 days, and the changes of the components according to the fermentation time were measured. .

As a result, the total sugar content increased with the fermentation time until the 9th fermentation and then decreased. The uronic acid content of acidic sugars also showed a similar tendency to the total sugar content. . Changes in polyphenol content increased slightly during fermentation and decreased slightly at 15 days. The number of bacteria involved in fermentation during surface fermentation increased rapidly after 3 days fermentation and then gradually decreased, but there was no significant difference between fermentation times. However, at 15 days of fermentation, the number of bacteria decreased. In the case of rice ginseng, fermentation tended to be faster because of its surface area wider than that of main ginseng.

Claims (9)

Preparing ginseng;
Inoculating the ginseng with Lactobacillus brevis M2 (Accession No. KCTC11390BP) to solid culture on the surface of the ginseng; and
Method of producing a fermented red ginseng with increased content of Rg3 and Compound K, comprising the step of producing red ginseng by increasing the solid cultured ginseng.
The method of claim 1,
The ginseng is a production method, characterized in that at least one selected from the group consisting of main ginseng and misam.
The method of claim 1,
The culture method is characterized in that it is carried out for 2 to 30 days.
The method of claim 1,
The fermented red ginseng is characterized in that the ginsenoside content is increased compared to the red ginseng not fermented.
The method of claim 1,
The fermented red ginseng is characterized in that the content of Rh2, Rg3 and compound K is increased compared to the red ginseng not fermented.
Fermented red ginseng prepared by the method of any one of claims 1 to 5.
Food composition comprising a fermented red ginseng prepared by the method of any one of claims 1 to 5 as an active ingredient.
8. The method of claim 7,
A food composition comprising fermented red ginseng prepared from main ginseng and fermented red ginseng prepared from misam.
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