KR102750984B1 - A Monascus fermented composition of mountain-cultivated ginseng sprout with increased ginsenoside Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxadiol and protopanaxatriol and a preparation method thereof - Google Patents

Abstract

The present invention discloses a fermented composition containing ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol, and protopanaxadiol, which are produced by fermenting wild ginseng sprouts with red ginseng fungus, and having significantly enhanced antioxidant activity, alpha-glucosidase inhibitory activity, and pancreatic lipase inhibitory activity, and a method for producing the same.
The fermented composition of wild ginseng sprouts and Honggukgyun according to the present invention contains a high content of ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol, and protopanaxadiol, and has excellent antioxidant activity and diabetes and obesity improvement effects, so that it can be used as a material for functional foods and cosmetics.

Description

{A Monascus fermented composition of mountain-cultivated ginseng sprout with increased ginsenoside Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxadiol and protopanaxatriol and a preparation method thereof}

The present invention relates to a fermented composition of wild ginseng sprouts and Hong Aspergillus oryzae, which is enhanced in ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol, and a method for producing the same. More specifically, the present invention relates to a fermented composition prepared by fermenting wild ginseng sprouts with Hong Aspergillus oryzae, which is enhanced in ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol, and which has significantly enhanced antioxidant activity, alpha-glucosidase inhibitory activity, and pancreatic lipase inhibitory activity, and a method for producing the same.

Ginseng is classified as ginseng, mountain ginseng, or sheep ginseng depending on the differences in artificial and natural growth depending on the cultivation environment, as well as morphological differences. In particular, sheep ginseng belongs to the Araliaceae family and refers to the seeds or young ginseng of ginseng ( Panax ginseng ), a perennial herb, that have grown naturally in the wild under mountain forests, and these are called sheep ginseng. In the case of ginseng, most have 3-7 heads, but sheep ginseng often has more depending on age. In addition, the roots of ginseng are thick and short, but sheep ginseng is long and thin. Cultivated ginseng refers to a plant of the genus Panax in the family Acanthopanax that is cultivated in a mountain area as defined in Article 2, Paragraph 1 of the Mountain Area Management Act and that has passed the quality inspection as defined in Article 18, Section 4 of the Forestry and Mountain Village Development Promotion Act. Cultivated ginseng is a species of ginseng of the Korean ginseng family and is called Panax ginseng CA Mey. Its scientific name is Panax, and its common name, Panax, means a cure-all.

Ginsenosides are unique saponins that exist only in plants of the ginseng genus in the dammarane series of triterpenoids. They have a structure in which sugars such as glucose, xylose, and rhamnose are bonded to carbon positions 3, 6, and 20 of the basic skeleton, and they are divided into protopanaxadiol, protopanaxatriol, oleanolic acid, and octillol series, and are known to have very mild pharmacological properties, no toxicity due to overdose, and almost no hemolytic effect. The protopanaxadiol series includes ginsenosides Rb1, Rb2, Rb3, Rc, Rd, and Rg1, and the protopanaxatriol series includes ginsenosides Re, Rf, Rg1, and Rg2. These ginsenosides are known to have various efficacies. It has been reported to have anticancer, antidiabetic, and immune function regulating effects. Usually, only the root of ginseng is used for food, and this is because it becomes difficult to eat due to hardening of the leaves as the number of years of cultivation passes. However, it has been reported that ginseng leaves, which are related to photosynthesis, also contain a considerable amount of ginsenoside.

Mountain-cultivated ginseng sprouts refer to mountain-cultivated ginseng that is at least 3 years old and has leaves and stems grown in a forest at least 500 m above sea level between May and June, or mountain-cultivated ginseng that is at least 3 years old and has leaves and stems grown for 20 to 120 days in a facility with a controlled environment (smart farm or plant factory). Mountain-cultivated ginseng sprouts can be produced year-round in a facility with a controlled environment during cultivation, and because their short cultivation period has advantages in terms of industrial use, the development of processed products from them is demanded.

Monascus belongs to the genus Monascus in the family Hemiascomycetaceae, and about 70 species have been isolated and identified at present. It is characterized by producing a red pigment by culturing Monascus fungi in steamed rice, and has been used for the purpose of preservatives, coloring in marine products, and processed livestock products, and as a digestive aid in East Asian countries since about 600 years ago. An anti-inflammatory and immune-enhancing composition containing a fermented coffee bean product fermented using Monascus is disclosed (Korean Patent Registration No. 10-1492174).

However, not only is there no precedent for successful fermentation of wild ginseng sprouts using Hong Aspergillus oryzae, but in particular, a wild ginseng sprout fermentation composition with significantly increased ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol, and protopanaxadiol has not been developed.

Accordingly, the inventors of the present invention continued their research to meet the needs of the prior art, and as a result, they confirmed that when wild ginseng sprouts are fermented with Hongguk fungus, ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol are significantly increased, and thus completed the present invention.

Therefore, the purpose of the present invention is to provide a fermented composition of wild ginseng sprouts with Hongsugung fungus, which is enriched with ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol.

Another object of the present invention is to provide a method for producing a fermented composition of wild ginseng sprouts containing ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol.

Another object of the present invention is to provide a functional food containing a fermented composition of the above-mentioned wild ginseng sprouts and Hongguk fungus.

Another object of the present invention is to provide a cosmetic comprising a fermented composition of the wild ginseng sprouts of the present invention.

To achieve the above object, the present invention provides a fermented composition of wild ginseng sprouts with Hong Aspergillus oryzae, which is produced by fermenting a mixture of wild ginseng sprouts and soybeans with Hong Aspergillus oryzae, and is enriched with ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol, and protopanaxadiol.

In the present invention, wild ginseng sprouts refer to wild ginseng that is at least 3 years old and has leaves and stems grown in a forest at least 500 m above sea level between May and June, or wild ginseng that is at least 3 years old and has leaves and stems grown for 20 to 120 days in a facility (smart farm or plant factory) with a controlled environment. It is preferable to use wild ginseng that is at least 3 years old and has leaves and stems grown for 25 to 45 days in a forest at least 500 m above sea level in a facility with a controlled environment in May.

In the mixture of the present invention, it is preferable that the mountain ginseng sprouts are included in an amount of 90 to 94 wt% and the soybeans are included in an amount of 6 to 10 wt%, and most preferably, the mountain ginseng sprouts are included in an amount of 90 wt% and the soybeans are included in an amount of 10 wt%.

In the present invention, soybeans mainly serve as a nitrogen source necessary for the growth of red yeast, while also having the function of promoting the fermentation (substance conversion) of red yeast.

If the content of mountain ginseng sprouts is out of the above range, the contents of ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol, and protopanaxadiol decrease, and if the content of soybeans is out of the above range, the mycelial growth of Hong Aspergillus oryzae may not be sufficient. Considering both the growth degree of Hong Aspergillus oryzae and the contents of ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol, and protopanaxadiol, the weight ratio of mountain ginseng sprouts: soybeans is most preferably 9:1.

In the present invention, it is preferable to use the wild ginseng sprouts after washing, drying, cutting, and drying them at 50-55℃ for 2-3 days, and the soybeans are used after washing and removing moisture.

In the present invention, ‘Honggukgyun’ refers to a fungus of the genus Monascus .

In the present invention, the red aspergillus oryzae fungus that is mixed-cultured with the wild ginseng sprout substrate is not limited in type as long as it belongs to the genus Monascus and can produce monacolin K as a secondary metabolite. For example, the red aspergillus oryzae fungus that can be used in the present invention include Monascus pilosus , Monascus ruber , Monascus purpureus, Monascus kaoliang, Monascus barykery , Monascus anka , etc., and considering the high productivity of monacolin K and the low productivity of citrinin, it is preferable to select Monascus pilosus and/or Monascus purpureus .

In the present invention, 'fermentation' means fermentation using Hong Aspergillus oryzae as a starter. When a mixture of wild ginseng sprouts and soybeans was fermented with Hong Aspergillus oryzae, surprisingly, ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol, and protopanaxadiol were significantly increased (Table 1, Table 2).

In the present invention, the fermentation can be carried out by inoculating Honggukgyun in a mixture of wild ginseng sprouts and soybeans at a concentration of 3 to 10% (v/w) and fermenting at 20 to 30°C for 8 to 12 days. Most preferably, the fermentation is carried out for 10 days.

If the amount of Hong Guo Mung bean inoculation is less than 3% (v/w), the fermentation rate may be delayed, and if it exceeds 10% (v/w), the mycelial growth rate may be fast and the conversion rate may be low, and if the fermentation temperature is less than 20℃, the fermentation period may be long, which may lead to contamination by various bacteria, and if it exceeds 30℃, the growth of mycelia may be reduced, and if the fermentation period is less than 8 days, fermentation may not be sufficient, resulting in low production of physiologically active substances, and if it exceeds 12 days, the physiologically active substances may be decomposed due to overfermentation.

In the present invention, it is preferable to add 5 to 10 times (v/v) of water to the mixture of wild ginseng sprouts and soybeans before fermentation, hydrate them for 5 to 8 hours to adjust the moisture content to about 40 to 50%, and then steam them at 100 to 120°C or higher for 30 to 60 minutes. It is most preferable to steam them at 120°C for 30 minutes.

In the present invention, when the moisture content is 40% or less during the hydration treatment, the growth of the Hong Aspergillus mycelia is not smooth, and when the moisture content is 50% or more, not only the growth of the Hong Aspergillus mycelia but also the proliferation of bacteria may occur. On the other hand, when the steaming treatment is performed for less than 30 minutes, sterilization is not completely achieved, and abnormal fermentation may proceed due to the proliferation of bacteria in addition to the Hong Aspergillus mycelia, and when the treatment is performed for more than 60 minutes, the fermentation may proceed excessively, destroying nutrients.

The fermented composition of wild ginseng sprouts according to the present invention contains ginsenoside Rg2 of 1.34 mg/g or more, F1 of 1.3 mg/g or more, protopanaxadiol of 0.22 mg/g or more, Rd of 5.63 mg/g or more, Rd2 of 1.18 mg/g or more, F2 of 3.25 mg/g or more, Rg3 of 1.15 mg/g or more, and protopanaxadiol of 14.6 mg/g or more (Table 1).

The fermented composition of wild ginseng sprouts according to the present invention has contents of ginsenosides Rg2, F1, protopanaxatriol, Rd, Rd2, F2, Rg3 and protopanaxadiol increased by about 2.8 times or more, 1.6 times or more, 2.4 times or more, 2.1 times or more, 3.2 times or more, 4.2 times or more, 6.7 times or more and about 7.9 times or more, respectively, compared to the mixture before processing (Comparative Example 1) (Table 1).

In the fermented composition of wild ginseng sprouts according to the present invention, the contents of protopanaxatriol series, protopanaxadiol series, and total ginsenosides are 2.98 mg/g or more, 25.81 mg/g or more, and 28.79 mg/g or more, respectively (Fig. 3).

The fermented composition of wild ginseng sprouts according to the present invention increases the contents of protopanaxatriol series, protopanaxadiol series, and total ginsenosides by about 2.3 times or more, 4.5 times or more, and 4.1 times or more, respectively, compared to the mixture before processing (Comparative Example 1) (Fig. 3).

In addition, the fermented composition of wild ginseng sprouts according to the present invention has enhanced contents of physiologically active substances such as ginsenosides Rg2, F1, protopanaxatriol, Rd, Rd2, F1, Rg3, and protopanaxadiol, total phenolics and total flavonoids, thereby having enhanced antioxidant activity, antidiabetic activity and antiobesity activity (Tables 1 to 2, Figs. 3 and 4 to 8).

In order to achieve another object of the present invention, the present invention provides a method for producing a fermented composition of wild ginseng and Hong Aspergillus oryzae by fermenting a mixture of wild ginseng sprouts and soybeans with Hong Aspergillus oryzae, wherein ginsenosides Rg2, F1, protopanaxatriol, Rd, Rd2, F1, Rg3, and protopanaxadiol are increased.

Specifically, the above method

i) A step of mixing 90 to 96 wt% of wild ginseng sprouts and 6 to 10 wt% of soybeans, then mixing with water in a ratio of 5 to 10 times (v/w) and hydrating for 5 to 7 hours;

ⅱ) Steaming step at 100~120℃ for 30~60 minutes; and

ⅲ) It includes a step of inoculating Honggukgyun at a concentration of 3 to 10% (v/w) and fermenting at 20 to 30°C for 8 to 12 days.

In the manufacturing method of the present invention, the weight ratio of the mixture of wild ginseng sprouts and soybeans, hydration, steaming, red yeast and fermentation are as defined above.

To ensure smooth fermentation, water can be added to the mixture of wild ginseng sprouts and soybeans before fermentation to hydrate them.

Before fermentation, it can be steamed to sterilize. It is preferable to steam at 100~120℃ or higher for 30~60 minutes, and most preferably at 120℃ for 30 minutes.

According to another object of the present invention, the present invention provides a functional food comprising a fermented composition of wild ginseng sprouts and Hong Guk-myun, which is produced by the above-mentioned production method and has increased ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol.

The functional food according to the present invention has excellent antioxidant activity, diabetes improvement, and obesity improvement effects (Figs. 4a to 6).

According to another object of the present invention, a cosmetic is provided which includes a fermented composition of wild ginseng sprouts and Hong Guk-myun having enhanced ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol.

The food or cosmetic of the present invention can be manufactured by adding the fermented composition of the present invention or its extract as is or by mixing it with other food ingredients, and can be manufactured appropriately according to a conventional method.

In the present invention, the type of the food is not particularly limited, and may include, but is not limited to, pills, tablets, capsules, fermented tea, fermented foods (kimchi, pickles, etc.), fermented beverages (pouches, drinks, etc.). The type of cosmetics may include, but is not limited to, mask packs, skins, lotions, creams, etc.

The fermented composition of wild ginseng sprouts and Honggukgyun according to the present invention contains high contents of ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol, and can be used as a material for functional foods and cosmetics.

In addition, the functional food or cosmetic according to the present invention has a high content of ginsenoside Rg2, Rg3 F1, F2, Rd, Rd2, protopanaxatriol and protopanaxadiol, and further has excellent antioxidant activity, alpha-glucosidase inhibitory activity and pancreatic lipase inhibitory activity, and is useful for suppressing fat production, controlling weight, lowering cholesterol, improving hyperlipidemia, alleviating arteriosclerosis, improving diabetes, improving obesity, improving blood circulation, improving immunity, beautifying skin and improving female menopausal syndrome.

Figure 1 is an example of a manufacturing process diagram of the fermented composition of Honggukgyun of the present invention.
Figure 2 shows ginsenoside HPLC chromatograms. (A) is the ginsenoside HPLC chromatogram of the wild ginseng sprout raw material as a comparative example, (B) is the ginsenoside HPLC chromatogram of the fermented composition of Monascus perpreus as Example 1, and (C) is the ginsenoside HPLC chromatogram of the fermented composition of Monascus piloseus as Example 2.
Figure 3 shows the contents of protopanaxatriol series, protopanaxadiol series, and total ginsenoside of the fermented composition according to the present invention.
Figure 4 shows the DPPH radical scavenging activity of the fermentation composition according to the present invention.
Figure 5 shows the ABTS radical scavenging activity of the fermentation composition according to the present invention.
Figure 6 shows the reducing power (FRAP) of the fermentation composition according to the present invention.
Figure 7 shows the alpha-glucosidase inhibitory activity of the fermentation composition according to the present invention.
Figure 8 shows the pancreatic lipase inhibitory activity of the fermented composition according to the present invention.

The present invention is further described in detail by the following examples. These examples are intended to illustrate the present invention, and the scope of the present invention should not be limited by them.

Manufacturing Example 1: Manufacturing of fermentation composition

The wild ginseng sprouts were grown in the Baekjeon-myeon area of Hamyang-gun in 2020 and supplied by Jinseong Bio, an agricultural corporation. The soybeans (orial-tae soybeans) were purchased from a large mart in Jinju-si.

Monascus pilosus and Monascus purpureus were obtained from the Korean Microbiological Conservation Center and used after activation. Potato Dextrose broth/agar (PDB/PDA, BD-Difco, Sparks, MD, USA) was used for the subculture of Monascus.

The wild ginseng sprouts were washed three times in running water, cut into approximately 1 cm pieces, and dried at 55°C for 3 days to prepare them. A wild ginseng sprout mixture sample was prepared by mixing dried wild ginseng and soybeans in a weight ratio of 9:1.

A 10-fold (v/w) volume of purified water was added to a mixed sample of wild ginseng sprouts, and the mixture was hydrated for 6 hours to adjust the moisture content to approximately 40%. The mixture was steamed at 120°C for 30 minutes using an autoclave, and 5% (v/w) of a culture of Hong Guo Myeong was inoculated. Fermentation was performed at 25°C for 10 days to obtain a fermented composition (Example 1 (Fermentation with Monascus phyllosus), Example 2 (Fermentation with Monascus perpreus); Fig. 1).

For comparison, a mixed sample composition of dried wild ginseng sprouts (comparative example) was prepared by mixing dried wild ginseng sprouts and soybeans in a weight ratio of 9:1.

Reference example: Analysis sample

The fermentation compositions of Examples 1 and 2 and the composition of Comparative Example 1 were each dried at 55°C for 3 days and pulverized to prepare powder samples.

1 g of powdered sample was extracted for 12 hours by adding 20 times 50% methanol, completely concentrated, dissolved by adding 2 ml of 50% methanol, filtered through a 0.45 μm filter, and placed in an HPLC bale to analyze ginsenosides.

A portion of the 10 g of powdered sample, extracted for 12 hours by adding 20 times 50% alcohol, was filtered through a 0.45 μm filter and used for total phenolics and total flavonoids analysis. The remaining extract was filtered using a vacuum filter, completely concentrated using a vacuum evaporator, and then freeze-dried using a freeze-dryer (FD-1000, Tokyo, Rikakikai, Japan). 50% alcohol was added to the final freeze-dried sample to prepare sample concentrations of 0.25, 0.5, 1.0, and 2.0 mg/mL, which were then used in the following physiological activity test.

Test Example 1: Analysis of Ginsenoside Content

Ginsenoside analysis was performed by high pressure liquid chromatography (HPLC) using a modified method described in the red ginseng saponin analysis method of the functional food analysis method. The analysis column used was TSKgel ODS-100Z, the sample injection amount was 10 ㎕, the temperature was 30℃, the measurement wavelength was 203 nm, and the flow rate was 1.0 ㎖/min. HPLC water was used as the mobile phase for Solution A and acetonitrile for Solution B. The HPLC analysis conditions were as follows: at 0 min, 81% solution A : 19% solution B was flowed, at 15 min, 80% solution A : 20% solution B was flowed, at 40 min, 77% solution A : 23% solution B, at 40 min, 77% solution A : 23% solution B, at 42 min, 70% solution A : 30% solution B, at 75 min, 65% solution A : 35% solution B, at 80 min, 30% solution A : 70% solution B, and at 90 min, 10% solution A : 90% solution B. The HPLC chromatograms of ginsenosides of Examples 1, 2, and Comparative Examples are shown in Fig. 2, and the contents of the analyzed ginsenosides Rg2, Rg3 F1, F2, Rd, Rd2, protopanaxadiol (PPT), and protopanaxadiol (PPD) are respectively shown in Table 1. In addition, the contents of protopanaxadiol series (Rg2, F1, and PPT) ginsenosides, protopanaxadiol series (Rd, Rd2, F2, Rg3, and PPD) ginsenosides, and total ginsenosides were calculated and shown in Fig. 3.

Content (mg/g) Comparative Example 1
(raw material) Example 1
(Hong Guk-gyun-1) Example 2
(Hong Guk-gyun-2) Ginsenoside Rg2 0.48 1.37 1.34 Ginsenoside F1 0.66 1.78 1.30 Protopanaxtriol 0.14 0.22 0.34 Ginsenoside Rd 2.65 8.08 5.63 Ginsenoside Rd2 0.37 1.32 1.18 Ginsenoside F2 0.77 3.49 3.25 Ginsenoside Rg3 0.17 1.40 1.15 Protopanaxdiol 1.84 16.18 14.60 Hong Guk-gyun-1: Monascus purpureus .
Hong Guk-gyun-2: Monascus piolsus .
All experiments were repeated five times and the average value was displayed.

As shown in FIG. 2, in the composition of Comparative Example 1, the Rg2, Rg3 F1, F2, Rd, Rd2, protopanaxatriol and protopanaxadiol peaks were low, whereas in the Monascus phyllosus fermentation composition of Example 1 and the Monascus perpreus fermentation composition of Example 2 according to the present invention, it can be confirmed that the Rg2, Rg3 F1, F2, Rd, Rd2, protopanaxatriol and protopanaxadiol peaks were significantly high.

In addition, as shown in Table 1, the Rg2, F1, PPT, Rd, Rd2, F2, Rg3 and PPD contents of the unprocessed wild ginseng sprout mixed raw material of Comparative Example 1 are 0.48 mg/g, 0.66 mg/g, 0.14 mg/g, 2.65 mg/g, 0.37 mg/g, 0.77 mg/g, 0.17 mg/g, and 1.84 mg/g, respectively, and the Rg2, F1, PPT, Rd, Rd2, F2, Rg3 and PPD contents of the Monascus phyllosus fermented composition of Example 1 are 1.37 mg/g, 1.78 mg/g, 0.22 mg/g, 8.08 mg/g, 1.32 mg/g, 3.49 mg/g, 1.40 mg/g, and The contents of the Monascus perpreus fermented composition of Example 2 increased by about 2.9 times, 2.7 times, 1.6 times, 3.0 times, 3.6 times, 4.5 times, 8.2 times, and 8.8 times, respectively, to 16.18 mg/g, and the Rg2, F1, PPT, Rd, Rd2, F2, Rg3, and PPD contents of the Monascus perpreus fermented composition of Example 2 increased by about 2.8 times, 2.0 times, 2.4 times, 2.1 times, 3.2 times, 4.2 times, 6.7 times, and 7.9 times, respectively, to 1.34 mg/g, 1.30 mg/g, 0.34 mg/g, 5.63 mg/g, 1.18 mg/g, 3.25 mg/g, 1.15 mg/g, and 14.60 mg/g.

As shown in FIG. 3, the contents of protopanaxatriol series (Rg2, F1, and PPT) ginsenosides, protopanaxadiol series (Rd, Rd2, F2, Rg3, and PPD) ginsenosides, and total ginsenosides of Comparative Example 1, which is an unprocessed mixed raw material of wild ginseng, are 1.28 mg/g, 5.80 mg/g, and 7.08 mg/g, respectively, and the contents of protopanaxatriol series (Rg2, F1, and PPT) ginsenosides, protopanaxadiol series (Rd, Rd2, F2, Rg3, and PPD) ginsenosides, and total ginsenosides of the fermented composition of Monascus phyllosus of Example 1 are 3.37 mg/g, 30.47 mg/g, and 33.84 mg/g, respectively, which are about 2.6 times, 5.3 times, and The contents of protopanaxadiol series (Rd, Rd2, F2, Rg3, and PPD) ginsenosides, and total ginsenosides of the Monascus perpreus fermented composition of Example 2 were 2.98 mg/g, 25.81 mg/g, and 28.79 mg/g, respectively, indicating an increase of about 2.3 times, 4.5 times, and 4.1 times.

Therefore, it can be seen that the fermented composition of wild ginseng sprouts according to the present invention fermented with Honggukgyun has significantly enhanced contents of ginsenosides Rg2, F1, PPT, Rd, Rd2, F2, Rg3 and PPD.

Test Example 2. Analysis of the content of physiologically active ingredients

The content of total phenolics and total flavonoids, which are physiologically active components exhibiting antioxidant activity, were analyzed.

<Total phenol content>

The analysis sample was dispensed (0.5 ml) into a test tube, to which 0.5 ml of 25% Na ₂ CO ₃ solution was added, allowed to stand for 3 minutes, 0.25 ml of 2N Folin-Ciocalteu phenol reagent was added and mixed, allowed to stand for 1 hour at 30°C, and then the absorbance was measured at 750 nm using a spectrophotometer. At this time, the total phenolix content was calculated as the amount equivalent to gallic acid by obtaining the content from a standard curve created using gallic acid, and the results are shown in Table 2.

As shown in Table 2, the total phenol content was 3.28 mg/g in Comparative Example 1, 5.50 mg/g in Example 1, and 4.85 mg/g in Example 2, confirming that the fermented composition according to the present invention has an increased total phenol content.

Content (mg/g) Comparative Example 1
(raw material) Example 1
(Hong Guk-gyun-1) Example 2
(Hong Guk-gyun-2) Total Phenolix 3.28 5.50 4.85 Total flavonoids 1.36 2.86 2.75 Hong Guk-gyun-1: Monascus purpureus .
Hong Guk-gyun-2: Monascus piolsus .
All experiments were repeated five times and the average value was displayed.

<Total flavonoid content>

After dispensing 1.0 ml of diethylene glycol to 0.5 ml of the analysis sample, 0.01 ml of 1 N NaOH was added, and the sample was left in a constant temperature water bath at 37°C for 1 hour. The absorbance was measured at 420 nm using a spectrophotometer. At this time, the total flavonoid content was obtained from a standard curve created using rutin, and the results are shown in Table 2.

As shown in Table 2, the total flavonoid contents were 1.36 mg/g, 2.86 mg/g, and 2.75 mg/g in Comparative Example 1, Example 1, and Example 2, respectively, confirming that the fermented composition according to the present invention significantly increases the total flavonoid content.

Test Example 3. Antioxidant Activity Analysis

Antioxidant activity was analyzed by measuring DPPH radical scavenging activity, ABTS radical scavenging activity, and FRAP reducing power.

As in the reference example, the analysis samples of Example 1, Example 2, and Comparative Example 1 were prepared and used at concentrations of 0.25, 0.5, 1, and 2 mg/ml.

<DPPH radical scavenging activity>

Each sample (0.2 ml) was added with 0.8 ml of DPPH methanol solution (1.5×10 ^-4 M), stirred for 10 seconds, left in a dark room for 30 minutes, and then the absorbance was measured at 525 nm. The negative control group of DPPH radical scavenging activity was performed in the same manner using an extraction solvent instead of the sample, and the difference in absorbance was calculated as a percentage (%) by the following formula, and the results are shown in Fig. 4.

Radical scavenging activity (%) = [1-(absorbance of negative control group ÷ absorbance of experimental group)] × 100

As shown in Fig. 4, the DPPH radical scavenging activity of Comparative Example 1, Example 1, and Example 2 at a concentration of 1 mg/ml was 17.81%, 66.89%, and 64.03%, respectively, indicating that the fermented compositions of Example 1 and Example 2 exhibited significantly higher activity.

<ABTS radical scavenging activity>

7 mM ABTS ⁺ and 2.45 mM K ₂ S ₂ O ₈ were mixed in a 1:1 ratio and reacted in a darkroom for 12 to 16 hours, then mixed with methanol in a 1:88 ratio to adjust the absorbance value of the control group to 0.7±0.02 at 732 nm. 0.1 ml of each sample and 0.9 ml ^{of ABTS + solution} were added and mixed, and after standing for 3 minutes, the absorbance was immediately measured using a spectrophotometer at 732 nm. The negative control experiment was conducted by taking an extraction solvent instead of the sample, and the absorbance of the experimental group and the negative control group was calculated as a percentage (%) using the above formula, and the results are shown in Fig. 5.

As shown in Fig. 5, ABTS radical scavenging activity was 32.93%, 78.16%, and 72.24% for Comparative Example 1, Example 1, and Example 2, respectively, at a concentration of 0.5 mg/ml, indicating that the fermentation compositions of Example 1 and Example 2 exhibited significantly higher activity.

<FRAP Reduction Power Analysis>

In the FRAP reducing power analysis, 10 mM 2,4,6-tripyridyl-s-triazine (TPTZ, T1253, C ₁₈ H ₁₂ N ₆ , MW312.33) and 20 mM FeCl ₃ (F7134, MW 162.20, in DW) dissolved in 30 mM acetate buffer (pH 3.6) and 40 mM hydrochloric acid were prepared as the reaction solution. The acetate buffer, TPTZ solution, and FeCl ₃ solution were mixed in a ratio of 10:1:1 (v/v/v) and pre-reacted at 37 °C for 15 minutes. 0.95 ml of FRAP reagent was added to 0.05 ml of each sample, and the reaction was performed at 37 °C for about 15 minutes, and the absorbance was measured at 590 nm. The results are shown in Fig. 6.

As shown in Fig. 6, the FRAP reducing power of Comparative Example 1 was 0.789 OD _{593 nm} at a concentration of 2 mg/ml, while Examples 1 and 2 showed high activities of 1.566 OD _{593 nm} and 1.502 OD _{593 nm} , respectively.

From these results, it can be seen that the fermented composition according to the present invention has significantly enhanced antioxidant activity.

Test Example 4: Digestive Enzyme Inhibition Activity Test

Alpha-glucosidase inhibitory activity, which is an indicator of anti-diabetic (diabetes improvement) effect, was tested for the fermented composition of wild ginseng sprouts and Hongguk fungus according to the present invention, and pancreatic lipase inhibitory activity, which is an indicator of anti-obesity (obesity improvement) effect, was tested.

<Alpha-glucosidase inhibitory activity>

30 ㎕ of the analysis sample, 70 ㎕ of an enzyme solution of alpha-glucosidase (0.5 U/ml) and 50 ㎕ of a 200 mM sodium phosphate buffer solution (pH 6.8) were mixed and pre-reacted at 37℃ for 10 minutes. Then, 100 ㎕ of p-NPG (10 mM) dissolved in sodium phosphate buffer solution (pH 6.8) was added and reacted again at 37℃ for 10 minutes. After stopping the reaction by adding 750 ㎕ of Na ₂ CO ₃ (100 mM) to the reaction solution, the absorbance was measured using a spectrophotometer at 420 nm. For the negative control, an extraction solvent was used instead of the sample, and the difference in absorbance between the group with and without the analysis sample addition was expressed as a percentage (%), and the results are shown in Fig. 7.

As shown in Figure 7, the alpha-glucosidase inhibitory activity was 19.84% in Comparative Example 1 when treated at 2.0 mg/ml, but was increased to 45.66% in Example 1 and 40.58% in Example 2.

Therefore, it can be seen that the fermented composition according to the present invention has significantly enhanced alpha-glucosidase inhibitory activity, excellent blood sugar lowering activity, and thus enhanced diabetes improvement effect.

<Pancreatic lipase inhibitory activity>

30 ㎕ of the analysis sample was mixed with 70 ㎕ of an enzyme solution of pancreatic lipase (0.5 U/ml) and 50 ㎕ of a 200 mM sodium phosphate buffer solution (pH 6.8), and the mixture was pre-reacted at 37 °C for 10 minutes. After the reaction, 100 ㎕ of p-NPB (10 mM) dissolved in sodium phosphate buffer was added, and the mixture was reacted in the same manner for 10 minutes. Then, 750 ㎕ of 100 mM Na ₂ CO ₃ was added to terminate the reaction, and the absorbance was measured at 420 nm. For the negative control, an extraction solvent was used instead of the sample, and the difference in absorbance between the groups with and without the addition of the sample solution was expressed as a percentage (%), and the results are shown in Fig. 8.

As shown in Figure 8, the pancreatic lipase inhibitory activity was 15.46% in Comparative Example 1 when treated with 2.0 mg/ml, but it showed higher activity at 48.46% in Example 1 and 44.33% in Example 2.

Therefore, it can be seen that the fermented composition according to the present invention has enhanced pancreatic lipase inhibitory activity and thus an enhanced obesity improvement effect.

From the above activity (functionality) test results, it can be seen that the fermented composition according to the present invention not only increases the contents of ginsenosides Rg2, F1, PPT, Rd, Rd2, F2, Rg3 and PPD, but also has excellent antioxidant activity, anti-diabetic activity and anti-obesity activity, making it useful as a material for functional foods and cosmetics.

Claims (12)

delete delete delete delete delete delete A method for producing a fermented composition of wild ginseng sprouts enriched with ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol, wherein the method
i) A step of mixing 90 to 94 wt% of wild ginseng sprouts and 6 to 10 wt% of soybeans, then mixing with water in a ratio of 5 to 10 times (v/w) and hydrating for 5 to 7 hours;
ⅱ) Steaming step at 100~120℃ for 30~60 minutes; and
ⅲ) A method including a step of inoculating Honggukgyun at a concentration of 3 to 10% (v/w) and fermenting at 25 to 30°C for 8 to 12 days.
In claim 7, the Hong Guk Myeon fermentation composition of mountain ginseng sprouts is prepared by enhancing ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol, wherein the Hong Guk Myeon is at least one selected from the group consisting of Monascus pilosus and Monascus purpureus .
A fermented composition of wild ginseng sprouts with Hongsugung fungus, fortified with ginsenosides Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol, manufactured by a manufacturing method according to Article 8.
An antioxidant functional food containing a fermented composition of wild ginseng sprouts with red yeast according to Article 9.
Functional food for improving diabetes and obesity, containing a fermented composition of wild ginseng sprouts according to Article 9.
Cosmetics containing a fermented composition of wild ginseng sprouts with Hong Gukyeum according to Article 9.