KR20230065499A - A mushroom-mycelium fermented composition of mountain-cultivated ginseng sprout with increased ginsenoside F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxadiol and protopanaxatriol and a preparation method thereof - Google Patents

Abstract

The present invention discloses a fermentation composition of mushroom mycelium of wild ginseng sprouts enriched with ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol and a manufacturing method thereof. The fermentation composition of the mushroom mycelium of the wild ginseng sprouts according to the present invention has a high content of ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol, and in addition, has excellent antioxidant activity, alpha-glucosidase inhibitory activity, and pancreatic lipase inhibitory activity, making it useful as a material for functional foods and cosmetics.

Description

Ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol fermented composition of fermented mountain goat sprout mushroom mycelium and its manufacturing method {A mushroom-mycelium fermented composition of mountain- cultivated ginseng sprout with increased ginsenoside F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxadiol and protopanaxatriol and a preparation method thereof}

The present invention relates to a fermented composition of wild goat sprout mushroom mycelium enriched with ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol and a method for producing the same, in more detail ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol and antioxidant activity, alpha-glucose prepared by fermenting wild goat sprouts with mycelium of cordyceps or enoki mushrooms. It relates to a fermented composition with significantly enhanced sidase inhibitory activity and pancreatic-lipase inhibitory activity and a method for producing the same.

Ginseng is classified into ginseng, wild ginseng, or wild ginseng according to the difference between artificial growth and natural growth according to the cultivation environment and morphological differences. In particular, Sanyangsam belongs to the Ogapicaceae family, and Panax ginseng , a perennial plant, refers to wild ginseng seeds or young ginseng grown naturally in the wild under mountainous forests, which are artificially cultivated in the mountains, and they are called Sanyangsam. In the case of ginseng, most of them have 3-7 outcrops (head part), but sanyangsam has more than that depending on age, and the root of ginseng is thick and short, but sanyangsam is long and thin. Sanyangsam (山養蔘) is a ginseng genus of the ginseng family that has been cultivated in a mountainous area under Article 2, Item 1 of the 「Mountain Management Act」 and has passed the quality inspection in accordance with Article 18-4 of the 「Forestry and Mountain Village Promotion and Promotion Act」蔘屬) refers to plants. Wild ginseng is a ginseng species of Goryeo ginseng (高麗人蔘) and is called Panax ginseng CA Mey.

Ginsenoside is a unique saponin that exists only in plants of the genus ginseng of the dammarane family of triterpenoids. It has a structure in which sugars such as glucose, xylose, and rhamnose are linked to carbon 3, 6, and 20 positions of the basic skeleton, and these are protopanaxadiol, protopanaxatriol, and oleanol. It is divided into oleanolic acid and octillol, and is known to have very mild medicinal 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 like, and the protopanaxatriol series includes ginsenosides Re, Rf, Rg1, and Rg2. These ginsenosides are known for several efficacies. Representatively, it has been reported that it has anticancer, antidiabetic, and immune function regulating effects. In general, only the root of ginseng is used for food, and this is because it is difficult to eat due to the hardening of the leaves as the number of years of cultivation passes. However, it has been reported that the leaves of ginseng, which are related to photosynthesis, also contain a significant amount of ginsenosides.

Mountain-cultivated ginseng sprout (mountain-cultivated ginseng sprout) is a facility (smart farm or plant factory) that grows more than 3 years old wild ginseng leaves and stems between May and June in a forest at an altitude of 500m or more, or more than 3 years old wild ginseng. It refers to the growth of leaves and stems for 20 to 120 days. Wild ginseng sprouts can be produced year-round in facilities where the environment is controlled during cultivation, and the cultivation period is short, so there is an advantage in terms of industrial use, and the development of processed products is required.

Mushroom mycelium is a fungus that lives as a parasitic and byproduct in organic matter such as humus soil or dead wood in the form of a fluffy thin thread, and corresponds to the roots, stems, and leaves of plants. A method for increasing total phenolic compounds and beta-glucan by solid fermentation of bellflower with mushroom mycelium (Registration Patent No. 10-1884298) and Matsutake mushroom mycelium fermented composition of mixed grains having excellent flavor and enhanced functionality (Registration Patent No. 10-1889596 Ho) is disclosed.

However, there is no precedent for successful fermentation of wild goat shoots by the mycelium of Cordyceps sinensis or Enoki mushroom, especially ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol. This significantly improved fermented goat mushroom mycelium composition has not been developed.

Accordingly, the inventors of the present invention continued research to meet the needs of the prior art, and as a result, when fermenting goat ginseng sprouts with mycelium of cordyceps sinensis or enoki mushroom, ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, The present invention was completed after confirming that protopanaxatriol and protopanaxadiol were remarkably enhanced.

Therefore, an object of the present invention is to provide a mushroom mycelium fermented composition of wild goat sprouts enriched with ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol.

Another object of the present invention is to provide a method for producing a mushroom mycelium fermentation composition enriched in ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol.

Another object of the present invention is to provide a functional food containing the mushroom mycelium fermented composition of wild ginseng sprouts of the present invention.

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

In order to achieve the above object, the present invention is ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxa prepared by fermenting a mixture of goat shoots and soybeans into mushroom mycelium. Provides a diol-enhanced mushroom mycelium fermentation composition of wild ginseng sprouts.

In the present invention, wild ginseng sprouts grow more than 3 years old wild ginseng in a forest at an altitude of 500m or more from May to June, or grow more than 3 years old wild ginseng in a facility where a certain environment is controlled (smart farm or plant factory) 20 to 120 It refers to growing leaves and stems for days, preferably growing more than 3 years of wild ginseng in a forest at an altitude of 500m or more in May and in a facility where a certain environment is controlled for 25 to 45 days. It is preferable to use the grown leaves and stems do.

In the mixture of the present invention, 90 to 94% by weight of wild ginseng sprouts and 6 to 10% by weight of beans are preferably included, and most preferably, 90% by weight of wild ginseng sprouts and 10% by weight of soybeans are included.

When the content of wild goat sprouts is out of the above range, the content of ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol, and protopanaxadiol falls, and the content of soybeans falls within the above range. If it is out of range, mycelial growth of mushroom mycelium may not be sufficient. Considering both the degree of mycelial growth of mushroom mycelium and the contents of ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol, the weight ratio of wild goat sprouts: soybeans was 9: 1. It is most preferable to be

In the present invention, beans mainly serve as a nitrogen source necessary for the growth of mushroom mycelium, but also function to enhance the fermentation (material conversion action) of mushroom mycelium.

In the present invention, it is preferable to use goat ginseng sprouts after washing, drying, cutting, and drying at 50 to 55 ° C for 2 to 3 days, and beans are washed and dried.

In the present invention, the 'mushroom mycelium' is preferably a mycelium of Cordyceps militaris or Flammulina velutipes .

In the present invention, 'fermentation' means fermentation using the mycelium of Cordyceps sinensis or Enoki mushroom as a spawn. When the mixture of wild goat sprouts and soybeans was fermented with the mycelium of Cordyceps sinensis or Enoki mushroom, ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol were remarkably increased. (Table 1, Figures 2 and 3).

In the present invention, the mycelium of Cordyceps sinensis or Enoki mushroom can be performed by inoculating a mixture of wild goat sprouts and beans at a concentration of 3 to 10% (v / w) and fermenting at 20 to 30 ° C. for 8 to 12 days. Most preferably fermented for 10 days.

If the inoculation amount of mushroom mycelium of Cordyceps sinensis or Enoki mushroom 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, resulting in low conversion rate, and the fermentation temperature may be lowered. If the fermentation period is less than 20 ℃, the fermentation period is prolonged, resulting in contamination of various germs, and if it exceeds 30 ℃, the growth of mycelia may be reduced. If it exceeds 12 days, physiologically active substances may be decomposed by over-fermentation.

In the present invention, before fermentation, the mixture of wild ginseng sprouts and soybeans is hydrated for 5 to 8 hours by adding 5 to 10 times (v / v) of water to adjust the moisture to about 40 to 50%, and then ~60 min steaming is preferred. Most preferably, it is steamed for 30 minutes at 120°C.

In the present invention, when the moisture content is 40% or less during the hydration treatment, the growth of mycelia of cordyceps and enoki mushrooms is not smooth, and when the moisture content is 50% or more, not only the growth of mycelia of cordyceps and enoki mushrooms, but also the growth of bacteria, etc. It can be done. On the other hand, when steamed for 30 minutes or less, sterilization is not completely performed, so abnormal fermentation may proceed due to the proliferation of bacteria, etc. in addition to the mycelium of cordyceps and enoki mushrooms, and nutrients may be destroyed excessively when treated for 60 minutes or more.

The mushroom mycelium fermented composition of wild goat sprouts according to the present invention contains ginsenoside F3 of 0.23 mg/g or more, Rg2 of 1.31 mg/g or more, F1 of 1.27 mg/g or more, and protopanaxatriol of 0.28 mg/g or more. , Rd of 6.59 mg/g or more, Rd2 of 1.10 mg/g or more, F2 of 2.48 mg/g or more, Rg3 of 1.16 mg/g or more, and protopanaxadiol of 4.53 mg/g or more (Table 1) .

The mushroom mycelium fermented composition of wild goat sprouts according to the present invention contains 3.17 mg/g or more, 22.65 mg/g or more, and 25.82 mg/g or more of protopanaxatriol series, protopanaxadiol series, and total ginsenosides, respectively. contains (Fig. 3).

The mushroom mycelium fermented composition of wild goat sprouts according to the present invention has a content of protopanaxatriol series, protopanaxadiol series, and total ginsenosides of about 2.4 times or more, respectively, compared to the mixture before processing (Comparative Example 1). 3.9-fold or more and about 3.6-fold or more (FIG. 3).

In addition, the mushroom mycelium fermented composition of wild goat sprouts according to the present invention is ginsenosides F3, Rg2, F1, protopanaxatriol, Rd, Rd2, F1, Rg3, and protopanaxadiol, total phenolics and total flavonoids, etc. The content of physiologically active substances is enhanced, and it has enhanced antioxidant activity, antidiabetic activity, and antiobesity activity (Table 1, FIG. 3, FIGS. 4 to 5, and FIGS. 6 to 10).

In order to achieve another object of the present invention, the present invention is fermented mushroom mycelium of goat ginseng sprouts with enhanced ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol. A method for preparing the composition is provided.

Specifically, the method

i) hydrating for 5 to 7 hours by mixing 90 to 94% by weight of wild goat sprouts and 6 to 10% by weight of soybeans and then mixing with water 5 to 10 times (v / w);

ii) steaming for 30 to 60 minutes at 100 to 120°C; and

iii) inoculating the mycelium of Cordyceps sinensis or Enoki mushroom at a concentration of 3 to 10% (v/w) and fermenting at 20 to 30 ° C for 8 to 12 days.

In the production method of the present invention, the weight ratio, hydration, steaming, mushroom mycelium of cordyceps sinensis and enoki mushroom, and fermentation of the mixture of wild goat sprouts and soybeans are as defined above.

For smooth fermentation of the mycelium of Cordyceps sinensis and Enoki mushroom, water can be added to the mixture of wild goat shoots and soybeans before fermentation to hydrate them.

Prior to fermentation, it may be steamed to sterilize. It is preferable to steam for 30 to 60 minutes at 100 ~ 120 ℃, most preferably for 30 minutes at 120 ℃.

According to another object of the present invention, the present invention is a mushroom of wild ginseng bud with enhanced F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol prepared by the above production method It provides a functional food containing a mycelium fermentation composition.

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

According to another object of the present invention, cosmetics containing a fermented composition of mushroom mycelium of wild goat sprouts with enhanced ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol provides

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

In the present invention, the type of food is not particularly limited, and may include pills, tablets, capsules, fermented tea, fermented foods (kimchi, pickles, etc.), fermented beverages (pouches, drinks, etc.), but is not limited thereto. don't Types of cosmetics may include mask packs, toners, lotions, creams, and the like, but are not limited thereto.

The mushroom mycelium fermented composition of wild ginseng sprouts according to the present invention contains ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol in high content, making it suitable for functional foods and cosmetics. material can be used.

In addition, the functional food or cosmetic according to the present invention has a high content of ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol, as well as excellent antioxidant activity, alpha-glucose It has sidase inhibitory activity and pancreatic-lipase inhibitory activity, so it has anti-adipogenesis effect, weight control, cholesterol lowering, hyperlipidemia improvement, arteriosclerosis relief, diabetes improvement, obesity improvement, blood circulation improvement, immunity improvement, skin beauty, female menopausal syndrome useful for improvement

1 is an example of a manufacturing process diagram of a mushroom mycelium fermentation composition according to the present invention.
Figure 2 shows a ginsenoside HPLC chromatogram. Figure 2a is a ginsenoside HPLC chromatogram of a raw material of wild ginseng sprouts as a comparative example, Figure 2b is a ginsenoside HPLC chromatogram of the mycelium fermented composition of Cordyceps mushroom mycelium of Example 1, Figure 2c is the fermented enoki mushroom mycelium composition of Example 2 This is the ginsenoside HPLC chromatogram of
Figure 3 shows the protopanaxatriol series, protopanaxadiol series, and total ginsenoside contents of the fermented composition according to the present invention.
Figure 4 shows the total phenolics content of the fermentation composition according to the present invention.
Figure 5 shows the total flavonoid content of the fermentation composition according to the present invention.
Figure 6 shows the DPPH radical scavenging activity of the fermentation composition according to the present invention,
7 shows the ABTS radical scavenging activity of the fermentation composition according to the present invention.
8 shows the reducing power (FRAP) of the fermented composition according to the present invention.
Figure 9 shows the alpha-glucosidase inhibitory activity of the fermented composition according to the present invention.
10 shows the pancreatic-lipase inhibitory activity of the fermented composition according to the present invention.

The present invention is explained in more 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 thereto.

Preparation Example 1: Preparation of fermentation composition

Sanyang ginseng sprouts grown in Baekjeon-myeon, Hamyang-gun in 2020 were supplied from Ginseng Bio Co., Ltd., an agricultural corporation, and used. Beans (duck roe beans) were purchased and used at a large mart in Jinju City.

Mushroom mycelia were purchased from the Korea Center for Biological Resources, the Korea Center for Microbial Conservation, or the Gyeongsangnam-do Agricultural Research and Extension Services, and mycelia of Ganoderma lucidum, king oyster mushroom, oyster mushroom, cordyceps mushroom, and enoki mushroom were used after being activated. For subculture of mushroom mycelium, Potato Dextrose broth/agar (PDB/PDA, BD-Difco, Sparks, MD, USA) was used.

Wild ginseng sprouts were prepared by washing three times in running water, cutting them to about 1 cm, and then drying them at 55 ° C. for 3 days. A mixed sample of wild ginseng sprouts was prepared by mixing dried wild ginseng and soybeans at a weight ratio of 9:1.

After adding 10 times (v/w) purified water to the mixed sample of wild goat sprouts and hydrating for 6 hours to adjust the moisture content to about 40%, steaming was performed at 120 ° C for 30 minutes using a high-pressure sterilizer, and the mushroom mycelium prepared above After inoculating the culture medium at 5% (v/w), fermentation was performed at 25° C. for 10 days to obtain a fermentation composition (FIG. 1).

For comparison, a mixed sample composition (Comparative Example 1) was prepared by mixing dried wild ginseng and soybeans at a weight ratio of 9:1.

Reference example: analysis sample

Fermentation composition of Example 1 (fermentation with cordyceps mushroom mycelium), fermentation composition of Example 2 (fermentation with enoki mushroom mycelium), composition of Comparative Example 1 (mixed sample of wild goat sprouts), fermentation composition of Comparative Example 2 (Ganoderma lucidum mycelium Fermented with oyster mushroom), the fermentation composition of Comparative Example 3 (fermented with king oyster mushroom mycelium), and the fermentation composition of Comparative Example 4 (fermented with oyster mushroom mycelium) were dried at 55 ° C. for 3 days and pulverized to prepare powder samples.

50% methanol was added to 1 g of the powder sample 20 times, extracted for 12 hours, completely concentrated, 2 ml of 50% methanol was added, dissolved, filtered through a 0.45 μm filtration filter, and put on an HPLC veil to analyze ginsenosides.

50% alcohol was added 20 times to 10 g of the powder sample, extracted for 12 hours, filtered through a 0.45 μm filtration filter, and used for analysis of total phenolics and total flavonoids. The remaining extract was filtered using a vacuum filter and completely purified using a vacuum concentrator. After concentration, it was 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 0.5, 1.0 and 2.0 mg/mL, and then used in the following physiological activity test.

Test Example 1: Analysis of ginsenoside content

Ginsenoside analysis was analyzed by high pressure liquid chromatography (HPLC) by modifying the method described in the functional food analysis method for red ginseng saponin analysis. The analysis column was TSKgel ODS-100Z, and the sample injection amount was 10 μl, the temperature was 30 ° C, the measurement wavelength was 203 nm, and the flow rate was 1.0 ml / min. As the mobile phase, HPLC water was used as solution A and acetonitrile was used as solution B. . The HPLC analysis conditions are: 81% of A solution: 19% of B solution at 0 minutes, 80% of A solution: 20% of B solution at 15 minutes, 77% of A solution at 40 minutes: 23% of B solution at 40 minutes 77% of solution A: 23% of solution B, 42 minutes 70% of solution A: 30% of solution B, 65% of solution A at 75 minutes: 35% of solution B, 30% of solution A at 80 minutes: 70% of solution B, 90 minutes The mobile phase was flowed with A solution 10% : B solution 90%. The ginsenoside HPLC chromatograms of Example 1, Example 2 and Comparative Example 1 are shown in Figures 2a to 2c, respectively, and F3, Rg2, F1, Protopanaxatriol (PPT), Rd, Rd2, F2, Rg3, and protopanaxadiol (PPD) contents are shown in Table 1. In addition, for Comparative Examples 1 to 4, Examples 1 and 2, protopanaxatriol series (Rg2, F1, and PPT) ginsenoside, protopanaxadiol series (Rd, Rd2, F2, Rg3, and PPD) ) ginsenoside, and the total ginsenoside content was calculated and shown in FIG.

content (mg/g) Comparative Example 1
(Raw material) Example 1 Example 2 Dongjunghacho top Ginsenoside F3 nd 0.23 0.46 Ginsenoside Rg2 0.48 1.31 1.80 Ginsenoside F1 0.66 1.35 1.27 Protopanaxtriol (PPT) 0.14 0.28 0.62 Ginsenoside Rd 2.65 8.13 6.59 Ginsenoside Rd2 0.37 1.10 4.17 Ginsenoside F2 0.77 2.48 5.33 Ginsenoside Rg3 0.17 1.16 2.86 Protopanaxdiol (PPD) 1.84 9.78 4.53 All experiments were repeated 5 times and displayed as the average value.
nd: not detected.

As shown in Figures 2a to 2c, the composition of Comparative Example 1 showed low F3, Rg2, F1, PPT, Rd, Rd2, F2, Rg3, and PPD peaks, whereas Cordyceps sinensis of Example 1 according to the present invention It can be seen that the F3, Rg2, F1, PPT, Rd, Rd2, F2, Rg3, and PPD peaks in the mushroom mycelium fermentation composition and the enoki mushroom mycelium fermentation composition of Example 2 were significantly increased.

In addition, as shown in Table 1, in the case of Comparative Example 1, which is a mixed raw material for unprocessed goat buds, F3 was not detected, and the Rg2, F1, PPT, Rd, Rd2, F2, Rg3, and PPD contents were 0.48, respectively. 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, but Cordyceps mushroom mycelium fermented composition of Example 1 In the case of F3, Rg2, F1, PPT, Rd, Rd2, F2, Rg3 and PPD contents were 0.23 mg/g, 1.31 mg/g, 1.35 mg/g, 0.28 mg/g, 8.13 mg/g, 1.10 mg/g, respectively. g, 2.48 mg/g, 1.16 mg/g and 9.78 mg/g, and in the case of the enoki mushroom mycelium fermented composition of Example 2, F3, Rg2, F1, PPT, Rd, Rd2, F2, Rg3 and The PPD content increased to 0.46 mg/g, 1.80 mg/g, 1.27 mg/g, 0.62 mg/g, 6.59 mg/g, 4.17 mg/g, 5.33 mg/g, 2.86 mg/g and 4.53 mg/g, respectively. can confirm.

As shown in FIG. 3, protopanaxatriol series (F3, Rg2, F1, and PPT) ginsenosides, protopanaxadiol series (Rd, Rd2, F2, Rg3, and PPD) ginsenosides, and total ginsenoside contents were 1.28 mg/g, 5.80 mg/g, and 7.08 mg/g, respectively, but Protopanaxatri of the Cordyceps mushroom mycelium fermented composition of Example 1 All-type (F3, Rg2, F1, and PPT) ginsenosides, protopanaxadiol-type (Rd, Rd2, F2, Rg3, and PPD) ginsenosides and total ginsenoside contents were 3.17 mg/g, 22.65 mg / g and 25.82 mg / g, respectively, compared to Comparative Example 1, it can be confirmed that the enhancement was about 2.4 times, 3.9 times, and 3.6 times. Protopanaxatriol-based (F3, Rg2, F1, and PPT) ginsenosides, protopanaxadiol-based (Rd, Rd2, F2, Rg3, and PPD) ginsenosides of the fermented enoki mushroom mycelium composition of Example 2 And the total ginsenoside content was 4.15 mg / g, 23.48 mg / g, and 27.63 mg / g, respectively, compared to Comparative Example 1.

In addition, the protopanaxadiol series (Rd, Rd2, F2, Rg3, and PPD) ginsenosides and total ginsenoside contents of Comparative Examples 2 to 4 (12.02 to 14.06 mg/g and 14.68 to 18.39 mg/g, respectively) ), it can be confirmed that in the case of Example 1 and Example 2 according to the present invention, the improvement was about 1.6 times or more and about 1.6 times or more, respectively.

Therefore, it can be seen that the content of ginsenosides F3, Rg2, F1, PPT, Rd, Rd2, F2, Rg3 and PPD in the fermented wild ginseng sprout composition according to the present invention fermented with the mycelium of Cordyceps sinensis and Enoki mushroom is significantly enhanced.

Test Example 2. Analysis of bioactive component content

The total phenolics and total flavonoid contents, which are physiologically active components showing antioxidant activity, were analyzed.

<Total phenolic content>

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

As shown in FIG. 4, the total phenolics content was 3.28 mg/g in Comparative Example 1, 6.08 mg/g in Example 1 and 6.69 mg/g in Example 2, and the fermented composition according to the present invention had a total It can be seen that the phenolics content is significantly increased.

<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 mixture was left in a constant temperature water bath at 37° C. for 1 hour, and then the absorbance was measured with a spectrophotometer at 420 nm. At this time, the total flavonoids content was obtained from a standard curve prepared using rutin, and the results are shown in FIG. 5.

As shown in FIG. 5, the total flavonoid content was 1.36 mg/g in Comparative Example 1, 2.43 mg/g in Example 1 and 2.67 mg/g in Example 2, and the fermented composition according to the present invention had a total flavonoid content. It can be seen that the content of

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 Examples 1, 2, and Comparative Examples 1 to 4 were prepared and used at concentrations of 0.5, 1, and 2 mg/ml.

<DPPH radical scavenging activity>

0.8 ml of DPPH methanol solution (1.5×10 ^-4 M) was added to 0.2 ml of each sample, stirred for 10 seconds, left in the dark for 30 minutes, and then absorbance was measured at 525 nm. The negative control of DPPH radical scavenging activity was performed in the same way using an extraction solvent instead of the sample, and the difference in absorbance was calculated as a percentage (%) by the following equation, and the results are shown in FIG. 6.

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

As shown in FIG. 6, the DPPH radical scavenging activity at a concentration of 1 mg/ml was 17.81%, 58.01%, 62.18%, 66.33%, 72.05%, and 78.15% of activity, respectively, the fermented compositions of Examples 1 and 2 showed significantly higher activity.

<ABTS radical scavenging activity>

Mix 7 mM ABTS ⁺ and 2.45 mM K ₂ S ₂ O ₈ at a ratio of 1:1 and react in the dark for 12 to 16 hours, then mix with methanol at a ratio of 1:88 to obtain a control absorbance value of 0.7±0.02 at 732 nm. A conditioned ABTS ⁺ solution was used. 0.1 ml of each sample and 0.9 ml of ABTS ⁺ solution were added and mixed, and then allowed to stand for 3 minutes, and the absorbance was immediately measured at 732 nm using a spectrophotometer. 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 and calculated in percentage (%) by the above formula, and the results are shown in FIG. 7.

As shown in FIG. 7, the ABTS radical scavenging activity at a concentration of 0.5 mg/ml was 32.93%, 63.61%, 70.73%, 74.76%, 83.74%, and 90.75% of the activity, respectively, the fermented compositions of Examples 1 and 2 showed significantly higher activity.

<FRAP reducing power analysis>

In the FRAP reducing power assay, the reaction solution was 30 mM acetate buffer (pH 3.6) and 10 mM 2,4,6-tripyridyl-s-triazine (TPTZ, T1253, C1 ₈ H ₁₂ N ₆ , dissolved in 40 mM hydrochloric acid). MW312.33) and 20 mM FeCl ₃ (F7134, MW 162.20, in DW) were prepared, and acetate buffer, TPTZ solution and FeCl ₃ solution were mixed at 10:1:1 (v/v/v) at 37 °C. A pre-reaction was allowed for 15 minutes. 0.95 ml of FRAP reagent was added to 0.05 ml of each sample, reacted at 37° C. for about 15 minutes, and absorbance was measured at 590 nm. The results are shown in FIG. 8 .

As shown in FIG. 8, the FRAP reducing powers of Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4, Example 1 and Example 2 at a concentration of 1 mg/ml were 0.44, 0.81, 0.88, 0.98, 1.07, 1.21 OD of _{593 nm,} the fermented composition of Example 1 and Example 2 showed a remarkably high activity.

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

Test Example 4: Digestive enzyme inhibitory activity assay

Alpha-glucosidase inhibitory activity, which is an indicator of anti-diabetic (diabetic improvement) effect, was assayed for the mushroom mycelium fermented composition of wild goat sprouts according to the present invention, and pancreatic lipase inhibitory activity, which is an anti-obesity (obesity improvement) index, was assayed.

<Alpha-glucosidase inhibitory activity>

70 μl of alpha-glucosidase (0.5 U/ml) enzyme solution and 50 μl of 200 mM sodium phosphate buffer solution (pH 6.8) were mixed with 30 μl of the analysis sample, followed by a preliminary reaction at 37° C. for 10 minutes. Thereafter, 100 μl of p-NPG (10 mM) dissolved in sodium phosphate buffer (pH 6.8) was added and reacted at 37° C. for 10 minutes, and 750 μl of Na ₂ CO ₃ (100 mM) was added to the reaction solution to After stopping, absorbance was measured using a spectrophotometer at 420 nm. In the negative control group, an extraction solvent was taken instead of the sample, and the difference in absorbance between the sample added and not added was expressed as a percentage (%), and the results are shown in FIG. 9 .

As shown in FIG. 9, alpha-glucosidase inhibitory activity was 19.84 and 35.66 in Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4, Example 1 and Example 2, respectively, when treated with 2.0 mg/ml. , 38.42, 41.71, 48.27, and 52.33% of the activity, respectively, and the fermented compositions of Examples 1 and 2 showed significantly high enzyme inhibitory activity.

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

<Pancreatic-lipase inhibitory activity>

70 μl of pancreatic lipase (0.5 U/ml) enzyme solution and 50 μl of 200 mM sodium phosphate buffer (pH 6.8) were mixed with 30 μl of the analysis sample, followed by pre-reaction at 37° C. for 10 minutes. After the reaction, 100 μl of p-NPB (10 mM) dissolved in sodium phosphate buffer was added and reacted for 10 minutes in the same manner, and then the reaction was terminated by the addition of 750 μl of 100 mM Na ₂ CO ₃ and absorbance was measured at 420 nm. In the negative control group, the extraction solvent was taken instead of the sample, and the difference in absorbance between the added and unadded groups of the sample solution was expressed in percentage (%), and the results are shown in FIG. 10.

As shown in FIG. 10, the pancreatic lipase inhibitory activity of Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4, Example 1 and Example 2 were 15.46, 35.17, 40.88, 45.66, 51.54, and 57.68% of the activity, respectively, the fermented compositions of Examples 1 and 2 showed significantly high enzyme inhibitory activity.

Therefore, it can be seen that the fermented composition according to the present invention enhances the pancreatic lipase inhibitory activity, thereby enhancing the obesity improvement effect.

From the above activity (functionality) assay results, the fermented composition according to the present invention not only increases the content of ginsenosides F3, Rg2, F1, PPT, Rd, Rd2, F2, Rg3 and PPD, but also has excellent antioxidant activity, It can be seen that it has diabetes and anti-obesity activity and is useful as a material for functional foods and cosmetics.

Claims (12)

Ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, Protopanaxatriol and Protopa prepared by fermenting a mixture of 90-94 wt% of wild ginseng and 6-10 wt% of soybean with mushroom mycelium Naxadiol-enhanced mushroom mycelium fermented composition of wild goat sprouts.
The method of claim 1, wherein the mushroom mycelium is cordyceps mushroom mycelium or enoki mushroom mycelium, ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol are promoted Mushroom mycelium fermented composition of goat ginseng sprouts.
The method of claim 1, wherein the fermentation composition contains ginsenoside F3 at least 0.23 mg/g, Rg2 at least 1.31 mg/g, F1 at least 1.27 mg/g, and protopanaxatriol at least 0.28 mg/g. That is, ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol are enhanced mushroom mycelium fermented composition of wild goat sprouts.
The method of claim 1, wherein the fermented composition has Rd of 6.59 mg/g or more, Rd2 of 1.10 mg/g or more, F2 of 2.48 mg/g or more, Rg3 of 1.16 mg/g or more, and protopanaxadiol of 4.53 mg / g or more containing, ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol is enhanced mushroom mycelium fermented composition of goat sprouts.
According to claim 1, wherein the fermentation is inoculated with mushroom mycelium at a concentration of 3 to 10% (v / w) and fermented at 20 to 30 ° C for 8 to 12 days, ginsenosides F3, Rg2, F1, Rd, Mushroom mycelium fermented composition of wild goat ginseng with enhanced Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol.
The ginseno according to claim 1, wherein the mixture is hydrated for 5 to 7 hours by mixing 5 to 10 times (v/w) of water before fermentation, and then steamed at 100 to 120 ° C for 30 to 60 minutes. Sides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol and protopanaxadiol fermented mushroom mycelium composition of wild goat sprouts.
A method for producing a fermented wild goat sprout mushroom mycelium composition with enhanced ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol, and protopanaxadiol, the method comprising:
i) hydrating for 5 to 7 hours by mixing 90 to 94% by weight of wild goat sprouts and 6 to 10% by weight of soybeans and then mixing with water 5 to 10 times (v / w);
ii) steaming for 30 to 60 minutes at 100 to 120°C; and
iii) A method comprising inoculating mushroom mycelium at a concentration of 3 to 10% (v / w) and fermenting at 20 to 30 ° C. for 8 to 12 days.
The method of claim 7, wherein the mushroom mycelium is cordyceps mushroom mycelium or enoki mushroom mycelium, ginsenosides F3, Rg2, F1, Rd, Rd2, F2, Rg3, protopanaxatriol, and protopanaxadiol Method for producing an improved fermented wild goat sprout mushroom mycelium composition.
A functional food having enhanced antioxidant activity comprising the fermented composition according to claim 1.
A functional food for improving diabetes comprising the fermented composition according to claim 1.
A functional food for improving obesity comprising the fermented composition according to claim 1.
Cosmetics containing the fermented composition according to claim 1.

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