KR102545365B1 - Fermentation composition of mountain-cultivated ginseng having increased ginsenoside Rd, Rc and Protopanaxadiol by using Tricholoma matsutake mycelium and preparation method thereof - Google Patents

Abstract

In the present invention, a fermented composition with significantly enhanced ginsenosides Rd and Rc, protopanaxadiol, oleic acid and linoleic acid prepared by fermenting wild ginseng with matsutake mushroom mycelium and a method for producing the same are disclosed.
Wild ginseng matsutake mushroom mycelium fermented composition according to the present invention contains a high content of ginsenosides Rd, Rc and protopanaxadiol, oleic acid, and linoleic acid, so it can be used as a material for functional foods and cosmetics.

Description

Fermentation composition of mountain-cultivated ginseng having increased ginsenoside Rd, Rc and Protopanaxadiol by using Tricholoma matsutake mycelium and preparation method thereof }

The present invention relates to a fermented composition of wild ginseng matsutake mushroom mycelium enriched with ginsenosides Rd, Rc and protopanaxadiol and a method for producing the same, and more particularly, to a ginsenoside Rd prepared by fermenting wild ginseng with matsutake mushroom mycelium. , Rc and protopanaxadiol, oleic acid and linoleic acid are remarkably enhanced, and a method for preparing 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.

Saponin is a generic term for triterpene and steroid-based glycoside compounds widely distributed in the plant kingdom. Wild ginseng, which contains ginsenoside, a saponin of hemp, is highly evaluated for efficacy next to natural wild ginseng in herbal medicine prescriptions. In addition, it has been reported that the ginsenoside content is higher than that of cultivated ginseng, and it is known that the efficacy is superior to that of cultivated ginseng. However, known wild ginseng-related processed products do not contain rare ginsenosides Rd and Rc, or the content of ginsenosides is below the standard level or has pharmacological action.

Ginsenoside Rd has been reported to enhance immunity, improve brain cognitive function, improve neurodegenerative brain diseases, and improve wrinkles. Ginsenoside Rc is known to have antidiabetic and anti-inflammatory effects, and ginsenoside protopanaxadiol It is known for its sedative, immune enhancing, detoxifying, anticancer, and antioxidant effects (Patent No. 10-1198266).

Oleic acid is classified as a monounsaturated omega-9 fatty acid, and linoleic acid is a polyunsaturated omega-6 fatty acid and is one of the essential fatty acids for humans that must be consumed through food. Oleic acid and linoleic acid are reported to lower blood cholesterol levels, reduce low-density cholesterol, lower blood pressure, prevent heart disease, and have skin moisturizing effects.

However, a fermented composition in which ginsenosides Rd and Rc, protopanaxadiol, oleic acid and linoleic acid are significantly enhanced has not been developed.

Accordingly, as a result of continuing research to meet the needs of the prior art, the inventors of the present invention confirmed that when wild ginseng was fermented with matsutake mushroom mycelium, ginsenosides Rd, Rc, protopanaxadiol, oleic acid and linoleic acid were remarkably enhanced. invention was completed.

Therefore, an object of the present invention is to provide a fermented composition of matsutake mushroom mycelium of wild ginseng, enriched with ginsenosides Rd, Rc, protopanaxadiol, oleic acid and linoleic acid.

Another object of the present invention is to provide a method for producing a fermented composition of matsutake mushroom mycelium of wild ginseng, which is enriched with ginsenosides Rd, Rc, protopanaxadiol, oleic acid and linoleic acid.

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

Another object of the present invention is to provide a cosmetic product containing the fermented matsutake mushroom mycelium composition of wild ginseng of the present invention.

In order to achieve the above object, the present invention ferments the matsutake mushroom mycelium of wild ginseng with enhanced ginsenosides Rd, Rc and protopanaxadiol, oleic acid and linoleic acid prepared by fermenting a mixture of wild ginseng, soybeans and brown rice into matsutake mushroom mycelium. composition is provided.

In the present invention, wild ginseng refers to ginseng produced without installing artificial facilities such as shading in the production area defined in Article 2, Subparagraph 1 of the 「Mountain Management Act」, preferably grown between 3 and 5 years. .

In the mixture of the present invention, it is preferable to include 90 to 94% by weight of wild ginseng, 3 to 5% by weight of soybeans, and 3 to 5% by weight of brown rice, most preferably 90% by weight of wild ginseng, 5% by weight of soybeans, and brown rice. 5% by weight.

When the weight % of wild ginseng is out of the above range, the content of ginsenosides Rd, Rc and protopanaxadiol, oleic acid and linoleic acid falls, and when the weight % of beans and brown rice is outside the above range, mycelial growth of matsutake mushroom mycelium may not be enough. Considering the degree of growth of matsutake mushroom mycelium and the contents of ginsenosides Rd, Rc, protopanaxadiol, oleic acid and linoleic acid, the weight ratio of wild ginseng: soybean: brown rice is most preferably 90: 5: 5.

In the present invention, beans and brown rice mainly serve as nitrogen and carbon sources necessary for the growth of matsutake mushroom mycelium, but also function to enhance fermentation (material conversion action) of matsutake mushroom mycelium.

In the present invention, wild ginseng is preferably used after washing, drying, and cutting, and beans and brown rice are used after washing and drying.

In the present invention, 'matsutake mushroom mycelium' means the mycelium of pine mushrooms. Matsutake mushroom mycelium is available at any time regardless of harvest time and has effects such as anti-cancer, body fat reduction, blood cholesterol lowering and immunity enhancement effects similar to fruiting bodies. It produces various hydrolytic enzymes such as lyase and lipolytic enzyme.

In the present invention, 'fermentation' means fermentation using matsutake mushroom mycelium as a spawn. When a mixture of wild ginseng, soybean and brown rice was fermented with matsutake mushroom mycelium, surprisingly, ginsenosides Rd and Rc and protopanaxadiol, oleic acid and linoleic acid were significantly increased (Tables 1 and 2).

In the present invention, the matsutake mushroom mycelium can be performed by inoculating a mixture of wild ginseng, beans and brown rice at a concentration of 3 to 10% (v / w) and fermenting at 25 to 30 ° C for 8 to 12 days. Most preferably fermented for 10 days.

If the inoculated amount of matsutake mushroom mycelium is less than 3% (v/w), the fermentation rate may be delayed, and if it exceeds 10% (v/w), the mycelium growth rate is fast, and the conversion rate may be low. If the fermentation temperature is less than 25℃, fermentation If the period is prolonged, it causes contamination of various germs, and if the temperature exceeds 30 ℃, the growth of the mycelium may be reduced. If the fermentation period is less than 8 days, fermentation is not sufficient, resulting in low production of physiologically active substances. However, if it exceeds 12 days, physiologically active substances may be decomposed due to over-fermentation.

In the present invention, the mixture of wild ginseng, soybean, and brown rice is hydrated for 5 to 8 hours by adding 2 to 3 times (v/v) of water before fermentation to adjust the moisture to about 40 to 50%, and then to 30% at 100 ° C or higher. ~90 min steaming is preferred. Most preferably, it is cooked for 60 minutes at 121°C.

In the present invention, during the hydration treatment, the growth of matsutake mushroom mycelia with a moisture content of 40% or less is not smooth, and when the moisture content is 50% or more, not only the growth of matsutake mushroom mycelia but also the growth of bacteria can be achieved. On the other hand, when the steaming treatment is performed for less than 30 minutes, sterilization is not completely performed, so abnormal fermentation may proceed due to the proliferation of bacteria in addition to matsutake mushroom hyphae, and when treated for more than 90 minutes, nutritional components may be destroyed excessively.

The fermented matsutake mushroom mycelium composition of wild ginseng according to the present invention has ginsenoside Rd of 2.64 mg/g or more, ginsenoside Rc of 2.45 mg/g or more and ginsenoside protopanaxadiol of 7.1 mg/g or more, oleic acid 263 mg/100 g or more and 859 mg/100 g or more of linoleic acid (Table 1, Table 2).

In the fermented matsutake mushroom mycelium composition of wild ginseng according to the present invention, the contents of ginsenosides Rd, Rc, and protopanaxadiol were about 6.7 times or more, about 1.5 times or more, and about 3.2 times, respectively, compared to the mixture before processing (Comparative Example 1). It is promoted more than before fermentation (Comparative Example 2) by about 3.5 times or more, about 1.2 times or more, and about 3.2 times or more, respectively (Table 1).

In addition, in the fermented matsutake mushroom mycelium composition of wild ginseng according to the present invention, the content of oleic acid and linoleic acid is increased by about 2.1 times or more and about 1.8 times or more, respectively, compared to the mixture before processing (Comparative Example 1) and before fermentation (Comparative Example 2) ( Table 2).

In addition, the fermented matsutake mushroom mycelium composition of wild ginseng according to the present invention has enhanced content of physiologically active substances such as ginsenosides Rd, Rc, protopanaxadiol, oleic acid, linoleic acid, total phenolics and total flavonoids, and browning substances. antioxidant activity (Table 1, Table 2, Fig. 3a ~ Fig. 4d).

In addition, the fermented matsutake mushroom mycelium composition of wild ginseng according to the present invention has improved alpha-glucosidase inhibitory activity and pancreatic-lipase inhibitory activity, thereby improving diabetes and obesity improvement (Figs. 5 and 6).

In order to achieve another object of the present invention, the present invention fermented a mixture of wild ginseng, soybeans and brown rice with matsutake mushroom mycelium, and ginsenosides Rd, Rc and protopanaxadiol, oleic acid, and linoleic acid were enhanced. It provides a method for producing a mushroom mycelium fermentation composition.

Specifically, the method

i) mixing 90-94% by weight of wild ginseng, 3-5% by weight of soybeans and 3-5% by weight of brown rice, then mixing with water 2-3 times (v / w) and hydrating for 5-7 hours;

ii) steaming at 100°C or higher for 30 to 90 minutes; and

iii) inoculating matsutake mushroom mycelium at a concentration of 3 to 10% (v/w) and fermenting at 25 to 30 ° C for 8 to 12 days.

In the manufacturing method of the present invention, the weight ratio, hydration, steaming, matsutake mushroom mycelium and fermentation of the mixture of wild ginseng, soybean and brown rice are as defined above.

For smooth fermentation of matsutake mushroom mycelium, water can be added to the mixture of wild ginseng, soybean and brown rice before fermentation to hydrate it.

Prior to fermentation, it may be steamed to sterilize. It is preferable to steam for 30 to 90 minutes at 100 ° C or higher, and most preferably for 60 minutes at 121 ° C.

According to another object of the present invention, the present invention is a functional food comprising a fermented wild ginseng matsutake mushroom mycelium composition with enhanced ginsenosides Rd, Rc and protopanaxadiol, oleic acid, and linoleic acid prepared by the above production method to provide.

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, to provide a cosmetic comprising a fermented wild ginseng matsutake mushroom mycelium composition with enhanced ginsenosides Rd, Rc, protopanaxadiol, oleic acid, and linoleic acid.

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.

Wild ginseng matsutake mushroom mycelium fermented composition according to the present invention contains a high content of ginsenosides Rd, Rc and protopanaxadiol, oleic acid, and linoleic acid, so it 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 ginsenosides Rd, Rc, protopanaxadiol, oleic acid, and linoleic acid, and has excellent antioxidant activity, alpha-glucosidase inhibitory activity, and pancreatic-lipase inhibitory activity. It is useful for inhibiting fat production, controlling weight, lowering cholesterol, improving hyperlipidemia, relieving arteriosclerosis, improving diabetes, improving obesity, improving blood circulation, improving immunity, and improving female menopausal syndrome.

1 is an example of a manufacturing process diagram of the fermentation composition of the present invention.
Figure 2 shows a ginsenoside HPLC chromatogram. (A) is an HPLC chromatogram of 21 ginsenoside standard materials, (B) is a ginsenoside HPLC chromatogram of Comparative Example 1, (C) is a ginsenoside HPLC chromatogram of Comparative Example 2, ( D) is a ginsenoside HPLC chromatogram of the fermented composition according to the present invention.
Figure 3a shows the total phenolics content of the fermentation composition according to the present invention.
Figure 3b shows the total flavonoid content of the fermented composition according to the present invention.
Figure 3c shows the content of the browning material of the fermentation composition according to the present invention.
Figure 4 will show the antioxidant activity of the fermented composition according to the present invention. 4A shows DPPH radical scavenging activity, FIG. 4B shows ABTS radical scavenging activity, FIG. 4C shows hydroxyl radical scavenging activity, and FIG. 4D shows reducing power (FRAP).
Figure 5 shows the alpha-glucosidase inhibitory activity of the fermented composition according to the present invention.
Figure 6 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

Wild ginseng grown in Baekjeon-myeon, Hamyang-gun in 2020 was supplied and used by Ginseng Viro Co., Ltd., an agricultural corporation. Beans (duck roe beans) and brown rice were purchased from a large mart in Jinju City and used.

Matsutake mushroom mycelium was stored in the Wood Chemistry Laboratory, Department of Forestry Engineering, Gyeongsang National University, and was used. Potato Dextrose broth/agar (PDB/PDA, BD-Difco, Sparks, MD, USA) was used for subculture of matsutake mycelium.

Wild ginseng was prepared by washing three times in running water, cutting to about 1 cm, and then drying at 55 ° C for 3 days. A mixed sample of wild ginseng was prepared by mixing dried wild ginseng, beans, and brown rice at a weight ratio of 90:5:5.

After adding 2 times (v/w) purified water to the wild ginseng mixed sample and hydrating it for 6 hours to adjust the moisture content to about 40%, it was steamed at 121℃ for 1 hour using a high-pressure sterilizer, and the matsutake mushroom mycelium culture medium was 5 After % (v/w) inoculation, fermentation was performed at 25° C. for 10 days to obtain a fermentation composition (Example 1; FIG. 1).

For comparison, dried wild ginseng, soybean, and brown rice were mixed at a weight ratio of 9:0.5:0.5, and 2 times (v/v) purified water was added to the mixed wild ginseng sample composition (Comparative Example 1) and the mixed wild ginseng sample, and hydrated for 6 hours. After adjusting the moisture content to about 40%, a steamed sample composition (Comparative Example 2) was prepared by steaming at 121 ° C. for 1 hour.

Reference example: analysis sample

Powder samples were prepared by drying the fermented composition of Example 1, the composition of Comparative Example 1, and the composition of Comparative Example 2 at 55 ° C. for 3 days, respectively, and then pulverizing them.

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 and 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 ㎛ 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 0.25, 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, Comparative Example 1 and Comparative Example 2 are shown in FIG. 2, and the analyzed ginsenoside Rd, Rc and protopanaxadiol contents are shown in Table 1.

Content (mg/g d.w.) Comparative Example 1 Comparative Example 2 Example 1 Ginsenoside Rc (10) 1.55±0.0.8 2.07±0.10 2.45±0.12 Ginsenoside Rd (14) 0.39±0.02 0.74±0.04 2.64±0.13 Protopanaxadiol (21) 2.16±0.11 3.15±0.16 7.10±0.36

As shown in Figure 2, the compositions of Comparative Examples 1 and 2 showed low ginsenoside Rd peaks (14), Rc peaks (10) and protopanaxadiol peaks (21), whereas according to the present invention In the fermented composition of Example 1, it can be seen that the ginsenoside Rd peak (14), the Rc peak (10), and the protopanaxadiol peak (21) are significantly increased.

In addition, as shown in Table 1, the fermented composition of Example 1 had ginsenoside Rd of 2.64±0.13 mg/g, ginsenoside Rc of 2.45±0.12 mg/g, and ginsenoside protopanaxadiol of 7.10. ± 0.36 mg / g, respectively, about 6.7 times or more compared to Comparative Example 1 (0.39 ± 0.02 mg / g, 1.55 ± 0.0.8 mg / g, 2.16 ± 0.11 mg / g), which is an unprocessed mixed raw material from goat production. . It can be confirmed that the above and about 2.2 times or more were enhanced.

Therefore, it can be seen that the fermented wild ginseng composition according to the present invention fermented with matsutake mushroom mycelium has significantly enhanced contents of ginsenosides Rd, Rc and protopanaxadiol.

Test Example 2: Analysis of unsaturated fatty acid content

Analysis of unsaturated fatty acids (oleic acid, linoleic acid) was performed according to the fatty acid analysis method reported by Hwang et al. 1 g of each of the compositions of Example 1, Comparative Example 1, and Comparative Example 2 prepared in Preparation Example was accurately weighed in a test tube, 3 ml of 0.5 N methanolic NaOH was added thereto, and heat treatment was performed at 100 ° C. for 10 minutes to hydrolyze fatty acids and glycerol process was performed. Thereafter, 2 ml of boron trifluoride (BF ₃ ) was additionally added, stirred, and heat-treated again for 30 minutes to proceed with methyl esterification of fatty acids. After completion of the methyl esterification reaction, 1 ml of isooctane was added, shaken vigorously, and allowed to stand to recover only the isooctane layer, which was then dehydrated with anhydrous sodium sulfate, filtered through a 0.45 μm-membrane filter, and GC (nitrogen and hydrogen gas and SP-2560 capillary column ( 100 m×0.25 mm id, 0.25-μm film thickness, Sigma-Aldrich Co., St. Louis, MO, USA) The analysis results are shown in Table 2.

Content (mg/100 g) Comparative Example 1 Comparative Example 2 Example 1 Oleic acid (C18:1c) 124.4±6.22 124.4±6.22 263.4±13.17 Linoleic acid (C18:2c) 466.0±23.30 466.0±23.30 859.3±42.97

The contents of oleic acid and linoleic acid, which are unsaturated fatty acids, were 124.4 ± 6.22 mg / 100g and 466.0 ± 23.30 mg / 100g, respectively, in the compositions of Comparative Example 1 and Comparative Example 2, confirming that the content of unsaturated fatty acids was not increased by the steaming treatment. can

In contrast, the contents of oleic acid and linoleic acid in the fermentation composition of Example 1 were 263.4 ± 13.17 mg / 100 g and 859.3 ± 42.97 mg / 100 g, respectively, about 2.1 times and about 1.8 times compared to Comparative Examples 1 and 2, respectively. It can be confirmed that it has been enhanced.

Test Example 3. Analysis of bioactive component content

Total phenolics, total flavonoids content, and browning material content, 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 result is shown in FIG. 3a.

As shown in Figure 3a, the total phenolics content was 3,26 and 4.88 mg / g in Comparative Example 1 and Comparative Example 2, respectively, and 5.56 mg / g in Example 1, the fermentation composition according to the present invention It can be confirmed that the phenolics content is 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 content of total flavonoids was obtained from a standard curve prepared using rutin, and the results are shown in FIG. 3B.

As shown in FIG. 3B, the total flavonoid content was 0.35, 0.86 and 1.30 mg/g in Comparative Example 1, Comparative Example 2 and Example 1, respectively. A significant increase can be seen.

<Content of browning substances>

Browning substances were measured using a non-enzymatic browning method. After adding 10 times of tertiary distilled water to 1 g of the powder sample, extracting for 1 hour, filtering with a 0.45 μm filtration filter, taking 1 ml of the filtrate, measuring the absorbance at 420 nm with a spectrophotometer, and the results are shown in FIG. 3C.

As shown in Figure 3c, the browning material content, Comparative Example 1, Comparative Example 2, and Example 1, 0.385, 0.893, and 2.240 OD _{420 nm} , the fermentation composition according to the present invention also significantly increased the content of browning material can confirm that

Test Example 4. Antioxidant activity analysis

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

As in the Reference Example, the analysis samples of Example 1, Comparative Example 1, and Comparative Example 2 were prepared and used at concentrations of 0.25, 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 manner 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. 4a.

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

As shown in Figure 4a, the DPPH radical scavenging activity was 16.67%, 49.76%, and 54.41% of Comparative Example 1, Comparative Example 2, and Example 2, respectively, at a concentration of 1 mg/ml. showed the highest 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 12-16 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 as a percentage (%) by the above formula, and the results are shown in FIG. 4B.

As shown in FIG. 4B, the ABTS radical scavenging activity was 26.22%, 39.13%, and 47.60% in Comparative Example 1, Comparative Example 2, and Example 2, respectively, at a concentration of 0.5 mg/ml. showed the highest activity.

<Hydroxyl radical scavenging activity>

0.2 ml of 10 mM FeSO ₄ 7H ₂ O-EDTA, 0.2 ml of 10 mM 2-deoxyribose, and 0.2 ml of 10 mM H ₂ O ₂ were dispensed into 1.4 ml of each sample into a test tube and reacted at 37°C for 4 hours. . After adding 1 ml of 1% TBA and 2.8% TCA and developing color at 100° C. for 20 minutes, absorbance was measured at a wavelength of 520 nm. The negative control group was tested using a PBS buffer solution instead of the sample, and the absorbance of the experimental group and the negative control group was obtained and calculated as a percentage (%) by the above formula, and the result is shown in FIG. 4c.

As shown in Figure 4c, the hydroxyl radical scavenging activity of Comparative Example 1, Comparative Example 2, and Example 2 at a concentration of 1 mg/ml showed 35.57%, 45.36%, and 50.38% of the activity, respectively. The composition showed the highest activity.

<FRAP reducing power analysis>

In the FRAP reducing power assay, the reaction solution was 30 mM acetate buffer (pH 3.6), 10 mM 2,4,6-tripyridyl-s-triazine (TPTZ, T1253, C18H12N6, MW312.33) dissolved in 40 mM hydrochloric acid, and 20 mM FeCl ₃ (F7134, MW 162.20, in DW) was prepared, and the acetate buffer, TPTZ solution, and FeCl ₃ solution were mixed at a ratio of 10:1:1 (v/v/v), followed by a pre-reaction at 37 °C for 15 minutes. I let it go. 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. 4D.

As shown in FIG. 4d, the FRAP reducing power of Example 1 at a concentration of 1 mg/ml showed the highest activity of 1.029 OD _{593 nm} .

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 5: Digestive enzyme inhibitory activity assay

Alpha-glucosidase inhibitory activity, which is an indicator of antidiabetic (diabetic improvement) effect, was assayed for the fermented matsutake mushroom carcass composition of wild ginseng 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. The negative control group took the extraction solvent instead of the sample, and the difference in absorbance between the analysis sample added and unadded was expressed as a percentage (%), and the results are shown in FIG. 5.

As shown in FIG. 5, alpha-glucosidase inhibitory activity was 14.142% in Comparative Example 1 when treated with 2.0 mg/ml, but increased to 32.83% in Example 1.

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, an 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. 6.

As shown in FIG. 6, the pancreatic lipase inhibitory activity was 10.00% in Comparative Example 1 and 28.77% in Comparative Example 2 when treated with 2.0 mg/ml, but in Example 1, it showed a higher activity at 35.12%.

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 improves the contents of ginsenosides Rd, Rc and protopanaxadiol, oleic acid, and linoleic acid, but also has excellent antioxidant activity, antidiabetic and antiobesity It can be seen that it has activity and is useful as a material for functional foods and cosmetics.

Claims (8)

After mixing 2-3 times (v/w) of water with a mixture of 90-94% by weight of wild ginseng, 3-5% by weight of soybean, and 3-5% by weight of brown rice, hydrated for 5-7 hours, and heated to 100℃ or higher. After steaming for 30 to 90 minutes, matsutake mushroom mycelium was inoculated at a concentration of 3 to 10% (v/w) and fermented at 25 to 30 ° C for 10 to 12 days to produce ginsenosides Rd, Rc and protopanaxadiol. , A composition for fermenting matsutake mushroom mycelium of wild ginseng with enhanced oleic acid and linoleic acid,
The fermented composition contains ginsenoside Rd of 2.64 mg/g or more, ginsenoside Rc of 2.45 mg/g or more, ginsenoside protopanaxadiol of 7.1 mg/g or more, oleic acid of 263 mg/100g or more, and linoleic acid of 859 mg/100g or more. Fermentation composition characterized in that it contains.
delete delete delete delete delete A functional food for improving diabetes and obesity, comprising the fermented composition according to claim 1.
Cosmetics containing the fermented composition according to claim 1.

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