A composition fermented by mycelium of Flammulina velutipes of Fabaton soybean leaves with increased glycitein and formononetin and naringin and a preparation method thereof

Abstract

The present invention discloses a fermented composition of Favaton soybean leaf enoki mushroom mycelium enriched with glycytaine, follononetin, and naringin, and a method for producing the same. The fermented composition according to the present invention contains high amounts of glycytaine, follononetin, and naringin, and also contains high amounts of daidzein and genistein, so it can be used as a material for functional foods and cosmetics.

Description

Technology Field

[0001] The present invention relates to a fermented composition of Favaton soybean leaves and enoki mushroom mycelium enhanced with glycitein, follononetin, and naringin, and a method for producing the same. More specifically, the invention relates to a fermented composition in which the non-glycosidic isoflavones glycitein and follononetin and the flavonoid naringin are significantly enhanced by fermenting Favaton soybean leaves with enoki mushroom mycelium, and a method for producing the same. Background Technology

[0003] Soybean leaves contain isoflavones, which are representative functional components of soybeans, and their glycosylation inhibitory activity varies depending on the growth stage of the leaves. It has been suggested that they can be utilized as an excellent material for the development of functional foods for obesity, diabetes, etc. (Yuk et al., 2011, Food Chem.).

[0004] Recently, cultivation technology for soybean leaves (Favaton soybean leaves) with a high content of isoflavone derivatives was developed using molecular farming technology (Patent Registration No. 10-1451298). However, the isoflavones in these soybean leaves consist of glycoside isoflavones, specifically β-glycoside and malonyl-β-glycoside, accounting for about 98%, while non-glycoside isoflavones with excellent physiological activity are present in amounts of less than 2%, making them insufficient for use as functional foods. Therefore, there is a need for the development of a technology to convert the high content of isoflavone derivatives contained in Favaton soybean leaves into non-glycoside isoflavones, and for the development of processed foods made from Favaton soybean leaves with enhanced active ingredients through this process.

[0005] Isoflavones exist in either glycoside form, in which a beta-glycoside is linked, or aglycone form, in which the sugar has been removed. Aglycon isoflavones include genistein, daidzein, glycitein, and formononetin, while glycoside isoflavones include genistin, daidzin, glycitin, and ononin. In particular, various biological activities are reported to be attributed to aglycone isoflavones in their aglycone form.

[0006] Among non-glycoside isoflavones, genistein is known to be effective against breast and prostate cancers as well as having antioxidant activity, while daidzein is known to be effective against osteoporosis in menopausal women. In particular, glycitein is known to have a preventive effect against oral diseases, and formononetin is known to activate the process of removing apoptotic cells, promote bone formation and muscle differentiation, and prevent hair loss and promote hair growth (Patent Publication No. 10-2017-0139971, Patent Publication No. 10-2022-0091747, Patent Registration No. 10-2390871, Patent Registration No. 10-1664531).

[0007] The Ministry of Food and Drug Safety suggests that isoflavones can help with bone health after menopause as a notified ingredient for health functional foods, and recommends a daily intake of approximately 24 to 27 mg of the total amount of non-glycosidic isoflavone derivatives.

[0008] Naringin is a flavonoid compound found in large quantities in the peels of citrus fruits, known for its antioxidant and anticancer effects, and is also known to promote alcohol metabolism (Patent Registration No. 10-0375047).

[0009] However, no fermented composition of Pavaton soybean leaves and enoki mushroom mycelium has been developed in which the non-glycoside isoflavones folmonetin, daidzein, glycitein and genistein, and the flavonoid naringin are significantly enhanced. The problem to be solved

[0011] Accordingly, the inventors continued research to meet the requirements of the prior art and confirmed that when Favaton soybean leaves are fermented with enoki mushroom mycelium, a fermented composition can be obtained in which glycysteine, follononetin, and naringin are significantly enhanced, and also daidzein and genistein are significantly enhanced, thereby completing the present invention.

[0012] Therefore, the objective of the present invention is to provide a fermented composition of enoki mushroom mycelium from Favaton soybean leaves that is enhanced with glycytaine, follononetin, and naringin.

[0013] Another objective of the present invention is to provide a fermented composition of enoki mushroom mycelium from Favaton soybean leaves that is enhanced with daidzein, glycysteine, genistein, follononetin, and naringin.

[0014] Another objective of the present invention is to provide a method for preparing a fermented composition of enoki mushroom mycelium from Favaton soybean leaves, enhanced with glycytaine, follononetin, and naringin.

[0015] Another objective of the present invention is to provide a method for preparing a fermented composition of enoki mushroom mycelium from Favaton soybean leaves, enhanced with daidzein, glycysteine, genistein, follononetin, and naringin.

[0016] Another objective of the present invention is to provide a functional food containing the above-mentioned fermentation composition.

[0017] Another objective of the present invention is to provide a cosmetic product comprising the above-mentioned fermentation composition. means of solving the problem

[0019] To achieve the above objective, the present invention provides a fermented composition of enoki mushroom mycelium from Favaton soybean leaves enhanced with glycytaine, follononetin, and naringin.

[0020] In addition, the present invention provides a fermented composition of enoki mushroom mycelium from Favaton soybean leaves, enhanced with daidzein, glycysteine, genistein, follononetin, and naringin.

[0021] In the present invention, 'Favaton soybean leaves' refers to soybean leaves containing a high amount of isoflavone derivatives by treating the soybean leaves with a plant growth hormone, such as ethylene, an ethylene donor (ethephon), or an ethylene generator before and after cultivation. Favaton soybean leaves typically contain about 5,000 μg / g or more of isoflavone derivatives and are commercially available.

[0022] In this invention, 'enoki mushroom mycelium' refers to the mycelium of the enoki mushroom. Taxonomically, the enoki mushroom belongs to the genus Pleurotus, family Tricholomataceae, order Agaricales, phylum Basidiomycota. It is an edible mushroom highly popular in Asia due to its unique taste, aroma, and texture, and is divided into a fruiting body and a mycelium. Among these, the mycelium is the body of a fungus that parasitizes organic matter such as fallen leaves and humus in a cottony form; it possesses excellent nutritional value, including dietary fiber, essential amino acids, vitamins, and minerals, and also has functional effects that enhance immunity and anticancer effects by containing polysaccharides such as beta-glucan. Enoki mushroom mycelium can be used at any time regardless of the harvest season, and during mycelial growth, it produces various hydrolytic enzymes, including cellulose-degrading enzymes, proteolytic enzymes, and lipid-degrading enzymes, including β-glucosidase.

[0023] In the present invention, brown rice can be mixed as a nutrient source for the pine mushroom mycelium, such as a carbon source, during the fermentation of Favaton soybean leaves.

[0024] The weight ratio of the mixture of favaton soybean leaves and brown rice is preferably 9:1 to 7:3, and most preferably 8:2.

[0025] If the mixing weight ratio of Favaton soybean leaves to brown rice falls outside the above range, the content of daidzein, glycitein, genistein, and follononetin may decrease, or the mycelial growth of the enoki mushroom mycelium may not be sufficient. When considering both the degree of growth of the enoki mushroom mycelium and the content of daidzein, glycitein, genistein, and follononetin, the mixing weight ratio of Favaton soybean leaves to brown rice is most desirable at 8:2.

[0026] In the present invention, it is preferable to use the favaton soybean leaves after drying and grinding them into a powder, and it is also preferable to use brown rice after grinding it into a powder.

[0027] In the present invention, 'fermentation' refers to fermenting using enoki mushroom mycelium as a starter culture. When Favaton soybean leaves were fermented with enoki mushroom mycelium, surprisingly, daidzein, glycysteine, genistein, follononetin, and naringin were significantly enhanced (Tables 1–2).

[0028] In the present invention, the enoki mushroom mycelium can be inoculated at a concentration of 3 to 10% (v / w) of the total weight of Favaton soybean leaves and brown rice, which is a nutrient source, and fermented at 25 to 30°C for 8 to 12 days. Most preferably, it is fermented for 10 days.

[0029] If the amount of enoki mushroom mycelium inoculated is less than 3% (v / w), the fermentation speed may be delayed, and if it exceeds 10% (v / w), the mycelium proliferation speed may be fast and the conversion rate may be low. If the fermentation temperature is less than 25℃, the fermentation period may be prolonged, leading to contamination by unwanted microorganisms, and if it exceeds 30℃, the growth of the mycelium may be reduced. If the fermentation period is less than 8 days, fermentation may not be sufficient, resulting in poor production of physiologically active substances, and if it exceeds 12 days, physiologically active substances may be decomposed due to over-fermentation.

[0030] In the present invention, the Favaton soybean leaves and brown rice, which is a nutrient source, may be steamed before fermentation. It is preferable to steam at 100°C or higher for 30 to 90 minutes, and most preferably at 120°C for 60 minutes.

[0031] The reason for performing the steaming treatment in the present invention is to sterilize unwanted microorganisms, such as bacteria present in Favaton soybean leaves or brown rice. If the treatment is performed at 100°C or below or for 30 minutes or less, unwanted microorganisms (miscellaneous bacteria) are not completely sterilized, which may affect fermentation. If the treatment is performed at 120°C or above or for 90 minutes or more, excessive heat treatment may lead to significant destruction of nutrients and active ingredients.

[0032] The fermented composition according to the present invention contains at least 1277 μg / g of daidzein, at least 55 μg / g of glycysteine, at least 896 μg / g of genistein, at least 110 μg / g of folmonetin, and at least 4332 μg / g of naringin (Table 1).

[0033] The fermented composition according to the present invention has a newly produced content of daidzein, genistein, and follononetin, respectively, which is about 14.5 times, 10.9 times, and 4.7 times, respectively, compared to the mixture before processing (Comparative Example 1), and the content of daidzein, genistein, and follononetin is increased by about 4.4 times, 1.5 times, 5.9 times, and 3.2 times, respectively, compared to before fermentation (Comparative Example 2) (Table 1).

[0034] In addition, the naringin content of the fermented composition according to the present invention is significantly increased by about 4.4 times or more compared to the mixture before processing (Comparative Example 1) and about 3.8 times or more compared to before fermentation (Comparative Example 2) (Table 2).

[0035] In addition, the fermented composition according to the present invention has enhanced antioxidant activity by increasing the content of physiologically active substances such as daidzein, glycysteine, genistein, follononetin, naringin, total phenolics, and total flavonoids (Tables 1 and 2, Figs. 4 and 5). Specifically, the fermented composition according to the present invention has a total phenolics content of 11.64 mg / g or more and a total flavonoid content of 8.14 mg / g.

[0036] To achieve another objective of the present invention, the present invention provides a method for preparing a fermented composition with enhanced follononetin and naringin by fermenting favaton soybean leaves with enoki mushroom mycelium.

[0037] In addition, the present invention provides a method for preparing a fermented composition in which daidzein, glycysteine, genistein, follononetin, and naringin are enhanced by fermenting favaton soybean leaves with enoki mushroom mycelium.

[0038] Specifically, the above method

[0039] i) A step of mixing Favaton soybean leaf powder and brown rice powder in a weight ratio of 9:1 to 7:3, and then mixing with water at a ratio of 3 to 5 times (v / w);

[0040] ii) a step of steaming at 100℃ or higher for 60 to 120 minutes; and

[0041] ii) Includes a step of inoculating enoki mushroom mycelium at a concentration of 3~10% (v / w) and fermenting at 25~30℃ for 8~12 days.

[0042] In the manufacturing method of the present invention, the weight ratio of the Favaton bean leaves, the mixture, the enoki mushroom mycelium, and the fermentation are as defined above.

[0043] For smooth fermentation, water can be added to a mixture of favaton soybean leaves and brown rice to ferment it. It is preferable to add water in an amount 3 to 5 times (v / w) of the mixture of favaton soybean leaves and brown rice.

[0044] Before fermentation, a mixture of Favaton soybean leaves, brown rice, and water may be steamed to sterilize it. It is preferable to steam at 100°C or higher for 60 to 120 minutes, and most preferably at 121°C for 60 minutes.

[0045] According to another object of the present invention, the present invention provides a functional food comprising a fermented composition enhanced with glycitein, follononetin, and naringin produced by the above-described manufacturing method.

[0046] In addition, the present invention provides a functional food comprising a fermented composition enhanced with daidzein, glycysteine, genistein, follononetin, and naringin, prepared by the above-described manufacturing method.

[0047] The functional food according to the present invention has excellent antioxidant activity (Fig. 5).

[0048] According to another object of the present invention, a cosmetic product comprising a fermented composition enhanced with glycysteine, follononetin, and naringin is provided.

[0049] In addition, the present invention provides a cosmetic product comprising a fermented composition enhanced with daidzein, glycylatein, genistein, formononetin, and naringin.

[0050] The food or cosmetic of the present invention may be prepared by adding the fermented composition or extract thereof of the present invention as is or by mixing it with other food ingredients, and may be prepared appropriately according to conventional methods.

[0051] In the present invention, the types of food are not particularly limited and may include pills, tablets, capsules, fermented tea, fermented foods (kimchi, pickles, etc.), fermented beverages (pouches, drinks, etc.), but are not limited thereto. The types of cosmetics may include mask packs, skin toners, lotions, creams, etc., but are not limited thereto. Effects of the invention

[0053] The fermented composition according to the present invention contains high amounts of daidzein, glycysteine, genistein, follononetin, and naringin, and can be used as a material for functional foods and cosmetics.

[0054] In addition, the functional food or cosmetic according to the present invention has a high content of daidzein, glycylatein, genistein, follononetin, and naringin, and has a high total phenolic content and total flavonoid content, and also has excellent antioxidant activity, so it is useful for improving blood circulation, lowering cholesterol, improving hyperlipidemia, improving obesity, improving immunity, and improving menopausal syndrome in women. Brief explanation of the drawing

[0056] Figure 1 is an example of a manufacturing process diagram of a fermentation composition of the present invention. Figure 2a shows the isoflavone HPLC chromatogram of the raw material mixture of the fermentation composition of the present invention. Figure 2b shows the isoflavone HPLC chromatogram before fermentation after steaming the raw material mixture of the fermentation composition of the present invention. Figure 2c shows the isoflavone HPLC chromatogram of the fermentation composition of the present invention. Figure 3 shows the glycoside isoflavone and non-glycoside isoflavone content of the fermented composition (Example 1), raw material mixture (Comparative Example 1), and pre-fermentation composition (Comparative Example 2) according to the present invention. Figure 4 shows the total phenolics and total flavonoid content of the fermentation composition, raw material mixture, and pre-fermentation composition according to the present invention. Figure 5 shows the DPPH radical scavenging activity and ABTS radical scavenging activity of the fermentation composition, raw material mixture, and pre-fermentation composition according to the present invention. Specific details for implementing the invention

[0057] The present invention is further explained in detail by the following examples. These examples are intended to illustrate the invention and the scope of the invention should not be limited by them.

[0058] Materials and Methods

[0059] Ingredients

[0060] Favaton soybean leaves were cultivated in the Namhae-gun area of ​​Gyeongsangnam-do and supplied in a dried state by JCN Pharm Co., Ltd. Specifically, ethephon was sprayed twice at 24-hour intervals at a concentration of 200 μg / ml on soybean leaves grown for approximately 60 days (R3 growth stage: the period of maximum growth before hull formation) following the sowing of Daewon soybean seeds, ensuring sufficient runny liquid. The soybean leaves were harvested 96 hours after the first spraying, washed with water, and dried in a food dehydrator (35℃). Brown rice was purchased from a large supermarket in Jinju-si.

[0061] Enoki mushroom mycelium ( Flammulina velutipes KCCM 60233) was obtained from the Korean Culture Collection of Microorganisms and used. Yeast Mold broth / agar (PDB / PDA, BD-Difco, Sparks, MD, USA) was used for subculture of Enoki mushroom mycelium.

[0062] <Preparation of Analysis Samples>

[0063] 1 g of powder sample was extracted by adding 50 times the amount of 50% ethanol for 12 hours, filtered using a vacuum filter, and completely concentrated using a vacuum concentrator, followed by freeze-drying using a freeze-dryer (FD-1000, TOKYO RIKAKIKAI, Japan). An extract was prepared by adding 100 times the amount of 50% ethanol to 1 g of the final freeze-dried sample and used as an analytical sample.

[0064] Isoflavone Content Analysis

[0065] The isoflavone content was analyzed using HPLC chromatograms according to the method of Cho et al. (2011). The stationary phase column used for the analysis was Lichrophore 100 RP C18 (ichroCART 125-4, 5μm, 125mm×4mm, Merck KGaA, Darmstadt, Grmany), and the mobile phase solvents were 0.2% glacial acetic acid in water (solution A) and 0.2% acetic acid in acetonitrile (solution B). The mobile phase conditions were analyzed as 0 min-100%, 15 min-90%, 25 min-80%, 30 min-75%, 45 min-65%, and 50 min-65% based on solvent A. 20 µL of the sample was injected, and the mobile phase flow rate was maintained at 1 ml / min at 30°C. Isoflavones were quantified at an absorbance of 254 nm using a diode array UV detector (Agilent 1200 series, Agilent, USA), and the content was calculated by comparing it with the calibration curve of the standard.

[0066] Preparation Example: Preparation of a fermented composition

[0067] Dried, ground Favaton soybean leaf powder and brown rice powder were mixed in a weight ratio of 8:2, 5 times (w / w) purified water was added and hydrated, then steamed at 121°C for 1 hour, and 5% (v / w) of enoki mushroom mycelium liquid culture solution was inoculated and fermented at 25°C for 10 days to prepare a fermented composition according to the present invention (Example 1), and a photograph of the result is shown in Fig. 1.

[0068] For comparison, a raw material mixture was prepared by mixing dried, ground Favaton soybean leaf powder and brown rice powder in a weight ratio of 8:2 (Comparative Example 1). A pre-fermentation composition (Comparative Example 2) was prepared by mixing dried, ground Favaton soybean leaf powder and brown rice powder in a weight ratio of 8:2, adding 5 times (w / w) of purified water to hydrate the mixture, and then steaming it at 121°C for 1 hour.

[0069] Test Example 1: Analysis of Non-glycosidic Isoflavone Content

[0070] The isoflavone content of the raw material mixture of Comparative Example 1, the pre-fermentation composition of Comparative Example 2, and the fermentation composition of Example 1 prepared in the above preparation example was analyzed, and the respective HPLC chromatograms are shown in Figures 2a to 2c and Figure 3, and the respective non-glycosidic isoflavone content of the results is shown in Table 1.

[0071] As can be seen from the chromatograms in Figures 2a to 2c and the graph in Figure 3, in the raw material mixture of Comparative Example 1, the peaks of the glycosides daidzin, genistin, malonyldaidzin, malonylglycitin, malonylgenistin, and ononin were high (Figure 2a), while the peaks of the non-glycosides daidzein, genistein, and formononetin were hardly visible. In the composition before fermentation of Comparative Example 2, the peaks of the glycosides daidzin and genistin were still high, and the peaks of the non-glycosides were only slightly elevated (Fig. 2b), whereas in the fermented composition of Example 1, the peaks of the non-glycosides daidzein, glycitein, genistein, and formononetin were significantly elevated (Fig. 2c).

[0072] Content (µg / g) Comparative Example 1 Comparative Example 2 Example 1 Daidzein 88.15±1.88 287.06±4.16 1277.15±12.35 Glycitein nd 35.94±0.57 55.63±1.08 Genistein 82.01±1.43 151.97±3.80 896.40±9.14 Formonetin 22.98±0.28 33.97±0.61 110.82±1.21 total 193.14 508.94 2340.00

[0073] As shown in Table 1 above, in Comparative Example 1, daidzein, genistein, and folmonetin were found to be 88.15±1.88 μg / g, 82.69±1.43 μg / g, and 22.98±0.28 μg / g, respectively, and glycysteine ​​was not detected. In Comparative Example 2, daidzein, glycysteine, genistein, and folmonetin were found to be 287.06±4.16 μg / g, 35.94±0.57 μg / g, 151.85±3.80 μg / g, and 33.97±0.61 μg / g, respectively. In contrast, the fermented composition of Example 1 showed significantly increased levels of daidzein, glycysteine, genistein, and follononetin, at 1277.15±12.35 μg / g, 55.63±1.08 μg / g, 896.40±9.14 μg / g, and 110.82±1.21 μg / g, respectively. That is, compared to Comparative Example 1, daidzein, genistein, and follononetin in Example 1 increased significantly by approximately 14.5 times, 10.9 times, and 4.7 times, respectively, and glycysteine ​​was newly produced, and compared to Comparative Example 2, daidzein, glycysteine, genistein, and follononetin also increased by approximately 4.4 times, 1.5 times, 5.9 times, and 3.2 times, respectively.

[0074] As shown in Figure 3, Comparative Example 1 (raw material mixture) had a significantly higher glycoside isoflavone content of 4.73 mg / g, but a significantly lower non-glycoside isoflavone content of 0.19 mg / g. Comparative Example 2 (pre-fermentation composition) had a slightly decreased glycoside isoflavone content of 3.0 mg / g, and a non-glycoside isoflavone content of 0.51 mg / g, which increased by about 2.6 times. In contrast, Example 1 (fermented composition) had a significantly decreased glycoside isoflavone content of 0.13 mg / g, and a non-glycoside isoflavone content of 2.34 mg / g, which increased by more than 12.3 times.

[0075] Therefore, it can be seen that the fermented composition of Favaton soybean leaves according to the present invention, fermented with enoki mushroom mycelium, has significantly enhanced content of daidzein, glycysteine, genistein, and follononetin.

[0077] Test Example 2. Naringin Content Analysis

[0078] The naringin content was analyzed for each sample prepared as described above using HPLC (high performance liquid chromatography). An XBridge C18 column (4.6×250 mm, 5 μm, Waters Corp., Milford, MA, USA) was used for analysis. The system was operated at 30°C for 60 minutes at a rate of 1 ml per minute with a linear gradient of 0.5% glacial acetic acid (mobile phase solvent A) and 100% methanol (mobile phase solvent B) using a 0–100% linear gradient, and detection was performed at 270 nm using a UV detector. The results are shown in Table 2.

[0079] Content (μg / g) Comparative Example 1 Comparative Example 2 Example 1 Naringin 978.36±3.63 1139.47±11.71 4332.08±13.13

[0080] As shown in Table 2, the naringin content was 978.36±3.63 μg / g for Comparative Example 1, 1139.47±11.71 μg / g for Comparative Example 2, and 4332.08±13.13 μg / g for Example 1. It can be confirmed that the naringin content of Example 1 was increased by approximately 4.4 times and approximately 3.8 times compared to Comparative Example 1 and Comparative Example 2, respectively.

[0082] Test Example 3. Analysis of Bioactive Component Content

[0083] The content of total phenolics and total flavonoids, which are bioactive components, was analyzed.

[0084] <Total Phenolic Content>

[0085] 0.5 ml of the analytical sample was dispensed into a test tube, 0.5 ml of 25% Na2CO3 solution was added, and the mixture was allowed to stand for 3 minutes. Then, 0.25 ml of 2N Folin-Ciocalteu phenol reagent was added and mixed, followed by standing at 30°C for 1 hour. Afterward, the absorbance was measured at 750 nm using a spectrophotometer. The total phenolic content was determined from a standard curve prepared using gallic acid and calculated as an amount equivalent to gallic acid; the results are shown in Figure 4.

[0086] As shown in Figure 4, the total phenolic content was 3.48 mg / g and 5.76 mg / g in Comparative Example 1 and Comparative Example 2, respectively, and 11.64 mg / g in Example 1, confirming that the total phenolic content of the fermented composition according to the present invention is increased by more than twofold.

[0087] Total Flavonoid Content

[0088] 1.0 ml of diethylene glycol was dispensed into 0.5 ml of the analysis sample, followed by the addition of 0.01 ml of 1 N NaOH. After leaving the mixture in a 37°C water bath for 1 hour, the absorbance was measured at 420 nm using a spectrophotometer. The total flavonoid content was determined from standard curves prepared by setting the final concentration of rutin to 0, 0.25, 0.5, and 1.0 mg / ml, and the results are shown in Figure 4.

[0089] As shown in Figure 4, the total flavonoid content was 5.38 mg / g, 6.06 mg / g, and 8.14 mg / g in Comparative Example 1, Comparative Example 2, and Example 1, respectively, confirming that the total flavonoid content of the fermented composition according to the present invention is also significantly increased.

[0091] Test Example 4. Antioxidant Activity Analysis

[0092] Antioxidant activity was analyzed by measuring DPPH radical scavenging activity and ABTS radical scavenging activity.

[0093] <DPPH 라디칼 소거활성>

[0094] 0.2 ml of DPPH methanol solution (1.5 × 10⁻⁶) to each analytical sample -4 The test was performed by adding 0.8 ml of M, stirring for 10 seconds, leaving it in a dark room for 30 minutes, and measuring the absorbance at 525 nm. The negative control for DPPH radical scavenging activity was conducted in the same manner using the extraction solvent instead of the sample, and the difference in absorbance was calculated as a percentage (%) using the following formula, and the results are shown in Figure 5.

[0095] Radical scavenging activity (%) = [1 - (Absorbance of negative control ÷ Absorbance of experimental group)] × 100

[0096] As shown in Figure 5, the DPPH radical scavenging activities of Comparative Example 1, Comparative Example 2, and Example 1 were 43.42%, 49.31%, and 76.2%, respectively, and the fermented composition of Example 1 showed the highest radical scavenging activity.

[0097] <ABTS 라디칼 소거활성>

[0098] 7 mM ABTS + ABTS mixed with 2.45 mM K2S2O8 in a 1:1 ratio and reacted in a dark room for 12–16 hours, then mixed with methanol in a 1:88 ratio and adjusted so that the absorbance value of the control at 732 nm was 0.7±0.02. + 0.1 ml of each sample and ABTS solution were used. + After adding and mixing 0.9 ml of the solution and letting it stand for 3 minutes, the absorbance was immediately measured at 732 nm using a spectrophotometer. The negative control experiment was conducted using the extraction solvent instead of the sample, and the absorbances of the experimental group and the negative control were determined and calculated as a percentage (%) using the above formula, with the results shown in Fig. 5.

[0099] As shown in Figure 5, the ABTS radical scavenging activities of Comparative Example 1, Comparative Example 2, and Example 1 were 60.66%, 63.22%, and 91.56%, respectively, showing that the fermented composition of Example 1 had the highest activity.

[0100] From these results, it can be seen that the fermented composition according to the present invention has significantly enhanced antioxidant activity.

Claims

Claim 1 A fermented composition of enoki mushroom mycelium from favaton soybean leaves, prepared by fermenting a mixture of favaton soybean leaves and brown rice in a weight ratio of 9:1 to 7:3 with enoki mushroom mycelium, wherein the fermented composition contains glycitein 55 μg / g or more, enoki mushroom mycelium 110 μg / g or more, and naringin 4332 μg / g or more. Claim 2 delete Claim 3 delete Claim 4 In claim 1, the above mixture is a fermented composition of enoki mushroom mycelium from Favaton soybean leaves enhanced with glycytaine, follononetin, and naringin, which is steamed at 100°C or higher for 60 to 120 minutes prior to fermentation. Claim 5 delete Claim 6 delete Claim 7 delete Claim 8 delete Claim 9 A method for preparing a fermented composition of Favaton soybean leaves and enoki mushroom mycelium enhanced with glycytaine, follononetin, and naringin according to claim 1, wherein the method comprises: i) a step of mixing Favaton soybean leaf powder and brown rice powder in a weight ratio of 9:1 to 7:3 and then mixing with water at a ratio of 3 to 5 times (v / w); ii) a step of steaming at 100°C or higher for 60 to 120 minutes; and ii) a step of inoculating enoki mushroom mycelium at a concentration of 3 to 10% (v / w) and fermenting at 25 to 30°C for 8 to 12 days. Claim 10 delete Claim 11 A food having enhanced antioxidant activity comprising a fermented composition according to claim 1. Claim 12 Cosmetic having enhanced antioxidant activity comprising a fermented composition according to claim 1. Claim 13 delete Claim 14 delete

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