KR102696728B1 - Method for Preparing Fermented Ginseng with Increased Ginsenosides by enzyme or acid treatment and sterilization - Google Patents

Abstract

Disclosed is a method for preparing an enzyme or acid-treated and sterilized ginseng fermented product in which the content of ginsenoside Rg3, F2, or compound K is significantly increased by treating a product, obtained by fermenting ginseng with the genus bacillus microorganisms, with an enzyme having activity such as pectinase, hemicellulase, beta-glucanase, polygalacturonase, or adding citric acid and then sterilizing the product.

Description

Method for preparing fermented ginseng with increased ginsenoside content by enzyme or acid treatment and sterilization

The present invention relates to a method for producing fermented ginseng (ginseng root, Ginsen radix) having increased ginsenoside content and treated with enzymes or acids and sterilized.

Ginseng, a perennial herbaceous plant, belongs to the genus Panax , of which 6-7 species are known worldwide, and has been widely used as a medicinal herb and health tonic in East Asian countries since 2,000 years ago. The main component showing pharmacological activity in ginseng is ginsenoside, a triterpenoid saponin, and various physiological activities beneficial to the human body, such as anticancer and antidiabetic, have been reported (J. Ginsen Res. 27:129-134, 2003; Mol. Biol. Rep. 37:2975-2979, 2009). The major ginsenosides found in ginseng include Rb1, Rb2, Rc, Re, and Rg1. In addition to the major ginsenosides, minor ginsenosides exist in trace amounts and are small molecular weight substances created when the sugar portion of ginsenoside is hydrolyzed. They are also called minor ginsenosides and are known to have a higher absorption rate and superior physiological activity than the major ginsenosides (J. Ginseng Res. 27:129-134, 2003; J. Agric. Food Chem. 57:5777-5782, 2009; J. Agric. Food Chem. 51:2555-2558, 2003). Ginsenoside F2, a precursor of compound K, has been reported to have activity in improving degenerative brain diseases and enhancing memory by inhibiting damage to nerve cells (Korean Patent No. 10-1509056), and has also been reported to have excellent anticancer effects (Cancer Lett. 321:144-153, 2012; Food Anal. Methods 6:112-122, 2013). Ginsenoside Rg3 has been reported to have effects such as enhancing immunity, improving blood flow, recovering from fatigue, having anticancer effects, and improving liver function (Korean J. Medic. Crop Sci., 21, 401-404, 2013).

There are examples of increasing the contents of Rh2, Rg2, Rg3, etc. through fermentation of ginseng (Korean J. Medicinal Crop Sci. 20(2):117-123, 2012), and reports of increasing the contents of Rg3, etc. through fermentation of ginseng leaves (Korean Journal of Food Science and Nutrition 39(8):1194-1200, 2010), but studies that significantly increased the contents of other minor ginsenosides, such as compounds K or F2, are still rare.

Accordingly, the present invention discloses a method for producing fermented ginseng, which has increased contents of ginsenoside compond K, F2, and Rg3, and has been treated with enzymes or citric acid and sterilized .

Republic of Korea Patent Publication No. 10-2106469 (April 24, 2020).

Kim, Do-Yeon et al. 2016. Production of Minor ginsenoside F2 from ginseng saponin using Viscozyme®. J. Chitin. Chitosan 21(2):94-99, 2016.

The purpose of the present invention is to provide a method for producing an enzyme-treated ginseng fermented product having an increased ginsenoside content.

The purpose of the present invention is to provide a method for producing a fermented ginseng product (enzyme-treated) having an increased content of a specific ginsenoside component through enzyme treatment, citric acid treatment, or sterilization.

Other purposes or specific purposes of the present invention will be presented below.

The present inventors confirmed in the examples below that when a product fermented with ginseng (hereinafter, referred to as ginseng, unless otherwise specified, means ginseng radix) powder by Bacillus sp. microorganisms was treated with an enzyme, ginsenoside compounds K and F2 increased, and when sterilized or sterilized after adding citric acid, the content of ginsenoside Rg3 significantly increased.

The present invention is provided based on these experimental results, and relates to a method for producing fermented ginseng with increased ginsenoside content, which has been treated with enzymes or acids and sterilized. The method for producing fermented ginseng with increased ginsenoside content, which has been treated with enzymes or acids and sterilized, comprises (1) a step of producing fermented ginseng or an extract thereof, and (2) a step of treating the extract with an enzyme, a step of sterilizing, or a step of treating the extract with citric acid and then sterilizing. Here, ginsenoside refers to K, F2, Rg3 or Rb1 when referring to the examples below.

In the present invention, the ginseng powder may be a powder of ginseng (fresh ginseng) that has not been dried after harvesting, a powder of dried ginseng (white ginseng), or a powder of steamed ginseng (red ginseng). The powder may be a fine-grained powder, a fine-grained piece, or a mixture thereof.

In addition, in the present invention, the ginseng extract refers to one obtained by extracting the ginseng root, which is the target of extraction, with water, an organic solvent, or a mixed solvent thereof. The organic solvent refers to lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.), methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, etc., and is preferably ethanol. Any method such as cold steeping, reflux, heating, ultrasonic radiation, or supercritical extraction may be applied as the extraction method. After extraction, it may be preferable to remove the extraction residue (solid matter later) through filtration, centrifugation, standing for a certain period of time, or the like. In particular, in the present invention, it is preferable that the ginseng extract be a supernatant obtained by removing the extraction residue after hot water (60 to 90°C) extraction, adding ethanol to the supernatant from which the extraction residue has been removed, reacting for a certain period of time (20 to 240 minutes), and then centrifuging. When ethanol is added to react and centrifuged, sugar components other than ginsenoside present in the supernatant are removed, thereby increasing the concentration of ginsenoside in the supernatant after centrifugation.

In the present invention, the "medium containing ginseng extract" can be prepared by adding an appropriate amount of water (purified water or distilled water) to ginseng powder or extract, preferably 2-4 times the weight of the ginseng powder or extract.

In the present invention, "fermented ginseng" is a fermented product cultured by inoculating a Bacillus genus microorganism into a "medium containing ginseng pulverized material", and any Bacillus genus microorganism may be used. Preferably, it may be the Bacillus genus S10 strain (Accession No. KACC 92429P) or Bacillus genus S11 strain (Accession No. KCTC13946BP) used in the examples below.

In the present invention, the “enzyme-treated product” refers to a product obtained by treating fermented ginseng (using the above Bacillus genus microorganism) with an enzyme having cellulase, pectinase, or glucanase activity, and may preferably be the enzymes used in the examples below.

It may be desirable to inoculate these Bacillus microorganisms into a medium containing ginseng extract and culture them for 1 to 3 days to acclimatize them for use in ginseng fermentation. Here, the ginseng extract may be a water or hot water (50 to 80°C) extract, and ultrasonic treatment, etc. may be performed during the extraction process to extract ginsenosides at a higher yield. The medium containing the ginseng extract may be prepared by preparing a ginseng extract in powder form and adding water thereto, or by adding water to pulverized ginseng to extract it, filtering the extract to obtain the filtrate, or leaving it alone to obtain the supernatant.

After the above Bacillus genus microorganism is inoculated, it may be desirable to perform shaking culture at a temperature of 25 to 45°C, especially 30 to 35°C, for at least 1 day, preferably 1 to 10 weeks, especially 4 to 8 weeks.

In the present invention, it is preferable to sterilize the medium containing ginseng powder or extract to prevent unintended fermentation by other microorganisms. Sterilization can be performed using a method known in the art, such as high-temperature/high-pressure sterilization or ultraviolet irradiation.

Also, in the present invention, in order to produce a fermented ginseng product having a particularly high content of ginsenoside Rg3, F2 or compound K, sequential inoculation and cultivation of Bacillus genus S11 strain or Bacillus genus S10 strain may be preferable. Here, the sequential inoculation and cultivation may be specifically inoculation and cultivation of Bacillus genus S11 strain followed by inoculation and cultivation of Bacillus genus S10 strain. In this case, each sequential culture may be for 1 day or longer, preferably 1 to 8 weeks, particularly 2 to 4 weeks.

Meanwhile, since the fermented ginseng obtained by the manufacturing method of the present invention, which has an increased ginsenoside content and has been treated with enzymes or acids and sterilized, is known to have various physiological activities, it can be manufactured into products such as pharmaceutical compositions, health functional foods, and functional cosmetics. Specifically, the pharmaceutical compositions, health functional foods, and functional cosmetics can have uses or functionalities known in the art, such as immunity enhancement, fatigue improvement, bone health promotion, anti-diabetes, anti-hypertension, anti-hyperlipidemia, cardiovascular disease treatment or prevention, atopic dermatitis improvement, anti-obesity, vitality recovery, skin wrinkle improvement, and skin whitening, which are possessed by these ginsenoside components.

When the fermented ginseng having an increased ginsenoside content, obtained by the manufacturing method of the present invention, and treated with enzymes or acids and sterilized, is manufactured into a pharmaceutical composition and used, the pharmaceutical composition may contain, in addition to the fermented ginseng, a pharmaceutically acceptable carrier, and may be manufactured into an oral formulation or a parenteral formulation according to the administration route by a conventional method known in the art. Here, the administration route may be any appropriate route including a topical route, an oral route, an intravenous route, an intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be combined and used. An example of a combination of two or more routes is a case where two or more drug formulations according to the administration route are combined, for example, a case where one drug is administered intravenously first and another drug is administered topically second.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or formulation, and specific examples can be found in the pharmacopoeias of each country, including the “Korean Pharmacopoeia.”

When the pharmaceutical composition of the present invention is prepared as an oral dosage form, it can be prepared in the form of powder, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers, etc., using a suitable carrier and a method known in the art. Examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol; starches such as corn starch, potato starch, and wheat starch; celluloses such as cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, and hydroxypropylmethylcellulose; polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyols, vegetable oils, ethanol, and glycerol. When formulating, appropriate binders, lubricants, disintegrants, colorants, diluents, etc. may be included as needed. Appropriate binders include starch, magnesium aluminum silicate, starch peristalsis, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, waxes, etc. Lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talc, stearic acid, magnesium salts and calcium salts thereof, polyethylene glycol, etc. Disintegrants include starch, methyl cellulose, agar, bentonite, xanthan gum, starch, alginic acid or sodium salts thereof. Other diluents include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and glycine.

When the pharmaceutical composition of the present invention is prepared as a parenteral formulation, it can be formulated in the form of injections, transdermal administration, nasal inhalers, and suppositories using a suitable carrier according to a method known in the art. When formulated as an injection, an aqueous isotonic solution or suspension can be used as a suitable carrier, and specifically, an isotonic solution such as PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, or 5% dextrose can be used. When formulated as a transdermal administration, it can be formulated in the form of ointments, creams, lotions, gels, external solutions, pastes, liniments, aerosols, etc. For nasal inhalers, the composition can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, or carbon dioxide; for suppositories, the composition can be formulated as a carrier such as Witepsol, Tween 61, polyethylene glycol, cacao butter, laurin butter, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, or sorbitan fatty acid esters.

Specific formulations of pharmaceutical compositions are known in the art and can be found, for example, in Remington's Pharmaceutical Sciences (19th ed., 1995), which is incorporated herein by reference.

The preferred dosage of the pharmaceutical composition of the present invention may be in the range of 0.001 mg/kg to 10 g/kg per day, preferably in the range of 0.001 mg/kg to 1 g/kg per day, depending on the patient's condition, weight, sex, age, severity of the patient, and route of administration. Administration may be performed once a day or divided into several times. Such dosage should not be construed as limiting the scope of the present invention in any way.

In another aspect, when the fermented ginseng with increased ginsenoside content obtained by the manufacturing method of the present invention, treated with enzymes or acids and sterilized, is manufactured and used as a health functional food composition, the food composition can be manufactured in any form, and for example, can be manufactured in the form of beverages such as tea, juice, carbonated beverages, and ionic beverages, processed dairy products such as milk and yogurt, foods such as gum, rice cakes, Korean traditional confectionery, bread, cookies, and noodles, and health functional food preparations such as tablets, capsules, pills, granules, liquid, powder, flakes, paste, syrup, gel, jelly, and bars.

In addition, the food composition of the present invention may have any product classification as long as it complies with the laws and regulations in effect at the time of manufacturing and distribution in terms of legal and functional classification. For example, it may be a health functional food according to the Korean 「Health Functional Food Act」, or a confectionery, legume, tea, beverage, special-purpose food, etc. according to each food type according to the food code of the Korean 「Food Sanitation Act」 (KFDA Notice 「Food Standards and Specifications」).

The food composition of the present invention may contain food additives in addition to the fermented liquid. Food additives can be generally understood as substances that are added to, mixed in with, or infiltrated into food during the manufacturing, processing, or preservation of food, and since they are consumed with food every day and for a long period of time, their safety must be guaranteed. In the food additive code according to the laws of each country that regulate the manufacturing and distribution of food (in Korea, the “Food Sanitation Act”), food additives whose safety is guaranteed are limitedly stipulated in terms of ingredients or functions. In the Korean Food Additive Code (the “Food Additive Standards and Specifications” notified by the Ministry of Food and Drug Safety), food additives are stipulated by classifying them into chemical synthetic products, natural additives, and mixed preparations in terms of ingredients, and these food additives are classified into sweeteners, flavoring agents, preservatives, emulsifiers, acidulants, and thickeners in terms of functions.

Sweeteners are used to provide appropriate sweetness to foods, and both natural and synthetic sweeteners can be used in the composition of the present invention. Preferably, a natural sweetener is used, and examples of natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents can be used to improve taste or aroma, and both natural and synthetic ones can be used. It is preferable to use natural ones. When using natural ones, in addition to flavor, the purpose of enhancing nutrition can also be used. Natural flavoring agents can be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or from green tea leaves, Polygonum multiflorum, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, etc. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo nuts, etc. can be used. Natural flavoring agents can be liquid concentrates or solid extracts. In some cases, synthetic flavoring agents can be used, and synthetic flavoring agents can use esters, alcohols, aldehydes, terpenes, etc.

Preservatives that can be used include sodium calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc., and emulsifiers that can be used include acacia gum, carboxymethyl cellulose, xanthan gum, pectin, etc., and acidulants that can be used include citric acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, etc. In addition to the purpose of enhancing taste, acidulants can be added to the food composition to have an appropriate acidity for the purpose of inhibiting the growth of microorganisms.

As thickening agents, suspending agents, sedimentation agents, gelling agents, and bulking agents can be used.

In addition to the food additives described above, the food composition of the present invention may contain physiologically active substances or minerals known in the art and guaranteed to be safe as food additives for the purpose of supplementing and reinforcing functionality and nutrition.

Examples of such physiologically active substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, and dibenzoylthiamine, and examples of minerals include calcium preparations such as calcium citrate, magnesium preparations such as magnesium stearate, iron preparations such as ferrous citrate, chromium chloride, potassium iodine, selenium, germanium, vanadium, and zinc.

The food composition of the present invention may contain the food additives described above in an appropriate amount that can achieve the purpose of addition depending on the product type.

With regard to other food additives that may be included in the food composition of the present invention, reference may be made to the food codes or food additive codes of each country.

In another aspect, when the fermented ginseng, which has been sterilized and treated with enzymes or acids and has an increased ginsenoside content obtained by the manufacturing method of the present invention, is manufactured into a functional cosmetic composition and used, the cosmetic composition may contain, in addition to the fermented liquid, components commonly used in cosmetic compositions, for example, conventional auxiliary agents such as stabilizers, solubilizers, surfactants, vitamins, pigments, and flavorings, and carriers.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide can be used as a carrier component.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and particularly in the case of a spray, a propellant such as chlorofluorohydrocarbon, propane/butane or dimethyl ether may be additionally included.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizer or an emulsifier is used as a carrier component, and specifically, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, and the like can be used.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, etc. can be used as carrier components.

When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkyl amidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester may be used as a carrier component.

The cosmetic composition of the present invention can be manufactured according to a method for manufacturing a cosmetic composition commonly practiced in the art, except that it includes an enzyme-treated ginseng fermentation product with an increased ginsenoside content obtained by the manufacturing method of the present invention.

As described above, according to the present invention, a method for producing an enzyme-treated ginseng fermented product having an increased ginsenoside content from ginseng can be provided. Such an enzyme-treated ginseng fermented product having an increased ginsenoside content can be usefully used in health functional foods, pharmaceuticals, etc.

Figure 1 shows the clarification aspect and turbidity measurement values after enzyme treatment of ginseng extract (S10).
Figure 2 is a chromatogram of ginsenosides of enzyme-treated fermented ginseng (S10).
Figure 3 is a ginsenoside chromatogram of enzyme-treated fermented ginseng (S111+S10) and acid-treated or sterilized fermented ginseng.
Figure 4 shows the ginsenoside components and contents of enzyme-treated fermented ginseng (S11+S10).

Hereinafter, the present invention will be described with reference to examples. However, the scope of the present invention is not limited to these examples.

<Example> Enzyme or acid treatment and sterilization of fermented ginseng powder

1. Preparation of fermented ginseng dried powder

1.1 Cultivation of fermentation strains

To prepare the inoculation strains, first, Bacillus sp. S10 strain ( Bacillus sp. S10, KACC 92429P, hereinafter S10 strain) and Bacillus sp. S11 strain ( Bacillus sp. S11, KCTC13946BP, hereinafter S11 strain) were inoculated onto R2A agar medium (R-2A Agar, Millipore 17209-500G) and cultured for about 12 hours until colonies became clearly visible. Then, each strain prepared in the R2A medium was inoculated into 1 mL of a medium containing 10% (w/v) ginseng extract, and the culture was shaken (80 rpm) at 30°C for more than 24 hours to prepare the inoculation strains. The medium containing 10% (w/v) ginseng extract was prepared by suspending 10 g of ginseng powder obtained through a ball mill in 90 ml of distilled water, sonicating the suspension at 50°C for 30 minutes, extracting it, leaving it overnight at 4°C, recovering the supernatant, centrifuging the supernatant (8,000 rpm/30 min), and then sterilizing it (121°C, 15 min).

1.2 Fermentation and production of dried fermented ginseng powder

Twice the weight of distilled water was added to the dried powder of ginseng root (fresh ginseng), sterilized (121°C, 15 minutes), and 0.2 mL of the strain culture of Example 1.1 was inoculated. Afterwards, the culture was cultured for a total of 8 weeks at 30°C to obtain a fermented ginseng product. In the case of S10, only the S10 strain was inoculated and cultured for 8 weeks, and in the case of S11+S10, the S11 strain was inoculated, cultured for 4 weeks, and then the S10 strain was inoculated and cultured for 4 more weeks.

The fermentation product was dried in aqueous solution mode in an evaporator (Genvac Evaporator, SP Scientific) to obtain a powder.

2. Enzyme or acid treatment and sterilization of fermented ginseng

2.1 Production of enzyme-treated fermented ginseng

Fermented ginseng dried powder is suspended in distilled water at a concentration of 1% (w/v), and then sonicated for 30 minutes. 1% volume (v/v) of enzyme is added (5.05 ml) and the reaction is performed at 50℃ for 2 days at a speed of 80 rpm. The enzyme reaction solution is diluted 4 times, and the turbidity is measured at 600 nm using a spectrophotometer, and dried using an evaporator.

The types of processed enzyme products and enzyme activities are as shown in Table 1 below.

Commercial enzymes used Serial number Product Name active enzyme genesis manufacturing company E1 Celluclast® 1.5L Cellulase, naringinase Trichoderma reesei Novozyme E2 Pectinex® Ultra Passover pectinase, hemicellulase, β-glucanase A. niger, A. aculeatus Novozyme E3 Pectinex® Ultra Pulp Pectin lyase with arabinase Novozyme E4 Pectinex® Ultra SP-L Polygalacturonase A. aculeatus Novozyme E5 Viscozyme® β-glucanase, Cellulase, Xylanase Novozyme

As a result, as shown in Fig. 1, the clarifying activity of fermented ginseng was the best in E3 and E4. Although E2 had somewhat lower clarifying activity, the content of compound K in the product was the highest (Fig. 2).

2.2 Production of acid-treated and sterilized fermented ginseng

For the production of acid-treated or sterilized fermented ginseng, the dried powder of S11+S10 fermented ginseng is suspended at 1% (w/v) and then sonicated for 30 minutes.

Acid-treated fermented ginseng was mixed well with 1 drop of 10% citric acid solution and the pH was measured. The pH was measured to be pH 4.5 before adding citric acid and pH 3.3 after adding citric acid. The acid-treated fermented ginseng was sterilized at 121℃ for 10 minutes using a high-pressure steam sterilizer.

Sterilized fermented ginseng was sterilized using a high-pressure steam sterilizer at 121℃ for 10 minutes after suspension and ultrasonically crushing fermented ginseng.

3. Analysis of ginsenoside content in enzyme or acid treated and sterilized fermented ginseng

As in Examples 2.1 and 2.2 above, the fermented ginseng treated with enzymes or acids and sterilized was suspended in 2 mL of 70% (v/v) methanol, extracted using ultrasonic waves, centrifuged, and then 1 mL of the supernatant was taken and analyzed.

Ginsenoside content was measured using an Agilent 1100 series HPLC system (Agilent Technolgies, USA). HPLC analysis was performed using a YMC-Pack ODS AM (250 × 4.6㎜, 5㎛, YMC, Inc. USA) column under the mobile phase conditions shown in Table 2 below.

Mobile phase concentration and flow rate conditions for HPLC analysis hour Acetonitrile (%) water (%) 0 27 73 10 27 73 45 42 68 47 95 5 63 95 5

At this time, the flow rate of the mobile phase and the column temperature were set to 0.8 ml/min and 43°C, respectively, and the analysis was performed at a detection wavelength of 203 nm of the UV detector (Korean Journal of Medicinal Crop Science 16(6):446-454, 2008).

Ginsenoside standards were purchased from Chromadex (Santa Anna, USA), and the content of ginsenoside was quantified based on the calibration curve of the standard.

As a result of the analysis,

In the fermented ginseng treated with enzymes, compound K and ginsenoside F2 were the main components of fermented ginseng S10 when treated with E2 enzyme (Fig. 2), and F2 was the main component of fermented ginseng S11+S10 when treated with E4 enzyme (Fig. 3).

In sterilized S11+S10 fermented ginseng, ginsenoside Rg3 was the main component (Fig. 3).

In the sterilized S11+S10 fermented ginseng after acid treatment, ginsenoside Rg3 accounted for 35.5 mg/g, or 79% of the total saponin (45 mg/g) (Table 3, Fig. 3, Fig. 4).

Amount of ginsenosides (mg/g) analyzed from samples sterilized after enzyme treatment or sterilization and acid treatment of fermented ginseng (S11+S10). C, control; E1, Celluclast® 1.5L; E2, Pectinex® Ultra Passover; E3, Pectinex® Ultra Pulp; E4, Pectinex® SP-L; pH3.3 sterilization, sterilization after adding citric acid division A600 K Rh2 F1 Rh1 F2 Rg3 Rg1 Rf Re Rd Rb2 Rc Rb1 sum C 0.94 　 　 　 1.5 1.5 1.7 5.5 2.6 4.0 5.4 3.8 　 22.1 48.0 E1 0.77 　 　 　 　 　 1.5 3.7 2.2 3.3 21.6 4.5 1.8 　 38.7 E2 0.22 12.9 　 2.1 1.9 5.9 　 11.8 　 4.1 　 　 　 　 38.7 E3 0.22 4.8 2.9 　 2.5 7.0 2.4 8.9 2.1 4.0 8.5 2.6 　 　 45.6 E4 0.22 2.2 1.9 1.8 2.4 23.4 　 11.1 　 5.6 2.1 　 　 　 50.7 E5 0.38 1.6 1.6 　 2.4 11.4 　 9.2 2.1 5.6 8.0 2.5 　 　 44.6 Sterilization - 　 1.7 　 2.1 1.2 25.2 3.7 2.3 2.3 2.9 2.1 　 9.2 52.6 pH3.3, sterilized - 　 2.5 　 3.6 1.2 35.4 0.6 1.8 　 　 　 　 　 45.0

In this way, when fermented ginseng is treated with enzymes or acids and sterilized, the content of low-molecular-weight saponins, including compound K with high absorption rate, increases, and since there are no by-products, the usability of these products increases when using them in powder and liquid products. In addition, it can be used as a high value-added functional food and cosmetic material.

Claims (5)

In the production of enzyme-treated fermented ginseng with increased contents of ginsenoside F2 and compound K,
A step of inoculating dried ginseng root (fresh ginseng) powder with Bacillus sp. S10 (Accession No. KACC 92429P) and fermenting it to obtain fermented ginseng; and
A method for producing enzyme-treated fermented ginseng, characterized by including a step of adding an enzyme having a combined activity of hemicellulase, pectinase, and β-glucanase to the fermented ginseng to produce an enzyme reactant, thereby obtaining enzyme-treated fermented ginseng.
In the production of enzyme-treated fermented ginseng with increased content of ginsenoside compound K,
A step of inoculating dried ginseng root (fresh ginseng) powder with Bacillus sp. S11 (Accession No. KCTC13946BP) and fermenting it, and then inoculating it with Bacillus sp. S10 (Accession No. KACC 92429P) and further fermenting it to obtain fermented ginseng; and
A method for producing enzyme-treated fermented ginseng, characterized by comprising the step of adding an enzyme having combined activities of hemicellulase, pectinase and beta-glucanase to the fermented ginseng to produce an enzyme reactant, thereby obtaining enzyme-treated fermented ginseng.
In the production of enzyme-treated fermented ginseng with increased content of ginsenoside F2,
A step of inoculating dried ginseng root (fresh ginseng) powder with Bacillus sp. S11 (Accession No. KCTC13946BP) and fermenting it, and then inoculating it with Bacillus sp. S10 (Accession No. KACC 92429P) and further fermenting it to obtain fermented ginseng; and
A method for producing enzyme-treated fermented ginseng, characterized by including a step of producing an enzyme reactant by adding an enzyme having polygalacturonase activity to the fermented ginseng to produce enzyme-treated fermented ginseng.
In the production of sterilized fermented ginseng with increased content of ginsenoside Rg3,
A step of inoculating dried ginseng root (fresh ginseng) powder with Bacillus sp. S11 (Accession No. KCTC13946BP) and fermenting it, and then inoculating it with Bacillus sp. S10 (Accession No. KACC 92429P) and further fermenting it to obtain fermented ginseng; and
A method for producing sterilized fermented ginseng, characterized by manufacturing the same through a step of sterilizing the above-mentioned fermented ginseng using a high-pressure steam sterilizer.
In the production of acid-treated fermented ginseng with increased content of ginsenoside Rg3,
A step of inoculating dried ginseng root (fresh ginseng) powder with Bacillus sp. S11 (Accession No. KCTC13946BP) and fermenting it, and then inoculating it with Bacillus sp. S10 (Accession No. KACC 92429P) and further fermenting it to obtain fermented ginseng; and
A step of adding citric acid to the above fermented ginseng to obtain acid-treated fermented ginseng;
A method for producing acid-treated sterilized fermented ginseng, characterized by manufacturing the same through a step of sterilizing the acid-treated fermented ginseng using a high-pressure steam sterilizer.