KR20120134166A - A method of preparing ginseng extract comprising minor saponin in high concentration - Google Patents
A method of preparing ginseng extract comprising minor saponin in high concentration Download PDFInfo
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- KR20120134166A KR20120134166A KR1020110052408A KR20110052408A KR20120134166A KR 20120134166 A KR20120134166 A KR 20120134166A KR 1020110052408 A KR1020110052408 A KR 1020110052408A KR 20110052408 A KR20110052408 A KR 20110052408A KR 20120134166 A KR20120134166 A KR 20120134166A
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- red ginseng
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
Abstract
The present invention comprises the steps of preparing a ginseng extract by adding at least one solvent selected from the group consisting of water and ethanol to the ginseng and performing a solvent extraction process; Inoculating and fermenting lactic acid bacteria to the ginseng extract to prepare a fermented product of ginseng extract; And relates to a method for producing a ginseng extract with an increased content of trace ginsenosides comprising the step of heat-treating the fermented product of the ginseng extract, further comprising the step of performing a chromatography method from the fermented product of the heat-treated ginseng extract It may include. In addition, the present invention relates to an anti-inflammatory composition comprising the ginseng extract excellent in anti-inflammatory activity prepared by the production method and the ginseng extract as an active ingredient.
According to the preparation method of the present invention, ginsenoside Rg3, ginsenoside Rk1, ginsenoside Rg5 and compound K (Compoun K, CK), which have been found to exhibit various physiological activities, are saponins contained in a small amount or little in ginseng. Ginseng extract can be produced with a significant increase in the amount of c), and the anti-inflammatory effect is remarkably improved. Therefore, it can contribute to the improvement of ginseng quality and the standardization of ginseng products, and is widely used in various industries related to ginseng. In addition, it can contribute to the profitability of producers related to the ginseng industry, including ginseng farmers.
Description
The present invention relates to a method for preparing ginseng extract containing a high concentration of ginsenosides.
Ginseng is a perennial herbaceous genus belonging to the ginseng of the Ogapi family, and there are about 11 species of ginseng (ginseng, Panax) grown in the Korean Peninsula. ginseng CA Meyer) is known to have excellent efficacy. It is reported that ginseng contains saponin, phenolic component, polyacetylene component, alkaloid component, polysaccharide, and the like.
Commonly referred to as ginseng refers to ginseng or white ginseng (about 75% moisture) dried white ginseng (about 12% moisture), red ginseng strictly steamed four to six years old ginseng, steamed and dried without peeling It refers to pale yellow brown or light red brown ginseng. Red ginseng obtained through the steaming process, which is a traditional method, is known to increase the efficacy of ginseng due to the use of more saponin than white ginseng.
Among the efficacy of ginseng known to date, there is an antioxidative activity, antimicrobial activity anticancer activity or anti-inflammatory activity and the like, the efficacy is known to be different for each person.
The main function of the ginseng is known to be due to ginsenosides contained in ginseng. The ginsenoside is named by specially distinguishing only the saponin component of ginseng among the various saponins present in the plant, and the ginsenoside has been reported to have various pharmacological effects. For example, the ginsenoside has been reported to have anti-cancer, anti-allergic, anti-inflammatory, mental stability, analgesic, memory improvement, liver function enhancement, anti-diabetic, anti-stress, antioxidant, immunoregulatory and anti-aging activity .
However, ginsenosides of ginseng are linked to a polymer component, so they are not easily absorbed by the body after ingestion. For example, red ginseng is specific to ginsenosides such as ginsenoside Rg3, ginsenoside Rh1, or ginsenoside Rh2, which are present in ginsenosides. It has ingredients The specific components of the red ginseng are ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, or ginsenoside Rd, which are originally included in the fresh ginseng. Saponins, such as (ginsenoside Rd), are components in the form of prosaponins produced by hydrolysis of the ginseng saponin glycosides by the heat applied during the preparation of red ginseng.
Therefore, the difference in the efficacy of the ginseng is different from person to person because the total number and activity of E. coli bacteria is different, the degree of degradation by the intestinal microorganisms of ginsenosides contained in the ingested ginseng is different, This is because the degree of absorption in the body is different.
Therefore, in order to enhance the efficacy of ginseng, that is, to increase the content of ginsenoside that is easily absorbed and to manufacture excellent ginseng or ginseng products, ginseng such as ginseng is not included or contained in trace amounts, but it is known to have various pharmacological effects. There is a need for development of methods for improving the content of ginsenoside Rg3, ginsenoside Rk1, ginsenoside Rg5, and compound K (Compoun K, CK).
The methods developed to date have limitedly increased the content of certain ginsenosides, such as Rg2, Rg3 or Rh2, and there is a need for the development of methods for increasing the content of various ginsenosides reported to have various physiological activities. do.
In addition, the innate immune response is the body's defense mechanism, which is when immune cells responsible for immune function induce a series of inflammatory reactions that promptly repair the damaged area when surrounding cells or tissues are damaged.
The immune cells move to damaged areas through blood vessels with the help of histamine, nitric oxide (NO) or prostaglandin E2 (PGE2) to initiate an inflammatory response. The immune cells that migrated to the damaged area were cytokines such as tumor necrosis factor-α, interleukin-1β, or interleukin-6, MIP-1, IL Chemokines such as -8 or MCP-1 secrete chemoine to destroy direct external invaders or to gather other immune cells to initiate an inflammatory response.
Inflammatory substances such as interferon-γ, lipoteichoic acid, and lipopolysaccharide (LPS) that cause the inflammatory response or when exposed to various inflammatory-induced cytokines, iNOS (inducible Nitric Oxide synthase) and COX-2 (cyclooxygenase-2) expression As a result, excess nitric oxide (NO) and prostaglandin E2 are produced. These various inflammation initiators (iNOS, COX-2, TNF-α, IL-6, etc.) are promoted transcription by activated NF-κB, which causes the expansion of shock-induced vasodilation, It causes tissue damage, mutagenesis, and nerve tissue damage caused by inflammatory reactions. In addition, COX-2 over-expression causes an inflammatory response.
In order to solve the problems of the prior art as described above, the present invention is a method for preparing a ginseng extract containing a high concentration of ginsenoside, ginseng extract prepared by the method and an anti-inflammatory comprising the ginseng extract as an active ingredient It is an object to provide a composition.
In order to achieve the above object, the present invention provides a method for producing a ginseng extract with an increased content of trace ginsenosides. More specifically, ginseng with increased content of ginsenoside Rg3, ginsenoside Rk1, ginsenoside Rg5, and compound K (Compoun K, CK), which are saponins that are not present or present in trace amounts in nature, ginseng collected in nature It provides a method for preparing an extract.
In addition, the present invention provides a ginseng extract prepared by the above method.
The present invention also provides an anti-inflammatory composition comprising the ginseng extract as an active ingredient.
Hereinafter, the present invention will be described in more detail.
The present inventors found that ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, or ginsenoside, which is a saponin component (Major Saponin) contained in a large amount collected in nature, ginsenoside Rb1 In the case of Rs (ginsenoside Rd), it is not easily absorbed by the body after ingestion, and in fact, it gives ginsenoside Rg3, which is a saponin that is not present in traces of ginseng collected in nature, or is present in a small amount. It was recognized that ginseng saponin glycosides such as ginsenoside Rk1, ginsenoside Rg5, and compound K (Compoun K, CK) are components of the prosaponin form produced by hydrolysis.
The inventors of the present invention while researching for the high value of the processed ginseng such as ginseng or ginseng extract on the basis of the above recognition, ginseng extract corresponding to the lactobacillus which is a GRAS microorganism, accession number KCTC 11377BP to Lactobacillus brevis BJ10 ( Lactobacillus brevis BJ20 KCTC 11377BP) using a fermentation process, and when the heat treatment process with it, it was confirmed that the anti-inflammatory effect is improved compared to the conventional ginseng extract, and furthermore, if the chromatographic method is performed after the heat treatment process, Ginseng extract, especially ginsenoside Rg3, ginsenoside Rk1 ginsenoside Rg5 and compound K (Compoun) K, CK) confirmed that ginseng extract including can be prepared to complete the present invention.
The present invention relates to a method for preparing ginseng extract with an increased content of trace ginsenosides.
More specifically, the present invention comprises the steps of preparing a ginseng extract by adding at least one solvent selected from the group consisting of water and ethanol to the ginseng and performing a solvent extraction process; Lactobacillus genus ( Lactobacillus) in the ginseng extract sp.) adding a lactic acid bacterium and carrying out a fermentation process to prepare a fermented product of ginseng extract; And the ginseng extract with an increased content of trace ginsenosides comprising the step of heat-treating the fermented product of the ginseng extract carried out by heating the fermented product of the ginseng extract at 100 ℃ to 150 ℃ for 60 minutes to 120 minutes It relates to a manufacturing method.
The ginseng is a medicinal plant of the family Elmaceae, and depending on the processing method, there are unprocessed ginseng, dried ginseng (white ginseng), dried red ginseng and steamed ginseng, etc., depending on the cultivation method, artificially cultivated in the ginseng field There is a cultivated ginseng, ginseng seed in the mountain sprinkled in the natural state, wild ginseng grown in the natural state, etc., in the present invention ginseng is used to include all kinds of ginseng. In this respect, the ginseng is Korean ginseng (ginseng, Panax ginseng CA Meyer), ginseng, white ginseng, hwagisam, jeonchisam, bamboo shoot ginseng, three leaf ginseng, Himalayan ginseng, red ginseng, wild ginseng, camphor ginseng, misam, wonsam and wild ginseng may be one or more selected from the group consisting of, for example, red ginseng, ginseng and It may be one or more selected from the group consisting of white ginseng.
The trace ginsenosides refer to ginsenosides or proginsenosides that are not included in trace ginseng or white ginseng or trace amounts, and are preferably ginsenosides Rg3, ginsenosides Rk1 ginsenosides Rg5 and compounds K ( Compoun K) may be one or more selected from the group consisting of, more preferably ginsenoside Rg3, ginsenoside Rk1, ginsenoside Rg5 and compound K (Compoun K).
The preparing of the ginseng extract may be performed by adding at least one solvent selected from the group consisting of water and an organic solvent to the ginseng, preferably at least one solvent selected from the group consisting of water and ethanol and extracting the solvent. .
The solvent extraction may be performed by a method such as heat extraction, reflux extraction, ultrasonic extraction, high pressure extraction, but is not limited thereto. In addition, the solvent extraction may be carried out at 50 ℃ to 100 ℃, or 60 ℃ to 90 ℃, or 70 ℃ to 80 ℃.
The ginseng may be one or more selected from the group consisting of ginseng root, ginseng roots and ginseng stems, for example, may be one or more selected from the group consisting of ginseng root and ginseng rice. The ginseng root and ginseng rice are 5: 1 (ginseng root: ginseng rice) to 1: 5 (ginseng root: ginseng rice) or 3: 1 (ginseng root: ginseng rice) to 1: 3 (ginseng root: Ginseng rice) can be used by mixing.
In addition, the extraction process may further include at least one selected from the group consisting of a concentration process and a drying process, the concentration process may be a reduced pressure concentration process as an example, the drying process is a freeze drying process, for example Can be.
Preparing the fermented product of the ginseng extract may be carried out by inoculating and fermenting the lactic acid bacteria in the ginseng extract. The lactic acid bacteria are lactic acid bacteria of the genus Lactobacillus, lactic acid bacteria of Bifidobacterium, lactic acid bacteria of the genus Leukonostock, lactic acid bacteria of the genus Pediococcus and lactic acid bacteria of the genus Lactococcus.
The lactic acid bacterium is preferably of the genus Lactobacillus sp.) Lactobacillus, more preferably may be Lactobacillus brevis ( Lactobacillus brevis ), and even more preferably may be Lactobacillus brevis BJ10 ( Lactobacillus brevis BJ20 KCTC 11377BP), Accession No. KCTC 11377BP. Lactobacillus brevis BJ10 ( Lactobacillus brevis BJ20) is a microorganism that ensures safety as a GRAS microorganism, and as a result of the present inventors confirmed that the fermentation ability is excellent, and as a result of performing the fermentation process of ginseng extract, it was confirmed that the glycoside decomposition ability of saponin is also excellent, Selected as strain. The strain was found to have excellent fermentation ability against seaweeds, and was deposited on August 19, 2008 at the Korea Institute of Bioscience and Biotechnology, Biotechnology Center, 111, Gwahak-ro, Yuseong-gu, Daejeon, Korea.
Preparing the fermented product of the ginseng extract is lactic acid bacteria, preferably Lactobacillus genus ( Lactobacillus) in ginseng extract sp.) by adding or inoculating the lactic acid bacteria and fermentation at 33 ℃ to 39 ℃, or 35 ℃ to 38 ℃ for 12 hours to 96 hours or 24 hours to 72 hours. The lactic acid bacteria may be one that includes the ginseng extract, pre-cultured in a medium to which 0.1% ginseng extract is added. In order to cultivate the fermented product of the ginseng extract, lactic acid bacteria, specifically, Lactobacillus genus ( Lactobacillus sp.) The lactic acid bacterium should contain the nutrients that can be grown, and in this aspect the fermented product of the ginseng extract may be the fermentation product of the ginseng extract to the medium in which the lactobacillus lactic acid bacteria can grow. .
The medium is not particularly limited as long as the Lactobacillus genus Lactobacillus is grown, and the nutrient component capable of fermenting the ginseng extract added to the medium component and the medium is not particularly limited, for example, MRS medium or edible The possible natural medium, dilution of ginseng extract or diluent of ginseng extract may be added to the nutrients such as oligosaccharides, sugars, proteins, amino acids.
The step of heat-treating the fermented product of the ginseng extract may be performed by a method of heat-treated the fermented product of the ginseng extract. The heat treatment may be performed by heating at 100 ° C. to 150 ° C., or 110 ° C. to 140 ° C., or 120 ° C. to 130 ° C. for 10 minutes to 240 minutes or 30 minutes to 180 minutes or 60 minutes to 120 minutes.
The fermentation step or the heat treatment step may further perform a filtration process. The fermentation may be performed before or after the fermentation step or before or after the heat treatment step, and may be performed by filtration using filter paper or by separating only the supernatant after centrifugation. For example, after centrifugation, the supernatant may be separated, and then filtered using filter paper.
The manufacturing method of the ginseng extract with an increased content of the trace ginsenoside may further include a step of purifying the fermented product of the heat-treated ginseng extract.
The purification process is a resin for adsorption, preferably polystyrene-based adsorption resin, polystyrene-divinylbenzene-based adsorption resin, diaion HP series adsorption resin, Amberlite XAD series adsorption resin (Amberlite XAD-2, Amberlite XAD-4, Amberlite XAD-7, Amberlite XAD-8, Amberlite XAD-16, Amberlite XAD-1180 and Amberlite XAD-1600), 2MG adsorption resin and Sepabead SP-850 adsorption resin can be performed using one or more selected from the group consisting of. . In addition, the purification may be performed by chromatography or adsorption.
The purification may be a method of performing an adsorption process and a desorption process using a column filled with the resin for adsorption, or performing a chromatography.
In order to purify, the process of adsorbing the fermented product of the heat-treated ginseng extract to the resin for adsorption may be carried out by loading with distilled water, the process of desorption to remove the polar material is 20% to 40 The aqueous ethanol solution or 25% to 35% aqueous ethanol solution may be injected into the resin for adsorption to desorb the polar material.
In the purification step, the obtained heat-treated fermented red ginseng extract is filled into a column filled with the resin for adsorption, 90% to 99.9% ethanol aqueous solution or 93% to 97% ethanol aqueous solution is injected, and a method of obtaining a fraction. It can be done with Before the injection of the 90% to 99.9% ethanol aqueous solution in the purification process to remove the polar material, specifically 20% to 40% ethanol aqueous solution or 25% to 35% ethanol aqueous solution is injected into the resin for adsorption, The method may further include removing the ethanol aqueous solution from which the polar material is desorbed.
In addition, the present invention may be a ginseng extract excellent in anti-inflammatory activity prepared by the above method.
The ginseng extract may include ginsenoside Rg3, ginsenoside Rk1 ginsenoside Rg5, and a large amount of compound K. More specifically, the content of the ginsenoside Rg3, ginsenoside Rk1, ginsenoside Rg5 and compound K is 100 parts by weight of the total weight of the ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc and ginsenoside Rd. 400 to 700 parts by weight or 450 to 600 parts by weight or 500 to 550 parts by weight based on the reference.
In addition, it may be an anti-inflammatory composition comprising the ginseng extract as an active ingredient.
The anti-inflammatory composition means a composition used to alleviate, ameliorate or treat inflammation.
The inflammation is a local protective reaction caused by injury or destruction of tissue, which means the action of destroying, attenuating or masking both the injury causing material and the injured tissue.
The inflammation is characterized by perforation of microvascular vessels, leakage of blood components into niche spaces, and migration of leukocytes to inflammatory tissues. In macroscopic terms, familiar clinical features such as erythema, edema, hyperalgesia (sensitivity) and pain are common. It is accompanied by symptoms.
The inflammatory response is any response that is characterized by inflammation as defined above. It is well known in medical practice that the inflammatory response causes many physical discomforts (eg loss of pain and function) that are associated with different diseases and injuries. Therefore, administering pharmaceutical agents that reduce the physical discomfort of the inflammatory response is a common medical practice. Agonists with these properties are classified as anti-inflammatory.
In this respect, anti-inflammatory agents or anti-inflammatory compositions are used for the treatment of a wide range of diseases, and the same agents are often used for the treatment of different diseases. Treatment with anti-inflammatory drugs is not about disease, but mostly about symptoms (ie inflammation).
The anti-inflammatory composition may include the active ingredient in an amount of 0.0001 to 10.0 weight ratio (%) based on the total weight of the composition. The anti-inflammatory composition may include additional ingredients so long as the anti-inflammatory activity thereof is not inhibited.
The anti-inflammatory composition is processed using ginseng used for a long time in edible or medicinal water, lactic acid bacteria which is guaranteed safety, ethanol (alcohol) or GRAS microorganisms, because the anti-inflammatory effect is very excellent, the anti-inflammatory composition is anti-inflammatory, irritant It can be widely used in medicine, cosmetics, food, etc. to achieve the purpose of sedation, etc. Specifically, the present invention may be used in products related to skin diseases with inflammatory reactions such as atopic dermatitis, seborrhea, psoriasis, acne, and the like, but is not limited thereto. .
In this aspect, the present invention may be a cosmetic, food or medicine containing the anti-inflammatory composition as an active ingredient. The present invention also provides cosmetics, foods, pharmaceuticals processed with the anti-inflammatory composition.
The drug is not particularly limited in the formulation thereof, and may be formulated in a conventional oral, parenteral, and topical administration formulation according to the purpose.
In the preparation for oral administration of powders, granules, tablets, capsules and the like, binders, suspending agents, disintegrating agents, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, flavoring agents and the like can be used.
Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents, water and liquid paraffin. In the case of injections, buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers and the like can be used in combination. In addition, it may be formulated into an ointment, a spray, a lotion, a patch, a pad, a cream.
The food includes sweets, beverages, alcoholic beverages, fermented foods, canned food, milk processed foods, land processed foods. In addition, the food may be a health food (functional food, health supplement food).
The health food (functional food, health supplement food) may be prepared in the form of tablets, capsules, powders, granules, liquids, pills and the like.
In addition, the cosmetics are not particularly limited in the formulation or purpose except for using the composition as an active ingredient, for example, formulations such as lotion, nutrition lotion, nutrition cream, massage cream, essence, pack, etc. May have In addition, in each formulation, other components other than the active ingredient may be arbitrarily selected and blended according to the formulation or purpose of use of other cosmetics.
Ingredients that can be blended into the cosmetics include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, antioxidants, pH adjusters, alcohols, pigments, fragrances and the like.
The fat or oil component may be an ester fat, a hydrocarbon fat, a silicone fat, a fluorine fat, an animal fat, a vegetable fat, or the like.
The moisturizing agent may be a water-soluble low molecular moisturizer, a fat-soluble low molecular moisturizer, a water-soluble polymer moisturizer, a fat-soluble polymer moisturizer and the like.
Examples of the emollient agent include long-chain acyl glutamic acid cholesteryl ester, hydroxy stearic acid cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid, lanolin fatty acid cholesteryl ester, and the like.
The surfactant may be a nonionic surfactant, anionic surfactant, cationic surfactant, amphoteric surfactant, or the like.
The alcohol may be used cetearyl alcohol and the like.
Compared to the conventional method, the present invention was confirmed to exhibit various physiological activities, but ginsenoside Rg3, ginsenoside Rk1, ginsenoside Rg5 and compound K (Compoun K, CK), which are saponins contained in a small amount or little in ginseng Ginseng extract can be produced with a significant increase in the amount of c), and the anti-inflammatory effect is remarkably improved. Therefore, it can contribute to the improvement of ginseng quality and the standardization of ginseng products, and is widely used in various industries related to ginseng. In addition, it can contribute to the profitability of producers related to the ginseng industry, including ginseng farmers.
1 is a graph showing the results of performing HPLC to confirm the ginsenoside composition and content of the extract and red ginseng extract of the present invention, Figure 1a shows the result of the red ginseng extract, Figure 1b is a fermentation of the present invention The results of the red ginseng tablets are shown.
Figure 2 is a graph confirming the cytotoxicity of the red ginseng extract, fermented red ginseng extract and fermented red ginseng purified according to an embodiment of the present invention, RG of the graph means red ginseng extract, FRG means fermented red ginseng extract, PFRG Means fermented red ginseng purified product, and the vertical axis of the graph represents cell viability (%).
FIG. 3 is a photograph confirming the anti-inflammatory activity of the red ginseng extract, fermented red ginseng extract and fermented red ginseng tablets according to an embodiment of the present invention, the amount of NF-Κb produced, N of the photo treated LPS It is not a negative control, P in the picture is a positive control treated with LPS, the time means the time elapsed after the LPS treatment, RG in the picture means red ginseng extract, FRG is fermented red ginseng Means extract, and PFRG means fermented red ginseng purified product.
Figure 4 is to confirm the anti-inflammatory activity of the red ginseng extract, fermented red ginseng extract and fermented red ginseng purified according to an embodiment of the present invention, the phosphorylation degree of I-κB-α of I-κB-α and pI-κB-α A photograph confirmed by the amount of production, N in the picture is a negative control without LPS treatment, P in the picture is a positive control treated with LPS, the time means the time elapsed after the LPS treatment, RG in the picture means red ginseng extract, FRG means fermented red ginseng extract, PFRG means fermented red ginseng purified product.
5 is a photograph confirming the expression of iNOS in order to confirm the anti-inflammatory activity of the red ginseng extract, fermented red ginseng extract and fermented red ginseng purified according to an embodiment of the present invention, N in the picture is a negative control not treated with LPS , P of the picture is a positive control treated with LPS, RG of the picture means red ginseng extract, FRG means fermented red ginseng extract, PFRG means fermented red ginseng purified, number is the throughput of the sample ( ug / ml).
6 is a photograph confirming the expression of iNOS in order to confirm the anti-inflammatory activity of the red ginseng extract, fermented red ginseng extract and fermented red ginseng purified according to an embodiment of the present invention, N in the picture is a negative control not treated with LPS , P of the picture is a positive control treated with LPS, RG of the picture means red ginseng extract, FRG means fermented red ginseng extract, PFRG means fermented red ginseng purified, number is the throughput of the sample ( ug / ml).
Figure 7 is a photograph confirming the anti-inflammatory activity of the red ginseng extract, fermented red ginseng extract and fermented red ginseng purified according to an embodiment of the present invention, the mRNA production of TNF-α, N of the photo treated LPS It is not a negative control, P in the picture is a positive control treated with LPS, RG in the picture means red ginseng extract, FRG means fermented red ginseng extract, PFRG means fermented red ginseng purified, number Means the throughput of the sample (ug / ml).
8 is a graph quantified by confirming the expression level of the TNF-α in order to confirm the anti-inflammatory activity of the red ginseng extract, fermented red ginseng extract and fermented red ginseng purified according to an embodiment of the present invention, N is LPS Untreated negative control, P of the graph is a positive control treated with LPS, RG of the graph means red ginseng extract, FRG means fermented red ginseng extract, PFRG means fermented red ginseng purified, The numbers refer to the throughput of the sample (ug / ml). The vertical axis of the graph represents the expression level of TNF-α.
9 is a graph quantified by confirming the degree of generation of NO in order to confirm the anti-inflammatory activity of the red ginseng extract, fermented red ginseng extract and fermented red ginseng purified according to an embodiment of the present invention, N of the graph does not process LPS N is a negative control, P of the graph is a positive control treated with LPS, RG of the graph means red ginseng extract, FRG means fermented red ginseng extract, PFRG means fermented red ginseng purified, number is Mean throughput of the sample (ug / ml). The vertical axis of the graph indicates the degree of generation of NO.
Hereinafter, the present invention will be described in more detail with reference to production examples and examples. However, the following Preparation Examples and Examples are merely illustrative examples for the purpose of facilitating understanding of the present invention, and can be modified into various other forms, and the scope of the present invention is not limited to the following Examples.
Example ; Preparation Method of Ginseng Extract
The red ginseng used to confirm the change and anti-inflammatory effect of ginsenosides contained in the extract by the method of this experiment was obtained from Geumsan Red Ginseng, Chungcheongnam-do, Korea.
First, red ginseng extract (RG) is added to 500 g of red ginseng pulverized product prepared by mixing and grinding the red ginseng root and red ginseng rice in a 1: 1 ratio, and adding 500 ml of an aqueous 70% ethanol solution as an extraction solvent and 60 at 75 ° C. Reflux extraction was performed for minutes. The reflux extraction process was repeated three times. The extracts obtained by the reflux extract were collected and concentrated under reduced pressure at 45 ° C. to remove all extractant components and freeze-dried to obtain a powder (RG).
Fermented red ginseng extract (Fermented Red Ginseng, FRG, Example 1) was prepared by fermenting the prepared red ginseng extract. The fermentation process was performed using lactic acid bacteria, specifically Lactobacillus brevis BJ20 (KCTC 11377BP). More specifically, the Lactobacillus brevis BJ20 (KCTC 11377BP) was precultured at 37 ° C. for 48 hours in MRS medium to which 0.1% red ginseng extract obtained as the powder was added. The pre-cultivated fermentation strains were inoculated in MRS medium containing 10% of the red ginseng extract and fermented at 37 ° C. for 72 hours.
After the fermentation was finished, the fermentation broth was centrifuged for 10 minutes at 3000 rpm, and the supernatant was separated. The suspended solids were completely removed from the separated supernatant through a filter filter (0.45 um). The fermentation broth from which the suspension was separated was heat-treated at 125 ° C. for 90 minutes to obtain a final fermented red ginseng extract (FRG).
The fermented red ginseng extract was purified to prepare fermented red ginseng purified product (Purified Fermented Red Ginseng, PFRG, Example 2). More specifically, the obtained fermented red ginseng extract was injected into a column filled with HP-20 resin, loaded with distilled water, and then injected with 30% ethanol aqueous solution to remove the polar substance. After removing the polar substance, 95% ethanol aqueous solution was injected, the eluate was collected, and lyophilized to prepare fermented red ginseng purified product.
Experimental Example One: Gin Senocide Determination of changes in the content of ingredients
The ginsenoside composition and content of the prepared red ginseng extract and fermented red ginseng purified were confirmed by HPLC method.
More specifically, the red ginseng extract and fermented red ginseng tablets were dissolved in methanol at a concentration of 1 mg / mL, and then HPLC was performed under the conditions of Table 1 to confirm the ginsenoside composition and content. The confirmation results are shown in Table 2 and FIG. 1. Major saponin of Table 2 means Rb1, Rb2, Rc and Rd, minor saponin means Rg3, Rk1, Rg5 and CK, the numerical value of Table 2 is contained in the red ginseng extract or fermented red ginseng purified Mean ginsenoside total content (mg / g) corresponding to each.
Condition
B) Acetonitrile: Water (9: 1)
100-105 min 85 B%, 106-120 min 15 B%
(mg / g)
As shown in Table 2, the components mainly contained in ordinary red ginseng are Rb1, Rb2, Rc and Rd, such as ginseng, and the content of major saponin including all ginsenosides of Rb1, Rb2, Rc and Rd is 21.5691 mg. / g, while the content of minor saponin including Rg3, Rk1, Rg5, and CK, all of which were identified as the causative agents of various physiological activities, was 3.6234 mg / g. It was confirmed to be.
On the other hand, in the fermented red ginseng purified product of Example 2 of the present invention, while the content of major saponin including all ginsenosides of Rb1, Rb2, Rc, and Rd is 2.8869 mg / g, the cause of various physiological activities The content of minor saponin including all of Rg3, Rk1, Rg5 and CK was significantly increased to 18.2365 mg / g, and specifically, about 68% of the major saponin was found to be converted to minor saponin.
In addition, as can be seen through Figures 1a and 1b, Example 2 of the present invention is not only Rg3 and Rk1 which is typically increased by heat treatment, but also a significant increase in the CK component of which the increase in content is limited only by heat treatment. It was confirmed.
According to the method of the present invention from the above results, ginseng containing significantly more minor saponin compared to the red ginseng known to have excellent medicinal effects, including minor saponins such as Rg3, Rk1, Rg5 and CK having various physiological activities compared to conventional ginseng Extract can be prepared, the ginseng extract was expected to be excellent in terms of various functionalities.
Experimental Example 2: Determination of anti-inflammatory activity
In Experimental Example 1, the fermented red ginseng purified product (Example 2) and fermentation expected to have excellent effects in terms of various functionalities include a large amount of minor saponins such as Rg3, Rk1, Rg5 and CK having various physiological activities. The anti-inflammatory activity of the red ginseng extract (Example 1) was compared with the red ginseng extract.
Experimental Example 2-1. Sample and Cell
Experiments were performed using the following samples and cells to measure the anti-inflammatory activity. The acquisition destination of the said sample is as follows.
Dulbecco's modified Eagle's medium (DMEM) was purchased from Hyclone (Thermo Scientific Inc., Germany). Fetal bovine serum (FBS) was purchased from Sigma Aldrich Co. (USA). Penicillin and streptomycin were purchased from Gibco / BRL (USA). In relation to the antibody, Anti-iNOS monoclonal antibody was purchased from Calbiochem (USA), Anti-β-actin monoclonal antibody was purchased from Sigma Aldrich Co. (USA), and Anti-NF-κB p65 monoclonal antibody, Anti-COX -2 monoclonal antibody, anti-IκB monoclonal antibody, anti-phospho-IκB monoclonal antibody and peroxidase-conjugated secondary antibody were purchased from Santa Cruz Biotechnology Inc. (USA).
In addition, RT-PCR premix kit was purchased from Solgent Co. (Korea) and TNF-α ELISA Kits were purchased from Pierce Endogen (USA).
In addition, 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide (MTT), sulfanilamide, lipopolysaccharide (LPS) and all other reagents were purchased from Sigma Aldrich Co.
The mouse macrophage RAW264.7 cells used in the experiment were used as a product from ATCC (USA). The RAW264.7 cells were cultured in an incubator at 37 ° C. and 5% CO 2 conditions using medium containing 10% FBS, 100 U / ml penicillin and 100 ug / ml streptomycin added to the DMEM.
The anti-inflammatory effect was confirmed by measuring the amount of mRNA, protein expression, and NO production associated with the inflammatory response using the sample, device, and cells. The results of the following experiments for confirming the anti-inflammatory effect were analyzed using ANOVA and independence t-test and expressed as mean ± SEM. The statistical significance of each experiment was defined as * p <0.05, ** p <0.01 or *** p <0.001.
Experimental Example 2-2. Confirmation of Cytotoxicity
In order to confirm the cytotoxicity of red ginseng extract, fermented red ginseng extract (Example 1) and fermented red ginseng purified product (Example 2), cell survival rate was confirmed by MTT assay using RAW264.7 cells.
More specifically, after dispensing the RAW264.7 cells at 5 × 10 5 cells / ml in a 96 well plate and stabilizing the cells for 24 hours, the red ginseng extract, the fermented red ginseng extract and the fermented red ginseng tablets were respectively 1 ug / Treated at a concentration of ml and 10 ug / ml and incubated for 1 hour. After the incubation, LPS was treated at a concentration of 0.1 μg / ml and incubated in an incubator at 37 ° C. and 5% CO 2 for 20 hours. After the incubation, the cell culture was removed and treated with MTT at a concentration of 1 mg / ml, followed by incubation for 2 hours. After the incubation, the supernatant was removed by centrifugation, and formazan crystals were dissolved by adding 150 ul of DMSO, and then absorbance was measured at 570 nm using an ELISA microplate reader (TECAN, Switzerland) to measure cell viability. Obtained. The measurement results are shown in FIG. 2, and the viability of the cells shown in FIG. 2 is expressed as a percentage of a control cell to which no sample is added.
As shown in FIG. 2, all of the red ginseng extract, the fermented red ginseng extract, and the fermented red ginseng tablet were confirmed to be safe. More specifically, the survival rate of RAW264.7 cells was about 100% in most of the experiments, and the cell viability of more than 80% was confirmed even in the group treated with 10 ug / ml of the fermented red ginseng purified as the most toxic. Only slight cytotoxicity was identified.
Experimental Example 2-3. Immunoblot analysis ( Western blotting ) One
INOS, COX-2 or TNF-α or the like induced by LPS is expressed by the transcription factor NF-κB. The NF-κB is inactivated in the cytoplasm with IκB complex, but when I-κB-α is phosphorylated by I-κB-α kinase, the bound complex is decomposed and activated by NF-Κb to move into the nucleus ( trnslocation).
Therefore, in order to confirm the anti-inflammatory activity of the red ginseng extract, fermented red ginseng extract (Example 1) and fermented red ginseng purified product (Example 2), Western blotting for NF-Κb, specifically related to inflammation The degree of expression of the protein was confirmed by analysis by immunoblot analysis (Immunoblot analysis).
More specifically, after dispensing the RAW264.7 cells at 5 × 10 5 cells / ml in a 96 well plate and stabilizing the cells for 24 hours, the red ginseng extract, the fermented red ginseng extract and the fermented red ginseng tablets were respectively 1 ug / The cells were treated at a concentration of ml and 10 ug / ml, and after 1 hour, the LPS was treated with 0.1 ug / ml and incubated for 24 hours.
After the incubation, cells were obtained and the cells thus obtained were washed twice with PBS, then placed in a microtube with PBS, centrifuged for 5 minutes under conditions of 12,000 x g, and PBS was removed. After removing the PBS, 200 ul of hypotonic buffer containing 10 mM HEPES (pH 7.9), 10 mM KCl, 0.1 mM EDTA, 0.5% Nonidet P-40, 1 mM DTT, 0.5 mM phenylmethylsulfonylfluoride (PMSF) After placing on ice for a minute, it was centrifuged for 5 minutes at 15,000 xg. Subsequently, the supernatant was removed and 50 μl of highsalt buffer containing 20 mM HEPES (pH 7.9), 400 mM NaCI, 1 mM EDTA, 10 mM DTT, 1 mM PMSF was added to the pellet and vortexed every 10 minutes. The mixture was left on ice for 40 minutes and centrifuged at 16,000 xg for 10 minutes to obtain a supernatant containing a nuclear fraction. The supernatant containing the obtained nuclear fraction was used for the experiment to quantify the protein content.
First, the same amount of cell lysate was mixed with 5X sample buffer, and boiled at 95 ° C. for 5 minutes, followed by SDS-PAGE. Electrophoresis of the gel gel (gel) was moved on the nitrocellulose membrane, the reaction was blocked for 2 hours at room temperature with a blocking buffer (5% skim milk) to inhibit the non-specific antibody binding. After inhibiting the antibody binding, the reaction solution was prepared by diluting the antibody against NF-κB in a blocking solution at 1: 1000, and then reacted with the reaction solution at 4 ° C. overnight. After the reaction, the reaction was washed with TBS-T for 30 minutes and reacted with a horse-radish peroxidase-conjugated secondary antibody for 1 hour.
The membrane reacted with the secondary antibody (nitrocellulose membrane) was washed with TBS-T for 30 minutes, and analyzed by FluorChem Imaging System (Cell Biosciences, USA) using an ECL kit (Amersham, GE healthcare, UK). The degree of formation was confirmed, and the results are shown in FIG. 3. As shown in FIG. 3, the translocation of NF-κB was increased in the LPS-treated group P compared to the group N not treated with LPS. On the other hand, when the red ginseng extract (RG), fermented red ginseng extract (FRG) and fermented red ginseng extract (PFRG) were treated, it was confirmed that the movement of NF-Κb is reduced, compared to the red ginseng extract fermented red ginseng extract, fermented red ginseng It was confirmed that the fermented red ginseng purified water was reduced compared to the extract.
Experimental Example 2-4. Immunoblot analysis ( Western blotting ) 2
In order to confirm the anti-inflammatory activity of the red ginseng extract and the fermented red ginseng extract (Example 1) and the fermented red ginseng tablet (Example 2), Western blotting of other proteins related to inflammation, in particular, proteins related to inflammation The expression level of was confirmed by analyzing by immunoblot analysis (Immunoblot analysis).
More specifically, after dispensing the RAW264.7 cells at 5 × 10 5 cells / ml in a 96 well plate and stabilizing the cells for 24 hours, the red ginseng extract, the fermented red ginseng extract and the fermented red ginseng tablets were respectively 1 ug / The cells were treated at a concentration of ml and 10 ug / ml, and after 1 hour, the LPS was treated with 0.1 ug / ml and incubated for 24 hours.
After incubation, the cells were washed with PBS, and buffer (50 mM HEPES (pH 7.4), 150 mM NaCl, 1% deoxycholate, 1 mM EDTA, 1 mM PMSF, 1 ug / ml aprotinin) was added to each cell. Then, vortexed and lysed at 4 ° C. for 30 minutes. After the lysis, the cell content was removed by centrifugation at 13,000 rpm for 30 minutes to remove cell debris, and the protein content was quantified using the obtained cell lysate.
First, the same amount of cell lysate was mixed with 5X sample buffer, and boiled at 95 ° C. for 5 minutes, followed by SDS-PAGE. Electrophoresis of the gel gel (gel) was moved on the nitrocellulose membrane, the reaction was blocked for 2 hours at room temperature with a blocking buffer (5% skim milk) to inhibit the non-specific antibody binding. After inhibiting the antibody binding, the reaction solution was prepared by diluting the antibody against COX-2, iNOS, IκB, p-IκB and actin, lamin 1: 1000 in a blocking solution, and then reacting with the reaction solution at 4 ° C. Overnight was performed. After the reaction, the reaction was washed with TBS-T for 30 minutes and reacted with a horse-radish peroxidase-conjugated secondary antibody for 1 hour.
The membrane reacted with the secondary antibody (nitrocellulose membrane) was washed with TBS-T for 30 minutes, and analyzed by FluorChem Imaging System (Cell Biosciences, USA) using an ECL kit (Amersham, GE healthcare, UK). The degree of formation was checked.
Of the experimental results, the experimental results for p-I-κB-α is shown in FIG. As shown in FIG. 4, the expression level of pI-κB-α was reduced (30 minutes) in the LPS treated group (P) compared to the group not treated with LPS (N), and then recovered (60 minutes). It was confirmed. In general, phosphorylation of I-κB-α induced by LPS (P) continues to increase from 15 minutes to 60 minutes, but the red ginseng extract (RG), fermented red ginseng extract (FRG) and fermented red ginseng purified products ( PFRG), the amount of phosphorylated I-κB-α (pI-κB-α) is reduced, especially at 60 minutes, the expression level was found to be significantly different, the degree of red ginseng Compared with the extract, the fermented red ginseng extract was confirmed to be larger than the fermented red ginseng extract, and the fermented red ginseng extract or the fermented red ginseng purified product was able to inhibit the phosphorylation of I-κB-α compared to the red ginseng extract. It became.
Of the above experimental results, the experimental results for the iNOS is shown in FIG. As shown in FIG. 5, the expression level of iNOS was increased in the LPS treated group (P) compared to the group (N) not treated with LPS. On the other hand, when the red ginseng extract (RG), fermented red ginseng extract (FRG) and fermented red ginseng purified product (PFRG) were treated, the amount of iNOS was reduced in proportion to the treatment concentration, and the degree of fermented red ginseng extract compared to the red ginseng extract The fermented red ginseng extract was found to be larger than the fermented red ginseng extract, and it was confirmed that the fermented red ginseng extract or the fermented red ginseng purified product inhibited the expression level of iNOS compared to the red ginseng extract.
In addition, the medium of RAW 264.7 cells was changed to FBS (-) medium and after 30 minutes, the red ginseng extract, fermented red ginseng extract and fermented red ginseng purified product were treated at a concentration of 1 ug / ml or 10 ug / ml. One hour after the sample treatment, the LPS was treated at a concentration of 0.1 ug / ml, and after 4 hours of incubation, the expression level of COX-2 was confirmed. The results are shown in FIG. 6.
As shown in FIG. 6, the amount of expression of COX-2 was significantly increased in the LPS treated group (P) compared to the group (N) not treated with LPS. On the other hand, when the red ginseng extract (RG), fermented red ginseng extract (FRG) and fermented red ginseng purified product (PFRG) were treated, the amount of COX-2 was reduced in proportion to the treatment concentration, the degree of fermentation compared to the red ginseng extract It was confirmed that the red ginseng extract was larger in fermented red ginseng extract than the fermented red ginseng extract, and the fermented red ginseng extract or fermented red ginseng purified product inhibited the expression level of COX-2 as compared to the red ginseng extract.
Experimental Example 2-5. TNF Whether to generate -α
In order to confirm the anti-inflammatory activity of the red ginseng extract, fermented red ginseng extract (Example 1) and fermented red ginseng purified product (Example 2), it was confirmed by analyzing the mRNA production inhibition of TNF-α and the expression amount of TNF-α.
The mRNA generation degree of the TNF-α was performed by the following method.
More specifically, after dispensing RAW264.7 cells into a 60 mm dish at 1 × 10 6 cells / ml and stabilizing the cells for 24 hours, red ginseng extract (RG), fermented red ginseng extract (FRG) and fermented red ginseng tablets Water (PFRG) was treated at 1 ug / ml and 10 ug / ml, respectively, and after 1 hour, LPS was treated at a concentration of 0.1 ug / ml, followed by 2 hours of incubation, and the cells were harvested. The harvested cells were centrifuged at 12,000 rpm for 3 minutes to obtain cells (cell pellet), 200 ul TRIzol ® was added to the obtained cell pellet and allowed to stand at room temperature for 5 minutes. Thereafter, 40 ul of CHCl 3 was added thereto, shaken for 15 seconds, allowed to stand at room temperature for 3 minutes, and then centrifuged for 15 minutes at 12,000 × g and 4 ° C. to separate only the supernatant. 100 ul of isopropanol was added to the supernatant and allowed to stand at room temperature for 10 minutes. Thereafter, the supernatant was removed by centrifugation for 10 minutes at 12,000 xg and 4 ° C, and 200 ul of ice-cold 75% ethanol was added and vortexed. Thereafter, the supernatant was removed after centrifugation for 5 minutes at 7500 xg and 4 ° C and air dried at room temperature for 10 minutes. After drying, RNase free water was added to the resultant precipitate and dissolved, and the mixture was left at 60 ° C. for 10 minutes and stored at −20 ° C.
Quantify the isolated RNA by less than 100 ng and forward TNF-α primer (5'-GGC CTT CCT ACC TTC AGA CC-3 ') and reverse TNF-α primer (5'-AAG CAA AAG AGG AGG CAA CA-3 ') And Actin forward actin primer (5'-TTC TTT GCA GCT CCT TCG TT-3') and Actin reverse actin primer (5'-CTT CTC CAT GTC GTC CCA GT-3 '), respectively, and RT-PCR ( annealing Tm = 60 ° C., cycle = 23). The PCR product subjected to RT-PCR was electrophoresed on a 2% agarose gel and stained with EtBr to confirm the result, and is shown in FIG. 7.
As shown in FIG. 7, the amount of mRNA produced by TNF-α was significantly increased in the LPS treated group (P) compared to the non-LPS treated group (N). On the other hand, when the red ginseng extract (RG), fermented red ginseng extract (FRG) and fermented red ginseng purified product (PFRG) were treated, the amount of mRNA of TNF-α was reduced in proportion to the treatment concentration, and the extent of the red ginseng extract In comparison, the fermented red ginseng extract was confirmed to have a larger fermented red ginseng extract than the fermented red ginseng extract, and it was confirmed that the fermented red ginseng extract or the fermented red ginseng purified product inhibited the expression level of TNF-α mRNA.
In addition, the expression level of TNF-α was quantified using ELISA Kits (Pierce Endogen, USA). The measurement results are shown in FIG. 8.
As shown in FIG. 8, it was confirmed that the anti-inflammatory activity of the fermented red ginseng tablets was the best. More specifically, in the group (N) not treated with LPS, about 50% of TNF-α was expressed, whereas the amount of NO production was significantly increased in the LPS treated group (P). On the other hand, when the red ginseng extract (RG), fermented red ginseng extract (FRG) and fermented red ginseng purified product (PFRG) were treated, the amount of NO was reduced in proportion to the treated concentration, the degree of fermented red ginseng extract compared to the red ginseng extract The fermented red ginseng extract was confirmed to be larger than the fermented red ginseng extract, and the effect difference was confirmed for each kind.
Experimental Example 2-6. Nitric oxide ( NO Generation and quantification
In order to confirm the anti-inflammatory activity of the red ginseng extract and fermented red ginseng extract (Example 1) and fermented red ginseng purified product (Example 2), it was confirmed by analyzing the inhibition of NO production.
First, RAW264.7 cells were dispensed in a 60 mm dish at 1 × 10 6 cells / ml, and after stabilizing the cells for 24 hours, the red ginseng extract, the fermented red ginseng extract and the fermented red ginseng tablets were respectively 1 ug / ml and Treatment was performed at a concentration of 10 ug / ml, and after 1 hour, 0.1 ug / ml of the LPS was incubated for 24 hours, and the culture solution was collected. The culture solution was centrifuged at 12,000 rpm for 3 minutes, 100 ul of the supernatant was quantified using Griess reagent. Specifically, 100 ul of the supernatant obtained above and 150 ul of Griess reagent (1% sulfanilamide in 5% phosphoric acid and 1% α-naphthylamide in H 2 0) were mixed in a 96 well plate and reacted for 5 minutes at room temperature. Absorbance was measured with an ELISA microplate reader at nm. After absorbance was measured with sodium nitrite (NaNO 2 ) to obtain a standard curve, the concentration of NO was calculated from the measured absorbance. The r 2 value of the standard curve for the standard NO (sodium nitrite) was more than 0.99. The calculated NO value is shown in FIG. 9.
As shown in FIG. 9, it was confirmed that the anti-inflammatory activity of the fermented red ginseng tablets was the best. More specifically, in the case of the group (N) not treated with LPS, NO was hardly measured, whereas the amount of NO produced was significantly increased in the LPS treated group (P). On the other hand, when the red ginseng extract (RG), fermented red ginseng extract (FRG) and fermented red ginseng purified product (PFRG) were treated, the amount of NO was reduced in proportion to the treated concentration, the degree of fermented red ginseng extract compared to the red ginseng extract The fermented red ginseng extract was confirmed to be larger than the fermented red ginseng extract, and the effect difference was confirmed for each kind.
From the above results, it was confirmed that the fermented red ginseng extract or the fermented red ginseng purified product inhibited the amount of NO in comparison with the red ginseng extract.
As described above, the present invention fermented red ginseng extract or fermented red ginseng purified product was not only recognized safety in experiments using cytotoxicity, but also the expression of iNOS protein, the amount of COX-2, the amount of NF-kB, the amount of TNF- The anti-inflammatory effect of fermented red ginseng extract or fermented red ginseng tablets was significantly superior to conventional red ginseng extracts as a result of α-m-RNA production and nitric oxide production. These results indicate that the content of ginsenoside Rg3, ginsenoside, Rk1 ginsenoside Rg5 and compound K (Compoun K, CK) contained in the fermented red ginseng purified water was significantly increased compared to the red ginseng extract. Is evaluated.
The ginseng produced in Korea is called Korean ginseng and its value is recognized worldwide, but it occupies about 3% of the world ginseng market. In this respect, in relation to ginseng processed products, the technology of the present invention that can produce high value-added ginseng products by processing Korean ginseng, which is an excellent raw ginseng containing various ginsenosides, is a product of ginseng cultivated farms and ginseng related industries. It is expected to contribute to the high value added.
Claims (8)
Lactobacillus genus ( Lactobacillus) in the ginseng extract sp.) adding a lactic acid bacterium and carrying out a fermentation process to prepare a fermented product of ginseng extract; And
Heat-treating the fermented product of the ginseng extract, which is carried out by heating the fermented product of the ginseng extract at 100 ° C. to 150 ° C. for 60 to 120 minutes.
Method for producing a ginseng extract with an increased content of trace ginsenosides comprising a.
The ginseng is a method of producing ginseng extract with increased content of trace ginsenosides of red ginseng.
Lactobacillus genus sp.) Lactobacillus was deposited with Lactobacillus brevis BJ10 ( Lactobacillus) brevis BJ20 KCTC 11377BP) A method for preparing ginseng extract with increased content of trace ginsenosides.
Preparing the fermented product of the ginseng extract is Lactobacillus genus ( Lactobacillus) in the ginseng extract sp.) A method for preparing a ginseng extract with an increased content of trace ginsenosides, which is added to lactic acid bacteria and performed at 33 ° C to 39 ° C for 12 to 96 hours.
The trace ginsenoside is ginsenoside Rg3, ginsenoside Rk1 ginsenoside Rg5 and compound K (Compoun K) The method of producing a ginseng extract with an increased content of the trace ginsenoside.
Method for producing a ginseng extract with an increased content of trace ginsenosides, characterized in that further performing the process of purifying the fermented product of the heat-treated ginseng extract.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20150113651A (en) * | 2014-03-31 | 2015-10-08 | 송하석 | Method for Preparing Ginsenoside R3-rich Fermented Red Genseng Using Bio-conversion |
| KR20150140550A (en) * | 2014-06-05 | 2015-12-16 | 동국대학교 경주캠퍼스 산학협력단 | Ginseng preparations containing abundant ginsenoside using a beauveria bassiana and method thereof |
| KR20160065245A (en) * | 2014-11-28 | 2016-06-09 | (주)지에프씨 | Composition of culture medium for cultivation of Lactobacillus, ginseng berry-fermented extracts comprising thereof and cosmetic composition for improving skin conditions |
| KR20190012299A (en) * | 2017-07-26 | 2019-02-11 | 이상호 | Composition and method for increasing saponin content of ginseng |
| KR101963978B1 (en) * | 2017-11-10 | 2019-04-01 | 농업회사법인 드림팜 유한회사 | Composition of fermented sprout ginseng having increased ginsenosides Rh2 and compound K, bioactive components, and preparation method thereof |
| KR20190114315A (en) * | 2018-03-29 | 2019-10-10 | 주식회사 화진바이오코스메틱 | Method for producing fermentative product of cultured mountain ginseng extract with increased small molecule ginsenoside content and fermentative product of cultured mountain ginseng extract with increased small molecule ginsenoside content produced by the same method |
| KR20200077060A (en) * | 2018-12-20 | 2020-06-30 | 경남과학기술대학교 산학협력단 | Composition for anti-inflammation containing active mountain-cultivated ginseng and preparation method thereof |
| KR20200121196A (en) * | 2019-04-15 | 2020-10-23 | 주식회사 리바이오 | Method for Preparation of Bioconverted Red Ginseng with Enhanced compound K content |
| KR20220132152A (en) * | 2021-03-23 | 2022-09-30 | 재단법인 진안홍삼연구소 | A validation method of compound k analysis method |
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2011
- 2011-05-31 KR KR1020110052408A patent/KR20120134166A/en not_active Application Discontinuation
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20150113651A (en) * | 2014-03-31 | 2015-10-08 | 송하석 | Method for Preparing Ginsenoside R3-rich Fermented Red Genseng Using Bio-conversion |
| KR20150140550A (en) * | 2014-06-05 | 2015-12-16 | 동국대학교 경주캠퍼스 산학협력단 | Ginseng preparations containing abundant ginsenoside using a beauveria bassiana and method thereof |
| KR20160065245A (en) * | 2014-11-28 | 2016-06-09 | (주)지에프씨 | Composition of culture medium for cultivation of Lactobacillus, ginseng berry-fermented extracts comprising thereof and cosmetic composition for improving skin conditions |
| KR20190012299A (en) * | 2017-07-26 | 2019-02-11 | 이상호 | Composition and method for increasing saponin content of ginseng |
| KR101963978B1 (en) * | 2017-11-10 | 2019-04-01 | 농업회사법인 드림팜 유한회사 | Composition of fermented sprout ginseng having increased ginsenosides Rh2 and compound K, bioactive components, and preparation method thereof |
| KR20190114315A (en) * | 2018-03-29 | 2019-10-10 | 주식회사 화진바이오코스메틱 | Method for producing fermentative product of cultured mountain ginseng extract with increased small molecule ginsenoside content and fermentative product of cultured mountain ginseng extract with increased small molecule ginsenoside content produced by the same method |
| KR20200077060A (en) * | 2018-12-20 | 2020-06-30 | 경남과학기술대학교 산학협력단 | Composition for anti-inflammation containing active mountain-cultivated ginseng and preparation method thereof |
| KR20200121196A (en) * | 2019-04-15 | 2020-10-23 | 주식회사 리바이오 | Method for Preparation of Bioconverted Red Ginseng with Enhanced compound K content |
| KR20220132152A (en) * | 2021-03-23 | 2022-09-30 | 재단법인 진안홍삼연구소 | A validation method of compound k analysis method |
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