KR20170001397A - Novel Paenibacillus sp. RMKS 72, fermented ginseng using the same and composition having anti-oxidant activity comprising extract of fermented ginseng using the same - Google Patents

Abstract

The present invention relates to a novel Paenibacillus sp. RMKS 72 strain (microorganism donation number KACC92078P), a fermented ginseng using the same, and a composition having an antioxidant activity and containing an extract of the fermented ginseng. A culturing liquid of the Paenibacillus sp. RMKS 72 strain (microorganism donation number KACC92078P) has an excellent effect in converting ginsenoside Rb1 in a ginseng into Rd, and a fermented ginseng produced by using the strain or an extract thereof has an excellent removing function with respect to active oxygen species. Accordingly, the fermented ginseng or the extract thereof can be useful as an antioxidant and anti-aging cosmetic composition, a health functional food, or a pharmaceutical composition.

Description

[TECHNICAL FIELD] [Technical Field] The present invention relates to novel RMKS 72 strain of Fanny Bacillus, a fermented ginseng using the same, and a fermented ginseng extract using the composition, which has antioxidant activity {Novel Paenibacillus sp. RMKS 72, fermented ginseng using the same and composition having anti-oxidant activity comprising fermented ginseng using the same}

The present invention relates to a novel strain of the Fanny Bacillus strain RMKS 72 ( Paenibacillus sp. RMKS 72, Microorganism Accession No. KACC92078P) and fermented ginseng using the same. Further, the present invention relates to a composition having an antioxidative activity containing an extract of fermented ginseng using the above-mentioned Fannibacillus sp. RMKS 72 strain.

Ginseng (Panax ginseng) has been widely used as a drug or health food in the East, including the country the past few thousand years ago, a herbaceous perennial belonging to the genus Araliaceae senticosus (Aralaiaceae) ginseng. In addition, ginseng can be used in various forms such as ginseng and red ginseng. Especially, it is known that red ginseng processed by steaming steam ginseng contains various ginsenosides useful for human body.

Saponin, which represents the major efficacy of ginseng, is composed of more than 30 kinds of ginsenosides. Ginsenosides are used to enhance immunity, antiinflammatory action, antiallergic action, anticancer effect, effect on erectile dysfunction, Action, anticholesterol action, antithrombotic action, adult disease and aging.

Saponin of ginseng is a neutral glycoside in which glucose, arabinose, xylose, rhamnose and the like are bonded to a triterpenoid-based dammarane skeleton . When ginseng is extracted with water and alcohol, it is confirmed that Rb1, Rb2, Rc, Rg1 and the like exist in a large amount among the ginsenosides of ginseng. However, these ginsenosides are not well absorbed in the body and are not suitable for use as a pharmaceutical composition. The ginsenosides in the ginseng are converted into ginsenosides which are not present in the natural world through the process of processing them with red ginseng or black ginseng, so that they are well absorbed in the body and can be suitably used as a pharmaceutical composition of various diseases have.

That is, the glycosyl residues are separated from Rb1, Rb2, Rc, Rg1 and the like among the ginsenosides of ginseng to produce ginsenosides Rg2, Rg3, Rh2, Rh1, Rk5, Rk1, Rg6, Rk3, F4, and Rh4 are produced through the dehydration process of the water.

In addition, the ginsenosides are hydrolyzed by the addition of an organic acid and the sugar is removed and the side chain portion is cyclized to form Panaxatriol or Panaxadiol (Chem. Pharm Chem. Pharm Bull., 22: 2407-2421, 1974; Chem. Pharm Bull., 22: 421-428, 1974).

On the other hand, when ginsenosides enter the body, ginsenosides such as Rb1, Rb2, Rc, and Rg1 are degraded by intestinal microorganisms and can be absorbed (Planta Med. 62. 453-457, 1996) . The ginsenosides are sequentially degraded by intestinal microorganisms, and Rb1 is sequentially converted to Rd, Compound K and protopanaxadiol, but the presence or absence of microorganisms and the degree of retention The absorption by the conversion of the metabolite is considerably different and can also vary considerably in its efficacy (J. Pharmacol. 50, 1155-1160, 1998).

Therefore, in order to eliminate the difference between the efficacy and the absorption rate in the body by the conversion of the metabolite, the ginsenoside absorption rate of the body is increased through the conversion of the ginsenoside in the ginseng by the microorganism, There is a need for development of fermented ginseng which is strengthened en masse.

On the other hand, skin exists in the outermost part of the body and is an important organ that protects the body from the outside. Every organ of all organisms is aging as it gets older, and skin is no exception. In general, skin aging is divided into extrinsic aging and extrinsic aging such as photoaging. Intrinsic aging means that aging occurs naturally with age, and photoaging refers to aging caused by sunlight.

Endogenous aging reduces the secretion of various hormones that regulate metabolism, decreases the function of immune cells and the activity of cells, reduces the biosynthesis of immune proteins and biologically constitutive proteins necessary for the living body, and exogenous aging destroys the ozone layer As the amount of ultraviolet rays reaching the surface of the sunlight increases and the environmental pollution is further intensified, free radicals and active noxious oxygen are increased, thereby decreasing the thickness of the skin, increasing the wrinkles and decreasing the elasticity Skin color is not good, skin troubles occur frequently, and spots, freckles and black blots also increase.

Among endogenous and exogenous extrinsic aging, they are known to be deeply involved with active oxygen. In the endogenous aging, active oxygen generated in the metabolism process occurs mainly in the living cells. In the case of exogenous aging, This is the main cause.

Therefore, it is necessary to develop an antioxidant capable of removing active oxygen to make a pharmaceutical composition for preventing skin aging or a skin aging delay or improvement cosmetic. As an antioxidant used conventionally, there is an enzyme called superoxide dismutase (SOD) and vitamin C, vitamin E (tocopherol) and flavonoid. However, since these antioxidants are unstable, they are applied to cosmetics having a long shelf life There is a problem that it is not easy and it is possible to cause side effects on the skin when using excessive amount.

Accordingly, the present inventors have continuously studied to identify microorganisms capable of converting ginsenosides into ginsenosides, which can increase the absorption rate of ginsenosides among a number of fermenting microorganisms. As a result, it has been found that Rb1 in ginseng can be selectively converted to Rd Gt; glucosidase < / RTI > enzyme. Also, it has been confirmed that the extract of fermented ginseng fermented with the microorganism has excellent antioxidative activity, but is not useful as a natural product and thus can be effectively applied as a pharmaceutical composition, a health functional food or a cosmetic composition.

As prior arts, Korean Patent No. 1285681 and Korean Patent No. 472964 disclose fermented ginseng having enhanced absorption rate in the body using lactic acid bacteria having saponin bioconversion ability, Korean Patent Publication Nos. 2013-0063959 and Korea Patent No. 1182741 discloses that microorganism-derived β-glucosidase has a function of converting ginsenoside Rb1 into a type of ginsenoside Rd that is easy to use in the body. These techniques, however, A strain completely different from the strain is used.

Korean Patent No. 1343434, Korean Patent No. 1343410 and Korean Patent No. 1345226 disclose that lactobacillus plantarum, Enterococcus faecalis, and Fanny Bacillus subtilis 213, which have ginseng extracting activity, Discloses a method for producing fermented ginseng using the strain. However, since these prior arts also produce a fermented ginseng using a strain completely different from the present invention, have.

Korean Registered Patent No. 472964 (Fermented Ginseng Containing Saponin Decomposition and Preparation Method Thereof, Registered on Feb. 15, 2005) Korean Registered Patent No. 1017378 (Method of producing fermented ginseng, fermented red ginseng, and Bacillus subtilis strain derived from chungkukjang, which is suitable for this, registered on Feb. 17, 2011) Korean Registered Patent No. 1182741 (Effective Bioconversion Method and Its Product of Ginsenoside Albi 1, Registered on Sep. 07, 2012) Korean Patent No. 1285681 (a strain having saponin biosynthesis ability and a method for producing fermented ginseng having enhanced absorption rate in the body using the same, 2013.07.04. Registered) Korean Registered Patent No. 1343434 (New Enterococcus faecalis CRNB-A3 [KCTC11930 BP] strain having ginseng extract efficacy and saponin conversion method using the same, registered on December 13, 2013) Korean Registered Patent No. 1343410 (New Lactobacillus plantarum CRNB-22 [KCTC11931 BP] strain having ginseng extract efficacy and saponin conversion method using the same, registered on December 13, 2013) Korean Registered Patent No. 1345226 (Composition for preventing or treating atopic dermatitis containing novel fermented ginseng and fermented ginseng extract using the same, 213 strain of Fanny Bacillus genus 213, registered on December 19, 2013) Korean Patent Laid-Open Publication No. 2013-0063959 (Effective Conversion Method of Ginsenoside Albi 1 Using an Enzyme Produced by Aspergillus Usami KCTC 6954 and Its Product, Published on Mar. 17, 2013)

David Kiefer et al., Panax ginseng. American Family Physician 68 (8), 1539-1542, 2003. Shibata S. et al. Studies on the constituents of Japanese and Chinese crude drugs. XI. Panaxadiol, a sapogenin of ginseng roots. Chem. Pharm. Bull. 11,759-761,1963. Shibata S. et al., Studies on the saponins of Ginseng. Ⅱ. Structures of ginsenoside-Re, -Rf and -Rg2. Chem. Pharm. Bull. 22, 2407-2421, 1974. Shibata S. et al., Studies on the saponins of Ginseng. I. Structures of ginsenoside-Ro, -Rb1, -Rb2, -Rc and -Rd. Chem. Pharm. Bull. 22, 421-428, 1974. Hasegawa H. et al., Main ginseng saponin metabolites formed by intestinal bacteria. Planta Med. 62 (5), 453-457, 1996. Akao T. et al., Intestinal bacterial hydrolysis is required for the appearance of compound K in rat plasma after oral administration of ginsenoside Rb1 from Panax ginseng. J Pharm Pharmacol. 50 (10), 1155-1160,1998. Chang-Su Park, et al., Biotransformation of ginsenosides by hydrolyzing the sugar moieties of ginsenosides using microbial glycosidases. Microbiol Biotechnol. 89, 9-19, 2010. Bong-Gwan Kim. et al., Changes in ginsenosides in Korean red ginseng (Panax ginseng) fermented by Lactobacillus plantarum M1. Process Biochemistry. 45, 1319-1324, 2010. Le-Qin Cheng. et al., Conversion of major ginsenoside Rb 1 to ginsenoside F 2 by Caulobacter leidyia. Biotechnol Lett. 28, 1121-1127, 2006. Le-Qin Cheng, et al., Conversion of major ginsenoside Rb1 to 20 (S) -ginsenoside Rg3 by Microbacterium sp. GS514. Phytochemistry. 69, 218-224, 2008.

It is an object of the present invention to provide a novel strain of RMKS 72 of the genus Fanny Bacillus ( Paenibacillus sp. RMKS 72, Microorganism Accession No. KACC92078P) and fermented ginseng using the same.

It is also an object of the present invention to provide a composition having an antioxidative activity containing an extract of fermented ginseng using the strain Fannibacillus RMKS 72.

The present invention relates to a novel strain of the Fanny Bacillus strain RMKS 72 ( Paenibacillus sp. RMKS 72, Microorganism Accession No. KACC92078P).

The present invention also relates to fermented ginseng using the strain of Fanny Bacillus sp. RMKS 72.

The fermented ginseng is prepared by mixing 100 parts by weight of ginseng, 200-1000 parts by weight of water and 1 to 25 parts by weight of a culture solution of a strain of FMKS 72 of the Fanny Bacillus strain And may be fermented at 25 to 40 DEG C for 3 to 15 days.

The present invention also relates to a cosmetic composition having an antioxidative activity containing an extract of fermented ginseng using a strain of Fannibacillus RMKS 72.

The extract of the fermented ginseng may be an extract obtained by extracting the fermented ginseng using the strain of Fanny Bacillus strain RMKS 72 with water, C1 to C4 lower alcohol, or a mixed solvent thereof.

The present invention also relates to a health functional food having an antioxidative activity containing an extract of fermented ginseng using a strain of Fanny Bacillus sp. RMKS 72.

The present invention also relates to an anti-aging pharmaceutical composition containing an extract of fermented ginseng using the strain Fannibacillus RMKS 72.

Further, the present invention provides a method for converting ginsenoside Rb1 into ginsenoside Rd using a strain of the genus RMK72 of the genus Fannibacillus, comprising the steps of: obtaining a culture of a strain of the genus RMK72 of the genus Fannibacillus; Mixing 0.01 to 10 mg / ml of ginsenoside Rb1 and the culture solution in a volume ratio of 1: 1 to 5; And reacting at 25 to 40 占 폚 and a pH of 5.0 to 9.0 for 3 to 15 days.

The present invention also provides a method for producing fermented ginseng having enhanced ginsenoside Rd using the strain of the genus RMKS 72 of the genus Fannibacillus, comprising the steps of: obtaining a culture of a strain of the Fanny Bacillus strain RMKS 72; 100 parts by weight of ginseng, 200-1000 parts by weight of water and 1 to 25 parts by weight of a culture of a strain of Fanny Bacillus strain RMKS 72; And reacting at 25 to 40 占 폚 and a pH of 5.0 to 9.0 for 3 to 15 days.

Hereinafter, the present invention will be described in more detail.

The fermentation broth of RMKS 72 is an anaerobic, gram-positive, motile, and sporulating bacterium. The temperature at which it can grow is 25 to 40 ° C, preferably 30 to 37 ° C .; the viable pH is 5.0 to 8.0, To 7.5, and a suitable time for culturing is 36 to 96 hours, preferably 48 to 96 hours, and most preferably 48 to 72 hours. The strain does not grow well under conditions of less than 25 캜, more than 40 캜, less than pH 5.0, and a pH exceeding 8.0.

The most suitable medium for culturing is TS medium (trypticase soy broth), BCP medium (Bromo Cresol Purple-Deoxycholate-Citrate-Lactose-Sucrose broth) , GYP medium (Glucose-Yeast-Peptone broth).

The FMANBASUS strain RMKS 72 has β-glucosidase activity and selectively converts ginsenoside Rb1 to Rd without acting on other ginsenosides. In particular, the RMKS 72 strain of the genus Fanny Bacillus has a high conversion rate to ginsenoside Rd.

The fermented ginseng using the RMKS 72 strain of the Fanny Bacillus strain may be fermented by mixing 100 parts by weight of ginseng, 200-1000 parts by weight of water and 1 to 25 parts by weight of a culture medium of the Fanny Bacillus strain RMKS 72. At this time, the fermentation progresses even if water is not added or only a very small amount is added, but the time for fermentation progresses until the conversion effect of ginsenoside appears. Therefore, it is preferable that the water content is 200 parts by weight or more, and if the content of water exceeds 1000 parts by weight, fermentation can proceed but this also increases the time of fermentation. In addition, when the culture broth is less than 1 part by weight based on 100 parts by weight of ginseng, the fermentation time is increased, and the culture broth may be more than 25 parts by weight. However, It is not necessary to add more.

The fermentation is preferably carried out at a temperature of 25 to 40 DEG C for 3 to 15 days. If the temperature is lower than 25 占 폚 or exceeds 40 占 폚, the fermentation does not work well. If the fermentation period is less than 3 days, the fermentation is not sufficiently advanced and it may be fermented for over 15 days. However, since the conversion of ginsenoside is sufficiently performed within 15 days, The period should be within 15 days. Therefore, the fermented ginseng having the best fermentation effect can be produced within the range of the fermentation temperature and the fermentation period.

The fermented ginseng according to the present invention is a fermented ginseng which has been converted into a form suitable for absorption and utilization in the body, and in particular, the fermented ginseng having an enhanced content of ginsenoside Rd It is ginseng.

On the other hand, it is also possible to obtain ginsenoside Rd from ginsenoside Rb1 by applying the FMANBASUS strain RMKS72 strain of the present invention to the ginsenoside Rb1 alone. To this end, the cultured medium of the strain FMKS 72 belonging to the Fanny Bacillus strain was centrifuged, 0.01 to 10 mg / ml of ginsenoside Rb1 dissolved in water and the centrifuged culture broth were mixed at a volume ratio of 1: 1 to 5, 40 ° C, pH 5.0 to 9.0 for 3 to 15 days. At this time, centrifugation of the culture liquid is suitably performed at 5000 rpm to 10,000 rpm.

The concentration of ginsenoside Rb1 can be used in any concentration, but 0.01 to 10 mg / ml is most appropriate in terms of conversion efficiency of ginsenoside. At this time, when the ginsenoside Rb1 and the centrifuged culture broth are mixed, it is possible to switch to ginsenoside Rd even when the volume ratio is 1: 1-5. However, Side conversion time is long, which is uneconomical.

Further, in the reaction condition of 25 to 40 캜, pH 5.0 to 9.0, and 3 to 15 days, which is a ginsenoside conversion condition, the conversion of ginsenoside is not performed well under 25 캜 or above 40 캜 It may be time-consuming, and it may be undesirable for the growth of the strain even under the condition of pH 5.0 or below, and the conversion of ginsenoside may be difficult or time-consuming. In addition, even if the reaction period is less than 3 days, the conversion of ginsenoside does not sufficiently take place, or it may take a long time. However, even if it exceeds 15 days, the conversion of ginsenoside is good, but the conversion is completed within 15 days and most of the ginsenoside is completed.

The extract of fermented ginseng using the strain FMKS 72 in the Fanny Bacillus strain was prepared by adding 2 to 20 times (w / w) water of the fermented ginseng to the fermented ginseng (on a dry matter basis) using the strain Fanny Bacillus strain RMKS 72, Of a lower alcohol or a mixed solvent thereof. The extract of fermented ginseng using the strain of Fannibacillus sp. RMKS 72 may be used as it is when it is prepared as a water extract or may be used in the form of a concentrate or a dried product in any case where it is extracted using other types of solvents such as water have.

The extract of fermented ginseng using the above-mentioned RMKS 72 strain of the genus Fanny Bacillus may be extracted with an organic solvent (alcohol, ether, acetone, etc.), an organic solvent (n-hexane, ethyl acetate, n-butanol) Column chromatography, and known methods used for the separation and extraction of plant components, alone or in combination, and further purified, and further purified according to the conventional method, if necessary.

The chromatography used in the present invention includes silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, thin layer chromatography thin layer chromatography, medium pressure liquid chromatography, and high performance liquid chromatography may be used.

Ginseng is a perennial plant belonging to the genus Panax (Panax) for use in the production of the fermented ginseng, ginseng (Panax ginseng), hwagisam (Panax quinquefolia , Panax notoginseng , Panax japonicus), Himalayan cedar (Panaxa pseudoginseng). Vietnamese Panax vancomycin , vietnamensis , Panax elegatior , Panax wangianus and Panax bipinratifidus , Panax angustifolium , and the like. But is not limited thereto. These ginsengs may be used alone or in combination of two or more.

It is preferable that the ginseng is roots of ginseng 4-6 years old.

The present invention also provides a cosmetic composition having an antioxidative activity comprising a fermented ginseng extract using a strain of Fanny Bacillus sp. RMKS 72.

The fermented ginseng extract using the RMKS 72 strain of the Fanny Bacillus strain may contain 0.001 to 90% by weight, preferably 0.01 to 50% by weight, more preferably 0.1 to 30% by weight based on the total weight of the cosmetic composition .

The cosmetic composition may contain, in addition to the fermented ginseng extract using the RMKS 72 strain of the Fanny Bacillus genus, all components commonly used in cosmetics. For example, it may contain common auxiliary components such as emulsifiers, thickeners, emulsions, surfactants, lubricants, alcohols, water soluble polymers, gelling agents, stabilizers, vitamins, inorganic salts, emulsifiers and perfumes. The above-mentioned components can be selected within a range that does not impair the effect inherent in the cosmetic, depending on the purpose of formulation or use.

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used in the form of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a flexible lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a hair tonic, a shampoo, a rinse, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray and a powder .

The cosmetic composition of the present invention can be used everyday or can be used for an unspecified period. Preferably, the amount of use, the frequency of use, and the period of time can be adjusted according to the age, skin condition or skin type of the user, and concentration of the fermentation extract.

The present invention also provides an anti-aging pharmaceutical composition containing an extract of fermented ginseng using a strain of Fannibacillus RMKS 72. The aging is preferably skin aging, but is not limited thereto.

The pharmaceutical composition of the present invention can be formulated in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or the like oral preparation, external preparation, suppository and sterilized injection solution according to a conventional method .

Examples of carriers, excipients and diluents that can be contained in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. The solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like. These solid preparations can be prepared by adding to the extract of fermented ginseng using the Fannibacillus RMKS 72 strain at least one excipient such as starch, Calcium carbonate, sucrose, lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The dosage of the extract of fermented ginseng using the strain Fannibacillus RMKS 72 varies depending on the age, sex, body weight, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the administration route and the judgment of the prescriber will be. Dosage determinations based on these factors are within the level of ordinary skill in the art and generally the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day on a dry basis. A more preferable dosage is 0.1 mg / kg / day to 500 mg / kg / day. The administration may be carried out once a day or several times. The dose is not intended to limit the scope of the invention in any way. The extract of fermented ginseng using the RMKS 72 strain of the Fanny Bacillus strain of the present invention can be administered to mammals such as rats, livestock, and humans in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection. Further, since the extract of fermented ginseng of the present invention is derived from a natural product, it has little toxicity and side effects and can be safely used for prolonged use even for prophylactic purposes.

The present invention also provides a health functional food having an antioxidative activity containing an extract of fermented ginseng using a strain of Fannibacillus sp. RMKS 72.

Food-acceptable food-aid additives may be added to the health functional food. The health functional food may be in the form of tablets, capsules, pills or liquids. The health functional foods include dairy products such as drinks, meat, sausage, bread, candy, snacks, noodles, It may include nutritional products, including beverages, alcoholic beverages and vitamin complexes, including ionic beverages.

In addition, the fermented ginseng extract of the Fanny Bacillus strain RMKS 72 may be directly commercialized as a health functional food, and various additives may be blended and commercialized to enhance flavor and fragrance. Examples of additives include various saccharides, emulsifiers, sweeteners, acidulants, juices, and the like. Specific examples thereof include saccharides such as glucose, sucrose, fructose, and acacia, sugar alcohols such as sorbitol, xylitol, erysuricol, lactitol, and paratinide, fatty acid esters such as glycerin fatty acid esters, and emulsifiers such as lecithin In addition, various vitamins such as vitamin A, vitamin B, vitamin C and vitamin E, plant extract, cereal ingredient and vegetable ingredient can be used as additives. On the other hand, the extract of the fermented ginseng may be provided as a form that is easily sterilized and packaged in a pouch form.

The present invention provides a novel antioxidant composition comprising a novel fermented ginseng and an extract of the fermented ginseng using the same, wherein the fermented ginseng is selected from the group consisting of Paenibacillus sp. RMKS 72 (microorganism accession number: KACC92078P).

The RMCS 72 strain of the Fanny Bacillus strain is excellent in converting ginsenoside Rb1 having a low bioavailability into Rd. Also, the fermented ginseng or its extract prepared by using the RMKS 72 strain has excellent scavenging activity against reactive oxygen species, and is useful as a cosmetic composition for preventing oxidation and aging, a health functional food, or a pharmaceutical composition Lt; / RTI >

FIG. 1 is a sequence chart (bottom, unknown 72 is RMKS 72) showing the nucleotide sequence (top) and phylogenetic position of 16s DNA gene of FannBacillus sp. RMKS 72 strain.
FIG. 2A shows a TLC analysis result of the reaction of ginsenoside Rb1 for 7 days at a temperature (30, 35, 40 DEG C) in a culture solution of a strain belonging to the genus RMKS 72 of the genus Fanny Bacillus.
FIG. 2B shows the results of TLC analysis of Rb1 reacted for 7 days at pH 4.0 (pH 4.0, 5.0, 6.0, 7.0, 8.0, and 9.0) in the culture medium of the FMKS 72 strain of the genus Fanny Bacillus.
FIG. 2C shows the results of TLC analysis of reaction of ginsenoside Rb1 at pH 7.0 and 35.degree. C. for 3 to 14 days in the culture medium of the strain FMKS 72 belonging to the genus Fanny Bacillus.
FIG. 2D shows the TLC analysis results of fermented ginseng obtained by reacting ginseng at pH 7.0 and 35 ° C for 3 to 14 days in a culture solution of RMKS 72 strain of the genus Fanny Bacillus.
FIG. 3 shows HPLC analysis spectra of Rb1 reacted with a culture medium of F. vulnificus RMKS 72 at pH 7.0, 35 ° C. for 3 to 14 days.
FIG. 4 shows HPLC analysis spectra of fermented ginseng fermented at pH 7.0, 35 ° C., and 3 to 14 days in the culture medium of Fannibacillus sp. RMKS 72.
FIG. 5 shows the result of measuring the reactive oxygen species scavenging activity of the extract of fermented ginseng using the strain RMKS 72 of the genus Fanny Bacillus.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, the intention is to provide an exhaustive, complete, and complete disclosure of the principles of the invention to those skilled in the art.

<Experimental Example 1 Isolation and Identification of New Microorganisms>

Bromocresol purple-deoxycholate-Citrate-Lactose-Sucrose agar was used to isolate 44 kinds of microorganisms from the EM culture medium. Among them, colonies having β-glucosidase activity were cultured in an Esculin agar medium and black colonies (β-glucosidase activity positive) colonies were separated to obtain 16S DNA Respectively.

The strain (colony) isolated from the Esculin agar medium is anaerobic, gram positive, motile, and sporulated bacterium. When cultured using a TS liquid medium, the growth temperature is 25 to 40 ° C, preferably 30 to 37 ° C , And the viable pH was 5.0 to 8.0, preferably 6.0 to 7.5. The time for culturing the strain was 36 to 96 hours, preferably 48 to 96 hours, and most preferably 48 to 72 hours.

On the other hand, DNA of the isolated strain was extracted and 16S DNA was analyzed by Solgent (Daejeon). The strain DNA was extracted and PCR was performed using a universal primer (5'-GAGTTTGATCCTGGCTCAG-3 'and 5'-AGAAAGGAGGTGATCCAGCC-3') to amplify 16S rDNA (16S rRNA coding gene). The amplification was carried out at 96 ° C for 2 minutes, followed by denaturation at 96 ° C for 10 seconds, annealing at 50 ° C for 5 seconds, extension at 60 ° C for 25 minutes, and final extension at 60 ° C for 2 minutes. Amplified 16S rDNA was purified using a PCR kit (Product Purification Kit, Qiagen) and analyzed using genetic analyzer 377 (Perkin-Elmer). Molecular phylogenetic analysis based on the nucleotide sequence of 16S rDNA revealed that the 16s DNA of Fanny Bacillus sp. Y412MC10 ( Paenibacillus sp. Y412MC10) as a strain belonging to the phylogenetic group including Paenibacillus genus was identical to 1,059 bp among 1,089 bp And 97%, respectively (see the bottom of FIG. 1). Thus, the strain Fannie Bacillus were identified as strains (Paenibacillus sp), Fannie Bacillus RMKS 72 strains (Paenibacillus sp. RMKS 72) as named by the National Academy of Agricultural Sciences Agricultural Genetic Resources Center in the year 06 2,015 05 microorganisms accession number KACC92078P. &Lt; / RTI &gt;

< Experimental Example  2. Fanny Bacillus  genus RMKS  72 strains Gin Senocide  Check your conversion>

Experimental Example 2-1. Preparation of strain culture medium

The RMKS 72 strain of the genus Fanny Bacillus was cultured in a TS liquid medium (trypticase soy broth, pH 7.3) at a culturing temperature of 30 ° C for 24 hours. After culturing, the mixture was centrifuged at 7,500 rpm for 15 minutes, and the centrifuged supernatant (liquid phase except supernatant and strain pellet) was used as a culture solution to convert ginsenosides.

Experimental Example 2-2. Identify conditions for ginsenoside conversion

In order to confirm the conditions for the conversion of ginsenoside, the centrifuged culture was mixed with 0.2 mg / ml Rb1 of ginsenoside at a ratio of 1: 1 (v / v), and the mixture was filtered with a 0.2 탆 filter. The filtered samples were fermented at 35 ° C and 190 rpm in shaking incubator for 3, 7, 10 and 14 days.

The pH was adjusted to pH 4, 5, 6, 7, 8, and 9 by mixing 0.2 mg / ml Rb1 at a ratio of 1: 1, filtered through a 0.2 μm filter, 7 days. The temperature was adjusted to pH 4, 5, 6, 7, 8, and 9 by mixing with 0.2 mg / ml Rb1 at a ratio of 1: 1 and filtered through a 0.2 μm filter. 7 days.

Experimental Example 2-3. Confirmation of conversion of Rb1 using TLC and HPLC

After the reaction, the conversion of ginsenoside Rb1 was analyzed using TLC and HPLC.

For TLC analysis, 3 μg each was collected at 0.8 cm intervals on a TLC plate sheet (Silica gel 60 F ₂₅₄ , Aluminum sheets), and the mixture was allowed to stand in the reactor, and chloroform / methanol / water (65/35/10 , v / v / v). 10% sulfuric acid (H ₂ SO ₄ ) was sprayed on the reaction TLC plate and heated at 110 ° C to develop color. The TLC analysis results are shown in FIG.

For HPLC analysis, the instrument used was Agilent Technologies 1260 infinity and the columns were ACE 5-C ₁₈ Column (250 x 4.6 mm) was used, and the conditions under which the components of saponin were analyzed are shown in Table 1 below.

Component analysis conditions of saponin -Agilent Technologies 1260 infinity column ACE 5-C ₁₈ (250 x 4.6 mm) flux
(Flow rate) 1.0 ml / min Detector
(Detector) UV detector, 205nm Solvent A; H ₂ O, B; CH ₃ CN
(35% B), 85- 105 minutes (50% B), 105-135 minutes (65% B), 135-145 minutes (85% B) 145-155 minutes (100% B), 155-160 minutes (100% B), 160-163 minutes (20% B), 163-165 minutes (20% B) Temperature 40 ℃

Referring to FIG. 2A, when ginsenoside Rb1 was reacted with FMCS 72 culture medium at 30, 35, and 40 ° C for 7 days, most of the ginsenosides Rb1 changed to ginsenoside Rd, As the temperature increased, the remaining band of ginsenoside Rb1 was found to be slightly soft. Therefore, the activity of RMKS 72 culture medium of Fanny Bacillus was the highest at 35 ~ 40 ℃.

 On the other hand, through preliminary experiments, it was confirmed that 5 to 10% of ginsenosides were converted at 25 ° C., and it was confirmed that conversion of ginsenoside was not observed at 20 ° C. or lower and 45 ° C. or higher.

Referring to FIG. 2B, to determine the optimum reaction pH of the culture medium of the Fansubacillus sp. RMKS 72 effective for the conversion of ginsenoside, the ginsenoside Rb1 was adjusted at pH 4.0, 5.0, 6.0, 7.0, After 7 days of reaction, pH 4.0 showed almost no conversion of ginsenoside, and the conversion of ginsenoside Rb1 to Rd was better than pH 5.0, 8.0, and 9.0 at pH 6.0 and 7.0. Therefore, the optimal pH for the operation of the culture medium of Fanny Bacillus in RMKS 72 was 6.0 ~ 7.0.

On the other hand, preliminary preliminary experiments confirmed that almost no conversion of ginsenoside occurs even at pH 10.0.

FIG. 2C shows the results of reaction of ginsenoside Rb1 for 14 days by adjusting the culture medium of F. vulnificus RMKS 72 to an optimum temperature of 35 ° C and an optimal pH of 7.0 as shown in FIGS. 2A and 2B. From the TLC results of FIG. 2C, it can be seen that the conversion from ginsenoside Rb1 to Rd significantly increased with time as the reaction time became longer at 3, 7, 10, and 14 days. In particular, on day 14, it was found that ginsenoside Rb1 was mostly converted to Rd, and that of Fanny Bacillus RMKS 72 was found to be very effective in converting ginsenoside Rb1 to ginsenoside Rd.

In addition, FIG. 2D shows the result of the reaction of the ginseng roots for 14 days by adjusting the culture medium of Fanny Bacillus subtilis RMKS 72 to an optimum temperature of 35 ° C and optimum pH of 7.0 as shown in FIGS. 2A and 2B. As can be seen from the TLC results of FIG. 2D, the conversion from ginsenoside Rb1 to Rd significantly increased with time as the reaction time became longer at 3, 7, 10 and 14 days. On day 14, ginsenoside Rb1 in ginseng was largely converted to Rd. Thus, the Fansubsillus genus RMKS 72 was found to be highly effective in converting ginsenoside Rb1 to ginsenoside Rd.

In addition, referring to FIG. 3, it was confirmed that the HPLC result of the reaction of ginsenoside Rb1 after 14 days of reaction with the culture medium of Fanny Bacillus subtilis RMKS 72 at 35 ° C and pH 7.0 was found to have a pattern similar to the TLC result. On day 0, the ginsenoside Rb1 peak was observed. On the third day of reaction, ginsenoside Rd was found to be generated in ginsenoside Rb1. On the 7th day and 14 days after the reaction, the peak of ginsenoside Rb1 gradually increased While the peak of ginsenoside Rd was getting bigger.

<Examples and Comparative Examples: Preparation of fermented ginseng and fermented ginseng extract>

< Example  One. Fanny Bacillus  genus RMKS  Preparation of fermented ginseng using 72 culture broth>

Five-year-old ginseng roots were pulverized using a pulverizer, and 400 g of water was added to 100 g of the ground ginseng. The water-added ginseng crushed product was heat-treated at 75 캜 for 10 minutes with a hot water bath, cooled to 30 캜, and then inoculated with 15 g of a culture medium of F. vulnificus RMKS 72 (cultured in TS liquid medium at 30 캜 for 72 hours) And fermented until the 14th day. Unlike the ginsenoside conversion experiment (Experimental Example 2), the cultures of the Fansubsillus genus RMKS 72 strain were directly used for culturing immediately after the culture without centrifugation.

< Example  2. Fanny Bacillus  genus RMKS  Preparation of Fermented Ginseng Water Extract Using 72 Culture Media>

The 14-day fermented ginseng prepared in Example 1 was dried to have a moisture content of 14%, and then 1 kg of the dried fermented ginseng was added to 10 kg of water. The resulting mixture was extracted at 90 ° C for 8 hours. The filtrate with the solid content removed was lyophilized, Water extract was prepared.

< Example  3. Fanny Bacillus  genus RMKS  Preparation of 100% Ethanol Extract of Fermented Ginseng Using Culture Medium of 72 Strain>

The 14-day fermented ginseng prepared in Example 1 was dried to a moisture content of 14%, 1 kg of the dried fermented ginseng was added to 4 kg of 100% ethanol, and the mixture was extracted at 50 ° C for 8 hours. Then, the filtrate with the solid content removed was distilled under reduced pressure A 100% ethanol extract of fermented ginseng was prepared.

<Comparative Example 1: Preparation of ginseng fermented using the Companion Bacillus strain culture MBT213>

Fermented ginseng was prepared using the same method as in Example 1 except that the culture medium of Fanny Bacillus MBT213 (TS liquid medium, which was disclosed in Korean Patent No. 1345226, for 72 hours at 30 ° C in the culture medium of Fannibacillus sp. RMKS 72, ), The ginseng was fermented for 14 days

< Comparative Example  2. Fanny Bacillus  genus MBT213  Preparation of fermented ginseng extract using a strain culture solution>

The 14-day fermented ginseng prepared in Comparative Example 1 was dried to a moisture content of 14%, and then 1 kg of the dried fermented ginseng was added to 4 kg of 100% ethanol, and the mixture was extracted at 50 ° C. for 8 hours. The filtrate with the solid content removed was distilled under reduced pressure A 100% ethanol extract of fermented ginseng was prepared by MBT213 strain.

<Experimental Example 3> Confirming the conversion effect of ginsenoside of fermented ginseng>

For the HPLC analysis of the fermented ginseng (Example 1) using the culture broth of Fanny Bacillus sp. RMKS 72, fermented ginseng of each fermentation date on the 3rd, 7th and 14th days after inoculation of the same culture broth to the same ginseng crushed product And ethanol treatment. At this time, each of the fermented ginseng and 95% ethanol aqueous solution was mixed at a weight ratio of 1: 4 (20 g in total), and then centrifuged at 10,000 × g for 10 minutes, and the supernatant was lyophilized. Thereafter, the lyophilized sample was dissolved in distilled water and treated with butanol. When the butanol was treated, the sample dissolved in the water and the water-saturated butanol were mixed at a ratio of 1: 1 (v: v) (total 2 ml), and the butanol layer was extracted and lyophilized, The conversion of side Rb1 to Rd was analyzed and the results are shown in Fig. HPLC was performed in the same manner as in the experiment for converting ginsenoside of Experimental Example 2-3.

Referring to the HPLC results in FIG. 4, the peak of ginsenoside Rb1 (peak at the 71th minute) was observed on day 0, but the peak of ginsenoside Rd was weak (peak of 81 minutes). However, as the ginsenoside Rd was produced over the fermentation period, the peak of ginsenoside Rb1 was decreased and the peak of ginsenoside Rd was found to be large on the 14th day of fermentation.

On the other hand, the fermented ginseng of Comparative Example 1 using the culture medium of FT-MBT213 strain was subjected to HPLC analysis in the same manner as in HPLC analysis of Example 1, and the content of Rd after fermentation is shown in Table 2 below. Referring to Table 2, it was confirmed that the fermented ginseng of Example 1 had a much higher conversion rate of ginsenoside Rb1 to Rd than the fermented ginseng of Comparative Example 1, which was twice as high.

division Peak No. Ret Time [min] Type Area [mAU * s] Amount [mg / ml] The fermented ginseng of Example 1  8 82.015 VV 3068.44238 0.123406 The fermented ginseng of Comparative Example 1  8 81.735 VV 2000.27380 0.070014

< Experimental Example  4. Active oxygen species of fermented ginseng extract (Reactive Oxygen Species, ROS ) Removal ability evaluation DCF -DA assay) &gt;

The 2'7'-Dichlorofluorescein diacetate (DCF-DA) assay was used to evaluate the removal of active oxygen by comparing the amount of active oxygen produced in the cells stimulated with silica after treatment with the test substance It is the test method used.

First, Raw 264.7 cells cultured for 24 hours were washed once with PBS, and cells were harvested. The cells were suspended in 15 ml of PBS, and 15 μl of a 20 mM DCF-DA solution (Sigma-Aldrich) was added and reacted for 1 hour. Centrifuged at 1200 × g for 2 minutes, washed once with PBS, diluted to 1 × 10 ⁵ cells / ml, treated with each test group and positive control sample, and allowed to react for 10 minutes. At this time, each treatment concentration was 200 mg / ml of the fermented ginseng water extract of Example 2, the fermented ginseng ethanol extract of Example 3, and the fermented ginseng ethanol extract of Comparative Example 2 as the test group, respectively, and the ascorbic acid (Ascorbic acid) was 100 ㎍ / ㎖, positive control group 2 was N-acetyl-L-cysteine 1 mM. Thereafter, 50 쨉 l of 20 mg / ml of silica was treated and further reacted for 1 hour. Cell pellets obtained by centrifugation at 6000 x g for 5 minutes were dispersed in 200 μl of PBS and inoculated on a 96-well plate. The fluorescence was measured at 485 nm excitation / 535 nm emission using a fluorescence microplate reader (Bio-Rad) to determine the ROS level. The results were expressed as means ± standard deviation from three independent experiments. P <0.05 was verified by Student's t-test and the significance of the results is shown.

To evaluate the ability of each sample to remove ROS, Raw 264.7 cells were treated with silica and a sample material, and the amount of reactive oxygen present was tested by measuring the luminescence of DCF fluorescent substance capable of binding to ROS.

sample Relative luminous power (%) of DCF fluorescent material Negative control 1 100% Negative Control 2 (silica treated group) 183.6% The fermented ginseng water extract of Example 2 137.7% The fermented ginseng ethanol extract of Example 3 144.7% The fermented ginseng ethanol extract of Comparative Example 2 177.4% Positive control 1 (ascorbic acid) 130.2% Positive control 2 (N-acetyl-L-cysteine) 111.5%

As shown in FIG. 5 and Table 3, when the negative control group 1 (the group not treated with silica) and the negative control group 2 (the silica treated group: active oxygen induction group) And the amount increased to more than 80%. In addition, it was confirmed that when the fermented ginseng extract of Example 2 and Example 3 was treated, the increased active oxygen species was reduced to 45.9% and 38.9%, respectively. On the other hand, as in Comparative Example 2, it was confirmed that the extract of Comparative Example 2 using the other species strains of the Fanny Bacillus genus had a lower reducing effect than that of the extract of the present invention.

< Experimental Example  5. Toxicity and skin irritation test>

Experimental Example  5-1. Acute toxicity

The toxicity of the extract of fermented ginseng (lyophilized product of the extract of Example 2) in the culture medium of Fanny Bacillus sp. RMKS 72 of the present invention to the animal body in an acute (within 24 hours) , And this experiment was performed to determine mortality. Twenty ICR mice were injected into each of the control and experimental groups. In the control group, only the PEG-400 / tween-80 / ethanol (8/1/1, v / v / v) was administered and in the experimental group, the extract of fermented ginseng using the culture medium of Fanny Bacillus sp. / tween-80 / ethanol (8/1/1, v / v / v) for oral administration. After 24 hours of administration, the mortality rate of each mouse was found to be alive in the control group and the experimental group administered with the extract of fermented ginseng using the culture medium of RMKS 72 of the Fanny Bacillus sp. At a concentration of 2 g / kg / day.

Experimental Example  5-2. Human body skin irritation confirmation experiment

The cream of the following formulation 1-3 containing the extract of fermented ginseng (lyophilized product of the water extract of Example 2) using the culture medium of the Fanny Bacillus sp. RMKS 72 of the present invention was subjected to a patch test on 10 volunteers , All showed no skin irritation.

&Lt; Preparation Example 1: Preparation of cosmetic &

1-1. Production of flexible lotion

Using the fermented ginseng extract (lyophilized product of water extract) using the culture medium of Fanny Bacillus sp. RMKS 72 according to Example 2, a flexible lotion was prepared by a conventional method according to the composition shown in Table 4 below.

Raw material name Content (% by weight) The fermented ginseng extract of Example 2 4.0 Butylene glycol 3.5 glycerin 2.5 Polyoxyethylene hardened castor oil 0.1 ethanol 2.5 Betaine 1.0 Citric acid 0.01 Sodium citrate 0.03 antiseptic Suitable amount Spices Suitable amount Purified water to 100

1-2. Manufacture of nutrition essences

Using the fermented ginseng extract of Example 2, nutritional essences were prepared according to the compositions shown in the following Table 5 by a conventional method.

Raw material name Content (% by weight) The fermented ginseng extract of Example 2 7.0 Cetostearyl alcohol 1.0 Self emulsifying monostearic acid 1.0 Wax 0.5 Squalane 5.0 Isocetyl octanoate 3.0 Dimethylsiloxane 0.3 Sorbitan monostearate 0.5 Polyethylene glycol monostearate 8.0 glycerin 4.0 Propylene glycol 0.2 Carboxy polymer 0.22 Triethanolamine 0.25 antiseptic Suitable amount Spices Suitable amount flash Suitable amount Purified water to 100

1-3. Manufacture of cream

Using the fermented ginseng extract of Example 2, a cream was prepared according to the composition shown in Table 6 below by a conventional method.

Raw material name Weight% (w / w) The fermented ginseng extract of Example 2 7.0 Cetostearyl alcohol 3.0 Self emulsifying monostearic acid 1.5 Chin type monostearate 1.5 Wax 0.5 Liquid paraffin 8.0 Squalane 7.0 Isocetyl octanoate 4.0 Refined jojoba oil 4.0 Dimethylsiloxane 0.3 Sorbitan monostearate 1.0 Polyethylene glycol monostearate 1.2 glycerin 6.0 Propylene glycol 4.0 Betaine 4.0 Xanthan gum 0.06 Triethanolamine 0.10 antiseptic 0.25 Spices Suitable amount flash Suitable amount Purified water to 100

1-4. Manufacture of packs

Using the fermented ginseng extract of Example 2, packs were prepared according to the compositions shown in the following Table 7 by a conventional method.

Raw material name Content (% by weight) The fermented ginseng extract of Example 2 8.0 glycerin 7.0 1,3-butylene glycol 3.0 Squalene 3.0 Dimethicone 3.0 Sodium hydrulonate 0.1 Glycine 2.0 Polyacrylamide 5.0 Triethanolamine 0.1 EDTA 0.1 Purified water to 100

< Formulation example  2. Pharmaceutical preparations>

2-1. Preparation of hydrophilic ointment

Using the fermented ginseng extract of Example 2, a hydrophilic ointment was prepared by a conventional method according to the composition shown in Table 8 below.

Raw material name Content (% by weight) The fermented ginseng extract of Example 2 8.0 White vaseline 35.0 Stearyl alcohol 30.0 Ethyl (or methyl) p-oxybenzoate a very small amount Propylene glycol 20.0 Sodium lauryl sulfate 3.4 Propyl p-oxybenzoate a very small amount

2-2. Manufacture of tablets

Using the fermented ginseng extract of Example 2, the components listed in Table 9 below were mixed, and tablets were prepared by tableting according to a conventional tablet production method.

Raw material name Weight (mg) The fermented ginseng extract of Example 2 100 mg Corn starch 100 mg Lactose 100 mg Magnesium stearate 2 mg

< Formulation example  3. Food Manufacturing>

3-1. Manufacture of cooking seasonings

The fermented ginseng extract of Example 2 was added to the cooking seasoning at 1 wt% to prepare a cooking sauce for health promotion.

3-2. Manufacture of flour food products

The fermented ginseng extract of Example 2 was added to wheat flour at 0.1 wt%, and bread, cake, cookies, crackers and noodles were prepared using the mixture to prepare a health improving food.

3-3. soup  And gravies

The health-promoting soup and juice were prepared by adding the fermented ginseng extract of Example 2 to the soup and juice at 0.1 wt%.

3-4. Manufacture of dairy products

The fermented ginseng extract of Example 2 was added to milk in an amount of 0.1 wt%, and various dairy products such as butter and ice cream were prepared using the milk.

3-5. Vegetable juice  Produce

0.5 g of the fermented ginseng extract of Example 2 was added to 1,000 ml of tomato juice or carrot juice to prepare health promotion vegetable juice.

3-6. Fruit juice  Produce

0.1 g of the fermented ginseng extract of Example 2 was added to 1,000 ml of apple juice or grape juice to prepare health promotion fruit juice.

National Academy of Agricultural Sciences KACC92078P 20150605

<110> THE INDUSTRY & ACADEMIC COOPERATION IN CHUNGNAM NATIONAL UNIVERSITY <120> Novel Paenibacillus sp. RMKS 72, fermented ginseng using the same          and composition having anti-oxidant activity comprising extract          of fermented ginseng using the same <130> DPC-15-0051 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 1098 <212> DNA <213> Paenibacillus sp. <400> 1 acaattctcc cacttcggcg gctggctccc ttgcgggtta ccccaccgac ttcgggtgtt 60 gtaaactctc gtggtgtgac gggcggtgtg tacaagaccc gggaacgtat tcaccgcggc 120 atgctgatcc gcgattacta gcaattccga cttcatgcag gcgggttgca gcctgcaatc 180 cgaactgaga ccagctttga taggattggc tccatctcgc gacttcgctt cccgttgtac 240 tggccattgt agtacgtgtg tagcccaagt cataaggggc atgatgattt gacgtcatcc 300 ccgccttcct ccggtttgtc accggcagtc attctagagt gcccacccga agtgctggca 360 actaaaatca agggttgcgc tcgttgcggg acttaaccca acatctcacg acacgagctg 420 acgacaacca tgcaccacct gtcacctctg tcccgaaggc cgcctctatc tctagaggat 480 tcagagggat gtcaagactt ggtaaggttc ttcgcgttgc ttcgaattaa accacatact 540 ccactgcttg tgcgggtccc cgtcaattcc tttgagtttc agtcttgcga ccgtactccc 600 caggcggaat gcttaatgtg ttaacttcgg caccaagggt atcgaaaccc ctaacaccta 660 gcattcatcg tttacggcgt ggactaccag ggtatctaat cctgtttgct ccccacgctt 720 tcgcgcctca gcgtcagtta cagcccagag agtcgccttc gccactggtg ttcctccaca 780 tatctacgca tttcaccgct acacgtggaa ttccactctc ctcttctgca ctcaagtcac 840 ccagtttcca gtgcgaacca aggttgagcc ttggccttaa acaccagact taagtgaccg 900 cctgcgcgcg ctttacgccc aataattccg gacaacgctt gccccctacg tattaccgcg 960 gctgctggca cgtagttagc cggggctttc ttctcaggta ccgtcactcc ggtagcagtt 1020 actctaccgg acgtcttccc tggcaacaga gctttacgat ccggaaacct tcatcactca 1080 cgcggcgttg ctccggtc 1098

Claims (12)

A new strain of the Fanny Bacillus strain RMKS 72 ( Paenibacillus sp. RMKS 72, Microorganism Accession No. KACC92078P). Fermented ginseng using RMKS 72 strain ( Paenibacillus sp. RMKS 72, Microorganism Accession No. KACC92078P) in Fanny Bacillus. 3. The method of claim 2,
The fermented ginseng is prepared by mixing 100 parts by weight of ginseng, 200-1000 parts by weight of water, and 1 to 25 parts by weight of a culture of a strain of Fanny Bacillus strain RMKS 72 Wherein the fermented ginseng is fermented at 25 to 40 DEG C for 3 to 15 days. A cosmetic composition having an antioxidative activity, characterized by containing an extract of fermented ginseng using RMCS 72 strain ( Paenibacillus sp. RMKS 72, microorganism accession number KACC92078P) of Fanny Bacillus. 5. The method of claim 4,
The fermented ginseng is prepared by mixing 100 parts by weight of ginseng, 200-1000 parts by weight of water and 1 to 25 parts by weight of a culture of the strain FMKS 72 in the Fanny Bacillus strain Wherein the composition is fermented at 25 to 40 占 폚 for 3 to 15 days. 5. The method of claim 4,
Wherein the extract of the fermented ginseng is obtained by extracting fermented ginseng using a strain of Fanny Bacillus sp. RMKS 72 with water, C1 to C4 lower alcohols, or a mixed solvent thereof. A health functional food having an antioxidative activity, characterized by containing an extract of fermented ginseng using RMCS 72 strain ( Paenibacillus sp. RMKS 72, microorganism accession number KACC92078P) of Fanny Bacillus. 8. The method of claim 7,
The fermented ginseng is prepared by mixing 100 parts by weight of ginseng, 200-1000 parts by weight of water, and 1 to 25 parts by weight of a culture of a strain of Fanny Bacillus strain RMKS 72 And fermented at 25 to 40 占 폚 for 3 to 15 days. 8. The method of claim 7,
Wherein the extract of the fermented ginseng is obtained by extracting fermented ginseng using the strain of Fanny Bacillus strain RMKS 72 with water, C1 to C4 lower alcohol, or a mixed solvent thereof. A pharmaceutical composition for preventing aging characterized by containing an extract of fermented ginseng using RMCS 72 strain ( Paenibacillus sp. RMKS 72, microorganism accession number KACC92078P) of Fanny Bacillus. A method for converting ginsenoside Rb1 to ginsenoside Rd using the strain of the Fanny Bacillus genus RMKS 72 described in claim 1,
Obtaining a culture of a strain of Fanny Bacillus strain RMKS 72;
Mixing 0.01 to 10 mg / ml of ginsenoside Rb1 and the culture solution in a volume ratio of 1: 1 to 5; And
Reacting the resulting ginsenoside Rb1 at 25 to 40 DEG C and a pH of 5.0 to 9.0 for 3 to 15 days. A method for producing fermented ginseng having enhanced ginsenoside Rd using the strain of the Fanny Bacillus genus RMKS 72 according to claim 1,
Obtaining a culture of a strain of Fanny Bacillus strain RMKS 72;
100 parts by weight of ginseng, 200-1000 parts by weight of water and 1 to 25 parts by weight of a culture of a strain of Fanny Bacillus strain RMKS 72; And
Wherein the fermented ginseng is reacted at 25 to 40 캜 at a pH of 5.0 to 9.0 for 3 to 15 days.

Images