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    • 专利摘要:
    The present invention relates to a lactic acid bacteria fermentation product of ginseng prepared by fermenting ginseng with lactic acid bacteria, ginseng yogurt containing lactic acid bacteria fermentation product of ginseng, and lactic acid bacteria strains used to obtain the lactic acid bacteria fermentation product of the ginseng.
    公开号:KR20040049217A
    申请号:KR1020020077081
    申请日:2002-12-05
    公开日:2004-06-11
    发明作者:김동현；한명주；추민경
    申请人:홍림통산(주)；
    IPC主号:
    专利说明:

    Lactic acid bacteria fermented product of ginseng, ginseng yogurt containing it and lactic acid bacteria strain used therein {GINSENG FERMENTED BY LACTIC ACID BACTERIUM;
    [1] The present invention relates to a lactic acid bacteria fermentation product of ginseng prepared by fermenting ginseng with lactic acid bacteria, ginseng yogurt containing lactic acid bacteria fermentation product of ginseng, and lactic acid bacteria strains used to obtain the lactic acid bacteria fermentation product of the ginseng.
    [2] Ginseng is a perennial ripening root belonging to the genus Oga ginseng according to the plant taxonomy, and about 11 species are known on the earth. For example, the ginseng grows in the Far East of Asia (North 33 ~ 48: Korea, North Manchuria, and Russia) Excellent ginseng from Panax ginseng CAMeyer; American ginseng (Panax quinquefolium L.) growing and growing in the United States and Canada; Panax notoginseng F.H.Chen, wild or cultivated in southwestern Guangxi province from southeastern Yunnan Province of China; And Panax japonicus C.A.Meyer distributed in Japan, southwestern China and Nepal.
    [3] Ginseng is not only stored as a commodity in new agricultural plants, but has been used as a valuable medicine since ancient times. Many pharmacological experiments to date have revealed that ginseng enhances the nonspecific resistance of the body to stress and has an acidic action. In addition, it has been shown to improve hypertension, enhance insulin action, hypoglycemic effect in ALLOXAN diabetic mice, liver RNA synthesis in rats, protein synthesis, sugar and lipid metabolism, and anticancer.
    [4] The ginseng has been used for various diseases such as psychiatric diseases, diseases of the nervous system, and diabetes in the form of herbal medicine mainly in Asian countries such as Korea, China, Japan, and saponins, the main components of the ginseng, are tonic, gangjeong, It is known to have an effect on sedation molding and antihypertension.
    [5] Currently, the use of ginseng is in the form of red ginseng, which is produced by heating white ginseng or ginseng as it is grown and harvested at room temperature, at 98-100 ° C., or ginseng prepared by heating at 120-180 ° C. It is used.
    [6] Meanwhile, the root of ginseng contains about 5.22% of ginseng saponin, which is a mixture of 13 or more ginsenosides, among which the content of ginsenosides Rb1, Rc and Rg1 is relatively high. to be.
    [7] The components of ginsenosides contained in ginseng and their pharmacological efficacy are shown in Table 1 below.
    [8] Kinds of Ginsenosides efficacy Ginsenoside-Rb1 Central and mental stability, central feeding, aggression, analgesia, anticonvulsion, anti-anxiety, adrenal cortex stimulating hormone and corticosterone secretion, cholesterol biosynthesis, memory improvement, high cholesterol and triglycerides Lowering of fatty acids, promoting neuronal survival, protecting liver injury, promoting DNA, RNA, protein and lipid synthesis of bone marrow cells, promoting the release of acylcholine, vasodilation, inhibiting platelet aggregation, inhibiting lipid peroxidation, promoting cholesterol metabolism, anti-inflammatory, Activation of phagocytosis, inhibition of renal glomeruli. Ginsenoside-Rb2 Promoting sugar and fat metabolism, antidiabetic action, maintaining nitrogen metabolism equilibrium, promoting protein and lipid synthesis, hypocholesterolemic and anti-arteriosclerosis, antagonism of cancer toxin hormone, inhibition of smooth muscle cell proliferation, DNA, RNA, corticosteroids and Promotes corticosterone secretion, improves stress appetite loss, suppresses tumor angiogenesis, promotes the production of antioxidant active substances, activates ATP supply of liver tissues, regulates cholesterol, promotes metabolism of cholesterol, promotes hepatocyte proliferation and DNA synthesis, suppresses platelet aggregation, and analgesic effect Ginsenoside-Rc Promotes liver, serum cholesterol and RNA synthesis, promotes bone marrow cell DNA, RNA, protein and lipid synthesis, analgesics, corticosteroid secretion, prostacyclin biosynthesis, renal glomerular hypertrophy Ginsenoside-Rd Adrenal cortex stimulating hormone and corticosteroid secretion, renal glomeruli hypertrophy Ginsenoside-Re Stimulation of adrenal cortical stimulation hormone and corticosteroid, analgesic, vasodilation, anti-hot stress, smooth muscle cell proliferation, myeloid cell DNA, RNA, protein and lipid synthesis, protection of liver injury, cholesterol metabolism Ginsenoside-Rg1 Immune function, platelet aggregation inhibition, antithrombin, good activation, memory and learning function, anti-fatigue, anti-stress, central excitement, vasodilation, anti-inflammatory, anti-nephritis and renal blood flow, high temperature environment and endogenous fever Protective action against harmful stimuli, improvement of stress slow-acting disorder, promotion of neuronal survival rate, promotion of hepatocyte proliferation and DNA synthesis, promotion of adrenal cortex stimulating hormone secretion, promotion of cholesterol metabolism, protection of liver damage Ginsenoside-Rh1 Experimental anti-inflammatory effect, tumor cell differentiation promotion, platelet aggregation inhibition, glandogenic activation Ginsenoside-Rh2 Inhibits cancer cell proliferation, promotes cancer cell regeneration, inhibits cancer cell infiltration, inhibits tumor growth, and enhances anticancer activity of anticancer agents Compound K Potent tumor angiogenesis and cancer cell metastasis inhibition, type IV collagenase secretion, anti-angiogenic activity and platelet aggregation inhibition, antiallergic effect
    [9] As is currently known, ginsenosides, the main component of the pharmacological efficacy of ginseng, are saponins of Rb1, Rb2 and Rc. However, the component K (20-0-β-D-glucopyranosyl-20 (S) -protopa, which is substantially contained in ginseng, has a component that substantially inhibits anticancer action or cancer metastasis of cancer cells or antiallergic action. incident diol), ginsenoside Rh1 and Rh2 seed, and △ 20 - it is known that the saponin component of the seeding ginsenoside Rh2.
    [10] Therefore, for use for anticancer action, antiallergic action, immune enhancing action, etc. of ginseng, compounds K, ginsenosides Rh1 and Rh2 contained in the ginseng, and Δ 20 -ginsenoside Rh2 (ginsenosides) It is desirable to increase the content of the saponin component of Rk2 and Ginsenoside Rh3.
    [11] The present inventors also obtained from ginseng K, ginsenoside seed Rh1 and Rh2, and △ 20 - binary result of the studies to obtain a saponin component of ginsenoside seed Rh2 more efficiently, it is obtained sikimeuroseo fermented ginseng with lactic acid bacteria leading to the binary found that a substantial amount of ginsenoside Rh2 oxide and completed the present invention in lactic acid bacteria fermentation of ginseng compound K, ginsenoside Rh1 and Rh2 seed, and △ 20.
    [12] Therefore, it is an object of the present invention to provide a lactic acid bacteria fermented product of ginseng.
    [13] In addition, another object of the present invention to provide a ginseng yogurt containing the lactic acid bacteria fermented product of the ginseng.
    [14] In addition, another object of the present invention is to provide a lactic acid bacteria used to obtain the lactic acid bacteria fermented product of the ginseng.
    [15] The present invention is useful for obtaining (1) lactic acid bacteria fermented product of ginseng obtained by fermenting ginseng into lactic acid bacteria, (2) ginseng yogurt containing lactic acid bacteria fermented product of the ginseng, and (3) the lactic acid bacteria fermented product. It provides a lactic acid bacteria strain.
    [16] Hereinafter, the present invention will be described in more detail.
    [17] According to the present invention, when ginseng is fermented with lactic acid bacteria, saponin components such as ginsenosides Rb1, Rb2 and Rc, which are mainly contained in ginseng before fermentation, are present in compound K (20- 0-β-D- glucopyranosyl -20 (S) - Prototype wave incident diol), ginsenoside Rh1 and Rh2 seed, and △ 20 - is biotransformation as ginsenoside Rh2 seed. According to conventional knowledge in the art, compound K, ginsenoside seed Rh1 and Rh2, and △ 20 - Gene saponin of ginsenoside seed Rh2 is ginsenoside seed Rb1, more excellent antitumor activity than the saponin, such as Rb2 and Rc , Antiallergic and cancer metastatic activity (Bae et al., Biol. Pharm. Bull., 25, 743-747 2002; Bae et al., 25, 58-63, 2002; Wakabayashi et al., Oncol Res., 9, 411-417, 1998; Saiki et al., Proceedings of the 8th international symposium on Ginseng (Seoul, Korea Ginseng Society), 305-316, 2002; Hasegawa and Saiki, Proceedings of the 8th international symposium on Ginseng (Seoul, Korea Ginseng Society), 317-334, 2002).
    [18] On the other hand, the ginseng raw materials that can be used for the fermentation of lactic acid bacteria is not particularly limited, and any of ginseng itself and ginseng processed products can be used, and more specifically, ginseng, red ginseng, white ginseng, rice ginseng, ginseng leaf, ginseng Any one or more of the extract and ginseng powder may be used. By processing, ginseng in dry powder form, acid treated ginseng, high temperature treated ginseng and pressurized ginseng may be preferably used.
    [19] In addition, the powdered degree of the ginseng in the dry powder form is not particularly limited, and may be powdered to the extent that the lactic acid bacteria can penetrate into the ginseng tissue or fibrin with high efficiency, and the powdered degree may be easily made by those skilled in the art. It can be appreciated that powdering methods are also commonly known in the art. On the other hand, acid-treated ginseng, hot-treated ginseng and pressurized ginseng may be prepared from ginseng itself, but it is preferable for the ginseng treatment effect and subsequent fermentation effects to be obtained from the ginseng in the dry powder form described above. Also in this case, the powdering degree of the ginseng in dry powder form is not specifically limited.
    [20] Acid treated ginseng may be selected from powders of any one or more of ginseng, red ginseng, white ginseng, rice ginseng, ginseng leaf, ginseng extract and ginseng powder (hereinafter referred to as 'ginseng raw material'), preferably acetic acid, lactic acid, stric acid, It can be obtained by adding butyric acid, tartaric acid or propionic acid, hydrochloric acid, then incubating at about 60 ° C. for about 5 hours and neutralizing with calcium salt. According to the present invention, when the acid-treated ginseng is fermented with lactic acid bacteria, the content of ginsenosides Rh1 and Rh2 components is particularly increased in the lactic acid bacteria fermented product of ginseng.
    [21] High-temperature treated ginseng can be obtained by heating the powder of the ginseng raw material at about 100 ° C. for about 2 hours. According to the present invention, the content of compound K and ginsenosides Rh1 and Rh2 is increased in the lactic acid bacteria fermented product of the ginseng obtained by fermenting the hot-treated ginseng with lactic acid bacteria.
    [22] Pressurized ginseng may be obtained by heat-treating the powder of the ginseng raw material under pressure at about 110 to 130 ° C. for about 2 hours. According to the present invention, these by pressure treatment fermented ginseng with Lactobacillus lactic acid bacteria fermentation of ginseng obtained water, even though the lowered content of the compound K, ginsenoside seed Rh1 and Rh2 component, and △ 20 - Jin content of ginsenoside seed Rh2 Will increase.
    [23] On the other hand, the method for obtaining ginseng fermentation is not particularly limited, and fermentation may be carried out under conditions known to be suitable for the fermentation conditions of lactic acid bacteria used for fermenting ginseng using conventional methods in the art. For example, the raw material of ginseng is suspended in water and then lactic acid bacteria are added, and the bioconversion process of obtaining lactic acid bacteria fermented product of ginseng by incubating at about 48 hours to 72 hours at an appropriate fermentation temperature of the lactic acid bacteria, and The lactic acid bacteria fermentation may be obtained by a method including centrifugation and concentration of the lactic acid bacteria fermentation product of ginseng by filtering only the supernatant.
    [24] On the other hand, as lactic acid bacteria that can be used for lactic acid bacteria fermentation of ginseng, if the conversion efficiency of the saponin components in ginseng to the compound K, ginsenosides Rh1 and Rh2, and △ 20 -ginsenoside Rh2 is not particularly limited, For example, strains of the genus Lactobacillus, strains of the genus Streptococcus or strains of the genus Bifidobacterium may be used, and more specifically, Bifidobacterium K-103 (Kim, Dong-Hyun Kim, Kyung Hee University, Laboratory, Arch. Pharm. Res. , 21, 54-61, 1998), Bifidobacterium K-506 (Professor, Dong-Hyun Kim, Pharmacy, Kyung Hee University, Arch. Pharm.Res . , 21, 54-61, 1998), Bifidobacterium K. One or more strains of -1 (KCCM-10364), Bifidobacterium minimum KK-2 (KCCM-10365) can be used.
    [25] Among them, Bifidobacterium cholerium KK-1 and Bifidobacterium minimum KK-2 are microorganisms developed by the present inventors to obtain lactic acid bacteria fermentation product of ginseng, and Bifidobacterium KK-1 is Accession No. KCCM-10364 (March 22, 2002), Bifidobacterium KK-2 has been deposited with the Korea Culture Center of Microorganisms, Accession No. KCCM-10365 (March 22, 2002).
    [26] Bifidobacterium of the present invention KK-1 and KK-2 strains are Gram-positive bacteria, bacilli, anaerobic strains, and are fructose 6-phosphate phosphoketolase positive and have the following sugar availability.
    [27] KindsSugar availabilityKinds Bifidobacterium KK-1Bifidobacterium KK-2 Amygdalin arabinose arabinose cellobiose dextrinin escurin fructose galactose gluconate glucose glycogen inositol inulin lactose maltose mannitol mannitol mannose melangeose (melezitose) melibiose raffinose ribose salicycin sorbitol starch sucrose trehalose xylose--- +-+-+ /-+ / --- ++ /-+ /-+++ /-++ /-+ ----+ /-+ /-+-++-++-+ /-+ /-+++ --- + /-+-++ /-+ /-
    [28] Meanwhile, the Bifidobacterium KK-1 and KK-2 have the same general taxonomic characteristics compared to the strains belonging to the same species, but when used in lactic acid bacteria fermentation of ginseng, a higher content of compound K, ginsenosides Rh1 and Rh2, and Δ20It is possible to provide a lactic acid bacteria fermentation product of ginseng containing ginsenoside Rh2.
    [29] On the other hand, the lactic acid bacteria fermented product of ginseng according to the present invention, containing more content of compound K, ginsenosides Rh1 and Rh2, and △ 20 -ginsenoside Rh2 through lactic acid bacteria fermentation, more specifically, lactic acid bacteria of ginseng In compound: (compound K + ginsenoside Rh1), (ginsenoside Rh2 + ginsenoside Rh1), (ginsenoside Rh2 + Δ 20 -ginsenoside Rh2 + ginsenoside Rh1) or (compound K + ginsenoside Rh1) The total amount of the fermentation components of the seed Rh1 + ginsenoside Rh2) is 0.1 or more relative to (ginsenoside Rc + ginsenoside Rd + ginsenoside Rb1 + ginsenoside Rb2 + ginsenoside Re + ginsenoside Rg1). It is preferable to contain as.
    [30] The present invention also provides a ginseng yogurt containing the lactic acid bacteria fermentation product of the ginseng described above.
    [31] Wherein the ginseng yogurt, because it contains a lactic acid bacteria fermentation of ginseng, or compound K, ginsenoside seed Rh1 and Rh2, and △ 20 according to the invention a large amount of binary fermentation of ginseng saponin of ginsenoside seed Rh2 obtained water because it contains the compound K, ginsenoside Rh1 and Rh2 seed, and △ 20 - with a high bioactive seed of ginsenoside Rh2, more specifically, it is a functional yogurt having such anti-cancer effect, an anti-allergic effect and immune enhancement effect.
    [32] The production of such ginseng yogurt can be prepared by simply adding, for example, 1 to 10% by weight of the lactic acid bacteria fermentation product of the ginseng. However, such a manufacturing method is not preferable in terms of industrial economics, because it is required to ferment the yogurt raw milk and lactic acid bacteria fermentation of ginseng once again to obtain the lactic acid bacteria fermentation product of ginseng in order to prepare yogurt. Therefore, the ginseng yoghurt according to the present invention can be prepared by the ginseng yogurt according to the present invention by simultaneously fermenting the yogurt raw oil and 1 to 10% by weight of ginseng. The reaction for the fermentation may further contain vitamin C to promote the growth of lactic acid bacteria.
    [33] On the other hand, the lactic acid bacteria used to produce the ginseng yogurt described above is not particularly limited as long as the lactic acid bacteria can be fermented simultaneously with yogurt raw oil and ginseng, for example, strains of the genus Lactobacillus, Streptococcus Strains or strains of the genus Bifidobacterium may be used, more preferably Bifidobacterium K-103, Bifidobacterium K-506, Bifidobacterium cholrium KK-1, Bifidobacterium minimum KK One or more strains of -2 can be used.
    [34] The above yoghurt raw milk is not particularly limited, and milk, goat's milk, sheep's milk, skim milk, whole milk, and the like can be used.
    [35] After the fermentation of lactic acid bacteria for yogurt production, the lactic acid bacteria may or may not be removed. However, since the lactic acid bacteria have beneficial functions such as improving the intestinal environment of the body, it is preferable not to remove the lactic acid bacteria. Therefore, the yoghurt according to the present invention can also perform a function of improving the intestinal environment.
    [36] Meanwhile, powdered ginseng yogurt may also be obtained by adding milk powder or the like to the obtained yogurt. Therefore, the yoghurt in the formulation thereof is not particularly limited, and may have various forms known in the art, which are obtained through the fermentation of lactic acid bacteria of yogurt raw milk such as milk.
    [37] Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto, and modifications and the like apparent to those skilled in the art can be performed. Will recognize it self-explanatory.
    [38] <Example 1>
    [39] Fresh ginseng was washed well with hot water and dried and finely powdered 1 g of fresh ginseng powder and 0.1 g of vitamin C were suspended in 100 ml of milk, and then cultured in advance, Lactobacillus Bifidobacterium KK-1 and Bifidobacterium KK. 1 ml (about 10 9 cells / ml) of each of -2 strains were transplanted and incubated at 37 ° C for 24 hours to obtain yogurt.
    [40] <Example 2>
    [41] 2 g powder and 0.1 g of vitamin C made by well-dried fresh ginseng or rice ginseng were suspended in 100 ml of milk, and then, pre-cultivated lactic acid bacteria Bifidobacterium K-103 and Bifidobacterium K-506 were respectively obtained. Ml (about 10 9 cells / ml) were transplanted and incubated at 37 ° C. for 24 hours to obtain yogurt.
    [42] <Example 3>
    [43] 1 g of dried ginseng powder and 0.1 g of vitamin C were added to 100 ml of milk and suspended before lactobacillus vulgaris, Streptococcus thermophilus, Bifidobacterium KK-1 and Bifidobacterium 1 ml (about 10 9 cells / ml) of the terium KK-2 strains were transplanted and incubated at 37 ° C. for 12 hours to obtain yogurt.
    [44] <Example 4>
    [45] 1 g of white ginseng powder and 0.1 g of vitamin C were suspended in 100 ml of milk, and then 1 ml (about 10 lactic acid bacteria Lactobacillus vulgaris, Streptococcus thermophyllus, and Bifidobacterium KK-1 strains, respectively, were cultured. 9 cells / ml) and incubated at 37 ° C. for 24 hours to obtain yogurt.
    [46] Example 5
    [47] 1 g of dried ginseng powder and 0.1 g of vitamin C were suspended in 100 ml of milk, and then 1 ml of each of the previously cultured lactic acid bacteria Bifidobacterium KK-1 and Bifidobacterium KK-2 strains (about 10 9 cells / Ml) were transplanted and cultured at 37 ° C. for 24 hours to obtain ginseng yogurt.
    [48] <Example 6>
    [49] Lactic acid was added to dry ginseng powder to 0.5%, treated at 60 ° C. for 5 hours, neutralized and dried, and then 1 g of ginseng powder (acid-treated ginseng) and 0.1 g of vitamin C were placed in 100 ml of milk and suspended. Cultured lactic acid bacteria Bifidobacterium KK-1 and Bifidobacterium KK-2 strains were transplanted 1 ml (about 10 9 cells / ml), respectively, and cultured at 37 ° C for 24 hours to obtain an acid-treated ginseng yogurt.
    [50] <Example 7>
    [51] 1 g of ginseng (heated ginseng) powder and 0.1 g of vitamin C obtained by suspending dried ginseng powder in distilled water and steaming at 100 ° C. for 2 hours and drying were put in 100 ml of milk and suspended, and then incubated with lactic acid bacteria Bifidobacterium KK. -1 and Bifidobacterium KK-2 strains were transplanted with 1 ml (about 10 9 cells / ml), respectively, and cultured at 37 ° C for 24 hours to obtain heated ginseng yogurt.
    [52] <Example 8>
    [53] 1 g of ginseng powder (pressurized ginseng) and 0.1 g of vitamin C obtained by pressing and drying the dried powder at 120 ° C. for 2 hours and suspended in 100 ml of milk, and then pre-cultivated lactic acid bacteria Bifidobacterium KK-1 And Bifidobacterium KK-2 strains were transplanted 1 ml (about 10 9 cells / ml), respectively, and cultured at 37 ° C. for 24 hours to obtain pressurized ginseng yogurt.
    [54] Example 9
    [55] 1 g of dried ginseng powder and 0.1 g of vitamin C were suspended in 100 ml of milk, and then 1 ml of each of the previously cultured lactic acid bacteria Bifidobacterium KK-1 and Bifidobacterium KK-2 strains (about 10 9 cells / Ml) were transplanted and cultured at 37 ° C. for 24 hours to obtain ginseng yogurt.
    [56] <Example 10>
    [57] Lactic acid was added to dry ginseng powder to 0.5%, treated at 60 ° C. for 5 hours and neutralized. Then, 1 g of dried ginseng (heated ginseng) powder and 0.1 g of vitamin C were added to 100 ml of milk, suspended and incubated in advance. One lactic acid bacteria Bifidobacterium KK-1 and Bifidobacterium KK-2 strains were transplanted 1 ml (about 10 9 cells / ml), respectively, and cultured at 37 ° C for 24 hours to obtain heated ginseng yogurt.
    [58] <Example 11>
    [59] The dried ginseng powder was suspended in distilled water, steamed at 100 ° C. for 2 hours, dried, and 1 g of ginseng powder (heated ginseng) and 0.1 g of vitamin C were placed in 100 ml of milk, suspended, and then cultured in advance. LK Bifidobacterium KK- 1 and Bifidobacterium KK-2 strains were transplanted 1 ml (about 10 9 cells / ml), respectively, and cultured at 37 ° C. for 24 hours to obtain heated ginseng yogurt.
    [60] <Example 12>
    [61] 1 g of ginseng (pressurized ginseng) powder obtained by pressing the dried powder at 120 ° C. for 2 hours and dried and 0.1 g of vitamin C were suspended in 100 ml of milk, and then pre-cultivated lactic acid bacteria Bifidobacterium KK-1 And 1 ml (approximately 10 9 cells / ml) of Bifidobacterium KK-2 strains were transplanted, respectively, and cultured at 37 ° C. for 24 hours to obtain pressurized ginseng yogurt.
    [62] <Examples 13 to 24>
    [63] In Examples 1 to 12, instead of 100 ml of milk, the same procedure as in Examples 1 to 12 was repeated except that 100 ml of water was used to obtain respective lactic acid bacteria fermentation products.
    [64] Experimental Example 1: Content Analysis of Saponin Components
    [65] Commercial ginseng commercially available (Nonghyup Korea Ginseng Geumsansan 4-6 years old purchased from Gyeongdong Market), and its dry ginseng powder, acid-treated ginseng, high-temperature ginseng and pressurized ginseng (prepared by the same method as in the above example), respectively. After adding 0.1 g of vitamin C to 100 ml of milk, 1 g of lactic acid bacteria Bifidobacterium KK-1 and Bifidobacterium KK-2 strains were added thereto, incubated at 37 ° C. for 72 hours, and concentrated under reduced pressure. , High temperature treated ginseng fermented yogurt, acid treated ginseng fermented yogurt, and pressurized ginseng fermented yogurt were obtained. Next, ginseng, and its dried ginseng powder, acid-treated ginseng, hot-treated ginseng, pressurized ginseng, and each ginseng fermented yogurt obtained above were taken 2 g each, extracted three times with 100 ml of methanol, concentrated, and then suspended in water. The mixture was extracted three times with 100 ml of ether. Then, the extract was extracted three times with 100 ml of butanol, and the butanol fraction was concentrated, and the obtained concentrate was dissolved in methanol and analyzed by TLC (CHCl 3 : MeOH: H 2 O = 65:35:10 as solvent. 5% sulfuric acid solution in MeOH as a coloring reagent; Shimadzu TLC scanner CS-9301PC was used as a detector), and the results shown in Table 2 were obtained. Component content was calculated by the content of each component contained in 100% of the final extraction fraction.
    [66] Ingredient NameIngredient Content (%) Ginseng (Example 5)Ginseng Fermented Yogurt (Example 5)High Temperature Ginseng (Example 7)Fermented yogurt of hot ginseng (Example 7) Ginsenoside Rb115.11.65.12.7 Ginsenoside Rb28.21.13.52.1 Ginsenoside Rc9.50.53.82.9 Ginsenoside Re10.72.77.84.5 Ginsenoside Rg3<1<114.68.5 △ 20 -ginsenoside Rg 3<1<1<1<1 Compound K028.64.52.9 Ginsenoside Rh2<1<1<13.2 △ 20 -ginsenoside Rh2<1<1<1<1 Ginsenoside Rh1<11.20.51.8 Protopananacidio Diol<12.1<12.1
    [67] Ingredient NameIngredient Content (%) Acid Ginseng (Example 6)Acid Treated Fermented Yogurt (Example 6)Pressurized Ginseng (Example 8)Fermented Yogurt of Pressurized Ginseng (Example 8) Ginsenoside Rb12.51.22.51.2 Ginsenoside Rb220.91.20.8 Ginsenoside Rc1.81.11.51.0 Ginsenoside Re6.85.23.92.9 Ginsenoside Rg325716.14.5 △ 20 -ginsenoside Rg 3<1<16.52.1 Compound K02.201.2 Ginsenoside Rh20.210.2<14.5 △ 20 -ginsenoside Rh20.11.8<13.8 Ginsenoside Rh10.52.12.12.5 Protopananacidio Diol<12.5<12.8
    [68] Examples 1 to 4, and Examples 9 to 12 also showed similar results to Table 2, the lactic acid bacteria fermented products of Examples 13 to 24 showed a saponin content similar to the fermented yogurt.
    [69] Experimental Example 2 Analysis of Anticancer Effect
    [70] HepG2 (human liver cancer cell line; KCLB-10023), A-549 (human lung cancer cell line; KCLB-10185), P-388 (mouse lymphoid neoplasm cell line; KCLB-10046), L-1210 (mouse lymphocyte leukemia cell line; KCLB- 10219) was incubated with RPMI 1640 medium supplemented with 10% FBS, 1% antibiotic-antimycotics (GIBCO, USA) and 2.2 g / L sodium bicarbonate.
    [71] The HepG2 and A-549 were treated with 0.25% trypsin to remove the cells from the flask, and 180 μl was added to each well of the 96 well plate at a cell number of 3 × 10 4 cells / well at 37 ° C. for 24 hours. The culture was incubated in a CO 2 incubator saturated with 5% CO 2 . P-388 and L-1210 were adjusted to the number of cells to be 4 × 10 4 / well, and 180 μl of each well was incubated in a CO 2 incubator saturated with 5% CO 2 for 2 hours. Subsequently, each of butanol extract of white ginseng and butanol fractions of lactic acid bacteria fermented ginseng shown in Table 3 was autoclaved under high pressure, and then 20 µl per well was added to 10 mg / ml, and samples were added. Cultured in a CO 2 incubator saturated with% CO 2 . Then, 50 μl of 20 mg / ml MTT reagent was added per well, reacted in a CO 2 incubator for 4 hours, then the medium was removed, 100 μl of DMSO was added to the precipitate, and the absorbance was absorbed at 580 nm using an ELISA reader. Was measured to determine the cytotoxicity. The measurement results are shown in Table 3 below.
    [72] divisionED 50 (μg / ml) P388L1210A549HepG2 Common Ginseng Extract> 100> 100> 100> 100 Lactic Acid Bacteria Fermentation of White Ginseng985016096 Lactic acid bacteria fermented product of acid-treated ginseng825110295 Lactic Acid Bacteria Fermentation of High-Temperature Ginseng78458791 Lactic Acid Bacteria Fermentation of Pressurized Ginseng855210592
    [73] Experimental Example 3: Effect on E. coli HGU-3 and Intestinal Hazard Enzymes
    [74] 24 hours a cultured E. coli HGU-3 (Kim Dong - Hyun Kyung Hee University College of Pharmacy, Professor laboratory, nearly identical to the E. coli general characteristics) and Bifidobacterium KK-1 or Bifidobacterium KK-2 respectively, so that the dog 10 7 After transplanting into 5 ml of GAM culture medium and incubating for 24 hours, the enzyme activities of beta glucuronidase and tryptophanase, which are enzymes causing colon cancer and liver injury, were measured.
    [75] β-glucuronidase enzyme activity was measured by adding 0.02 ml of 10 mM p-nitrophenyl-β-D-glucuronide and 0.1 ml of enzyme solution to 0.38 ml of 0.1 M phosphate buffer (pH 7.0). The reaction was terminated by adding 0.5 ml of 0.5 N NaOH, 1 ml of distilled water was added, followed by centrifugation (2000 x g, 20 minutes), and the absorbance at 405 nm was measured for the supernatant.
    [76] Tryptophanase enzyme activity was obtained by adding 0.2 ml of 0.02M tryptophan and 0.1 ml of enzyme solution by adding 30 ml of complete reaction mixture (0.1 M bisine (pH 8.0), 4% pyridoxal 5-phosphate, 20% calf serum albumin) After the reaction was completed, 2 ml of a colorant (14.7 g of p-dimethylaminobenzaldehyde, 948 ml of 95% ethanol, and 52 ml of CH 2 SO 4 ) was added to terminate the reaction, followed by centrifugation (2000 x g, 20 minutes), followed by supernatant. Absorbance was measured at 550 nm.
    [77] The measurement results are shown in Table 4 when the Bifidobacterium KK-1 was transplanted and Table 5 when the Bifidobacterium KK-2 was transplanted.
    [78] Inhibition Rate (%) Beta glucuronidaseTryptophanase Control00 Ginseng Extract3245 Lactic Acid Bacteria Fermentation of Ginseng8585 Lactic acid bacteria fermented product of acid-treated ginseng7578 Lactic Acid Bacteria Fermentation of High-Temperature Ginseng8987 Lactic Acid Bacteria Fermentation of Pressurized Ginseng7885
    [79] Inhibition Rate (%) Beta glucuronidaseTryptophanase Control00 Ginseng Extract3245 Ginseng Biotransformation Ginseng7681 Acid-treated bioconversion ginseng6788 High Temperature Treated Bioconversion Ginseng8268 Pressurized Bioconversion Ginseng7178
    [80] As shown in Table 4 and Table 5, the lactic acid bacteria fermented product of ginseng inhibited the anticancer effect and the activity of β-glucuronidase and tryptophanase produced by enterobacteriaceae when absorbed into the human body, thereby causing colon cancer and liver damage. It can be seen that the effect can be prevented.
    [81] As described above, compound K, ginsenoside seed Rh1 and Rh2, and △ 20 that lactic acid bacteria fermented product is contained a very small amount in the carrot raw material or little is the ginseng according to the present invention it contains ginsenoside seed Rh2 much , Anti-cancer, anti-allergic and intestinal environment can be found to be effective. Therefore, the yoghurt containing the lactic acid bacteria fermented product of the ginseng as an active ingredient can be usefully used as a functional yoghurt that provides the physiological activity indicative of the above-mentioned saponin components.
    权利要求:
    Claims (11)
    [1" claim-type="Currently amended] Lactic acid bacteria fermented product produced by fermenting ginseng with lactic acid bacteria.
    [2" claim-type="Currently amended] The lactic acid bacteria fermentation product of ginseng according to claim 1, wherein the ginseng is selected from the group consisting of dry ginseng powder, acid treated ginseng, high temperature treated ginseng, and pressurized ginseng.
    [3" claim-type="Currently amended] The method according to claim 1 or 2, wherein the ginseng fermented product is Compound K (20-0-β-D-glucopyranosyl-20 (S) -protopanaxadiol), ginsenoside Rh1, ginsenoside Rh2 seed and △ 20 - Gene lactic acid fermentation of carrot, characterized in that the selected containing at least one component from the group consisting of fermentation seed ginsenoside Rh2.
    [4" claim-type="Currently amended] The compound according to claim 3, wherein (compound K + ginsenoside Rh1), (ginsenoside Rh2 + ginsenoside Rh1), (ginsenoside Rh2 + Δ 20 -ginsenoside Rh2 + ginsenoside Rh1) or (compound K + ginsenoside Rh1 + ginsenoside Rh2), the total amount of the fermentation component is ginsenoside Rc + ginsenoside Rd + ginsenoside Rb1 + ginsenoside Rb2 + ginsenoside Re + ginsenoside Rg1 Lactic acid bacteria fermentation product of ginseng, characterized in that contained in the fermentation product of the ginseng at a ratio of 0.1 or more.
    [5" claim-type="Currently amended] The lactic acid bacterium fermentation product according to claim 1 or 2, wherein the lactic acid bacteria are lactic acid bacteria capable of bioconverting ginsenoside components contained in ginseng.
    [6" claim-type="Currently amended] 6. The method according to claim 5, wherein the lactic acid bacteria are at least one lactic acid bacteria selected from the group consisting of Bifidobacterium K-103, Bifidobacterium K-506, Bifidobacterium KK-1, and Bifidobacterium KK-2. Lactic acid bacteria fermented product of ginseng to be.
    [7" claim-type="Currently amended] Ginseng yogurt containing lactic acid bacteria fermentation product of ginseng according to claim 1 or 2.
    [8" claim-type="Currently amended] 8. The ginseng yogurt according to claim 7, wherein the lactic acid bacteria fermented product is contained in the ginseng yoghurt by simultaneously fermenting yogurt raw oil and ginseng with lactic acid bacteria.
    [9" claim-type="Currently amended] According to claim 8, wherein the lactic acid bacteria are Bifidobacterium K-103, Bifidobacterium K-506, Bifidobacterium KK-1 (KCCM-10364) and Bifidobacterium KK-2 (KCCM-10364) Ginseng yogurt, characterized in that at least one lactic acid bacteria selected from the group consisting of.
    [10" claim-type="Currently amended] Bifidobacterium KK-1 (KCCM-10364).
    [11" claim-type="Currently amended] Bifidobacterium KK-2 (KCCM-10365).
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    同族专利:
    公开号 | 公开日
    CN1738905A|2006-02-22|
    JP2006508660A|2006-03-16|
    AU2003284785B2|2009-12-03|
    US20060127379A1|2006-06-15|
    EP1567660A1|2005-08-31|
    WO2004050892A1|2004-06-17|
    CA2507443A1|2004-06-17|
    CN100378226C|2008-04-02|
    US7754252B2|2010-07-13|
    AU2003284785A1|2004-06-23|
    KR100497895B1|2005-06-29|
    EP1567660A4|2006-11-22|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    法律状态:
    2002-12-05|Application filed by 홍림통산(주)
    2002-12-05|Priority to KR10-2002-0077081A
    2004-06-11|Publication of KR20040049217A
    2005-06-29|Application granted
    2005-06-29|Publication of KR100497895B1
    优先权:
    申请号 | 申请日 | 专利标题
    KR10-2002-0077081A|KR100497895B1|2002-12-05|2002-12-05|Ginseng fermented by lactic acid bacterium, yoghurt containing the same, and lactic acid bacteria used in the preparation thereof|
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