KR101773081B1 - Dolwoe fermented extracts comprising saponins as an active functional food ingredient and preparation method thereof - Google Patents
Dolwoe fermented extracts comprising saponins as an active functional food ingredient and preparation method thereof Download PDFInfo
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- KR101773081B1 KR101773081B1 KR1020150190943A KR20150190943A KR101773081B1 KR 101773081 B1 KR101773081 B1 KR 101773081B1 KR 1020150190943 A KR1020150190943 A KR 1020150190943A KR 20150190943 A KR20150190943 A KR 20150190943A KR 101773081 B1 KR101773081 B1 KR 101773081B1
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- South Korea
- Prior art keywords
- extract
- saponin
- fermentation
- aspergillus
- fermented
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
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Abstract
The present invention relates to a method for treating Gynostemma pentaphyllum in Aspergillus ( Aspergillus oryzae) oryzae , Aspergillus awamori), Aspergillus Kawachi (Aspergillus kawachii), Lactobacillus brevis M2 (Lactobacilus brevis M2) and Lactobacillus plantarum P2 ( Lactobacilus plantarum P2) to produce an off-shore fermented product; And extracting the extracellular fermentation product with hot water. The present invention also relates to a method for producing an over-fermented extract having an increased content of saponin or low-molecular saponin.
Description
More particularly, the present invention relates to a method for enhancing the content of total saponin and the content of low-molecular-weight saponin by hot-water extraction of the outer fermented product as a strain, Lt; RTI ID = 0.0 > fermentation < / RTI >
Gynostemma pentaphyllum ( Gynostemma pentaphyllum ), a dicotyledonous plant belonging to the genus Cucurbitaceae , is a transgene and is a perennial vine plant. Distribution areas are high humidity places such as coastal areas and riverside, and they are naturally grown or cultivated in mountainous areas of Manchuria, Taiwan, Japan, Malaysia and India, and in southern part of Korea, Jeju Island and Ulleungdo.
The Japanese Pharmacy Association has announced that it contains more saponin components than ginseng in addition to stones. Since then, interest and research on the function of extra saponin and extramarchine extract have been actively carried out in Korea, Anti-inflammatory, anti-ulcer, and decreased body fat [(a) Korean Ginseng Journal 8 (2) 1984, 172-177; (b) Korean Patent No. 0930580; (c) Korean Patent No. 1450571].
Other than stone, saponin of dammarane type including various gypenoside is contained according to the native region, and gynosaponin, promeveroside, bisdesmocide bisdesmoside, sophoriside, gentiobioside, rutinoside and the like, and steroids and sugars. Extra saponin has been known to exhibit various health functionalities such as lipid metabolism improving action, cardiovascular disease protective action, hypoglycemic action, central nervous system action, anticancer action, platelet aggregation inhibition action and tonic action, It is reported that ginsenoside is an active ingredient of Korean ginseng (Panax ginseng CA Meyer). It has been reported that both ginseng and off-shore contain saponin in common with fermane [Chemistry and pharmacology of Gynostemma pentaphyllum phytochemistry Reviews, 4: 2005, 197-219].
Reduction of superoxide anion content and anti-vascular endothelial cell function, cardiovascular function improvement, cholesterol lowering, immune function regulating action, cancer cell growth inhibiting action, anti-diabetic action, liver function protecting action, anti- Inflammation (chronic bronchitis and gastric ulcer). Extra-saponin increases the AMP-activated protein kinase (AMPK) activity in muscle cells and adipocytes, facilitating lipolysis It has been reported that lipid peroxidation is inhibited and treated with liver tissue cells, resulting in an increase in beta -oxidation activity in mitochondria resulting in a decrease in body fat.
In addition, treatment of extracorporeal extracts with muscle cells increases glucose uptake into the cells, thereby regulating blood glucose levels. When treated with muscle cells, the activity of IKK-β and JNK, which inhibits signal transduction of insulin, is inhibited to improve insulin resistance And it has been reported that it decreases corticosterone which is a stress hormone in blood and it shows improvement of anxiety and depression symptoms by preventing reduction of dopamine and serotonin which are neurotransmitters in the brain [ , Vol. 42, No. 1, 32p-37p]. It has been found that ethanol extract of stones exerts a protective action against dopaminergic neurons to help improve symptoms of Parkinson's disease patients.
The extracellular extracts were reported to regenerate reduced splenocyte proliferative capacity by oral administration of cadmium when administered orally. Polysaccharide components outside the stones accelerated the secretion of nitric oxide by activating peritoneal macrophages, and TNF-α Secretion of the cells was also stimulated in a concentration-dependent manner.
In the case of some saponins, it can be converted into an active active ingredient and be absorbed through enzymatic conversion in the human body. Most of the stones, by themselves, are not bioavailable with the sugar being modified. The effect can be maximized when using Metabolite through bio-converting. Accordingly, there is a need for a new method for increasing the content of low molecular weight saponin, which is a useful physiologically active ingredient of the extracorporeal extract, in order to maximize an excellent health functional substance from the outside.
The object of the present invention is to provide an over-fermented extract having an increased content of saponin or low-molecular saponin and a process for producing the same.
Another object to be solved by the present invention is to provide a health functional food or pharmacological effect which is effective for the improvement or treatment of depression of immune function, obesity, diabetes or hyperlipidemia caused by stress comprising an overflow fermented extract having an increased amount of saponin as an active ingredient Compositions.
Another object of the present invention is to provide a cosmetic composition which has an efficacy for improving whitening activity or wrinkle, which comprises an extracellular extract having an increased content of low-molecular saponin as an active ingredient.
In order to achieve the above object, the present invention is the Aspergillus duck material (Aspergillus doloe (Gynostemma pentaphyllum) oryzae), Aspergillus awamori (Aspergillus awamori), Aspergillus Kawachi (Aspergillus kawachii , Lactobacillus brevis M2 and Lactobacillus < RTI ID = 0.0 > P2 & plantarum P2) to produce an off-shore fermented product; And extracting the extracellular fermentation product with hot water to obtain an extracellular fermentation extract having an increased content of saponin or low molecular saponin.
The saponin according to the present invention may be any one or two or more selected from the group consisting of fumaran type saponin, zinosaponin, ginsenoside, primeverose, smalloside, bisdesmoside, genothiobiose and rutinoside, And low molecular saponin.
The low molecular weight saponin according to the present invention may specifically be any one or two or more selected from Rg3, Rg5, Rk1, Rh2, Ziphenocide L and Ziphenocide LI.
The fermentation according to the present invention may be carried out at 25 to 45 ° C for 6 to 240 hours.
According to the present invention, there is provided a method for producing fermented mushroom extract, which comprises extracting fermented mushroom husk, which is extracellular fermented product extracted with hot water, at 30 to 130 ° C in water or a C 1-4 alcohol aqueous solution of 20 to 70% And mixing the hydrolyzate extract of the fermented stem with the hydrothermal extract of the fermented product of step (1).
The extracorporeal extract according to the present invention may further include a pretreatment step of ultrasonic treatment at 15 to 25 kHz and 500 to 800 watts for 1 to 20 minutes before fermentation.
The present invention relates to a process for producing Aspergillus oryzae , Aspergillus awamori), Aspergillus Kawachi (Aspergillus kawachii), Lactobacillus brevis M2 (Lactobacilus brevis M2) and Lactobacillus plantarum P2 ( Lactobacilus plantarum P2) (hereinafter referred to as " off-axis fermentation extract ").
The extracellular fermentation extract may contain 5 to 85 mg / g of Gypenoside L and 4 to 70 mg / g of Gypenoside LI.
The extracellular fermentation extract according to the present invention is effective for the improvement or treatment of immune function deteriorated by obesity, diabetes, hyperlipidemia and stress, and thus is used for the pharmaceutical composition or health functional food for the prevention or treatment of the disease .
The out-of-body extract according to the present invention exhibits an effect of improving whitening or wrinkles, and thus can be used as a cosmetic composition for the same.
The present invention has been made to overcome the disadvantages and disadvantages of the prior art, as compared with the conventional method of producing an out-of-cell extract, in that the total amount of saponin as a useful physiologically active ingredient is increased, Which is superior in the activity of activating Lipid peroxidase and Lipid peroxidase activity. Particularly, there is provided a method for producing an extract in which the content of low-molecular saponin which is rarely found or present in trace amounts in the extracellular matrix, specifically, liposomes L, LI and ginsenosides Rg3, Rg5, Rk1 and the like is increased. The extracellular fermentation extract according to the present invention not only shows an effect on improving or treating immune function deteriorated by diabetes, hyperlipidemia and stress, but also exerts an excellent effect on skin beauty such as whitening or wrinkle improvement.
FIG. 1 is a graph showing changes in AMPK, ACC, and FAS activity in T3-L1 cells of low molecular weight saponin derived from an over-fermented fermentation extract according to an embodiment of the present invention.
FIG. 2 is a graph showing changes in AMPK, ACC, and FAS activity in T3-L1 cells of low molecular weight saponin derived from an extramarleous fermentation extract according to an embodiment of the present invention.
FIG. 3 is a graph showing the effect of inhibiting fat accumulation of low-molecular-weight saponin derived from fermentation in-vitro extract according to an embodiment of the present invention.
Hereinafter, the present invention will be described in more detail.
The present invention provides an over-fermented extract having an increased content of saponins, which is a useful physiologically active ingredient, and an increased content of low-molecular-weight saponins,
Doloe (Gynostemma pentaphyllum) the Aspergillus duck material (Aspergillus oryzae), Aspergillus awamori (Aspergillus awamori), Aspergillus Kawachi (Aspergillus kawachii), Lactobacillus brevis M2 (Lactobacilus brevis M2) and Lactobacillus plantarum P2 ( Lactobacilus plantarum P2) to produce an off-shore fermented product; And extracting the extracellular fermentation product with hot water to obtain an extracellular fermentation extract having an increased content of saponin or low molecular saponin.
In the present invention, the stone disturbance includes the above-ground portion, the ground portion of the Gynostemma sp.
According to the present invention, the saponin contained in the over-fermented extract produced by the above method is selected from among fermane-base saponin, zinosaponin, primeverose, tenoside, bisdesmoside, genothiobiose and rutinoside May be any one or two or more. The fumarate based saponin according to the present invention may include ziphenocide.
The saponins contained in the extracellular extract include, for example, ginsenosides Rb1, Rb2, Rc, Rd, Rg3, Rg5, Rk1, Rh and the like and saponins V, IX, XVI, XXXVI, LXXIII, LXVII, LXX, But are not limited to, chicomoride IV, damulin A, fektronine, protopanaxadiol, hydroxycyclo saponin d, mazoroside F5, pannicolin, cano floside VI, and the like.
The extracellular fermentation extract prepared according to the present invention is particularly useful for the production of ginsenosides Rg3 and Rg5, which are physiologically active substances easily absorbable in the body, gypenoside L and gypenoside LI, ) Of the present invention is greatly increased.
First, Rg3 and Rg5 in order to obtain a form of a low molecular weight saponin and jipe higher the content of the furnace side L and LI extract doloe (Gynostemma pentaphyllum) the Aspergillus duck material (Aspergillus oryzae), Aspergillus awamori (Aspergillus awamori), Aspergillus Kawachi (Aspergillus kawachii , Lactobacillus brevis M2 and Lactobacillus < RTI ID = 0.0 > P2 & plantarum P2). Preferably, the fermentation is carried out at 25 to 45 DEG C for 6 to 240 hours.
The extracted extract may be filtered to remove impurities, concentrated, and then dried or pulverized according to the intended use.
According to the invention, the hot-water extract doloe C 1 of the fermentation the fermentation is water stone sleep out the water or function of 20 to 70% to prepare a fermented stone sleepovers extract extracted from 4 to aqueous alcohol solution from 30 to 130 ℃; And mixing the hydrolyzate extract of the fermented stem with the hydrothermal extract of the fermented product of step (1).
Doloe Since the extracted open the fermented product, and there is still Saponin residue remaining foil, C 1 of the water or function of 20 to 70% from the foil-using 4 Alcohol Solutions human usability component, preferably extracted saponin or low molecular weight saponin Could. Therefore, the above step is preferable because it can further increase the total saponin content and the low-molecular-weight saponin content of the fermentation in-vitro extract, which is the final extract.
According to one embodiment of the present invention, the stone may be collected and used as it is, but it may be ground to further increase the ginsenoside content of the extraneous extract, and may be dried and powdered. When the above-mentioned stones are ground or pulverized, the surface area is improved to facilitate fermentation and extraction.
According to the present invention, it is possible to further include an ultrasonic treatment before fermentation of the off-shore into a strain. According to one embodiment of the present invention, when the extracorporeal extrasefactor is fermented into a strain, the extracted total saponin content is greatly increased. In particular, the content of low molecular weight ginsenosides such as Rg3 and liposomes L and LI is greatly increased .
In addition, the inventors of the present invention have confirmed through experimentation that the total saponin content is further increased when ultrasonic treatment is performed in the pre-fermentation step before the fermentation, compared with the case where ultrasonication is performed in the post fermentation stage. According to the present invention, it is preferable that the ultrasonic irradiation is performed at 15 to 25 kHz and 500 to 800 watt for 1 to 20 minutes. If the irradiation energy and the irradiation time of ultrasonic waves or microwaves are less than the above range, there is almost no increase in total saponin by irradiation. If the irradiation energy is longer than the above range, the increase of the total saponin content in the extract may be insignificant or may be decreased.
According to the present invention, in the extracted C 1 - 4 alcohol it may be any one selected from methanol, ethanol, propanol, butanol, isopropanol, may preferably be ethanol.
According to one embodiment of the present invention, when subjected to hot water extraction after fermentation, gypenoside L, gypenoside L, LI) and ginsenosides Rg3, Rg5, and the like were increased. Especially, in the extracellular material, gypenoside L and gypenoside LI, which have little or no trace, LI) was contained in a high content.
According to an embodiment of the present invention, the extraction is carried out for 1 to 8 hours in a temperature range of 90 to 125 DEG C, preferably 110 to 121 DEG C, so that the total saponin content is increased, Saponins such as Lipid, Lipid, Lipid, Lipid, Lipid, Lipid, Lipid, Lipid, and Lipid.
The extragranular fermentation extract according to the present invention obtained an extract having an increased total saponin content when fermented with the Aspergillus or Lactobacillus sp. Strain and then extracted with hot water, Extracts with increased contents of low molecular saponin Rg3 and Gypenoside L and Gypenoside LI were obtained.
It is preferable to sterilize the off-shore flour before fermentation as a strain in the fermentation process. The sterilization method may be any method conventionally practiced in the art, and may include, for example, high pressure sterilization, high temperature sterilization, and low temperature sterilization.
The extract of the present invention includes not only the extract obtained by the above-mentioned extraction solvent, but also an extract obtained by further performing a conventional purification process. For example, fractions obtained through various purification methods such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value and separation by chromatography (size, ion, hydrophobicity or solvent affinity) are also included in the category of the extract of the present invention .
According to the present invention, the 'off-shore extract' obtained by fermenting outdoors with Aspergillus or Lactobacillus sp. Strain showed melanin synthesis inhibitory activity, and at the same time, promoted collagen synthesis. Specifically, the melanin synthesis inhibitory activity was increased by more than 40%, collagen synthesis promoting activity was increased by more than 30%, and cytotoxicity was lower than that of the extract which was not fermented. Therefore, the out-of-body extract can be used as an active ingredient of a cosmetic composition for whitening activity and wrinkle improvement.
According to the present invention, the 'off-shore extract' obtained by fermenting outdoors with Aspergillus or Lactobacillus sp. Strain according to the above-described method has an effect of restoring immune function deteriorated by stress. Specifically, the extracorporeal extract according to the present invention was orally administered to a model of immunocompromised mice that induced stress by dexamethasone administration, and cytokine production capacity was restored. Therefore, the extracellular extract according to the present invention can be applied as a functional food for health promotion or a pharmaceutical composition for restoring immune function.
When the extracorporeal extract according to the present invention is used as a health functional food for immunomodulation, the type thereof is not particularly limited and may be in the form of a powder, granule, tablet, capsule or beverage containing the extracorporeal extract as an active ingredient, And the amount of the active ingredient to be mixed may be appropriately determined depending on the purpose of use such as prevention, health or treatment.
When the extracorporeal extract according to the present invention is used as a pharmaceutical composition for enhancing immunity, an oral preparation such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories and sterilized injection solutions , And the amount of the active ingredient to be mixed may be appropriately determined depending on the purpose of use such as prevention, health, or treatment.
When the extracellular extract according to the present invention is formulated into powder, granule, tablet, or capsule form, it may further include appropriate carriers, excipients and diluents conventionally used in food manufacturing. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, 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.
Solid preparations for oral administration can be prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin and the like in the above extract. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.
Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions, syrups and the like. In addition to water and liquid paraffin which are commonly used diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .
The health functional food or the pharmaceutical composition comprising the extracorporeal extract according to the present invention as an active ingredient can be used as a nutrient, a vitamin, an electrolyte, a flavoring agent, a colorant, a pectic acid and its salt, an alginic acid and its salt, an organic acid, a protective colloid thickener, a pH adjusting agent, a stabilizer, a preservative, a glycerin, an alcohol, and a carbonating agent used in a carbonated beverage. These components may be used independently or in combination. The ratio of such additives is not limited, but is generally selected in the range of 0.01 to 0.1 parts by weight based on 100 parts by weight of the total composition of the present invention.
Hereinafter, the present invention will be described in more detail with reference to preferred embodiments. It will be apparent, however, to those skilled in the art that these embodiments are for further explanation of the present invention and that the scope of the present invention is not limited thereby.
Example
Example 1.
end. culture
The stone outer leaves were washed, dried and crushed. The water content in the pulverized doloe 100 g packed in a flask then added to the 50% that after sterilization in an autoclave, and the reaction mixture was allowed to cool in a clean bench, Aspergillus duck material (Aspergillus oryzae ) were inoculated and fermented at 28 ° C for 7 days.
I. extraction
The fermented stone outer leaves were subjected to hot extraction at 121 DEG C for 6 hours. Filtered and separated into fermented hot-water extracts and foams. The foil was reextracted with 70 vol.% Ethanol aqueous solution and filtered to obtain a fermented strawberry extract. The total saponin content, the type and the content of each saponin were measured for each extract, and the extracts were mixed and named as the extracellular fermentation extract.
Example 2.
Aspergillus oryzae ) instead of aspergillus Awamori ( Aspergillus awamori ) was used in place of the above-mentioned extract.
Example 3.
Aspergillus oryzaeAspergillus oryzae) Instead of Aspergillus kawachi (Aspergillus kawachii) Was used instead of the above-mentioned extract.
Example 4.
Aspergillus oryzae) doloe the extract was prepared in place of the Lactobacillus bracket bis same manner as in Example 1 except for using the M2 (Lactobacilus brevis M2).
Example 5
Aspergillus oryzae), and is instead a doloe extract in the same manner as in Example 1 except that Lactobacillus Planta room P2 (Lactobacilus plantarum P2) was prepared in the.
Comparative Example 1
The extracellular extract was prepared in the same manner as in Example 1 except that the non-fermented stone outer leaves were used.
Comparative Example 2
Extracts were obtained by extracting for 6 hours at 80 ℃ using non-fermented stone leaves.
Test Example 1. Comparison of the amount of saponin in the marine saponin of the outer leaves of the leaves
(A), Fujian (B), Geochang (C), Yeosu (D), and Jeju (D) areas in China to determine the content of saponin in the cultivated area , And Ulleungdo (F) were purchased from the Korean plant extract bank.
Sample samples were prepared by hot extrusion at 110 ° C for 12 hours.
HPLC analysis
Based on the analytical methods for the three indicator substances (dammulin A, Gp L, and GP LI) set by the inventors of the present invention, the HPLC analysis conditions were examined, and 11 kinds of standard substances (Gp L , GP LI, dammulin A, ginsenoside Rb1, Rb2, Rc, Rd, Rg3, Rg5, Rk1 and Rh2) were finally determined under the condition that simultaneous analysis was performed. HPLC results of 11 kinds of standard substances are shown in Table 1 below.
The analysis was performed using an HPLC Infinity 1260 system (Agilent), and the analysis conditions were as follows. The column was XSelect HSS T3 (100 x 2.1 mm, 2.5 쨉 m, Waters, USA). The sample injection volume was detected with a 2,, VWD (Variable Wavelength Detector) detector at 204 nm. Mobile phase A was prepared by adding 1 ml of formic acid to 999 ml of tertiary pure water and adding 1 ml of acetonitrile 999 ml of formic acid. The concentration of the mobile phase B solution was maintained at 20% from 0 to 10 minutes and gradually increased to 29% until 39 minutes, increased to 41% at 67 minutes, 47% at 70 minutes, and 71% at 90 minutes And then decreased to 20% concentration for up to 95 minutes and maintained for up to 115 minutes. The moving speed of the solvent was kept constant at 0.3 ml / min.
Q- TOF analysis
In order to identify the components of the extract from the stony leaves, the 11 kinds of standard substances were analyzed using Q-TOF. The components other than the standard substances were identified using the TCM (Traditional Chinese Medicine) Library (2014, Agilant. Respectively. The Q-TOF analysis conditions are as follows.
Drying gas (N2) flow rate: 17 L / min
Drying gas (N2) temperature: 225 ℃
Nebulizer pressure: 45 psi
Sheath gas temperature: 350 ℃
Sheath gas flow: 11 L / min
Capillary voltage: 3,500 V
Nozzle voltage: 2,000 V
Fragmentor: 400 V
skimmer: 65 V
OCT 1 RF Vpp: 750 V
Collision Energy: 25 V
Ion polarity: negative
HPCL, Q-TOF, and TCM Library, the content of extracellular saponin in each region was analyzed and shown in Table 2 below. Saponin contents were different in each mountainous area and saponin contents were different from those in the mountainous areas of China. The saponin content was twice as high as those of the outer areas of Fujian province in the Guangxi province, Yeosu, Geochang, and Ulleungdo.
Saponin concentration (mg / ml)
From the above results, it was confirmed that there were differences in the kinds and contents of saponins according to the regions.
Test Example 2
Evaluation of extraction time
3, 4, 5, 6, 7, 8, 9, 10, and 24 at 121 ℃, respectively, were used to compare the components of total saponin content After extraction for a period of time, saponin detection components and contents were compared. STD is the result of confirming several kinds of the standard substances (gypenoside L, gypenoside LI, ginsenoside Rg3).
extraction
time
(h)
As shown in Table 3, the content of gypenoside L, LI and ginsenoside Rg3 was decreased from 7 hours after the extraction time.
Extraction temperature related evaluation
Extracts were obtained at 85, 95, 102, 115, 121, and 130 ℃ for 4 hours using the unfermented extracorporeal extractions to compare the components of the extragranular extracts and the total saponin content according to the extraction temperature. , ginsenoside Rg3 were investigated.
extraction
Temperature
(° C)
* ND: not detecting
As shown in Table 4, when the extraction temperature was increased to 121 ° C, the content of major saponin components tended to increase. The total saponin content decreased when the extraction temperature exceeded 121 ℃.
Specifically, the content of gypenoside L, gypenoside LI and ginsenoside Rg3 in extracts having an extraction temperature of 121 ° C was significantly higher than that at 85 ° C.
Test Example 3. Comparison of saponin content after fermentation
Rb1, Rb2, Rc, Rd, Rg3, Rg5, Rk1, Rh and the like were analyzed by HPLC and Q-TOF analysis. Zincomide IV, damulin A, fektorinaline, protopanaxadiol, hydroxycyclo saponin d, mazoroside F5, pannicola V, IX, XVI, XXXVI, LXXIII, LXVII, LXX, L, Choline and canoyl fluoride VI were detected. The main saponin and total saponin contents are shown in Table 5 below.
STD
As shown in Table 5, the contents of total saponin and low-molecular-weight saponin in the extracts of Examples 1 to 5 according to the present invention were greatly increased compared with the comparative examples. Especially, the extracts of Aspergillus kawachii ) strains.
Before fermentation, the extruded dry pulverized material was processed by ultrasonic wave generation at 25 ° C, 20 kHz, 750 watt and 20 amplitude using an ultrasonic wave extractor (SEEC-SONIC II, UL-Tech, Korea). In order to keep the temperature constant, a circulating constant temperature water bath was connected and used. As a result of ultrasonication, total saponin was not increased in less than 1 minute, and when the ultrasonic irradiation time exceeded 20 minutes, the number of detected saponin was decreased and the total saponin content was also decreased. In addition, the total saponin content was increased by about 13% in the section irradiated with ultrasonic waves for 5 to 15 minutes, compared with the case without ultrasonic irradiation, and the content of gypenoside L and gypenoside LI Was increased by about 15%.
Test Example 4.
The changes of AMPK, ACC, and FAS activity in 3 T3-L1 cells according to saponin of the extract of the present invention are shown in FIGS. 1 and 2. AMPK is an enzyme involved in fatty acid oxidation and inhibition of adipocyte differentiation, and it is confirmed that it phosphorylates ACC (inactivated) through AMPK phosphorylation (p-AMPK) to inhibit fatty acid biosynthesis and participate in fatty acid burning. FAS It is a fatty acid synthesis enzyme that promotes fatty acid synthesis.
In the case of Gypenoside LI and ginsenoside Rg3 of the present invention, p-AMPK and p-ACC were activated in a concentration dependent manner and inhibited the expression of FAS (Fatty Acid Synthase), which was effective in inhibiting lipid differentiation and fatty acid synthesis Respectively.
Test Example 5
The result of the fat accumulation change according to the saponin of the extracellular fermentation extract of the present invention was confirmed. As shown in FIG. 3, Gypenoside L, Gypenoside LI and ginsenoside Rg3 derived from the extracellular fermentation extract according to the present invention were found to have inhibitory effect on fat accumulation to control, respectively.
Claims (10)
(2) extracting the over-fermented fermented product with hot water; and (2) extracting the over-fermented fermented product by increasing the content of two or more kinds of low-molecular saponins and total saponins selected from ginsenoside Rg3, ginsenoside Rg5, ≪ / RTI >
Wherein the fermentation is carried out at 25 to 45 DEG C for 6 to 240 hours.
Preparing a fermentation extract stone sleep out by extraction at 30 to 130 ℃ to 4 alcohol aqueous solution to hot water extraction of the fermentation is water doloe fermentation stone C 1 sleep out of the water or function of 20 to 70%; And
Further comprising a step of mixing the fermented stem outer skin extract with the hot water extract of the outer fermented product of step (1).
Further comprising a pre-treatment step of ultrasonic treatment at 15 to 25 kHz and 500 to 800 watt for 1 to 20 minutes before fermentation.
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| KR20210017108A (en) * | 2019-08-07 | 2021-02-17 | 옥민 | Composition for skin whitening comprising extract of fermented black rice with improved antioxidant activity as effective component |
| WO2023017868A1 (en) * | 2021-08-10 | 2023-02-16 | (주)메디언스 | Composition for enhancing energy containing dipsaci radix extract as active ingredient |
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| KR101969062B1 (en) * | 2017-07-21 | 2019-04-15 | 주식회사 비티씨 | Preparation Method of Gynostemma Pentaphyllum leaves Extract for Increasing small molecular effective Saponin contents, and Decreasing Benzopyrene, And Gynostemma Pentaphyllum Extract Using Thereof |
| US10639343B2 (en) | 2018-01-04 | 2020-05-05 | Btc Corporation | Preparation method of Gynostemma pentaphyllum leaves extract and Gynostemma pentaphyllum extract prepared by the method thereof |
| CN108484713B (en) * | 2018-03-23 | 2019-03-05 | 娄志春 | A method of preparing gypenoside L |
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| KR102239470B1 (en) | 2019-08-07 | 2021-04-12 | 옥민 | Composition for skin whitening comprising extract of fermented black rice with improved antioxidant activity as effective component |
| WO2023017868A1 (en) * | 2021-08-10 | 2023-02-16 | (주)메디언스 | Composition for enhancing energy containing dipsaci radix extract as active ingredient |
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