KR102100493B1 - A method of Compound K using Panax ginseng sprout and the composition having antioxidation and antibacterial activities comprising Compound K prepared therefrom - Google Patents

Abstract

The present invention relates to a method for preparing compound K using sprout ginseng and an antioxidant and antimicrobial composition comprising compound K prepared accordingly, in the present invention, sprout ginseng extract, sprout ginseng fraction or a mixture thereof (Sumyzyme) AC, Cellulase KN, Plantase TL, Plantase PE Conc, Plantase TCL, Plantase MAX, BrewMax GXL, Alphalase AP3, A variety of ginsenosides contained in bud ginseng are compounded through a simple process of processing an enzyme selected from the group consisting of alfasease NP, sumyzyme SPC, plantase AK, pluszyme 2000D and rohament CL Can be converted to K.

Description

A method of Compound K using Panax ginseng sprout and the composition having antioxidation and antibacterial activities comprising Compound K prepared therefrom}

The present invention relates to a method for preparing compound K using sprout ginseng and an antioxidant and antimicrobial composition comprising compound K prepared accordingly, more specifically, to extract the main extract or fraction thereof from sprout ginseng. Sumyzyme AC, Cellulase KN, Plantase TL, Plantase PE Conc, Plantase TCL, Plantase MAX, BrewMax GXL, Alphalase AP3, Alpha The enzymes selected from the group consisting of Raze NP, Sumizyme SPC, Plantase AK, Pluszyme 2000D and Rohament CL are treated and reacted to react with various ginsenosides contained in bud ginseng. It relates to a method for converting compound K to high efficiency and a composition for antioxidant and antibacterial comprising compound K prepared accordingly.

Wild ginseng is a perennial perennial plant belonging to the family Ogalpidae, and is a plant that uses fleshy roots resembling human shapes for medicinal purposes. Wild ginseng refers to ginseng that has been germinated and grown by natural seeds of wild ginseng in the natural environment in the forest. The names of wild and wild ginseng are Panax ginseng C.A. When looking at the etymology of Mayer, 'Pan' means 'everything' and 'Axos' means 'medicine', which means 'everything rule'. Goat ginseng has a variety of active ingredients and is rich in content, so it is traded at a high price, but bio-goat ginseng is generally not easy to commercialize as it is difficult to store it for more than a week. Therefore, the existing scientific research has been conducted mainly focusing on the pharmacological efficacy and ingredients of ginseng, and the pharmacological efficacy and physiological research on the wild ginseng has not been systematically established due to the scarcity value of goats. Saponins and non-saponin-based substances (Panacen, polysaccharides, amino acid derivatives, polyacetylene derivatives, phenolic compounds) contained in ginseng and acid (sheep) ginseng have excellent pharmacological activity, and have excellent efficacy in removing free radicals. It has excellent effects on anti-cancer, lowering blood pressure, lowering lipids, and liver toxicity. Ginseng saponins, which are well known as the main pharmacological components of ginseng, are divided into PPD (Protopanaxadiol) and PPT (Protopanaxatriol). PPD-based saponins have a structure in which various substituents are bonded to the basic structure PPD, and ginsenosides Rb1, Rb2, Rc, Rd, F2, Rg3, compound K, and the like are typical saponins. In the saponin of the PPT series, PPT is the basic structure, and ginsenoside Re, Rf, Rg1, Rh1, etc. are typical.

In general, many glycoside compounds present in nature show a tendency to increase physiological activity when sugar is decomposed to become a non-glycemic substance rather than itself (Med. Pharm. Soc. 1992, 9: 1-13). Ginsenosides Rg3, Rh1, Rh2, F2, CY and CK generated by hydrolysis of some sugars than ginsenosides Rb1, Rb2, Rd, and Re, in which three or more sugars are bound, are absorbed or absorbed in vivo. It is known to exhibit a much better effect in terms of activity, etc. (Ginseng Res. 2003, 27: 129-134). Ginseng saponin, which has a lot of sugar, is known to have a very small amount absorbed into our body in the small intestine. As a result of testing the hydrolysis ability of ginsenoside Rb1 of gut microorganisms extracted from human feces, 21 of gut microorganisms % Was found to have no decomposition ability, and it was confirmed that intestinal microorganisms of about 70% with decomposition ability had a large difference in the ability to decompose ginseng saponin (Planta Medica. 1998, 64: 696-700).

Hasegawa et al. Is a new substance with a higher activity, such as Prevotella ori s, which is a new substance with superior activity of ginseng saponin in the human body, such as immune enhancement, tumor angiogenesis inhibition, and cancer cell invasion. Compound K; FGM1, M1, IH-901, 20-O-β-D-glucopyranosyl-20 (S) -protopanaxadiol) was reported to be produced (Planta medica. 1996, 62: 453-457). Compound K is a substance that is known to have excellent pharmacological effects such as liver cancer suppression and immune-enhancing effects, but the amount present in nature is extremely small, and methods to manufacture it have been researched so far, such as heat treatment, acid treatment, and enzyme treatment. The method of is developed. However, the heat treatment or the acid treatment method has a disadvantage that the yield of compound K is very low due to the non-specific reaction of randomly hydrolyzing sugar, whereas when enzyme is used, only a specific portion of sugar is selectively hydrolyzed to produce compound K with high yield. (J. Microbiol. Biotechnol. 17 (12): 1937-1943). However, in the case of the enzyme conversion method, the enzyme activity is inhibited by the ginseng saponin used as a substrate, and thus a high concentration of the substrate cannot be used. In addition, the production of compound K by the microbial conversion method requires considerable incubation time and it is difficult to manufacture with high efficiency by inducing the killing of conversion bacteria due to the antibacterial properties of the produced compound K (J. Ginseng Res. 2008, 32 (3 ): 226-231).

Patent documents related to the production of compound K include beta-galactosidase, an enzyme of diol-based saponins (Korean Patent Publication No. 2003-94757), cellulase isolated from microorganisms of the genus Penicillium, or beta isolated from Aspergillus -Compound K is prepared by treating galactosidase (Korea Patent No. 377546), naringinase isolated from Penicillium or pectinase (Korea Patent No. 418604) isolated from Aspergillus. How to do is known.

Recently, Korean Patent Publication No. 2012-0035472 uses an edible polysaccharide degrading enzyme, Cellluclast and Rapidase TF, to convert the enzyme from the ginseng saponin to a high yield of Compound K by substrate. In order to solve the suppression, the production efficiency of compound K has been increased by introducing a continuous reaction method rather than a conventional batch reaction method.

Accordingly, the present inventors continued to study the method of producing compound K using sprout ginseng, and produced sprout ginseng in the form of extracts or fractions, and added them to Sumyzyme AC, Cellulase KN, and plantase. (Plantase) TL, Plantase PE Conc, Plantase TCL, Plantase MAX, BrewMax GXL, Alphalase AP3, Alpase NP, Sumizyme SPC, Plantase AK, Pluszyme (Pluszyme) The present invention was completed by discovering that various ginsenosides contained in bud ginseng can be converted to compound K with high efficiency by treating and reacting enzymes such as 2000D and Rohament CL.

Therefore, the technical problem to be solved in the present invention is to provide a method for manufacturing compound K using sprout ginseng.

In addition, another technical problem to be solved in the present invention is to provide a composition for anti-oxidation and antimicrobial containing compound K prepared according to the above manufacturing method.

In order to solve the above technical problem, in the present invention, ginseng extract, bud ginseng fraction or a mixture thereof, Sumyzyme AC, Cellulase KN, Plantase TL, Plantase PE Conc, Plantase TCL, Plantase MAX, BrewMax GXL, Alphalase AP3, Alpase NP, Sumizyme SPC, Plantase AK, Pluszyme 2000D and Rohament ) By providing an enzyme selected from the group consisting of CL and reacting at 30 to 60 ° C. for 20 to 50 hours, a method for preparing Compound K of the following Chemical Formula 1 from sprout ginseng is provided:

In addition, in the present invention, in order to solve the other technical problems described above, provides a composition for antioxidant and antimicrobial containing compound K prepared according to the above manufacturing method as an active ingredient.

In the present invention, bud ginseng extract, bud ginseng fraction or a mixture thereof, Sumyzyme (Sumyzyme) AC, Cellulase (Cellulase) KN, Plantase (Plantase) TL, Plantase PE Conc, Plantase TCL, Plantase Reactions by processing enzymes such as No. MAX, BrewMax GXL, Alphalase AP3, Alpase NP, Sumizyme SPC, Plantase AK, Pluszyme 2000D and Rohament CL By doing so, various ginsenosides contained in sprout ginseng can be converted to compound K with high efficiency through enzyme treatment.

The ginseng (Panax ginseng) used in the present invention is a perennial herbaceous plant belonging to the family of the elmaceae family, having a height of 60 cm and one main stem at the end of the root stem. Leaves are rotated 3 ~ 4 at the end, 5 long biplanes hang at the end of a long petiole, and small leaves have a pointed egg shape or an inverted egg shape with a small edge at the edge, and on the front veins of the leaves. There is a little hair. Flowers bloom in April, and as a family tree, light green mountain inflorescences hang at the end of the stem and at the end of a long stalk in the middle of the leaves, and the fruits ripen in red color, and in a flat, round shape, several run as mountain inflorescences. Ginseng has long been used in the prevention or treatment of various diseases in Korea and China, and is known to have excellent therapeutic effect and no side effects even after long-term use. In addition, it contains 27 types of ginsenosides called saponins, and is also known to be good for cancer, diabetes, hypertension, arteriosclerosis, dementia, osteoporosis prevention, anti-aging, brain activity promotion, climacteric disorders, and female skin care, mainly for herbal medicine Has been used as At this time, ginseng root, which is known for its pharmacological efficacy, has been mainly used, but it has been found that ginseng leaves, ginseng seeds, and ginseng flowers have pharmacological efficacy. However, the period of cultivation of ginseng takes an average of 4 years or more, and in order to obtain the highest-efficiency ginseng, it must be cultivated for 6 years. Therefore, it has a disadvantage that it is expensive and has a low preference for young people and foreigners due to its unique aroma and bitter taste, and is not suitable for use as a raw material for ginseng processed products other than herbal medicines.

In particular, the sprout ginseng used in the present invention is cultivated for a period of usually within 1 to 2 years, and has a feature that contains about 7 to 8 times more saponin components of leaves and stems than the root, and also hydroponics, Due to the possibility of year-round production and shortening of the cultivation period, it has the advantage of forming a lower price than the root ginseng used conventionally, and is easy to use as a raw material for processed ginseng products.

The typical saponin-specific efficacy of sprout ginseng includes: 1) Stimulant: Ginseng improves mental performance, especially in the elderly. Ginseng contains choline, a chemical in the brain that is essential for learning and memory retention (Rb1, Rg1); 2) Antioxidants: Ginseng contains antioxidants (R0, Rf, Rg2), substances that prevent cell damage caused by exposure to unstable molecules called oxidation, free radicals; 3) Anti-aging: Ginseng exhibited anti-aging effect, and induced the relaxation of aging-related symptoms in the group of middle-aged and old-age subjects (Rd, etc.); 4) Anti-cancer: The results of a number of studies found that unpurified saponin, a compound found in ginseng, inhibited the growth of cancer cells and actually converted diseased cells into normal cells. Ginseng also helps the body cope with the side effects of chemotherapy (Rb2, Re, Rg3, Rh1, Rh2, Rh3); 5) Adaptogenic: The excellent 'tonic' quality of ginseng (helps the body adapt to stress, fatigue and cold) was confirmed (Rb1, Rc, Rg1); 6) Menopause: Ginseng contains a compound that has a similar action on the female sex hormone estrogen (Rd); 7) Antidiabetic: Ginseng helps the body maintain normal blood sugar and cholesterol levels, and stimulates a wide range of immune systems and endocrine responses (Rb1, Rb2, Rc, Rb2, etc.).

In the present invention, "extract (extract)" refers to a preparation that is concentrated by squeezing the crude drug into an appropriate leachate and evaporating the leachate, but is not limited thereto, but extracts, dilutions or concentrates of extracts obtained by extraction treatment, and dried extracts It may be a dried product obtained, a crude product or a purified product thereof.

The sprout ginseng extract of the present invention can be prepared using general extraction methods, separation and purification methods known in the art. As the extraction method, a method such as hot water extraction, hot water extraction, cold saliva extraction, reflux cooling extraction, high temperature high pressure extraction, or ultrasonic extraction may be used. , Can be obtained as a reflux organic solvent extract, hot water extract or high temperature and high pressure extract. These extraction methods are general extraction methods known in the art and are not particularly limited.

The organic solvent is methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1 , 3-butylene glycol, propylene glycol, or a mixed solvent thereof, and can be extracted at room temperature or by heating in an environment where the active ingredient of the extract is not destroyed or minimized. Depending on the organic solvent to be extracted, the extraction degree and loss degree of the active ingredient of the extract may differ, so it is preferable to select and use a suitable organic solvent.

According to one embodiment of the present invention, the whole plant including leaves and roots of sprout ginseng can be cleaned and extracted using alcohol, and the concentration of alcohol may be 20 to 100% (v / v). have. In the present invention, it is possible to prepare a sprout ginseng extract by repeatedly refluxing at 50 to 100 ° C using alcohol.

According to one embodiment of the present invention, the sprout ginseng extract in the present invention is characterized in that it is produced by extracting sprout ginseng using 20 to 100% (v / v) alcohol.

In the present invention, the bud ginseng extract is further characterized by using column chromatography to prepare and use a fraction of the bud ginseng extract (hereinafter referred to as bud ginseng fraction).

According to one embodiment of the present invention, the sprout ginseng fraction of the present invention is characterized by being produced by performing column chromatography using the sprout ginseng extract as water or 20 to 100% (v / v) alcohol as a solvent. do.

According to one embodiment of the present invention, the bud ginseng extract, bud ginseng fraction or a mixture thereof is Sumyzyme AC, Cellulase KN, Plantase TL, Plantase PE Conc, Plantase TCL, Plantase MAX, BrewMax GXL, Alphalase AP3, Alpase NP, Sumizyme SPC, Plantase AK, Pluszyme 2000D and Rohament CL Bioconversion of various ginsenosides contained in the bud ginseng extract and fractions to compound K may be performed using one or more enzymes selected from the group consisting of.

In this case, the method of adding the enzyme is not particularly limited as long as the method of inactivating the enzyme does not occur.

In addition, the reaction temperature may be performed at a temperature at which the enzyme inactivation does not occur, and its temperature is not particularly limited as the temperature for each enzyme activity.

Specifically, the ginseng extract, bud ginseng fraction or a mixture thereof, the Sumyzyme (Sumyzyme) AC, Cellulase (Cellulase) KN, Plantase (TL), Plantase PE Conc, Plantase TCL, Plantase TCL, Plantase Lantase MAX, BrewMax GXL, Alphalase AP3, Alpase NP, Sumizyme SPC, Plantase AK, Pluszyme 2000D and Rohament CL enzymes were added respectively, Compound K can be prepared in high yield by reacting at 30 to 60 ° C. for 20 to 50 hours.

According to a preferred embodiment of the present invention, enzymes that bio-convert ginsenosides contained in the sprout ginseng into Compound K (Sumyzyme AC), Cellulase KN, Plantase (Plantase) TL, Plantase PE Conc, Plantase TCL, Plantase MAX, BrewMax GXL, Alphalase AP3, Alpase NP, Sumizyme SPC, Plantase AK, Pluszyme (Pluszyme) It is preferred to use one or more enzymes from the group consisting of 2000D and Rohament CL.

According to one embodiment of the present invention, the present invention is characterized in that it further comprises a process of adding an enzyme to react and then filtering to extract using an organic solvent.

According to one embodiment of the present invention, Sumizyme AC, Cellulase KN, Plantase TL, Plantase PE Conc, Plantase in the bud ginseng extract or bud ginseng fraction The group consisting of TCL, plantase MAX, BrewMax GXL, Alphalase AP3, Alfasease NP, Sumyzyme SPC, Plantase AK, Pluszyme 2000D and Rohament CL In addition, by reacting by adding one or more enzymes, ginsenosides contained in sprout ginseng can be bioconverted to produce compound K with high efficiency.

According to one embodiment of the present invention, it is possible to further convert ginsenosides contained in sprout ginseng with higher efficiency by further filtering the bioconverted product using the enzyme and extracting it with an organic solvent.

The bioconversion product of the sprout ginseng extract obtained according to the present invention, the bioconversion product of the sprout ginseng fraction, and the organic solvent extract of the bioconversion product may have an improved antioxidant content and a significant increase in antioxidant and antibacterial activity.

According to a preferred embodiment of the present invention, as a result of bioconversion of the ginseng extract and fractions using enzymes, the content of various useful substances (polyphenols, flavonoids, tannins) increased, and compound K, a high value-added component, It was confirmed that it was generated.

According to a preferred embodiment of the present invention, the production amount of Compound K bioconverted methanol extract of the 70% fraction of the bioconversion filtrate was about 10% of the total substrate.

According to a preferred embodiment of the present invention, the antioxidant activity in all samples was high overall, in particular, the highest alcohol 20% fraction, and showed high efficacy in the bioconverted alcohol 70% fraction (laboratory, factory). .

According to a preferred embodiment of the present invention, in the antimicrobial effect, the overall effect was high in the extract obtained by extracting the methanol-exchanged filtrate of the ginseng extract and the fraction. In particular, all the bioconverted samples from Staphylococcus epidermidis , a skin fungus, showed an effect, and in Enterobacter cloacae , methanol extract of the sprout extract and the bioconversion filtrate of the fraction was methanol-extracted. It was effective in all of the extracts.

According to a preferred embodiment of the present invention, Staphylococcus aureus and Pseudomonas auruginosa from the extract obtained by extracting the biotransformed filtrate of the bud ginseng extract and the fraction with methanol have antibacterial activity against Pseudomonas auruginosa . Shown.

In the present invention, the antibacterial activity that did not appear in the sprout ginseng extract appeared through bioconversion, and it was determined that it can be used as an antibacterial active material later by confirming that it is high in the methanol extract of the filtrate.

Therefore, according to a preferred embodiment of the present invention, in the case of the methanol extract of the filtrate that is bioconverted the main fraction of the sprout ginseng extract, the utilization value is high due to the metabolic production of compound K, a high value-added product, and the remaining fraction of the sprout ginseng extract In the case of the methanol extract of the bioconversion filtrate, it is desirable to use it as an antimicrobial material, and in the case of the extract-alcohol 20% fraction, to use it as an antioxidant active material.

On the other hand, the present invention provides a composition for antioxidant and antibacterial containing compound K prepared according to the above production method.

As described above, in the present invention, Sumizyme AC, Cellulase KN, Plantase TL, Plantase PE Conc, Plantase TCL, Plantase TCL, Selected from the group consisting of Plantase MAX, BrewMax GXL, Alphalase AP3, Alfasease NP, Sumizyme SPC, Plantase AK, Pluszyme 2000D and Rohament CL Various ginsenosides contained in sprout ginseng can be converted into compound K through a simple process of processing the enzyme. In addition, compound K prepared according to this method has excellent antioxidant and antibacterial activity, and thus can be usefully used as a composition for antioxidant and antibacterial.

Figure 1 discloses an example of a process for bioconverting ginsenosides contained in sprout ginseng in the present invention to compound K.
Figure 2 is a graph showing the results of measuring the electron donation capacity of each scale of Ginseng Ginseng Extract.
Figure 3 is a graph showing the results of measuring the electron donating capacity of the sprout ginseng extract and fraction bioconversion filtrate and filtrate methanol extract.
4 is a result of measuring the cation radical scavenging ability of fractions by laboratory ginseng extract scale (laboratory or factory).
5 is a result of measuring the cation radical scavenging ability of the sprout ginseng extract and fraction bioconversion filtrate and filtrate methanol extract.
6 is a result of measuring the reducing power of the extract of sprout ginseng.
7 is a result of measuring the reducing power of the sprout ginseng extract and fraction bioconversion filtrate and filtrate methanol extract.
8A to 8F show the results of compound K and ginsenoside F2 analysis of sprout ginseng extract and fractions, bioconversion filtrate and filtrate methanol extract.
9 is a result of verifying the antibacterial activity of the sprout ginseng extract and fractions for various strains.
10 is a result of verifying the antibacterial activity of the ginseng extract and fraction bioconversion filtrate for various strains.
11 is a result of verifying the antibacterial activity of the sprout ginseng extract and fraction bioconversion filtrate methanol extract for various strains.

Hereinafter, examples and the like will be described in detail to help understanding of the present invention. However, the embodiments according to the present invention can be modified in many different forms, and the scope of the present invention should not be construed as being limited to the following examples. The embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

<Example 1> Preparation of sprout ginseng extract and fractions

1-1. Preparation of sprout ginseng extract

After washing the leaves and roots of ginseng ginseng clean, the extracts (hereinafter referred to as Lab extracts) were prepared by repeating reflux extraction 3 times at 60 ° C for 3 hours using alcohol, and then producing sprout ginseng in bulk. A reflux repetitive extraction was repeated 3 times at a temperature of 60 ° C. or higher using alcohol to prepare a sprouted ginseng bulk extract (Bulk extract). After that, the extract was filtered using filter paper, and the filtered extract was concentrated using a rotary vacuum concentrator, and the concentrated samples were filtered using a sterile membrane filter and used in the following experiment.

1-2. Preparation of sprout ginseng fraction

Each of the sprout ginseng Lab extract and Bulk extract prepared in 1-1 was prepared using column chromatography (hereinafter, the fraction of Lab extract is called Lab fraction, and the fraction of Bulk extract is called Bulk fraction). When preparing the fraction, water and alcohol were used as solvents, and thus 10 fractions were obtained. Then, the obtained fraction was filtered using filter paper, and the filtered fraction was concentrated using a rotary vacuum concentrator, and the concentrated samples were filtered using a sterile membrane filter and used in the following experiment.

<Example 2> Bio-conversion of extracts and fractions

Samples were prepared by mixing Lab extracts and fractions of Bulk Ginseng of Example 1-2 with Bulk extracts and fractions in various cases, respectively, and preparing Sumizyme AC, Cellulase KN, and Plantase (Plantase). ) TL, Plantase PE Conc, Plantase TCL, Plantase MAX, BrewMax GXL, Alphalase AP3, Alphase NP, Sumizyme SPC, Plantase AK, Pluszyme ) Biosynthesis of various ginsenosides contained in bud ginseng extract and fractions into Compound K was carried out using 2000D and Rohament CL enzymes, respectively.

Specifically, after mixing 100 mL of the sample and 100 mL of distilled water, the sample for analysis (pre-reaction sample) was removed, and then the enzymes were added. Then, after shaking culture, it was filtered using filter paper, and 20 mL of the reaction filtrate was placed in a 50 mL tube and dried. The remaining reaction filtrate was concentrated under reduced pressure and then stored after scale down drying. What remained on the filter paper after filtration was collected after drying.

Then, the filtrate was extracted with MeOH. The lyophilized filtrate was analyzed by filtration through membrane filtration. After the reaction, analysis was conducted by sampling the same as before the reaction.

Figure 1 discloses an example of a process for bioconverting ginsenosides contained in sprout ginseng in the present invention to compound K.

The following experiment was conducted using a sample bioconverted with enzymes.

<Example 3> Antioxidant activity verification

3-1. Experimental method

(1) Condensation type tannin content measurement

To the total tannin content, ₂ mL of vanillin dissolved in H ₂ SO ₄ solution was added to 1 mL of the sample diluted in distilled water, and after standing for 15 minutes, absorbance was measured at 500 nm. As a standard, catechin was used, and after preparing a standard calibration curve, the total tannin content of the extract was expressed as μg CE (catechin equivalent) / g.

(2) Total polyphenol content measurement

To quantify the total polyphenol, add 25 µl of distilled water and 25 µl of Folin-Ciocalteu phenol reagent reagent to 25 µl of the sample diluted in distilled water, react for 6 minutes, add 100 µl of saturated solution Na ₂ CO ₃ , mix, and then mix at room temperature for 90 minutes. After standing and measuring absorbance at 765 nm, the polyphenol content was calculated by comparing the absorbance value of a standard curve previously prepared with gallic acid, a standard material.

(3) Total flavonoid content measurement

The total flavonoid content was added 1.25 mL and 75 μL of a 5% NaNO ₂ solution was added and left for 5 minutes. 150 μL of a 10% AlCl _{3 ·} 6H ₂ O solution was added and allowed to stand for another 6 minutes. After adding 500 μL of 1M NaOH and 275 μL of distilled water to the reaction solution, absorbance was measured at 510 nm. After preparing a standard calibration curve using quercetin as a standard, the total flavonoid content of the extract was expressed as μg QE (Quercetin equivalent) / g.

(4) Measurement of electron donation ability

The electron donating ability (EDA) was measured by applying the method of Blois. 60 μl of 0.45 mM 1,1-diphenyl-2-picrylhydrazyl (DPPH) was added to 120 μl of each sample solution, allowed to stand for 15 minutes, and absorbance was measured at 517 nm. The electron donating ability was represented by the rate of decrease in absorbance of the sample-added and non-added groups according to Equation 1 below.

(5) ABTS cationic radical scavenging ability measurement

Measurement of antioxidant power using ABTS cationic radicals It was measured by cationic radical decolorization analysis. 7 mM 2,2-azino-bis (3-ethyl-benzothiazoline-6-sulfonic acid) and 2.45 mM potassium persulfate were mixed and left at room temperature for 24 hours to form ABTS cationic radicals, diluted with ethanol, and then substrate 100 After adding 100 µl of sample to µl and leaving for 7 minutes, absorbance was measured at 734 nm. The erasing ability was calculated according to Equation 2 below.

(6) FRAP analysis

10 mM 1: 1 in 300 mM acetate buffer (pH 3.6), 10 mM TPTZ (2,4,6-tripyridyl-S-triazine) solution in 40 mM HCl and 20 mM FeCl ₃ · 6H ₂ O, respectively. (v / v / v) was mixed in advance and then heated in a 37 ° C water bath was used. 100 μL of the extracts were sequentially mixed, reacted at 37 ° C. for 4 minutes, and absorbance was measured at 593 nm using a UV spectrophotometer.

3-2. Experiment result

(1) Total polyphenol, flavonoid and condensed tannin content measurement results

The total polyphenol, flavonoid and condensed tannin content measurement results are shown in Table 1 below.

Sample name Polyphenols (GAE) ^a Flavonoids
(QE) ^b Tannins
(CE) ^c a 43.08 ± 0.83 290.67 ± 5.88 50.48 ± 0.33 b 27.16 ± 0.58 229.19 ± 41.70 53.82 ± 0.76 c 13.94 ± 0.06 144.00 ± 8.89 75.76 ± 0.54 d 38.84 ± 0.16 248.44 ± 1.57 57.87 ± 1.26 e 22.02 ± 0.27 183.63 ± 4.49 52.82 ± 0.44 f 12.87 ± 0.05 160.67 ± 26.71 65.21 ± 1.21 a-1 16.35 ± 0.35 1.66 ± 0.23 5.71 ± 0.33 b-1 15.12 ± 0.08 2.57 ± 0.23 5.97 ± 0.40 c-1 31.45 ± 0.26 19.54 ± 1.51 10.00 ± 0.29 d-1 23.91 ± 0.48 13.78 ± 0.57 11.35 ± 1.93 e-1 14.12 ± 0.06 5.06 ± 0.01 10.65 ± 0.29 a-2 12.48 ± 0.19 37.04 ± 1.17 52.63 ± 4.91 b-2 11.32 ± 0.15 30.18 ± 1.24 43.72 ± 0.62 c-2 35.91 ± 0.20 41.20 ± 2.62 16.74 ± 0.99 d-2 22.45 ± 0.25 13.93 ± 1.72 20.65 ± 0.73 e-2 8.02 ± 0.04 12.23 ± 1.12 19.43 ± 0.91

a: Sprout ginseng 70% alcohol extract-20% alcohol fraction (lab), b: Sprout ginseng 70% alcohol extract-70% alcohol fraction (lab), c: Sprout ginseng 70% alcohol extract-100% alcohol fraction (lab) , d: Sprout ginseng 70% alcohol extract-20% alcohol fraction (factory), e: Sprout ginseng 70% alcohol extract-70% alcohol fraction (factory), f: Sprout ginseng 70% alcohol extract-100% alcohol fraction (factory) )

a-1: sprout ginseng 70% alcohol extract bioconversion filtrate (laboratory), b-1: sprout ginseng 70% alcohol extract bioconversion filtrate (factory), c-1: sprout ginseng 70% alcohol extract-70% alcohol Fractional bioconversion filtrate (laboratory), d-1: 70% alcoholic extract of sprouted ginseng-70% alcoholic filtrate fraction (factory), e-1: 70% alcoholic extract of sprouted ginseng-100% alcoholic fractionation Liquid (laboratory + factory)

a-2: Sprout ginseng 70% alcohol extract bioconversion filtrate methanol extract (laboratory), b-2: Sprout ginseng 70% alcohol extract bioconversion filtrate methanol extract (factory), c-2: Sprout ginseng 70% alcohol extract -70% alcohol fraction bioconversion filtrate methanol extract (lab), d-2: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate methanol extract (factory) e-2: sprout ginseng 70% alcohol extract- 100% alcohol fraction bioconversion filtrate methanol extract (laboratory + factory)

As shown in Table 1, among the fractions extracted from the total polyphenol content, a: sprout ginseng 70% alcohol extract-20% alcohol fraction (laboratory) showed the highest content (43.08 ± 0.83), and then d: sprout ginseng 70% alcoholic extract-20% alcoholic fraction (plant) was the highest.

The total flavonoid content showed the same pattern as the polyphenol content (a: sprout ginseng 70% alcohol extract-20% alcohol fraction (laboratory): 290.67 ± 5.88, sprout ginseng 70% alcohol extract-20% alcohol fraction (factory): 248.44 ± 1.57), for condensed tannin content, c: 70% ethanol extract from sprout ginseng-100% alcohol fraction (laboratory): 75.76 ± 0.54, f: 70% ethanol extract from sprout ginseng-100% alcohol fraction (factory): 65.21 ± 1.21), a: Sprout ginseng 70% alcohol extract-20% alcohol fraction (laboratory) and b: Sprout ginseng 70% alcohol extract-70% alcohol fraction (laboratory) showed similar contents.

 The total polyphenol content in the 70% ethanol extract bioconverted filtrate was c-1: 70% ethanol extract from sprout ginseng-the 70% alcohol fraction bioconversion filtrate (lab) (31.45 ± 0.26) was the highest, followed by d-1: Sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate (factory) (23.91 ± 0.48) was high. The flavonoid content showed a similar pattern, the highest in the laboratory at 19.54 ± 1.51, and then the plant contained 23.91 ± 0.48. However, for the condensed tannin, 70% alcohol fraction bioconversion filtrate (laboratory: 10.00 ± 0.29, factory: 11.35 ± 1.93) and 100% alcohol fraction bioconversion filtrate (laboratory + factory: 10.65 ± 0.29) were similar.

The total polyphenol content in the methanol extract of 70% ethanol extract from sprout ginseng was similar to that of the extract and filtrate. Sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate methanol extract (Lab: 35.91 ± 0.20) was the highest, followed by the plant at 22.45 ± 0.25. However, in the total flavonoids, unlike other groups, the sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate methanol extract (laboratory: 41.20 ± 2.62) was the highest, but the laboratory: 37.04 ± 1.17, factory: 30.18 ± 1.24 It showed this highly appearing pattern. In the case of condensed tannin, the methanol extract (lab: 52.63 ± 4.91) with the highest percentage of sprouted ginseng 70% alcohol extract (Lab: 52.63 ± 4.91) was highest, and the plant was high at 43.72 ± 0.62. Through this, it was confirmed that a higher useful component can be secured by bioconversion of the same amount of sample. In particular, the methanol extract of the unfiltered portion after bioconversion showed a very high content.

(2) Result of measuring electron donation ability

Figure 2 is a graph showing the results of measuring the electron donation capacity of each scale of Ginseng Ginseng Extract. Where: a: sprout ginseng 70% alcohol extract-20% alcohol fraction (laboratory), b: sprout ginseng 70% alcohol extract-70% alcohol fraction (laboratory), c: sprout ginseng 70% alcohol extract-100% alcohol fraction (Lab), d: Sprout ginseng 70% alcohol extract-20% alcohol fraction (factory), e: Sprout ginseng 70% alcohol extract-70% alcohol fraction (factory), f: Sprout ginseng 70% alcohol extract-100% alcohol Fraction (plant), Vit.C: ascorbic acid.

As shown in FIG. 2, the electron donating ability showed the highest electron donating ability of about 90%, with the highest alcohol 20% fraction (laboratory) among the extracted fractions, followed by the highest alcohol 20% fraction (factory). Next, 70% and 20% were the highest, and the laboratory scale showed higher donation capacity than the factory.

Figure 3 is a graph showing the results of measuring the electron donating capacity of the sprout ginseng extract and fraction bioconversion filtrate and filtrate methanol extract. Here, a-1: sprout ginseng 70% alcohol extract bioconversion filtrate (laboratory), b-1: sprout ginseng 70% alcohol extract bioconversion filtrate (factory), c-1: sprout ginseng 70% alcohol extract- 70% alcohol fraction bioconversion filtrate (laboratory), d-1: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate (factory), e-1: sprout ginseng 70% alcohol extract-100% alcohol fraction Bioconversion filtrate (laboratory + factory), a-2: sprout ginseng 70% alcohol extract bioconversion filtrate methanol extract (laboratory), b-2: sprout ginseng 70% alcohol extract bioconversion filtrate methanol extract (factory), c-2: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate methanol extract (lab), d-2: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate methanol extract (factory), e-2: Sprout ginseng 70% alcohol extract-100% alcohol fraction bioconversion filtrate methanol extract ( + Laboratory plant), Vit.C: an ascorbic acid.

As shown in FIG. 3, in the 70% ethanol extract of sprouted ginseng extract and filtrate methanol extract, the 70% fraction bioconversion filtrate methanol extract (laboratory, factory) exhibited the highest electron donating capacity (80% or more). Next, the 70% alcohol fraction bioconversion filtrate (laboratory) and the 100% alcohol fraction bioconversion filtrate methanol extract (factory) were high.

In the case of electron donating ability, there were extracts, fractions and bioconversion filtrates and filtrate methanol extracts showing high activity, based on the concentration of 1000 μg / mL, and among them, the highest was the alcoholic 20% fraction (laboratory). In addition, in the case of bioconversion filtrate and filtrate methanol extract, alcoholic 70% fraction filtrate and filtrate methanol extract showed high activity. This resulted in significant results when compared to the content analysis.

(3) ABTS cationic radical scavenging ability measurement

4 is a result of measuring the cation radical scavenging ability of fractions by laboratory ginseng extract scale (laboratory or factory). Where: a: sprout ginseng 70% alcohol extract-20% alcohol fraction (laboratory), b: sprout ginseng 70% alcohol extract-laboratory 70% alcohol fraction (laboratory), c: sprout ginseng 70% alcohol extract-100% alcohol fraction ( Laboratory), d: sprouted ginseng 70% alcohol extract-20% alcoholic extract (factory), e: sprouted ginseng 70% alcoholic extract-70% alcoholic extract (factory), f: sprouted ginseng 70% alcoholic extract-100% alcoholic fraction (Factory), Vit.C: Ascorbic acid.

5 is a result of measuring the cation radical scavenging ability of the sprout ginseng extract and fraction bioconversion filtrate and filtrate methanol extract. Where a-1: sprout ginseng 70% alcohol extract bioconversion filtrate (laboratory), b-1: sprout ginseng 70% alcohol extract bioconversion filtrate (factory), c-1: sprout ginseng 70% alcohol extract-70 % Distillate fraction bioconversion filtrate (laboratory), d-1: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate (factory), e-1: sprout ginseng 70% alcohol extract-100% alcohol fraction Conversion filtrate (laboratory + factory), a-2: sprout ginseng 70% alcohol extract bioconversion filtrate methanol extract (laboratory), b-2: sprout ginseng 70% alcohol extract bioconversion filtrate methanol extract (factory), c -2: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate methanol extract (laboratory), d-2: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate methanol extract (factory), e -2: Sprout ginseng 70% alcohol extract-100% alcohol fraction bioconversion filtrate methanol extract (real Room + plant), Vit.C: an ascorbic acid.

The ABTS cationic radical scavenging activity showed the highest scavenging activity of about 90% of the alcohol fraction of 20% (laboratory), followed by the highest alcohol extraction fraction (factory). Next, 70% and 20% were the highest, and the laboratory scale showed higher scavenging ability than the factory. This shows a pattern similar to the electron donating ability.

Sprout ginseng 70% alcohol extract bioconversion filtrate and filtrate methanol extracts have the highest ABTS cationic radical scavenging ability of alcohol 70% fraction bioconversion filtrate methanol extract (laboratory, factory) and 70% alcohol fraction bioconversion filtrate (laboratory) (90% or more), and the ABTS cationic radical scavenging ability showed a pattern similar to the results of the electron donating ability and content analysis when viewed in terms of the concentration of 1000 μg / mL.

(4) FRAP analysis

6 is a result of measuring the reducing power of the extract of sprout ginseng. Where: a: sprout ginseng 70% alcohol extract-20% alcohol fraction (laboratory), b: sprout ginseng 70% alcohol extract-70% alcohol fraction (laboratory), c: sprout ginseng 70% alcohol extract-100% alcohol fraction (Lab), d: Sprout ginseng 70% alcohol extract-20% alcohol fraction (factory), e: Sprout ginseng 70% alcohol extract-70% alcohol fraction (factory), f: Sprout ginseng 70% alcohol extract-100% alcohol Fraction (plant), Vit.C: ascorbic acid.

7 is a result of measuring the reducing power of the sprout ginseng extract and fraction bioconversion filtrate and filtrate methanol extract. Here, a-1: sprout ginseng 70% alcohol extract bioconversion filtrate (laboratory), b-1: sprout ginseng 70% alcohol extract bioconversion filtrate (factory), c-1: sprout ginseng 70% alcohol extract- 70% alcohol fraction bioconversion filtrate (laboratory), d-1: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate (factory), e-1: sprout ginseng 70% alcohol extract-100% alcohol fraction Bioconversion filtrate (laboratory + factory), a-2: sprout ginseng 70% alcohol extract bioconversion filtrate methanol extract (laboratory), b-2: sprout ginseng 70% alcohol extract bioconversion filtrate methanol extract (factory), c-2: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate methanol extract (lab), d-2: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate methanol extract (factory), e-2: Sprout ginseng 70% alcohol extract-100% alcohol fraction bioconversion filtrate methanol extract (real Humsil + Factory), Vit.C: Ascorbic acid.

FRAP (reducing power) also showed a pattern similar to the previous electron donating ability, ABTS cationic radical scavenging ability, and content analysis results. Among the extracted fractions, the 20% alcohol fraction (laboratory) showed the highest reducing power, followed by the highest 20% alcohol fraction (factory). Next, 70% and 20% were the highest, and the laboratory scale showed higher scavenging ability than the factory. In addition, in the 70% ethanol extract of sprouted ginseng extract, and the methanol extract of filtrate, the 70% fraction bioconverted methanol extract (laboratory, factory) showed the highest reducing power, and in the case of reducing power, the concentration of 1000㎍ / mL Based on the criteria, it showed a pattern similar to other antioxidant activity verification experiments.

< Example  4> Ingredient analysis

4-1. Experimental method

(1) Quantitative and qualitative analysis using LC-MS and ELSD

LC / MS used was composed of 1200 series liquid chromatography / 6130 single quadrupole mass selective detector (Agilent Technologies, USA), and the ionization method of MS was electrospray ionization (ESI). It is a positive mode of the method, and was analyzed by reverse phase liquid chromatography using LC / MS with a quadrupole mass separation tube attached as an ion separation method. As a separation condition, a Shiseido capcell pak C18 MG-II (5 μm 4.6 mm * 250 mm) column was used, and the mobile phase was HPLC grade water (A) containing 0.5% formic acid and HPLC grade acetonitrile (B) containing 0.5% formic acid. B: 5% in 0-4 minutes, B: 5-25% in 4-10 minutes, B: 25-40% in 10-20 minutes, B: 40-60% in 20-26 minutes, 26 Gradient conditions such that B: 60 to 85% until -40 minutes, B: 85 to 100% until 40-50 minutes, B: 100% until 50-56 minutes, and B: 100 to 5% until 56-60 minutes Was used, and the flow rate was 1 mL / min. As an analysis condition of the electrospray ionization method, nitrogen (99.99%) was used as a dry gas at a flow rate of 12 L / min, and the vaporization temperature was 350 ° C., spraying a sample at a 35 psi injection pressure, and using 70 fragmentation voltage. . The capillary voltage was 3,000 V, and the detection method used a scan mode with a range of 100-1500. The mass spectrometer used 360 ELSD (Agilent Technologies, U.S.A), and was used as 1.60 SLM gas flow rate, 100% LED intensity, 30 ° C evaporation temperature, 30 ° C atomizer temperature. Quantification was quantified through each standard curve. The results are shown in Table 2 below.

sample Ginsenoside F2 Compound K Sprout ginseng extract (factory) 2.82 ㎍ / 80㎍ ㎍ / 80㎍ Sprout ginseng extract-alcohol 70% fraction (Lab) 0.45 Sprout ginseng extract-alcohol 70% fraction (factory) 0.62 Sprout ginseng extract-alcohol 100% fraction (laboratory + factory) 1.11 Sprout ginseng extract bioconversion filtrate (laboratory) - Sprout ginseng extract bioconversion filtrate (factory) - Sprout ginseng extract-alcohol 70% fraction bioconversion filtrate (laboratory) 0.44 Sprout ginseng extract-alcohol 70% fraction bioconversion filtrate (factory) 0.47 0.68 Sprout ginseng extract-100% fraction bioconversion filtrate (laboratory + factory) - 0 Sprout ginseng extract bioconversion filtrate methanol extract (laboratory) 0.91 0 Sprout ginseng extract bioconversion filtrate methanol extract (factory) 1.48 0.79 Sprout ginseng extract-alcohol 70% fraction bioconversion filtrate methanol extract (laboratory) 0.49 0.51 Sprout ginseng extract-alcohol 70% fraction bioconversion filtrate methanol extract (factory) 0.61 1.20 Sprout ginseng extract-alcohol 100% fraction bioconversion filtrate methanol extract (laboratory + factory) 0.94 7.25

From the results of quantitative and sexual analysis of sprout ginseng extract and fraction, bioconversion filtrate and filtrate methanol extract, it was confirmed that compound K was produced by performing bioconversion. As shown in Table 2, it could be confirmed that the sprout ginseng extract-alcohol 100% fraction bioconversion filtrate methanol extract (lab + plant) was approximately 10% converted to 7.25㎍ / 80㎍. Extracts and fractions, bioconversion filtrate and filtrate methanol extract Overall, ginsenoside F2 contains a trace amount, but among them, the ginseng extract (plant) showed the highest content of 2.82㎍ / 80㎍.

Through this, it was confirmed that compound K, a high value-added component, can be converted through bioconversion using enzymes. In addition, a process and an analysis method for generating it were established.

8A to 8F show the results of compound K and ginsenoside F2 analysis of sprout ginseng extract and fractions, bioconversion filtrate and filtrate methanol extract.

<Example 5> Antibacterial activity verification

 5-1. Experiment method

(1) Culture of bacteria

The pre-cultivation and liquid media for this culture are Staphylococcus epidermidis and Escherichia coli ), Enterobacter cloacae , Pseudomonas auruginosa ) is used for nutrient medium (NB) and nutrient agar as Staphylococcus aureus as Gifu anaerobic medium (GAM) and Gifu basic medium (GAM) agar. As a liquid and solid medium of Candida albicans , yeast peptone dextrose (YPD) medium and yeast peptone dextrose (YPD) agar medium were used. Incubated at 37 ° C in a BOD incubator.

(2) Measurement of growth clear zone

Antibacterial activity was measured by a paper disc agar diffusion method. That is, 1 platinum of each strain cultured on a plate medium is taken and activated by culturing in 10 ml of liquid medium for 18 to 24 hours, and then, 10 ml of liquid medium is inoculated with 0.1 ml of the bacterial solution for 3 to 6 hours. The cells were inoculated so that the number of bacteria was about 1X10 ⁷ cells and uniformly smeared with a sterile cotton swab. After placing the sterilized filter paper disc (8mm, Whatman, Japan) on a solid flat plate, the sample is absorbed by concentration so as to be 0.05mL / disc, and cultured at 37 ° C for 18 to 24 hours to grow it around the disc (clear zone). The diameter of (mm) was measured.

5-2. Experiment result

(1) Measurement results of antibacterial activity growth inhibition rings of bud ginseng extract and fractions for various strains

The antibacterial activity of 6 types of sprout ginseng extract and fractions was confirmed through 6 different strains. All sample concentrations were used at a concentration of 4 mg / disc, and the positive control material, Vit.C, was also treated at a concentration of 4 mg / disc.

9 is a result of verifying the antibacterial activity of the sprout ginseng extract and fractions for various strains. Here, A: Staphylococcus epidermidis , B: Candida albicans , C: Staphylococcus aureus , D: Enterobacter cloacae , E: Escherichia coli , F: Pseudomonas auruginosa , a: ascorbic acid, b: 70% sprouted ginseng extract (plant), c: 70% sprouted ginseng extract-20% Alcohol fraction (laboratory), d: sprout ginseng 70% alcohol extract-20% alcohol fraction (factory), e: sprout ginseng 70% alcohol extract-70% alcohol fraction (laboratory), f: sprout ginseng 70% alcohol extract-70 % Alcohol fraction (plant), g: sprout ginseng 70% alcohol extract-100% alcohol fraction (laboratory + plant).

As shown in Figure 9 and Table 3, when S. epidermidis strain was first treated with ginseng extract and fractions, growth inhibition rings of 4.0 ± 1.0 mm, 10.7 ± 1.5 mm, and size appeared in b and g, positive control material Phosphorus Vit.C showed growth inhibition of 11.0 ± 1.0 mm. In the case of C. albicans , a growth inhibition ring of 2.7 ± 0.6 mm was observed in g, but no antibacterial activity was observed in the positive control material. In the case of S. aureus , a growth inhibition ring of 5.7 ± 0.6 mm was observed in g, but no antibacterial activity was observed in the positive control material. In the case of E. cloacae , E. coli , and P. auruginosa strains, growth inhibition rings of 11.0 ± 1.0 ㎜, 5.0 ± 1.0 ㎜, and 15.3 ± 0.6 ㎜ were found in the positive control, but antibacterial bacteria were treated when the bud ginseng extract and fraction were treated. There was no activity.

(2) Measurement results of growth inhibition of antibacterial activity of bio-conversion filtrate of sprout ginseng extract and fractions for various strains

Similarly, through 6 different strains, the antibacterial activity of 6 types of sprout ginseng extract and fraction bioconversion filtrate was confirmed. All sample concentrations were used at a concentration of 4 mg / disc, and the positive control material, Vit.C, was also treated at a concentration of 4 mg / disc.

10 is a result of verifying the antibacterial activity of the ginseng extract and fraction bioconversion filtrate for various strains. Here, A: Staphylococcus epidermidis , B: Candida albicans , C: Staphylococcus aureus , D: Enterobacter cloacae , E: Escherichia coli , F: Pseudomonas auruginosa , a: Ascorbic acid, b: Sprout ginseng 70%, extract bioconversion filtrate (laboratory), c: Sprout ginseng 70% Extract bioconversion filtrate (factory), d: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate (laboratory), e: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate (factory) , f: Sprout ginseng 70% alcohol extract-100% alcohol fraction bioconversion filtrate (laboratory + factory).

10 and Table 4, when S. epidermidis strain was first treated with the ginseng extract and fraction bioconversion filtrate, 4.7 ± 1.2 mm, 4.0 ± 0.1 mm, 2.0 at b, c, d, e, f Growth inhibition rings of ± 0.1 ㎜, 4.3 ± 0.6 ㎜, 7.3 ± 1.5 ㎜, and size were shown. Vit.C, a positive control substance, showed growth inhibition rings of 10.7 ± 0.6 ㎜. In the case of C. albicans and S. aureus , antibacterial activity was not observed when treated with the positive control substance and the bud ginseng extract and fraction bioconversion filtrate. In the case of E. cloacae , growth inhibition rings of 5.3 ± 0.6 mm and 6.3 ± 0.6 mm were observed in d and e, and growth inhibition rings of 16.3 ± 1.2 mm were found in the positive control substance. In the case of E. coli and P. auruginosa strains, growth inhibition rings of 7.7 ± 0.6 mm and 8.0 ± 0.1 mm were found in the positive control material, but antibacterial activity was not observed when the bud ginseng extract and fraction bioconversion filtrate were treated.

(3) Measurement results of antibacterial activity growth inhibition rings of sprout ginseng extract and fraction bioconversion filtrate methanol extract for various strains

Similarly, through 6 different strains, the antibacterial activity of 6 types of sprout ginseng extract and fraction bioconversion filtrate methanol extract was confirmed. All sample concentrations were used at a concentration of 4 mg / disc, and the positive control material, Vit.C, was also treated at a concentration of 4 mg / disc.

11 is a result of verifying the antibacterial activity of the sprout ginseng extract and fraction bioconversion filtrate methanol extract for various strains. Here, A: Staphylococcus epidermidis , B: Candida albicans , C: Staphylococcus aureus , D: Enterobacter cloacae , E: Escherichia coli , F: Pseudomonas auruginosa , a: ascorbic acid, b: sprouted ginseng 70% alcohol extract bioconversion filtrate methanol extract (laboratory), c: sprouted ginseng 70 % Alcohol extract bioconversion filtrate methanol extract (factory), d: sprout ginseng 70% alcohol extract-70% alcohol fraction bioconversion filtrate methanol extract (lab), e: sprout ginseng 70% alcohol extract-70% alcohol fraction Conversion filtrate methanol extract (factory), f: sprout ginseng 70% alcohol extract-100% alcohol fraction bioconversion filtrate methanol extract (laboratory + factory).

As shown in Fig. 11 and Table 5, when S. epidermidis strain was first treated with a sprout ginseng extract and a fraction bioconversion filtrate methanol extract, 16.7 ± 0.6 mm, 9.0 ± 0.1 mm in b, c, d, e, and f , 2.7 ± 0.6 ㎜, 5.3 ± 0.6 ㎜, 15.7 ± 0.6 ㎜ growth inhibition ring, positive control material Vit.C showed 10.3 ± 0.6 ㎜ growth inhibition ring. In the case of C. albicans , antibacterial activity was not observed when treated with the positive control substance, the sprout ginseng extract and the fraction bioconversion filtrate methanol extract. In S. aureus , growth inhibition rings of 3.0 ± 0.1 mm, 2.0 ± 0.1 mm, and 2.0 ± 0.1 mm were observed in b, c, and f, but antibacterial activity was not observed in the positive control material. In the case of E. cloacae , growth inhibition rings of 3.7 ± 0.6 ㎜, 1.7 ± 0.6 ㎜, 5.0 ± 0.1 ㎜, 5.3 ± 0.6 ㎜, 3.7 ± 0.6 ㎜ in b, c, d, e, and f were observed, and positive control In the substance, growth inhibition of 16.3 ± 0.6 mm was observed. In the case of the E. coli strain, growth inhibition of 5.7 ± 0.6 mm was observed in the positive control material, but did not show antibacterial activity when treated with the sprout ginseng extract and the fraction bioconversion filtrate methanol extract. In the case of P. auruginosa , growth inhibition rings of 4.3 ± 0.6 mm and 3.3 ± 0.6 mm were observed in b and c, and growth inhibition rings of 8.7 ± 0.6 mm were found in the positive control substance.

Claims (5)

The extract of Sprout ginseng extract is extracted from the fraction by fractionation by performing column chromatography with a solvent.Syzyzyme AC, Cellulase KN, Plantase TL, Plantase PE Conc, Plantase Made of TCL, Plantase MAX, BrewMax GXL, Alphalase AP3, Alpase NP, Sumyzyme SPC, Plantase AK, Pluszyme 2000D and Rohament CL As a method for preparing a sprout ginseng extract, characterized by extracting with methanol after reacting at 30 to 60 ° C. for 20 to 50 hours by adding one or more enzymes selected from the group, the sprout ginseng extract is a compound of Formula 1 ( Compound) Method for preparing sprout ginseng extract containing K 7.25ug / 80ug:
Formula 1

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