KR20190135902A - Fermented product of garlic and Cirsium setidens nakai by lactic acid bacteria, method of manufacturing the same, and use of the same - Google Patents

Abstract

The present invention relates to a garlic and Cirsium setidens fermented product using lactic acid bacteria which ferments a mixture of a garlic extract and a Cirsium setidens enzyme treatment extract using lactic acid bacteria, to a manufacturing method thereof, and to an antioxidant and immune improving composition comprising the same. The fermented product using the lactic acid bacteria according to the present invention can be utilized as a material for health functional foods such as beverages and the like.

Description

Fermented product of garlic and Cirsium setidens nakai by lactic acid bacteria, method of manufacturing the same, and use of the same}

The present invention relates to garlic and gondry lactobacillus fermented product, a method for preparing the same, and a use thereof, and more particularly, to a garlic and gondola lactobacillus fermented product obtained by fermenting a mixture of garlic extract and gondre enzyme-treated extract with lactic acid bacteria, a method for preparing the same, and the same. It relates to a composition for antioxidant and immune improvement.

Due to food additives, pesticides, chemical fertilizers, medicines, irregular eating habits and westernized diets, the number of adult diseases such as frequent illnesses, high blood pressure, diabetes, obesity due to reduced immunity continue to increase, resulting in a stronger natural-oriented life. Interest in health, such as health and medical information continues to increase, and in recent years, a lot of interest in functional health foods, supplements have increased.

The functional activity of health foods is known to be derived from numerous bioactive compounds such as carotenoids, dietary fiber, polyunsaturated fatty acids, bioactive peptides and phenolic compounds (Trends in Food Science & Technology, 69, 214-219 (2017); Siscovick et al., Circulation, CIR (2017)).

Garlic ( Allium sativum L. ) has been reported to have antioxidant, anti-cancer, anti-aging, antimicrobial effects, blood circulation improvement, and good cardiovascular effects.It was included in the 10 health foods selected by the Times Magazine in 2002. It is known that its efficacy is excellent enough to select garlic first among 48 foods with anticancer activity.

Cirsium setidens nakai has been used as a traditional Chinese medicine for centuries. It is a perennial herb belonging to the Asteraceae family, distributed mainly in Gangwon-do. The gondre contains bioactive compounds such as hispidulin 7-O-neohesperidoside, pectolinarin, luteolin, and apigenin. Therefore, it has been used for the treatment of edema, bleeding and hemoptysis (Archives of pharmacal research, 34 (3), 455 (2011)).

In addition, Gondre is known to have antioxidant activity, hepatoprotective activity, and activity against nonalcoholic fatty liver disease (The American journal of Chinese medicine, 36 (01), 107-114 (2008); Biological and Pharmaceutical Bulletin, 31 ( 4), 760-764 (2008); Bioscience, biotechnology, and biochemistry, 77 (7), 1424-1429 (2013)).

Meanwhile, conventional techniques using garlic include food additives containing garlic fermented extract (Patent Publication No. 10-2018-0002046), cosmetics containing garlic fermented extract (Patent Publication No. 2018-0001765), garlic Disclosed is a composition for immunopotentiation comprising a fermentation extract (Patent Publication No. 10-2016-0060443) and the like.

In addition, the conventional technology using the gondre as a method for producing a gondry fermentation for inhibiting adipocyte differentiation through bioconversion (Publication Patent Publication No. 10-2017-0079448), a composition for enhancing blood sugar using a gondre extract as an active ingredient Patent Publication No. 10-2017-0058075), Bread for protecting brain nerve cells using Gondre extract (Patent Publication No. 10-2015-0092400) and the like are disclosed.

However, since the gondre excellent in antioxidant activity and immune activity is limited in the amount that can be collected or cultivated and supplied, development of a new antioxidant and immune enhancing composition through blending of garlic and gondre is urgently required.

The present invention has been made to solve the problems of the prior art as described above, it is an object of the present invention to provide a garlic and Gondre lactic acid bacteria fermentation.

Another object of the present invention is to provide a method for producing the garlic and gondre lactic acid bacteria fermentation.

Still another object of the present invention is to provide a composition having enhanced antioxidant and immune activity by including garlic and gondle lactic acid bacteria fermentation.

In order to achieve the above object, the present invention provides a garlic and gondry lactobacillus fermented product obtained by fermenting a mixture of garlic extract and gondre enzyme treatment extract with lactic acid bacteria.

In addition, the present invention is a garlic comprising the step of inoculating and fermenting lactic acid bacteria in the mixture of (a) garlic extract and gondre enzyme treatment extract, and (b) the mixture of the garlic extract and gondle enzyme treatment extract; Provided is a method of preparing gondre lactic acid bacteria fermentation.

In addition, the present invention provides a composition for antioxidant and immune improvement, including garlic and Gondre lactic acid bacteria fermentation.

According to the present invention, by using a blend of garlic and gondre and fermentation of lactic acid bacteria, it is possible to provide garlic and gondry lactobacillus fermented products having enhanced antioxidant and immune activity.

In addition, according to the present invention has the advantage that the lactic acid bacteria fermented product can be used as a material of health functional food, such as beverages.

Figure 1 shows a process for producing garlic and gondre lactic acid bacteria fermentation according to an embodiment of the present invention.
Figure 2 shows a manufacturing process of the gondre enzyme treatment extract according to an embodiment of the present invention.
Figure 3 shows 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging ability. Different alphabets in FIG. 3 represent statistically significant differences (p <0.05), and the same also applies to the following drawings.
Figure 4 shows the results of measuring Fluorescence recovery after photobleaching (FRAP) activity.
5 shows the results of cytotoxicity evaluation.
Figure 6 shows the DPPH radical scavenging ability of Gondre garlic fermentation.
Figure 7 shows the FRAP activity of Gondre garlic fermentation.
Figure 8 shows the cytotoxicity results of Gondre garlic fermentation.
Figure 9 shows the NO production ability of Gondre garlic fermentation.
Figure 10 shows the TNF-α producing ability of Gondre garlic fermentation.
Figure 11 shows the IL-1β production capacity of Gondre garlic fermentation.
Figure 12 shows the IL-10 production capacity of Gondre garlic fermentation.

The present invention relates to garlic and gondry lactobacillus fermented product, in which a mixture of garlic extract and gondre enzyme treatment extract is fermented with lactic acid bacteria.

In the present invention, the term "extract" has a meaning commonly used as a crude extract in the art, but broadly also includes a fraction additionally fractionating the extract.

That is, the extract includes not only one obtained by using an extraction solvent but also one obtained by additionally applying a purification process thereto. For example, fractions obtained by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity), etc. Fractions obtained through various purification methods are included in the extract.

As the extraction solvent, a polar solvent may be used. Suitable solvents for the polar solvent include purified water and alcohol (preferably methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, and 1-pentane). Allol, 2-butoxyethanol or ethylene glycol), acetic acid, dimethyl-formamide (DMFO) and dimethyl sulfoxide (DMSO), more preferably purified water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (methanol, Ethanol, propanol, butanol, and the like), and a mixed solvent of the lower alcohol and purified water, and most preferably, purified water, methanol, ethanol, and mixtures thereof.

In the present invention, the term "fermented product" refers to a product that has undergone a fermentation process using a lactic acid strain, and includes all solids that have not been removed or removed after fermentation.

In the lactic acid bacteria fermented product of the present invention, the mixture of the garlic extract and gondre enzyme extract is a volume ratio of 7: 3-9: 1 based on the solid content of the garlic extract and the gondle enzyme treated extract, preferably 7.5: 2.5 It may be mixed at a volume ratio of ~ 8.5: 1.5, more preferably 8: 2.

In the lactic acid bacterium fermentation product of the present invention, the enzyme may be biscozyme, but is not limited thereto.

In the lactic acid bacteria fermentation product of the present invention, the lactic acid bacteria are flow Pocono stock mesen teroyi des (Leuconostoc mesenteroides), Lactobacillus Planta room (Lactobacillus plantarum), Lactobacillus ash FIG pillar's (Lactobacillus acidophilus), Lactobacillus ramno suspension (Lactobacillus rhamnosus), Lactobacillus casei (Lactobacillus casei), Lactobacillus para casei (Lactobacillus paracasei), Lactobacillus ruteri (Lactobacillus reuteri), Lactobacillus salivarius (Lactobacillus salivarius), Bifidobacterium ronggeom (Bifidobacterium longum), BP Bifidobacterium breve ), Bifidobacterium lactis and Bifidobacterium bipiderum bifidum ) may be one or more selected from the group consisting of, but is not limited thereto.

In the lactic acid bacteria fermentation product of the present invention, the lactic acid bacteria may be leukonostock mesenteroides, and more preferably may be leukonostock mesenteroides KCTC 13302.

In addition, the present invention is a garlic comprising the step of inoculating and fermenting lactic acid bacteria in the mixture of (a) garlic extract and gondre enzyme treatment extract, and (b) the mixture of the garlic extract and gondle enzyme treatment extract; It relates to a production method of gondre lactic acid bacteria fermentation.

In the production method of the lactic acid bacteria fermentation product of the present invention, as the extraction solvent of the garlic extract and gondre enzyme treatment extract, a polar solvent may be used. Suitable as the polar solvent is purified water, alcohol (preferably methanol, ethanol, Propanol, butanol, normal-propanol, iso-propanol, normal-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol), acetic acid, dimethyl-formamide (DMFO) and dimethyl sulfoxide (DMSO) Preferably, purified water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), a mixed solvent of the lower alcohol and purified water, and most preferably, purified water, methanol , Ethanol and mixtures thereof.

In the method for producing the lactic acid bacteria fermentation product of the present invention, the garlic extract is immersed in 2 to 3 times, preferably 2.5 times, water by weight of garlic, and then heated to 80 to 100 ° C, preferably 85 to 95 ° C. Extraction is preferably performed for 3 to 5 hours, followed by concentration to 8 to 12 brix, preferably 9 to 11 brix.

Garlic extract manufacturing method according to an embodiment of the present invention is as shown in FIG.

In the production method of the lactic acid bacteria fermentation product of the present invention, the gondre enzyme treatment extract is immersed in 5 to 15 times, preferably 8 to 12 times, more preferably 9 to 11 times water by weight of dried gondre , 80-100 ° C., preferably 85-95 ° C., followed by extraction, preferably for 3-5 hours, followed by filtration; And the filtered extract is concentrated to 8-12 briquettes, preferably 9-11 briquettes, followed by enzymes at 40-55 ° C., preferably 45-55 ° C. for 5-8 hours, preferably 5-7 hours. Preferably, it may be prepared, including the step of treating the viscozyme.

The preparation method of gondre enzyme treatment extract according to an embodiment of the present invention is as shown in FIG.

In the production method of the lactic acid bacteria fermentation product of the present invention, the step (a) is based on the solid content of the garlic extract and gondre enzyme treatment extract 7: 3 ~ 9: 1, preferably 7.5: 2.5 ~ 8.5: 1.5 It may be mixed in a volume ratio, more preferably in a volume ratio of 8: 2.

In the method for preparing the lactic acid bacteria fermentation product of the present invention, the lactic acid bacteria is Leuconostoc mesenteroides , Lactobacillus plantarum , Lactobacillus acidophilus , Lactobacillus acidophilus , Lactobacillus rhamnosus (Lactobacillus rhamnosus), Lactobacillus casei (Lactobacillus casei), Lactobacillus para casei (Lactobacillus paracasei), Lactobacillus ruteri (Lactobacillus reuteri), Lactobacillus salivarius (Lactobacillus salivarius), Bifidobacterium ronggeom (Bifidobacterium longum) , Bifidobacterium breb breve ), Bifidobacterium lactis and Bifidobacterium bipiderum bifidum ) may be one or more selected from the group consisting of, but is not limited thereto.

In the method for preparing the lactic acid bacteria fermentation product of the present invention, the lactic acid bacteria may be leukonostock mesenteroides, and more preferably may be leukonostock mesenteroides KCTC 13302.

In the production method of the lactic acid bacteria fermentation product of the present invention, the fermentation may be immersed for 36 to 60 hours, preferably 46 to 50 hours, more preferably 48 hours at 36 ~ 38 ℃, preferably 37 ℃. .

The method for preparing the lactic acid bacteria fermentation product of the present invention may further include sterilizing, filtering and concentrating after the step (b). The sterilization may be preferably performed at 100 ° C. for 1 hour, and the concentration may be concentrated to 25 to 35 brix, preferably 30 brix, but is not limited thereto.

In addition, the present invention relates to an antioxidant and immune-improving composition comprising garlic and gondre lactic acid bacteria fermentation.

In the present invention, the term "antioxidation" means the action of inhibiting oxidation, and the term "immune improvement" means increasing the immune response or activity of the immune system in vivo.

In the present invention, the term "health functional food" (health functional food) means a supplementary food processed in the form of tablets, capsules, powders, granules, liquid, pills, etc. using a raw material having a useful function to the human body.

In the antioxidant and immune improving composition of the present invention, the composition may be a dietary supplement.

In the antioxidant and immune-improving composition of the present invention, the composition may be a beverage, but is not limited thereto.

The health functional food can be prepared by drinking the garlic and gondry lactobacillus fermented product of the present invention in the form of tea, juice and drink, or granulated, encapsulated and powdered.

In addition, it can be prepared in the form of a composition by mixing with the lactic acid bacteria fermented product of the present invention and known substances or active ingredients known to have antioxidant and immune function enhancing effects.

The health functional food of the present invention includes not only the lactic acid bacteria fermented product as an active ingredient, but also components commonly added during food production, and include, for example, proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. do.

Examples of such carbohydrates are monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, erythritol, and the like.

As the flavoring agent, natural flavoring agents (tautin; stevia extracts such as rebaudioside A, glycyrgin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) may be used, but are not limited thereto.

For example, when the health functional food composition of the present invention is a beverage composition citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, apple concentrate, pear concentrate, sugar concentrate, jujube concentrate in addition to the lactic acid bacteria fermented product of the present invention , Licorice concentrate, Schisandra chinensis concentrate, complex golden extract may be further included.

In the antioxidant and immune-improving composition of the present invention, the composition may be a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

In the antioxidant and immune-improving composition of the present invention, the pharmaceutical composition is in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, external preparations and sterile injectable solutions according to a conventional method. It can be used in the formulation, but is not limited thereto.

In the antioxidant and immune improving composition of the present invention, the composition may be a cosmetic composition.

In the antioxidant and immune improving composition of the present invention, the cosmetic composition may be prepared in any formulation commonly prepared in the art, for example, creams, lotions, skins, softening, converging cosmetics Formulations, such as foundation, cleansing, face wash, soap, essence, but are not limited thereto.

Preferred content of the lactic acid bacteria fermentation product in the antioxidant and immune-improving composition of the present invention is 0.01 to 50% by weight, but is not limited thereto.

Hereinafter, the present invention will be described in detail through examples. However, these examples are only for illustrating the present invention in detail, and the scope of the present invention is not limited by these examples.

Example 1 Establishment of Optimum Conditions for Garlic and Gondre Raw Materials

1. Experiment Method

1-1. sample

Garlic ( Allium sativum ) and enzyme-treated gondre were purchased from Erom company Limited, Korea. Leuconostoc mesenteroides ) KCTC 13302 was obtained from the Department of Food and Biotechnology, Kangwon National University. The strain was chosen because in the previous experiments it produced garlic products with the strongest biological activity.

1-2. Chemicals and Reagents

Trichloroacetic acid (TCA), gallic acid and Folin-Ciocalteau's reagents were sourced from Sigma-Aldrich, Inc. (Korea). Dinitrophenyl hydrazine (DNPH) is from ACROS Organics (New Jersey, USA), while hydrogen peroxide, methanol and FeCl ₃ are BDH Chemicals Ltd. Purchased from Poole, England, and thiourea, CuSO ₄ · 5H ₂ O, H ₂ SO ₄ , sodium carbonate, AlCl ₃ , potassium acetate, tris hydrochloric acid buffer, FeSO ₄ , potassium ferricyanide and ferric chloride were analyzed Grade, and water was glass distillation.

1-3. Strain culture

Leukonostock Mesenteroides KCTC 13302 was grown in MRS broth medium at 37 ° C. for 36 hours. The cultures were centrifuged to obtain cell pellets, which were then washed twice with double distilled water. Cells were then diluted with distilled water and stored at 4 ° C. until use.

1-4. Garlic and Gondre  Ready

The garlic was peeled, washed, dried in a laminar flow hood using a blender and ground using a blender. 100 g of garlic was weighed with an electronic balance and mixed with 250 mL of distilled water.

The mixture obtained above was steamed at 100 ° C. for 1 hour and cooled to 50 ° C. 0.5 mL of cellulase was added for 30 minutes, followed by 0.75 mL of protease for 1 hour. 1.25 mL of amylase was added and heated at 70 ° C. for 1 hour.

The enzyme was then deactivated by increasing the temperature at 90 ° C. for 30 minutes and the mixture was concentrated to give 7 brics. The sample was concentrated to 17%, adjusted to 10 brix and then lyophilized.

To prepare the gondres, the gondres originals were extracted with water twice at 90 ° C. and filtered. The sample was then concentrated to 10 brix and treated with 0.5% viscozyme (v / v) at 50 ° C. for 6 hours. The samples were then cooled and lyophilized and stored at minus 20 ° C. until use (see FIG. 2).

The ratio of garlic and gondre was mixed in volume ratios of 90:10, 80:20, 50:50, 100: 0 and 0: 100, respectively, and autoclaved.

1-5. Gondre garlic Fermented product  Produce

Garlic and gondles (volume ratios of 90:10, 80:20, 50:50) were inoculated with 0.1% Leuconostoc mesenteroides KCTC 13302 (v / v) with distilled water. Samples were incubated at 37 ° C. for 48 hours. The sample was then sterilized by heating at 100 ° C. for 1 hour and concentrated to 30 brics. Final pH was 5.4-5.7.

1-6. Samples for Bioactivity Assessment

Raw Gondre (S1), Enzyme Gondre (S2), Raw Garlic (S3), Fermented Garlic (S4), Enzyme Gondle 10% (0.1g) and Fermented Garlic 90% (0.9g) Mixture (S5), Enzyme Gondre 20% (0.2g) and 80% (0.8g) fermented garlic mixture (S6), fermented product of 90% garlic and 10% enzyme treated gondre (S7), fermented product of 80% garlic and 20% enzyme treated gondre (S8), the following biological activities were evaluated for the fermented product (S10) of 50% enzyme treated gondre and 50% fermented garlic (S9), and 50% garlic and 50% fermented gondle.

1-7. Determination of Total Phenolic Content

Total phenolic content was determined according to the method of Singleton, Orthofer, & Lamuela-Raventos, 1999 (Methods in enzymology, Vol. 299, pp. 152-178) with some modifications. A sample (sample: water = 1: 100) was taken from the aqueous extract, filtered, and 100 µl was oxidized in vitro with 2.5 mL of 10% Folin-Ciocalteau's reagent (v / v), followed by addition of 2.0 mL of 7.5% sodium carbonate. Neutralized.

The reaction mixture was incubated at 45 ° C. for 40 minutes and the absorbance was measured at 765 nm using a Biospectrometer (Eppendorf Biospectrometer ^® fluorescence, Eppendorf Korea, Korea). The total phenolic content was then expressed as gallic acid equivalent calculated from the standard curve of gallic acid absorbance.

1-8. Total Flavonoid Content

The total flavonoid content of the crude extract was determined by aluminum chloride colorimetric method (Chang, Yang, Wen, & Chern, 2002, Journal of food and drug analysis, 10 (3)).

In brief, 50 μL of sample (1 mg / mL ethanol) was made up to 1 mL with methanol, mixed with 4 mL of distilled water and then mixed with 0.3 mL of 5% NaNO ₂ solution. 0.3 mL of 10% AlCl ₃ solution was incubated for 5 minutes and then added, and the mixture was left for 6 minutes.

Then, 2 mL of 1 mol / L NaOH solution was added and the final volume of the mixture was brought to 10 mL with distilled water, the mixture was left for 15 minutes and the absorbance was measured at 510 nm.

Total flavonoid content was calculated from the calibration curve and the results are expressed in mg routine equivalents per gram of dry weight.

1-9. Antioxidant activity

1-9-1. 2,2-diphenyl-1- Picrylhydrazyl  Free radicals Scavenging power ( DPPH )

The hydrogen atom or electron donating ability of the sample was measured from bleaching of the purple DPPH methanol solution. The free radical scavenging ability of the extract against DPPH (1,1-diphenyl-2-picrylhydrazyl) free radicals was evaluated by the method of Gyamfi, Yonamine, & Aniya (The Vascular System, 32 (6), 661-667 (1999)). .

The dilution of the extract (1 mL) was mixed with 1 mL of 0.4 mM methanol solution containing DPPH radicals, and the mixture was left in the dark for 30 minutes, and then 516 nm using a Biospectrometer (Eppendorf Biospectrometer ^® fluorescence, Eppendorf Korea). Absorbance was measured at.

The radical scavenging ability (A ₁ ) of the sample was calculated as the percentage of DPPH free radicals inhibited by the sample compared to radical inhibition (A ₀ ) in water, the negative control used. DPPH inhibition was calculated by the following equation.

DPPH elimination effect (% inhibition) = ((A ₀ -A ₁ ) / A ₀ ) × 100

1-9-2. Reducibility

The reducibility of the extract was determined by assessing the ability of the extract to reduce the FeCl ₃ solution (Zhao et al., 2008, Food Chemistry, 107 (1), 296-304). 2.5 mL aliquot extracts were mixed with 2.5 mL 200 mM sodium phosphate buffer (pH 6.6) and 2.5 mL 1% potassium ferricyanide.

The mixture was incubated at 50 ° C. for 20 minutes, then 2.5 mL of 10% trichloroacetic acid was added. The mixture was centrifuged at 45 x g for 10 minutes, 5 mL of the supernatant was mixed with an equal volume of water and then 1 mL of 0.1% ferric chloride was added.

Absorbance was measured at 700 nm with a BioSpectrometer (Eppendorf Biospectrometer ^® fluorescence, Eppendorf Korea, Korea). Ascorbic acid was used as a positive control. The iron reducing antioxidation properties were then calculated.

1-10. Cytotoxicity Assessment

In order to confirm the cytotoxicity of the 10 samples prepared using Gondre and garlic, a cell viability assay (XTT assay) was performed in vitro.

The XTT Assay is a macrophage of 1 × 10 ⁴ Cells and wells were added to 96-well plates and incubated for 3 days with the ethanol extract dmf of the sample. 50 μl of XTT solution was added to each well and 4 hours of incubation followed by ELISA reader (Molecular Devices, CA, USA) was used to determine the absorbance at 450nm.

1-11. NO Generating ability  Measure

For the 10 samples prepared using gondre and garlic, each sample was treated at 50, 100, 200 μg / mL concentration in Raw 264.7 cells in order to see the effect of the samples on NO production by activating macrophages. NO (nitric oxide) production capacity was measured.

2. Experimental Results

2-1. Total Phenolic and Total Flavonoid Compound Analysis

10 kinds of samples prepared by using gondre and garlic, namely raw gondre (S1), enzyme-treated gondre (S2), fresh garlic (S3), fermented garlic (S4), fermented garlic 10% (0.1 g) and fermented garlic 90 % (0.9 g) mixture (S5), 20% (0.2 g) enzyme treated gondre (80 g) (0.8 g) fermented garlic mixture (S6), fermented product (S7) of 90% garlic and 10% enzyme treated gondre, Fermented product (S8) of a mixture of 80% garlic and 20% enzyme and gondre (S9), a mixture of 50% enzyme and 50% fermented garlic (S9) and a mixture of 50% garlic and 50% enzyme and gondre Table 1 shows the results of analyzing the total phenol and the total flavonoid compounds.

As shown in Table 1, the total phenol content was found to be high in the order of S1, S2, S9, S8, S5 for each sample, the total flavonoid content is S2, S9, S5, S6, S8 for each sample Appeared to be in the highest order.

The total phenolic content of the gondre-enzyme-treated raw material and the garlic fermented product was found to be higher than the individual raw materials of the garlic fermented product.

2-2. Antioxidant activity

2-2-1 2,2-diphenyl-1- Picrylhydrazyl ( DPPH Free radical Scavenging power

In order to confirm the radical scavenging ability of each sample, L-ascorbic acid (AsA) was prepared at a concentration of 10, 50, and 100 µg / ml, compared to a standard paper, and the results of comparing DPPH radical scavenging ability are shown in FIG. 3.

As shown in Figure 3, all samples were evaluated as excellent overall antioxidant activity, in particular S1, S2, S9, S6, and S8 samples were shown to have excellent DPPH radical scavenging ability.

2-2-2. Reducibility

FRAP activity was measured to evaluate the antioxidant capacity of each sample, and the results of measuring FRAP activity are shown in FIG. 4.

As shown in Figure 4, S1, S9, S2, S8 was found to have high antioxidant capacity in order. In addition, when the garlic fermentation and gondre were mixed than the garlic fermentation, the antioxidant power was confirmed to be higher, and the higher the gondre content, the higher the antioxidant activity.

Compared with the results of the analysis of total phenolic compounds and flavonoids and the antioxidant activity through DPPH radical scavenging activity and FRAP analysis, the antioxidant power was shown to be higher correlated with the total phenolic compound content than the flavonoid content.

2-3. Cytotoxicity

The cytotoxicity measurement results using the XTT assay are shown in FIG. 5.

As shown in FIG. 5, the 10 samples showed cell viability of 90% or more at 50, 100, and 200 μg / mL concentrations, and were evaluated as having no effect on cytotoxicity.

2-4. NO Generating ability

As a result of the experiments, the concentration of NO production increased with concentration of each sample treatment. In particular, S8 produced NO at concentrations of 5.35, 6.43, and 8.23 μM, indicating that the immune activity was significantly higher than that of other samples.

3. Consideration

As described above, the total phenolic compound and total flavonoid content, DPPH radical scavenging ability and antioxidant activity evaluation by FRAP analysis and macrophage NO production ability as an indicator was evaluated for immunological activity, for each sample of S1 to S10 All samples showed increased antioxidant activity and immunostimulating effect as treatment concentrations increased, especially when the two ingredients were mixed rather than garlic fermented or Gondre enzyme treatment alone.

In the case of gondre, the enzyme treatment was higher than that without the enzyme treatment, and the antioxidative and immunological activity increased significantly as the gondre content increased.

From the results of this study, it was confirmed that the optimum process of garlic and gondre was a mixing process of gondre enzyme treatment and garlic fermented product.

On the other hand, Gondre has a problem that the unit price of the raw material is very expensive, and when considering up to 50% in consideration of the results of the above study (S9), the economy is inferior, and in the case of the sample (S8) in which the Gondry fermented product is mixed at 20% Antioxidant and immune activity was not significantly reduced than S9.

Therefore, considering the physiological activity and economic efficiency of the raw materials, the optimal mixing ratio of the raw materials for the bioconversion of garlic and gondre was determined as the ratio of 80% garlic and 20% gondle.

Example 2 (Invitro Functional Evaluation of Gondre Garlic Ferment)

1. Functional Ingredient Analysis

(One). DPPH radical scavenging activity

In order to evaluate the antioxidant capacity of the gondre garlic fermented raw material (FCG) produced by applying the optimized process of Example 1, the DPPH radical scavenging ability as shown in Example 1 is shown in FIG.

As shown in FIG. 6, DPPH radical scavenging ability of the Gondre garlic fermentation was measured. When the raw materials were treated with cells at 0.1, 0.5, and 1 mg / mL, DPPH radical scavenging ability increased in a concentration-dependent manner.

In addition, when treated at a concentration of 500 μg / mL, it showed DPPH radical scavenging ability corresponding to L-ascorbic acid (AsA) at a concentration of 10 μg / mL.

(2). FRAP analysis

As shown in Example 1, the results of analyzing the FRAP activity of the gondre garlic fermentation are shown in FIG. 7.

As shown in FIG. 7, the FRAP activity was significantly increased when the lot mixture was treated by concentration, and the FRAP higher than the control L-ascorbic acid (AsA) 10 μg / mL when treated with a concentration of 500 μg / mL. Showed activity.

2. In Vitro Immune Activity Evaluation of Gondre Garlic Ferment

(One). Cytotoxicity

In order to confirm the cytotoxicity of the Gondre garlic fermented product (FCG) to which the selected raw material was applied, the results of measuring cytotoxicity using the XTT assay are shown in FIG. 8.

As shown in Figure 8, when treated with Gondre garlic fermentation at a concentration of 50, 100, 200 μg / mL all showed more than 90% cell viability, did not appear to have a significant effect on cytotoxicity.

(2). NO generation ability

In the same manner as in Example 1, the result of treating Gondre garlic fermented products at 50, 100, and 200 μg / mL concentrations is shown in FIG. 9.

As shown in FIG. 9, the Gondre garlic ferment produced NO of 4.83, 4.71, and 5.14 μM, which produced significantly higher NO compared to the non-treated group.

(3). TNF-α producing ability

TNF-α is an inflammatory cytokine secreted from activated macrophages or splenocytes. It enhances the elimination activity of pathogens and tumor cells that invade the human body, and interacts with lymphocytes to activate and grow lymphocytes. Adjust

Therefore, the results of observing the effect on TNF-α production while culturing by adding a test substance to macrophages to evaluate the immune enhancing effect is shown in Figure 10.

As shown in Figure 10, the TNF-α production of the cell culture cultured by treatment with gondre garlic fermentation showed an increase in all, Gondre garlic fermentation activates macrophages and promotes TNF-α production, immunity It was found to be effective in promoting the

(4). Interleukine Generating Ability

IL-1β is a cytokine produced by monocytes and macrophage cells and plays an important role in the cytokine network by activating T cells and enhancing the activity of cytokines such as IL-2 secreted from T cells.

In addition, IL-10 induces proliferation of B cells, increases production of lgM, lgG, and lgA antibodies, and plays a role in immunosuppression and partially promotes lymphocyte growth in vivo. Simultaneously plays the role of.

Therefore, the results of observing the effect on the production of IL-1β and IL-10 by culturing added to macrophages to evaluate the immune enhancing effect of the raw material is shown in Figure 11 and 12.

As shown in Figure 11, when treated with Gondre garlic fermentation by concentration in macrophages, showed a significantly higher IL-1β production capacity than the control.

In addition, as shown in Figure 12, IL-10 production ability was also significantly increased than the non-treated group was confirmed that there is an immune enhancing effect.

Example 3 (Preparation of Health Drink Containing Gondre Garlic Ferment)

The main ingredients are garlic gondre fermented products (garlic: gondre = 80:20) produced through the bioconversion process, and the main ingredients are various plant-derived natural concentrates, including unit price, taste, flavor and image of raw materials. Fruit ingredients such as pear concentrate, apple concentrate, and citron concentrate were selected, and herbal ingredients such as schizandra concentrate, ginger concentrate, donkey concentrate, jujube concentrate, and licorice concentrate were selected.

1. Pure Fermentation Concentration Concept of Gondre Garlic Ferment

By highlighting the image of pure garlic and gondry fermentation concentrate to prepare a recipe as shown in Table 2 using only the raw material for the sensuality (Group A). In Table 2 below, components other than the raw materials are purified water.

2. Fresh and delicious fermented beverage concept of juice type

Recently, according to the trend of young layer, adding the image of stamina given by garlic and the fruit juice and taste preferred by the young layer, a recipe as shown in Table 3 of the energy drink concept of non-carbonated or carbonated beverage type was prepared (B group). In Table 3 below, components other than the raw materials are purified water.

3. Herbal Fermented Beverage Concept

The product of the herbal sensory concept is a highly preferred sensory in various layers, and recently prepared a recipe as shown in Table 4 in which garlic gondry fermented product is mixed with herbal ingredients as the market is rapidly growing in liquid pouch or stick products. C group). In Table 4 below, components other than the raw materials are purified water.

Example 4 (Sensory Evaluation)

1. Evaluation method

Sensory evaluation was conducted on 15 sensory test panel trained for the nine goren garlic fermented beverages prepared in Example 3 by mixing the gondry garlic fermented product and the subsidiary materials in various ratios.

Samples were represented by three digit numbers, and after each sample was sampled, water at room temperature was provided to rinse the mouth. The evaluation was made with seven evaluation items (overall feeling, appearance, aroma, taste, mouth feel, aftertaste, and satisfaction). A seven-point scale (1: disliked, 7: very good) was used.

The analysis of variance (ANOVA) was performed to verify the significant differences between the samples on the sensory characteristics and palatability test results of nine beverages. Duncan's multiple range test was performed according to the results (α = 0.05) . For statistical analysis, SPSS statics 17.0 (SPSS Inc., Chicago, IL, USA) was used.

2. Evaluation result

Nine gondre garlic fermented beverages were divided into three groups according to the concept and the preference analysis and group ranking were confirmed.

The results of the analysis of the overall preference of three beverages of group A, which is the concentration of raw materials, are shown in Table 5 below.

Detailed preference results are shown in Table 6 below, A1 was evaluated as better than A2 and A3 in taste and aftertaste, and it was confirmed that the difference between samples was not apparent in appearance, aroma, and mouthfeel.

The results of the analysis of the comprehensive preference of three kinds of beverages of the Group B, which is the fruit addition concept, are shown in Table 7 below.

Detailed preference results are shown in Table 8 below, but there was no significant difference between the samples in appearance, aroma, taste, and mouth feeling, and in the aftertaste category, B3 was evaluated to have higher preference than B1 and B2.

The results of comprehensive preference analysis of three kinds of herbal group C beverages are shown in Table 9 below. C2 was evaluated higher than the rest of the samples in overall overall preference and overall preference, and there was no significant difference in satisfaction. There was no difference between samples in the satisfaction category, and B3 was higher than B1 and B2 in overall preference.

Detailed preference results are shown in Table 10 below. There was no difference between samples in appearance, aroma, mouthfeel, and aftertaste, and C2 was evaluated to be higher than C1 and C3 in the taste category.

In addition, the satisfaction and the comprehensive preference of A1, B3, and C2, which had high preference for each group, are shown in Table 11 below, and C2 was evaluated to be higher than A1, B3 in the overall preference and overall preference, and there was a significant difference between samples in satisfaction. There was no.

The detailed preference of A1, B3, and C2 is shown in Table 12 below. In taste, mouth feeling, C2 was evaluated to have higher preference than other samples, and there was no significant difference in appearance, aroma, and aftertaste.

3. Conclusion and Discussion

As a result of sensory evaluation of nine Gondre garlic fermented beverages, A1, B3, and C2 were found to have high overall preference and preference in each group, and compared with the preference of three samples (A1, B3, C2). , C2 was found to be the most sensory preference.

Example 5 (Preparation of Healthy Drink Containing Gondre Garlic Ferment)

Garlic and gondry fermented product prepared in 1-5 of Example 1 (garlic: gondry = 80: 20) ......................... 1 g

Vitamin C ........................ 15 g

Vitamin E (powder) ............ 100 g

Iron lactate ............... 19.75 g

Zinc Oxide ............... 3.5 g

Nicotinamide ......... 3.5 g

Vitamin A ............................ 0.2 g

Vitamin B1 ............ 0.25 g

Vitamin B2 ............. 0.3g

Water ................................... 1 L

After mixing the above components according to the conventional healthy beverage production method, and then stirred and heated at 85 ℃ for 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and refrigerated stored in a healthy beverage composition Was prepared.

Although the composition ratio is mixed with a component suitable for a favorite beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and use purpose.

Although described with reference to the preferred embodiment of the present invention as described above, those skilled in the art without departing from the spirit and scope of the present invention described in the claims below It will be understood that various modifications and variations can be made to the invention.

According to the present invention, by using a blend of garlic and gondre and fermentation of lactic acid bacteria, it is possible to provide garlic and gondry lactobacillus fermented products having enhanced antioxidant and immune activity, and the lactic acid bacteria fermented products of the health functional food such as beverages Since there is an advantage that can be utilized, it can be usefully applied to the technical field to which the present invention belongs.

Claims (14)

Garlic and gondre lactobacillus fermentation, in which a mixture of garlic extract and gondre enzyme treatment extract is fermented with lactic acid bacteria. According to claim 1, wherein the garlic extract and garlic gondola fermented product, characterized in that the gondre enzyme treatment extract is mixed in a volume ratio of 7: 3-9: 1 based on the solid content. The garlic and gondre lactobacillus fermentation according to claim 1, wherein the enzyme is biscozyme. According to claim 1, wherein the lactic acid bacteria Leuconostoc mesenteroides ( Luconostoc mesenteroides ), Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus acidophilus , Lactobacillus rhamnosus ( Lactobacillus rhamnosus ) Lactobacillus casei , Lactobacillus paracasei , Lactobacillus reuteri , Lactobacillus salivarius , Bifidobacterium longgum ( Bifidobacterium) longum ), Bifidobacterium breve , Bifidobacterium lactis lactis ) and Bifidobacterium bifidum Garlic and gondry lactobacillus fermentation, characterized in that at least one member selected from the group consisting of. (a). Mixing garlic extract and gondre enzyme treatment extract; And
(b). Lactic acid in the mixture of the mixed garlic extract and gondre enzyme treatment extract
Inoculating bacteria and fermentation, comprising the step of fermenting garlic and gondre lactic acid bacteria. The method of claim 5, wherein the garlic extract, after immersing the garlic in a water ratio of 2-3 times by weight ratio, extracting for 3 to 5 hours by applying a heat of 80 ~ 100 ℃, and then concentrated to 8 to 12 brix Garlic and gondre lactobacillus fermented product manufacturing method characterized in that it comprises a. The method according to claim 5, wherein the gondre enzyme treatment extract, after immersing the dried gondre in water by 5 to 15 times by weight ratio, extracting for 3 to 5 hours by applying 80 ~ 100 ℃ heat and then filtering; And
Concentrating the filtered extract to 8 to 12 Brix, garlic and gondre lactobacillus production method characterized in that it comprises the step of treating biscozyme (viscozyme) for 5-8 hours at 40 ~ 55 ℃ . The method of claim 5, wherein the step (a) is a garlic and gondre lactobacillus fermentation method, characterized in that the garlic extract and the gondre enzyme treatment extract is mixed in a volume ratio of 7: 3 ~ 9: 1 based on the solids content . The method according to claim 5, wherein the lactic acid bacteria are Leuconostoc mesenteroides , Lactobacillus plantarum , Lactobacillus acidophilus , Lactobacillus acidophilus , Lactobacillus rhamnosus Lactobacillus casei , Lactobacillus paracasei , Lactobacillus reuteri , Lactobacillus salivarius , Bifidobacterium longgum ( Bifidobacterium) longum ), Bifidobacterium breve , Bifidobacterium lactis lactis ) and Bifidobacterium bifidum ( Bifidobacterium bifidum ) A method for producing garlic and gondre lactobacillus fermented product, characterized in that at least one selected from the group consisting of. The method of claim 5, wherein the fermentation is a method of producing garlic and gondre lactic acid bacteria fermentation, characterized in that the immersion fermentation for 36 to 60 hours at 36 ~ 38 ℃. The method of claim 5, further comprising the step of sterilizing, filtering and concentrating after the step (b). Antioxidant and immune-improving composition comprising the garlic and gondry lactic acid fermentation of any one of claims 1 to 4. The composition for antioxidant and immune improvement of claim 12, wherein the composition is a dietary supplement. 13. The composition for antioxidant and immune improvement according to claim 12, wherein the composition is a beverage.

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