KR101902104B1 - Food composition for enhancing function of immunity comprising fermented extract from leek by lactic acid bacteria and preparation method thereof - Google Patents

Abstract

The present invention relates to a fermentation broth of fermented Lactobacillus acidus having an antioxidant function and blood flow improving effect. More particularly, the fermentation broth of fermented Lactobacillus lactobacillus prepared by adding dead bacteria to fermented broth of fermented Lactobacillus sp. Can be manufactured.

Description

TECHNICAL FIELD The present invention relates to an immunological activity-enhancing functional composition containing a fermentation broth of leek fermented lactic acid bacteria,

The present invention relates to a leek fermentation broth having an antioxidative activity and blood flow improvement using lactic acid bacteria. More particularly, the present invention relates to a leek fermentation broth having an active ingredient, And a manufacturing method thereof.

It is a perennial herb of lilies. In the spring, a line of thick leaves that are about 30cm long from the small scale stem of the earth are gathered together. A long flower stalk appears from August to September, and a small white flower blooms with a phloem flower. The fruit will bear capsules, and if it is ripe, it will burst itself, and black seed will come out. The stem of the scales should be written on the surface, the image, and the leaves edible. It is distributed in Korea and Japan as origin of China and India.

The beneficial effect of leek is rich in vitamins A, B, C, carotene, and iron, which smoothes blood circulation, strengthens digestive organs, stimulates autonomic nerves, and stimulates energy metabolism.

As a prior art related to the present invention, Korean Patent No. 10-1018765 discloses a pharmacological composition containing an extract of Leek extract as an active ingredient and a method for producing the same, and Korean Patent Publication No. 10-2014-0067242 discloses a fermented extract of lactic acid bacteria And an antidiabetic composition containing the same. However, to date, there has been no known method for producing an immunostimulatory functional composition prepared by adding dead bacteria to a fermentation broth of Lactobacillus lacunae and a preparation method thereof.

Accordingly, an object of the present invention is to provide a food composition containing as an active ingredient a fermentation broth of fermented Lactobacillus acidus having enhanced immunity activity by adding killed microbes to a fermented broth having antioxidative function and blood flow improving effect, and a method for producing the same.

The above object of the present invention is achieved by a method for preparing a fermented beverage comprising the steps of: (a) adjusting the concentration of the leek slurry to pH 6.0, preferably to 10 to 20 Brix, most preferably to 13 Brix; Adding 0 to 5% of yeast extract as a nutrient source to the leek slurry obtained in the above step, correcting it to pH 6.0, and sterilizing at 121 ° C for 15 minutes;

Separately, (b) sterilizing the lactic acid pre-culture medium at 121 占 폚 for 15 minutes; After cooling the medium obtained in the above step, the lactic acid bacteria Lactobacillus plantarum , Lactobacillus brevis , Enterococcus faecium or Pediococcus acidilactici at 37 ° C. for 24 to 32 hours, most preferably for 28 hours to produce lactic acid bacteria pre-culture;

(c) 1 to 5% by weight of the culture solution of lactic acid bacteria obtained in step (b) is inoculated into the sterilized leek slices obtained in step (a), and then fermented at 37 ° C for 68 to 76 hours, ; ≪ / RTI > The fermentation broth of the fermented broccoli fermented broth obtained by the step of obtaining the fermented broth fermented broth by adding 100 to 160 mg / g of fungus Enterococcus faecalis , most preferably 134 mg / g, And evaluating the effect of enhancing the immune activity with the material.

The present invention has an excellent effect in enhancing the immune activity of a composition obtained by fermenting lactic acid bacteria with fermented Lactobacillus fermentation broth which has antioxidative function and blood flow improving effect.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic view showing a process for producing a fermentation broth of submerged lactic acid fermentation broth perch of the present invention. FIG.
2 is a graph showing the total flavonoid content of the submerged lactic acid fermentation broth according to the present invention.
FIG. 3 is a graph showing the total polyphenol content of the submerged lactic acid fermentation broth of the present invention.
4 is a graph showing DPPH activity among the antioxidant activities of fermentation broth of submerged lactic acid fermentation broth per day of the present invention.
FIG. 5 is a graph showing SOD-like activity of the fermentation broth of submerged lactic acid fermentation broth of the present invention.
6 is a graph showing the results of HMG-CoA reductase inhibitory activity of the submerged lactic acid fermentation broth of the present invention.
7 is a graph showing the results of ACE inhibition activity of the submerged lactic acid fermentation broth of the present invention.
Fig. 8 is a graph showing the results of the fermentation of E. faecalis And the results of NO production by adding 134 mg / g of the strain.

The present invention provides a fermented broccoli fermentation broth composition having excellent antioxidative activity and blood flow improving effect by fermenting fermented Lactobacillus japonica with fermentation broth and improving its immunity.

According to the present invention, the lactic acid bacteria which can be used for the fermentation of the leek are preferably Lactobacillus , Enterococcus or The Pediococcus sp. Strain is most preferably selected from the group consisting of Lactobacillus plantarum , Lactobacillus brevis , Enterococcus faecium Or Pediococcus acidilactici Lt; / RTI >

The strain of Enterococcus faecalis is most preferably selected from the group consisting of 100-160 mg / g of insecticidal activity and immunological activity.

Hereinafter, the present invention will be described in detail with reference to Examples, Experimental Examples and Preparation Examples. The following Examples, Experimental Examples and Preparation Examples are for illustrating the present invention, and the scope of the present invention is not limited thereto.

Example 1. Manufacture of pre-culture of lactic acid bacteria

In order to prepare lactic acid bacteria pre-culture solution which can be used for the fermentation of submerged lactic acid fermentation broth per the present invention, lactic acid bacteria pre-culture medium was sterilized at 121 ° C for 15 minutes, and then lactic acid bacteria Lactobacillus plantarum , Lactobacillus brevis , Enterococcus faecium or Pediococcus acidilactici Was inoculated on the medium and cultured with shaking at 37 DEG C and 90 rpm for 28 hours to obtain a lactic acid bacteria pre-culture solution.

Example 2. The present invention relates to a fermentation broth of fermented lactic acid bacteria

The submerged lactic acid fermentation broth for use in the present invention was prepared as follows. First, the pH of the submerged saliva was adjusted to 6.0, and the concentration was adjusted to 10-20 ° Brix. 0 to 5% yeast extract was added to the leek slurry obtained in the above step and autoclaved at 121 ° C for 15 minutes. The fermented broth fermentation broth perch was obtained by cooling the autoclaved leek slurry obtained in the above step and incubating the fermentation broth obtained in Example 1 at 1 to 5% at 37 ° C and 90 rpm for 68 to 76 hours with shaking.

Example 3. The present invention provides a fermentation broth composition

To prepare the present invention encourage lactic acid fermentation liquor composition dead cells of lactic acid bacteria in the fermentation broth per immersion leek total weight obtained in Example 2 Enterococcus faecalis The strain was added in a range of 100 to 160 mg / g to obtain a fermented broth fermentation broth.

Experimental Example 1 Measurement of physiological activity of lactic acid fermentation broth of submerged lactic acid bacteria according to the present invention

To measure the total flavonoid content for the fermentation of the fermentation broth of the submerged lactic acid fermentation broth of the present invention, 1200 μL of distilled water was added to 300 μL of the submerged lactic acid fermentation broth perch of the present invention obtained in Example 2, and then 90 μL of 5% NaNO ₂ was added, After reacting for 5 minutes, adding 10% AlCl ₃ 6H ₂ O and reacting for 6 minutes, 300 μL of 1N NaOH was added and the absorbance was measured at 510 nm using UV-spectrophotometer. Rutin was used as a reference material. As a control group, raw leek was used in place of fermented broth of submerged lactic acid fermentation broth of the present invention.

As shown in FIG. 2, Lactobacillus plantarum , Lactobacillus brevis, Enterococcus faecium or Pediococcus acidilactici Inoculated with the strain All of the fermented broth of submerged lactic acid bacteria of the present invention showed a higher content of total polaronolide than that of green leek.

For measurement of the total polyphenol content, 1 mL of 1N Folin and Ciocalteu's phenol was added to 1 mL of the submerged lactic acid fermentation broth per pullulan of the present invention obtained in Example 2, reacted at room temperature for 3 minutes, and then 1 mL of 10% Na ₂ CO ₃ was added After reacting at room temperature for 1 hour, absorbance was measured at 700 nm. Gallic acid (Sigma co., USA) was used as a reference material. As a control group, raw leek was used in place of fermented broth of submerged lactic acid fermentation broth of the present invention.

As shown in FIG. 3, the total polyphenol content of the fermented broth of submerged lactic acid fermentation broth of the present invention was higher than that of green leek.

Experimental Example 2 Antioxidant Activity of Fermented Lactic Acid Bacteria from Pepper

The scavenging activity of DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity was measured in order to measure the antioxidative activity of the submerged lactic acid fermentation broth. To this end, 1 mL of 200 uM DPPH and 1 mL of the fermentation broth of the present invention obtained in Example 2 were mixed and reacted at room temperature and in a dark place for 30 minutes, and the absorbance was measured at 517 nm. As a control group, raw leek was used in place of fermented broth of submerged lactic acid fermentation broth of the present invention. The DPPH radical scavenging activity (%) was calculated by the following equation (1).

As shown in FIG. 4, the DPPH radical scavenging ability of the fermented broth of submerged lactic acid bacteria of the present invention was higher than that of green leek.

In addition, SOD-like activity was measured to measure the antioxidant activity of the submerged lactic acid fermentation broth of the present invention. For this, 3 μL of tris-HCl buffer (pH 8.5) and 200 μL of 7.2 mM pyrogallol were added to the fermentation broth of fermented lactic acid fermentation broth obtained in Example 2, and the reaction was carried out at 25 ° C. for 10 minutes. And the absorbance was measured at 420 nm. As a control group, raw leek was used in place of fermented broth of submerged lactic acid fermentation broth of the present invention. The difference in absorbance between the addition of the fermented vermiculite solution and the no addition was calculated by the following formula (2).

As shown in FIG. 5, the SOD-like activity of the fermented broth of the present invention was higher than that of the fermented broth.

Experimental Example 3. The inhibitory activity of HMG-CoA reductase in fermentation broth of submerged lactic acid bacteria

The inhibitory activity of HMG = CoA reductase was measured using the HMG-CoA reductase assay kit (Sigma-Aldrich) to confirm the blood flow improvement effect of the submerged lactic acid fermentation broth per pullulan of the present invention. For this, 1 uL of the submerged lactic acid fermentation broth of the present invention obtained in Example 2, 2 uL of HMG-CoA reductase, 4 uL of NADPH and 12 uL of HMG-CoA were added to 182 uL of the assay buffer obtained in Example 2, followed by reaction at 37 ° C for 10 minutes, Absorbance was measured. As a control group, raw leek was used in place of fermented broth of submerged lactic acid fermentation broth of the present invention.

As shown in FIG. 6, the inhibitory activity of HMG-CoA reductase was increased in the fermented broth of the present invention.

Experimental Example 4. ACE inhibitory activity of submerged lactic acid fermentation broth

Angiotensin converting enzyme (ACE) inhibitory activity was measured in order to measure the therapeutic effect of hypertension related diseases such as cardiovascular diseases and cerebrovascular diseases in fermented broth of submerged lactic acid bacteria according to the present invention. To this, 20 uL of substrate buffer and 20 uL of enzyme buffer solution were added to 20 uL of fermented lactic acid fermentation broth obtained from Example 2, and the mixture was incubated at 37 ° C for 1 hour, and 200 uL of indicator working solution was added thereto. Lt; / RTI > The absorbance was then measured at 450 nm. As a control group, raw leek was used in place of fermented broth of submerged lactic acid fermentation broth of the present invention.

As a result, as shown in FIG. 7, the ACE inhibitory activity was increased at a concentration of 15.6 uL / mL compared with that of the live leucocephalic broth of the present invention.

Experimental Example 5 Measuring the Immune Activity of the Fermented Lactic Acid Fermentation Solution of the Invention

To measure the immune activity of the present invention a fermentation broth of lactic acid bacteria L. plantarum leek obtained in Example 2 Inoculated with the strain The fermentation broth of submerged lactic acid bacteria in leek and the Enterococcus fungus obtained in Example 3 faecalis The amount of NO released was determined by using the fermented broth lactic acid fermentation broth composition containing the strain, most preferably 134 mg / g, to confirm immunological activity. RAW 264.7 cells were stimulated with LPS at a concentration of 100 ng / mL and then inoculated with L. plantarum The amount of nitric oxide (NO) released was measured at 0.1 ~ 1mg / mL concentration of lactic acid fermentation broth of leek fermented broth per day and fermented broth of lactic acid fermentation broth supplemented with the above killed bacteria. As a control group, LPS-untreated group was used.

As shown in FIG. 8, at the concentration of 1 mg / mL, the release of NO was 88.5%, compared with 81.7% of the submerged lactic acid fermentation broth of leek, 74.3% of leucine fermented broth composition containing dead bacteria NO was released to enhance the immune activity.

According to the present invention, the edible carrier, excipient or diluent which can be contained in the food composition containing the fermented Lactobacillus fermentation broth as an active ingredient include lactose, dextrose, sucrose, sorbitol, mannitol, starch, , Alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, cellulose, water, sugar syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc , Magnesium stearate and mineral oil are preferable.

In addition to the above components, the composition may further include any one of a commercially available lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, and a preservative.

Hereinafter, a specific composition of a health food composition preparation containing the fermentation broth of L. fermented liquor prepared according to the method of the present invention will be described.

Production Example 1 Manufacture of health supplements

The health supplement foods containing the fermentation broth of Lactobacillus acidus lactifera were prepared by the usual health food production method in the contents shown in Table 1 below.

Food ingredient content Name of material Content (% by weight) Leek fermented lactic acid bacteria 75.37 Vitamin A acetate 0.01 Vitamin E 0.08 Vitamin B1 0.01 Vitamin B2 0.01 Vitamin B6 0.04 Vitamin B12 0.00 Vitamin C 0.75 Biotin 0.00 Nicotinic acid amide 0.13 Folic acid 0.00 Calcium pantothenate 0.04 Ferrous sulfate 0.13 Zinc oxide 0.06 Magnesium carbonate 1.90 Potassium monophosphate 1.13 Dicalcium phosphate 4.15 Potassium citrate 6.78 Calcium carbonate 7.54 Magnesium chloride 1.87 Sum 100.00

Production Example 2 Manufacture of fermented beverage lactic acid bacteria

The beverage containing the fermented liquorice fermented liquor obtained according to Example 3 of the present invention was prepared by the ordinary beverage preparation method in the contents shown in the following Table 2.

Beverage ingredient content Name of material Content (% by weight) Leek fermented lactic acid bacteria 10.0 Liquid fructose (wt%) 12.0 Dextrin (wt%) 0.5 Citric acid (wt%) 0.13 Calcium lactate (wt%) 1.6 Vitamin C (wt%) 0.03 Sodium tetroxide (wt%) 0.02 Gel sagging (wt%) 0.03 Purified water (wt%) Balance Total (wt%) 100.0

As described above, since the lactic acid bacteria are added to the fermented broth, which has an antioxidative function and blood flow improvement effect using the lactic acid bacteria of the present invention, it is an extremely useful invention in the functional health food industry because it has an excellent effect in promoting the immune activity.

Claims (7)

(a) adjusting the submerged saliva to 10-20 [deg.] Brix; Adding yeast extract as a nutrient source to the leek slurry obtained in the above step and sterilizing at 121 ° C for 15 minutes;
Separately, (b) sterilizing the lactic acid pre-culture medium at 121 占 폚 for 15 minutes; Culturing the medium obtained in the above step by shaking after inoculation of the lactic acid bacteria after cooling the lactic acid bacteria;
(c) preparing a fermented broth of fermented lactic acid fermentation broth per spoon by inoculating the fermented broth fermented broth obtained in step (b) into the sterilized fermented broth fermented broth obtained in step (a); And adding the killed microbes to the fermentation broth obtained in the step (a).
Wherein the killed microbes are Enterococcus faecalis and the added amount thereof is in the range of 100 to 160 mg / g.
delete delete A fermented broth of fermented Lactobacillus sp. Cultured according to claim 1. 5. The immunogenic activity-enhancing food composition according to claim 4, wherein the lactic acid fermentation broth is contained. delete The food composition according to claim 5, wherein the food composition is one of a health supplement and a beverage.

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