KR101768817B1 - Food composition for anti-oxidation and inhanced immunity comprising chamomile flower extract short-term fermented by lactic acid bacteria and the method of preparation thereof - Google Patents

Abstract

The present invention relates to a method for manufacturing functional natural material composition having enhanced antioxidant and immune activity due to lactobacillus fermentation for water extract of a Matricaria chamomilla flower, in which a nutrient source is added to the water extract of the Matricaria chamomilla flower and a lactobacillus strain of Enterococcus sp. as a high-temperature short-term fermentation strain is injected and fermented, thereby having an excellent effect of providing a lactobacillus fermented liquor of the water extract of the Matricaria chamomilla flower having improved antioxidant and immune activity, and functional food for antioxidation and immunity improvement using the same.

Description

TECHNICAL FIELD The present invention relates to an antioxidant and immunosuppressive functional food composition containing a lactic acid fermentation broth of Chamomile flower and a method for preparing the same,

The present invention relates to an antioxidant and immunosuppressive functional food composition containing an edible fermented Chamomile extract. More particularly, the present invention relates to a food composition comprising a single raw material chamomile by adding distilled water to obtain a water extract, Which is obtained by inoculating and fermenting a lactic acid bacterium having an amino acid sequence of SEQ ID NO:

Chamomile (Matricaria recutita L.) belongs to Asteraceae, and is widely used as a herbal tea. For many years, chamomile has been used to relieve intestinal gas, headache, insomnia, back pain, neuralgia, rheumatism, skin diseases, indigestion and gout diseases. Recent studies have shown that chamomile essential oils are highly effective in antimicrobial and antioxidant (Natarajan et al., Phytotherapy Research, 27 (1) 118-125, 2013). Chamomile essential oils contain a variety of phytochemicals including flavonoids, some of which exhibit anti-inflammatory and antiallergic activity. Chamomile essential oils have long been studied in many fields, including the cosmetics industry, including pharmaceuticals and perfumes.

Fermentation of foodstuffs by microorganisms degrades the polymeric materials into low molecular weight substances to increase the digestion and absorption rate, to increase the palatability due to the generation of new flavor, and to produce physiologically active components. Recently, fermented foods have been actively studied.

Lactic acid bacteria are beneficial to ferment saccharides to obtain energy, to produce a large amount of lactic acid, and to prevent spoilage without producing more than 50% of the consumed sugar as a metabolic product. Lactic acid bacteria are Gram-positive bacteria and they are anaerobic or anaerobic microorganisms with no motility. Most of them are catalase-negative and require various vitamins, amino acids and peptides for growth. In general, the intake of lactic acid bacteria induces an increase in beneficial bacteria that help digestion, absorption, decomposition and excretion of food, thereby improving digestion and improving digestive system diseases (Rolfe, 2000, J Nutr, 130: 396 -402). In addition, it has nutrition and health promoting effect by synthesis of useful substance through fermentation of lactic acid bacteria, and has been reported to have nutritional effect such as effect of dressing, immunity enhancement, improvement of liver cirrhosis, anticancer effect, and skin care effect.

As a prior art related to the present invention, essential oil derived from chamomile leaf, especially Korean Patent No. 10-1465972, relates to a skin regeneration composition obtained by vacuum distillation extraction of a chamomile hydrosol using a chamomile leaf as a raw material, Lt; / RTI > Patent Document 10-2013-0125020 discloses an antioxidant-functional cosmetic composition containing chamomile extract of Chamomile leaf using ethanol and supercritical fluid CO ₂ not using an organic solvent.

However, there is no known technology for improving the antioxidant activity and immune activity by inoculating fermented Enterococcus sp. Lactobacillus strains having high-temperature fermentation characteristics for a short period of time to the water extract of chamomile flowers.

Accordingly, an object of the present invention is to provide a lactic acid fermentation broth of a water extract of Chamomile flower having remarkably enhanced antioxidant and immunopotentiating function by inoculating Enterococcus lactic acid bacteria having high-temperature and short-term high-temperature fermentation characteristics to a water extract of a single raw material Chamomile flower .

Another object of the present invention is to provide an antioxidant and immunosuppressive functional health food composition containing, as an active ingredient, a fermentation broth of a lactic acid bacterium having short-term high-temperature fermentation characteristics of the water extract of the chamomile flower.

The above object of the present invention can be achieved by a method for producing a chamois flower, comprising the steps of: collecting, washing, washing, drying and pulverizing chamomile flowers and soaking in distilled water corresponding to 15 times their weight; Extracting water at 30 to 90 DEG C for 1 to 3 hours and most preferably at 60 DEG C for 2 hours in an extraction condition having high antioxidant activity; Generally Recognized As Safe Lactic Acid Enterococcus with High Temperature and Short Term Fermentation Characteristics of Water Extract of Chamomile Flower faecium KCTC 13225, adding 0 ~ 5% of glucose as a nutrient source and 0 ~ 5% of yeast extract and culturing at 37 ° C for 30 ~ 48 hours to provide a fermentation broth of lactic acid bacteria .

The lactic acid fermentation broth of the Chamomile flower water extract of the present invention is not only excellent in antioxidant and immunity enhancing effects but also has no side effects on the human body and has an effect of being used as a material for health functional foods and does not contain an unpleasant organic acid due to high temperature short term fermentation There is an excellent effect of providing a highly functional fermentation broth.

1 is a graph showing the cytotoxicity of water extracts of Chamomile flowers and water extracts of fermented Chamomile flowers of lactic acid bacteria according to the present invention.
FIG. 3 is a graph showing the results of measuring the total polyphenol content of antioxidant activity of lactic acid fermentation broth for the water extract of Chamomile flowers prepared according to the present invention.
4 is a graph showing the amount of NO released from a fermentation broth of lactic acid bacteria in the water extract of Chamomile flowers according to the present invention.

The present invention provides a lactic acid fermentation broth for a water extract of Chamomile flower prepared by inoculating a water extract of Chamomile flower extracted from a Chamomile flower and fermenting it, and an antioxidative and immunostimulatory functional food composition containing the extract as an active ingredient.

Preferred lactic acid which can be used according to the invention Enterococcus (Enterococcus) genus, and most preferably from Enterococcus faecium KCTC 13225.

In addition, the method of producing the fermented lactic acid bacteria composition for the water extract of chamomile flowers according to the present invention can be used by (a) in the case of chamomile which is a raw material for fermentation, by blasting flowers or dried chamomile flowers. (b) the water extract of chamomile flowers comprises the steps of: adding 15 times distilled water to the chamomile flower powder; At 50 to 70 占 폚 for 1 to 3 hours, most preferably at 60 占 폚 for 2 hours. (c) Inoculating 1% (w / w) of lactic acid bacteria pre-cultured in the water extract of chamomile flowers obtained in the above step and adding 0 to 5% of glucose and 0 to 5% of yeast extract as a nutrient source. (d) adding the Enterococcus mixture to the mixture after steps a) and b) 24-72 hours was inoculated faecium (KCTC 13225) strain at 35 ~ 40 ℃, and most preferably comprises a step of fermentation at 37 ℃ for 40 to 48 hours.

The present invention also includes a step of evaluating the antioxidative and immunological activities of the fermentation broth in which the antioxidative activity and immune activity are enhanced by measuring the DPPH radical scavenging activity, the total phenol content and the NO release amount of the lactic acid fermentation broth of the chamomile flower.

The present invention also provides a result of measuring the number of lactic acid bacteria contained in a fermentation broth of a lactic acid bacterium in which a fermentation broth fermented with lactic acid bacterium has an antioxidative effect and an immune effect.

Hereinafter, the present invention will be described in detail with reference to examples. The following examples are intended to illustrate the invention and are not to be construed as limiting the scope of the invention.

Example 1. Preparation of water extract of Chamomile flower of the present invention

Although the chamomile flowers used in the present invention are used by pulverizing dried chamomile flowers, the flowers can be used. The water extract of the chamomile flowers was extracted with distilled water corresponding to 15 times (w / w) the weight of the dried chamomile flower powder at 60 ° C for 1 to 3 hours. The antioxidant activity and total polyphenol contents were the highest when extracted at 60 ℃ for 2 hours. After extraction, the water extract of Chamomile flower was used as a material for the filtrate recovered through vacuum filtration.

Example 2 Preparation of Lactic Acid Fermentation Liquid of Water Extract of Chamomile Flower

Enterococcus faecium KCTC 13225, which was cultivated separately, was inoculated in 10 ⁶ cfu / mL of the water extract of the test material Chamomile flower obtained in Example 1, followed by addition of 0 to 5% of yeast extract, 0 to 5% of glucose and 0 to 5% And most preferably fermented at 37 DEG C for 48 hours at a high temperature for a short time.

Example 3. Cell culture

RAW 264.7 cells were cultured in RPMI 1640 medium (Lonza Biowhittaker, Walkersville, MD, USA) supplemented with 10% fetal bovine serum (FBS), 100 units / mL penocillin and 100 ug / mL streptomycin And cultured in an incubator (5% CO2 / 95% air). Cells were washed with phosphate-buffered saline (PBS, pH 7.4) when cells were about 80% full, and cell culture medium was added to remove cells. Subsequently, cells were subcultured and the medium was changed every 2 days.

Experimental Example 1. Cytotoxicity measurement

The cytotoxicity of the cells obtained in Example 3 of the present invention when each of the cells obtained in Examples 1 and 2 was added was measured. For this purpose, RAW 264.7 cells were plated in 24-well plates at 5 × 10 ⁴ cells / well and incubated for 24 hours. The cultured cells were cultured in a cell culture medium containing 0, 10, 100, and 1,000 ug / mL of a fermented composition of a lactic acid bacterium of water extract of chamomile and chamomile flowers. Cells were cultured for 48 hours, and the number of viable cells was dissolved in isopropanol by MTT assay (Seanot F and Lang R. J Immunological Method 89: 271-277, 1985), and the absorbance was measured at a wavelength of 570 nm.

As a result, as shown in Table 1 and FIG. 1, it was confirmed that the cytotoxicity of the lactic acid bacteria fermented in Example 2 was lower than that of the water extract of Chamomile flower of Example 1.

Raw 264.7 cells Of chamomile flowers Water extract 0 g / mL 10 g / mL 100 g / mL 1,000 g / mL Viable cell numbers
(Absorbance at 570 nm) 2.000 0.091 1.555 0.036 ^** 1.331 0.040 ^*** 0.313 0.022 ^*** Raw 264.7 cells Of chamomile flowers Water extract  Fermented lactic acid bacteria composition 0 g / mL 10 g / mL 100 g / mL 1,000 g / mL Viable cell numbers
(Absorbance at 570 nm) 1.848 0.091 1.586 0.033 ^* 1.571 0.034 ^* 1.644 0.025

^{Note) * p <0.05, ** p} <0.01, *** p <0.001 significantly different

Experimental Example 2. Antioxidant activity by DPPH scavenging activity

DPPH (α, α-diphenyl-β-picrylhydrazyl) exists as a radical of the stabilized structure of nitrogen center in the compound. Antioxidant activity can be measured according to the degree of scavenging of the stabilized radical. In the present invention, the DPPH radical scavenging activity was measured by Biosis et al. (1958). 5 mL of DPPH was added to 0.5 mL of each of the samples obtained in Example 1 and Example 2, incubated at room temperature and dark for 30 minutes, and the absorbance at 517 nm was measured and the inhibition rate was calculated by the following equation (1). Vitamin C was used as a positive control.

As a result, the DPPH scavenging activity of the lactic acid bacteria fermented in Example 2 was increased and the antioxidative activity of DPPH scavenging activity was increased in comparison with the water extract sample of Chamomile flower of Example 1.

The lactic acid fermentation broth of Chamomile flower water extract prepared according to the present invention was fermented in a medium supplemented with 5% glucose and 5% yeast extract, and the antioxidative activity was increased by 30% or more by fermentation with lactic acid bacteria having high temperature and short term fermentation characteristics. The range of addition is not limited to the above 5%. Vitamin C was used as a positive control.

Experimental Example 3: Antioxidant activity by total polyphenol content

Total polyphenol content was determined by the Folin-Denis method (Zhou et al., 1980). 1 mL of each Folin-Ciocalteu's reagent was added to 1 mL of each of the samples of Example 1 and Example 2, and the mixture was allowed to stand for 3 minutes. 1 mL of 10% Na 2 CO 3 solution was added to the mixture and allowed to stand at room temperature for 1 hour. The polyphenol content was quantified.

As a result, as shown in FIG. 3, the total polyphenol content was increased in the samples of the lactobacillus fermentation according to Example 2 as compared with the samples of the chamomile flower water extract of Example 1.

The lactic acid fermentation broth of the Chamomile flower water extract prepared according to the present invention showed an increase in antioxidative activity of 65% or more by fermentation of lactic acid bacteria having high temperature and short term fermentation characteristics of the present invention in a medium supplemented with 5% glucose and 5% yeast extract, The range of addition is not necessarily limited to 5%.

Experimental Example 4. Measurement of NO emission

The amounts of NO released when the respective samples obtained in Examples 1 and 2 were added to the cells obtained in Example 3 of the present invention were measured. For this purpose, RAW 264.7 cells were cultured at a concentration of 1 x 10 ⁵ cells / mL, stimulated with LPS at a concentration of 100 ng / mL, and then the fermented composition of the lactic acid bacteria of the water extract of Chamomile flowers and the water extract of the chamomile flowers was added at a concentration of 1 mg / After treatment with 100 uL of griess reagent at a concentration of 0.04 g / mL, the reaction was allowed to proceed for 20 minutes, and the NO emission was measured at 540 nm absorbance. The NO emission amount was calculated by the following formula (2).

As a result, as shown in FIG. 4, compared to the water extract of the chamomile flowers of Example 1 on the basis of 100% of the LPS 100 ng / mL treated group, the NO emission of the lactic acid bacteria fermented according to Example 2 decreased, The activity was remarkably increased.

Production Example 1 Production of beverages containing lactobacillus fermented liquid of chamomile flower water extract

The beverage containing the lactobacillus fermentation broth of the chamomile flower water extract obtained according to Example 2 of the present invention was prepared as shown in Table 2 below.

Name of material content Lactic acid fermentation broth (wt%) of water extract of chamomile flower 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

The sensory evaluation was performed on each of 5 male and female randomly selected beverages according to the above Preparation Example using a panel. The control was a water extract of chamomile flowers. The sensory test was performed using a 7-point scaling method. The experimental results are shown in Table 3 below.

Experimental results showed that fermented Lactobacillus fermented by Enterococcus spp. Was the best in color, taste and overall acceptability.

division color flavor Comprehensive preference map Control 4.6 4.7 5.0 Experimental Section 6.0 6.3 6.3

As described above, the present invention provides a fermentation broth having an antioxidant activity and an improved immune activity by fermenting lactic acid bacteria having a high-temperature short-term fermentation characteristic after adding a nutrient and obtaining a water extract from the chamomile flower, It is a very useful invention in the health functional food industry because it has an excellent effect to provide.

Claims (2)

By the addition of distilled water for chamomile flowers powder in chamomile water extract extracted for 2 hours at 60 ℃ around the cultured Enterococcus faecium (KCTC 13225) inoculated a strain with a nutrient source, and prepared by fermentation 48 hours HTST at 37 ℃ chamomile flower water extract Lactic acid fermentation broth
An immunosuppressive food composition comprising the lactic acid fermentation broth of claim 1

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