KR20160072978A - A composite of chamomile fermented by microorganisms for functional foods with anti-oxidative effects - Google Patents

Abstract

The present invention relates to a composition having an antioxidative effect using chamomile. More specifically, the present invention relates to a composition having an antioxidative effect using chamomile. More specifically, the present invention relates to a composition having an antioxidant effect by using chamomile, And to a composition for health functional food which helps prevent various diseases caused by the disease.

Description

Technical Field [0001] The present invention relates to a composition for functional food having antioxidative effect containing fermented chomomile by microorganism,

The present invention relates to a composition for health functional food having a higher antioxidative effect by fermenting chomomile with a microorganism.

In modern society, wastes such as peroxides (peroxides) accumulate in the human body due to changes in diet, lack of exercise, and excessive mental stress in relation to a more complicated society, and accumulated peroxides accelerate aging and cause inflammation of the human body It causes immunity and cancer.

In other words, peroxides occur easily in the lipids present in the human body, especially in unsaturated fatty acids, and are promoted by various factors. These factors may be derived automatically from food, such as oxygen, heating, ultraviolet light, mechanical shock, or the presence of metal ions, but they may be produced by lipid oxidation enzymes such as lipooxygenase in the human body. These substances are very unstable and reactive, attacking and damaging intracellular DNA, attacking lipid-active cell membranes, causing chain fatty oxidation, promoting tissue aging, causing further cancer, or causing inflammation do.

Recently, studies on antioxidation have been actively conducted recently. In particular, polyphenols as natural materials are stabilized as resonance structure by absorbing free radicals into the phenol ring molecule itself, and free radical formation Which is known to inhibit various chain-oxidations. Polyphenols exist in a wide variety depending on the plant species, and a typical example thereof is flavonoid. Flavonoids are C ₆ -C ₃ -C ₆ , the main skeleton of which is present in nature as a glycoside linked to one or more sugars. Flavonoids are found mainly in fruit or flower parts of plants, and they also determine the color of fruits and flowers. Flavonol, flavanone, flavone, isoflavone, flavonols, fnthocyanidin, and flavonol are classified according to the structure according to the structure. . In addition to its strong antioxidant capacity, it has been studied that it has antimicrobial action, prevention of dementia, anti-inflammation and anti-cancer activity. It is known that the functionality is slightly different according to the small group of flavonoid (cited as Miles, EA, Zoubouli, P Calder, PC, Phil, D. (2005) Differential anti-inflammatory effects of phenolic compounds from extra virgin olive oil identified in human whole blood cultures. Nutrition 21 , 389-394).

For example, a cosmetic composition for anti-inflammation and antioxidation of green tea and pine mushroom extract has been filed (Korean Patent No. 10-2011-0045662), Kim et al. Have studied the antioxidative and anti-inflammatory activities of the fractions (2010) Kor J. Microbiol. Biotechnol . 38 , 434-442). Especially, polyphenols, especially flavonoids, among various antioxidants are known to have the strongest oxidizing power, and many studies have been conducted recently due to additional physiological functions.

However, flavonoids in natural products require various processing methods, such as extraction, concentration, or biotransformation through fermentation, because the amount of component is less than the effective concentration for the actual functionality. That is, most of the active ingredients present in natural products are present as glycosides or combined with other components. Therefore, in order to increase the extraction yield, enzymatic extraction with microorganisms or various solvents according to polarity are used. In practice, however, only ethanol is allowed in the use of solvents in food manufacturing, limiting the selection of solvents. However, in the case of the fermentation method, it is possible to separate and extract only the active ingredient from the glycoside efficiently by using the enzyme produced by the microorganism, and further physiological functioning can be expected by further progressing to the biotransformation. For example, a patent has been filed for further enhancing functionality through fermentation using Aspergillus fungi (Korean Patent No. 10-2014-0040611, Korean Patent No. 10-2014-0014025).

The present invention is characterized by a composition having an antioxidant ability superior to that of conventional ones by fermenting Chamomile (scientific name Matricaria recutica ).

Among the herbs, chamomile is a grass belonging to the dicotyledonous plant, and sometimes called camomile, chamomile, or chamomile. The country of origin is in England, but now it is found all over the world. It is 50 cm ~ 1 m in height, and features a lot of side branches on the straight stem. Flowers bloom from May to September and have an apple flavor. The aroma of chamomile is effective in alleviating stress, relieving pain such as headaches, and warming the body. It is currently being used as a medicinal vehicle for the treatment of gastrointestinal dysfunction and for the treatment of inflammatory diseases, fever, diarrhea, small bowel and liver tumors.

Flowers contain terpenes, flavonoids, coumarins, malicacids and minerals. Essential oils contain 28 kinds of terpenes (α-bisabolol, chamazulene and bisabolol oxide), 36 flavonoids (apigenin etc.) It is reported that about 120 kinds of compounds such as organic acids are contained. Among them, apigenin is known to have antioxidant effect. However, the concentration of the active substance in natural products is very small to exhibit functionalities in the human body, and the extraction rate is also very small in industry.

On the other hand, fermentation methods have been extensively studied for the purpose of increasing the extraction rate of various natural products and increasing physiological function through bioconversion. That is, a method for extracting enzymes from grains and plants using microorganisms, and a patent for manufacturing (Patent No. 10-2013-0018140) have been filed by the inventor. Joe et al. Have proposed ginsenoside ginsenosides as mold after the asbestos by biotransformation using the homogenizer's Aspergillus (Aspergillus niger) it reported that a greater and their physiological functional (citations: Jo, M., Jung, JE , Lee, JH, Park, SH Yoon, HJ Yoon, , Lim, KT, Paik, HD (2014) Cytotoxicity of the white ginseng extract and red ginseng extract treated with partially purified -glucosidase from Aspergillus usamii KCTC 6954. Food Sci. Biotechnol. 23 , 215-219). In addition, Kim et al. Studied the antioxidant activity of ginseng flowers using Bacillus subtilis. (Antioxidant activity of ginseng flower according to fermented microorganisms) (Kim, Kyunghee, Kim, Journal of the Korean Society of Food Science and Nutrition 42 , 5, 663-669).

Therefore, many chamomile flower parts having such active ingredients are considered to be harmless to the human body, that is, Bacillus polyfermenticus , KCCM10104, Lactobacillus plantarum , KCCM11613P and Lactobacillus acidilactici < RTI ID = 0.0 > After fermentation using KCCM11614P, it is confirmed that the antioxidant activity is improved as a function.

The fermented chamomile, which is a composition according to the present invention, is ingested by consumers in the form of a tablet, or a mixture of a mineral, a tea, or a mixture with an inert ingredient for synergistic effect on the antioxidant ability, thereby suppressing oxidation in the body, It is possible to obtain a higher effect than conventional natural chamomile in preventing or treating aging, cancer, inflammation and cardiovascular diseases derived from free radicals.

Fig. 1 shows (a) before and after fermentation of chamomile. Fig.

Example 1 Fermentation of Chamomile Samples

The strains of this experiment were Bacillus polyfermenticus ( Bacillus polyfermenticus ) KCCM10104 and Lactobacillus plantarum KCCM11613P and Pediococcus acidilactici KCCM11614P, which are representative lactic acid bacteria, were used, and cultivation was carried out by liquid culture. That is, in a 250 mL Erlenmeyer flask, 100 mL of trypticase soy broth (TSB) medium for Bacillus polyfermenticum, 250 mL of MRS medium for lactic acid bacteria such as Lactobacillus plantarum and Pediococcus acyldactyse And then cultured for 7 hours. To this culture, 20 g of chamomile, 500 mL of sterile distilled water, 2.5 g of peptone (total 0.5%, v / v) and 10 g of glucose (total 2% v / v) were added to a 1 L Erlenmeyer flask Lt; 0 > C, 1.5 atm and cooled. 10 mL (2%, v / v) of the previously cultivated strain was inoculated and fermented at 30 ° C for 72 hours, but in the case of bacillus 150 rpm, and lactobacillus was shaken at 50 rpm, respectively. After fermentation, the cells were sterilized at 121 ° C and 1.5 atm and lyophilized for 1 week. Fig. 1 shows the state before and after fermentation of the chamomile.

Example 2: Extraction of fermented chamomile

The lyophilized sample and 20 g of the control group were mixed with 200 mL (10-fold) of 70% ethanol and extracted at 50 ° C. for 24 hours. The extract was filtered with a filter paper (Watman No. 1) and concentrated using a rotary vacuum concentrator (EYELA, Tokyo Rikakikai Co., Japan) to obtain an ethanol extract. The concentrate was lyophilized and powdered and tested for its functionality (antioxidant effect) by dissolving it in 70% ethanol at an appropriate concentration.

Test Example 1: Total polyphenol content of chamomile

The total polyphenol content of the 70% ethanol extract as an effective substance of chamomile was measured colorimetrically using the Folin-Denis method. That is, 0.1 mL of sample and 2 mL of 2% aqueous sodium carbonate solution were mixed. After incubation at room temperature for 3 minutes, the mixture was mixed with 0.1 mL of 50% Folin-Ciocalteau reagent (Sigma Chemical Co., St. Louis, Mo., USA) and allowed to stand for 30 minutes. Absorbance was measured at 750 nm. The total polyphenol contents (unit: mg of gallic acid equivalent, mg GAE) were calculated using a standard curve of Gallic acid (0, 12.5, 25, 50, 100 ㎍ /

As a result, the total polyphenol content of the control group was 19.36 mg GAE / g, and the total polyphenol content of the chamomile fermented with the strains Bacillus polyfermenticus KCCM10104, Lactobacillus plantarum KCCM11613P, and Pediococcus acidiflutici KCCM11614P was 21.47 mg GAE / g , 24.23 mg GAE / g and 20.55 mg GAE / g, respectively. The fermented chamomile samples showed 1.10, 1.25, and 1.06 times higher than the control, respectively. Table 1 shows the yield and total polyphenol content of chamomile extract after fermentation with various microorganisms. The total yield was calculated as follows.

Total yield (%) = [sample weight after extraction (g) / dry weight (g) of sample before extraction)] × 100

Control group B. polyfermenticus KCCM10104 L. plantarum KCCM11613P P. acidilactici
KCCM11614P Dry weight (g) 20 20 20 20 Weight of extract (g) 9.26 10.40 11.2 9.6 Total yield (%) 46.3 52 58 48 Total phenol content
(mg GAE / g) 19.36 ± 0.09 21.47 + 0.34 24.23 + - 2.77 20.55 + 1.01

Test Example 2: Antioxidant effect of chamomile extract

1) DPPH free radical scavenging ability

As an antioxidant assay, the scavenging activity of free radicals was measured as follows. 0.2 mL of the sample and control was mixed with 1 mL of 100 μM DPPH (1,1-diphenyl-2-picrylhydrazyl), allowed to stand at room temperature for 15 minutes, and the optical density (OD) was measured at 517 nm. The antioxidative effect of DPPH free radical scavenging was analyzed by observing the color change of DPPH solution by removing DPPH free radical. The DPPH free radical scavenging activity was calculated as follows.

DPPH free radical scavenging ability (%) = [1- (OD of sample / OD of control)] × 100

The antioxidative effects of DPPH free radicals of the chamomile extracts fermented with each strain were confirmed, and the effect was confirmed to be higher than that of the control group. Table 2 shows the antioxidative effect of the fermented chamomile extract as a result.

Concentration (mg / mL) Control group B. polyfermenticus KCCM10104 L. plantarum KCCM11613P P.acidilactici
KCCM11614P 0.125 8.22 + 0.78 13.06 + 2.06 9.8 ± 1.04 12.19 ± 0.71 0.25 12.13 ± 1.19 18.79 + - 4.26 16.52 ± 3.28 17.86 + - 0.38 0.5 13.07 ± 2.91 30.40 ± 0.83 21.55 7.83 30.88 ± 3.81 1.0 34.38 ± 0.65 46.20 ± 2.34 42.31 + - 2.27 45.25 ± 1.99 2.0 44.47 ± 1.53 55.01 + - 0.96 48.29 ± 0.91 59.66 ± 1.62

2 ) Measurement of antioxidative effect by FRAP assay

Peuraep ahseyi (FRAP assay, ferric reducing antioxidant power ) method is that in principle be reduced at low pH as tripyridyltrizaine ferric (Fe ³⁺ -TPTZ) complex is tripyridyltriazine ferrous (Fe ²⁺ -TPTZ) by a reducing agent, the sample is Fe ³ when reduced to Fe ^{2+ +} to a method to measure the absorbance the Fe ²⁺ that indicates to evaluate the antioxidant activity. In the FRAP assay, DPPH and ABTS (2,2'-azinobis- (3-ethylbenzo-thiazoline-6-sulfonate)) radical scavenging activity assay was used as an experimental method with the assumption that most antioxidants have a reducing power It is also an antioxidant measurement method of other mechanisms.

As a method, FRAP reagent was prepared by heating 25 mL of acetate buffer (300 mM, pH 3.6) at 37 ° C. and adding 10 mM 2,4,6-tris (2-pyridyl) -s-triazine (TPTZ, Sigma Chemical Co.) and 2.5 mL of 20 mM ferric sulfate (FeSO ₄ ). 0.05 mL of the sample dissolved in the desired concentration was added to the prepared 0.95 mL FRAP reagent, reacted at 37 ° C. for 30 minutes, and absorbance was measured at 593 nm by colorimetry. Blank samples were measured by placing acetate buffer or distilled water in the same volume. At this time, the antioxidant activity was calculated as the amount of FeSO ₄ .7H ₂ O (unit: μM) corresponding to the OD value of the sample in the standard curve showing the relationship between the concentration of the standard reagent FeSO ₄ .7H ₂ O and the OD value of FRAP . The greater the measured value, the greater the reduction power.

As a result, the antioxidant effect through FRAP assay was as shown in Table 3. In other words, fermented chamomile showed much higher reduction ability of iron ion than non-bummed chamomile extract. Here, among the fermented products inoculated with the strain The antioxidant potency of Pediococcus acidiflucis KCCM11614P was higher than that of other strains.

Concentration (mg / mL) Control group B. polyfermenticus KCCM10104 L. plantarum
KCCM11613P P. acidilactici
KCCM11614P 0.125 - - - - 0.25 - - - - 0.5 32.13 + - 3.52 67.30 ± 8.45 42.85 + - 5.56 73.33 ± 7.12 1.0 158.0 + - 6.82 238.1 ± 1.51 180.7 ± 5.32 240.70 + - 4.33 2.0 364.8 ± 1.26 436.6 + - 7.03 374.1 ± 1.90 454.48 ± 1.90

According to the present invention, application examples of a functional food composition to which a composition containing fermented chamomile can be applied are as follows.

The composition of the present invention having an antioxidative effect can be used for the purpose of preventing or treating inflammation, aging, cancer and cardiovascular diseases which may occur due to lipid oxidation. Preferably, the concentration of the fermented chamomile extract is in the range of 2.0 mg / mL or more relative to the total composition and circulated.

Claims (1)

Composition for health functional food having antioxidative effect comprising chamomile fermented by microorganism and extract thereof as an active ingredient

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