WO2022045452A1 - Functional coffee composition and capsule coffee comprising same - Google Patents

Abstract

The present invention relates to a functional coffee composition and capsule coffee comprising same and, more specifically, to a coffee composition comprising a coffee bean extract including: an extract from coffee beans germinated with Jewhangsu; and a lactic acid bacterium-fermented Sparassis crispa extract, and having functionalities such as anticancer, anti-inflammatory, antibacterial, antioxidant activities and immunopotentiation, and capsule coffee comprising same.

Description

Functional coffee composition and capsule coffee comprising same

The present invention relates to a functional coffee composition and capsule coffee containing the same, and more specifically, to a coffee bean extract comprising a germinated coffee bean extract germinated using sulfuric water and a lactic acid bacterium fermented blossom mushroom extract, anticancer, anti-inflammatory, It relates to a coffee composition having functions such as antibacterial, antioxidant and immunity enhancement, and capsule coffee comprising the same.

In general, coffee beans are the seeds in the red fruit, and two seeds face each other in the fruit. After threshing, drying and refining the seeds are called coffee beans. These beans have a unique aroma and taste after the roasting process. Coffee contains various components such as water, protein, tannin, and caffeine, which have various physiological effects on the human body. It shows effects such as bronchodilation and gastrointestinal motility. In addition, it acts as a central nervous system stimulant, and about two cups of coffee is said to improve performance, reduce fatigue, and reduce aggression.

The various physiological activities of coffee are known through many studies. Among them, as a result of measuring the content of phenolic compounds in various beverages, the study on antioxidants showed that the phenolic content of coffee was higher than that of red wine. There are reports that it has the effect of preventing harmful activity and that consumption of coffee reduces LDL-cholesterol and MDA (Malondialdehyde) by reducing the sensitivity to oxidation of LDL.

However, if you drink coffee in excess of the daily intake, drink coffee made from oxidized coffee beans, or drink a mixture of instant mixed coffee additives such as sugar and cream, the pure function of coffee is lost and rather health problems are caused. can cause In addition, coffee contains about 1,000 chemical substances other than caffeine, and 20 of them have been confirmed as carcinogens as a result of a mouse experiment, and the World Health Organization (WHO) defines coffee as a level 3 carcinogen.

On the other hand, Sparassis crispa is a mushroom of the basidiomycete Minorus serrata family, and the fruiting body is fleshy and has a wavy edge.

Oyster mushroom contains a large amount of beta-glucan and is known to have anticancer effects (Ohno N, Miura NN, Nakajima M, Yadomae T. 2000. Antitumor 1, 3- β-Glucan from Cultured Fruit Body of Sparassis) crispa. Biol Pharm Bull 23: 866-872). In addition, oyster mushroom is known to be rich in phenolic compounds such as veratric acid as well as beta-glucan compared to other mushrooms (Kim MY, Seguin P, Ahn JK, Kim JJ, Chun SC, Kim EH, Seo). SH, Kang EY, Kim SL, Park YJ, Ro HM, Chung IM. 2008. Phenolic compound concentration and antioxidant activities of edible and medicinal mushrooms from Korea. J Agric Food Chem 56: 7265-7270).

Polysaccharides such as beta-glucan contained in mushrooms are not decomposed by digestive enzymes in the human body, and yield drops when hot water extraction is performed. One of the technologies that can compensate for this is fermentation.

Accordingly, the inventors of the present invention confirm that it is possible to prepare a coffee composition with functions such as anti-cancer, anti-inflammatory, antioxidant and immunity enhancement by including the flower mushroom extract extracted after lactic acid bacteria fermentation of the flower mushroom in the coffee bean extract, and came to the present invention.

[Prior art literature]

[Patent Literature]

Patent Document 1: Korean Patent Publication No. 10-1520332 (registration on May 8, 2015)

Patent Document 2: Korean Patent Publication No. 10-1183775 (Registered on September 11, 2012)

In order to solve the problems of the prior art, the present inventors include oyster mushroom to provide a functional coffee composition having more anti-cancer, anti-inflammatory, antioxidant and immune strengthening functions, but extract for oyster mushroom after lactic acid ferment for the oyster mushroom In addition, when including coffee bean powder and liquid extract germinated using sulfuric water, it is confirmed that the functionality is further enhanced while maintaining the flavor and aroma of coffee, and thus more anti-cancer, anti-inflammatory, antibacterial, and antioxidant And to provide a functional coffee composition with enhanced immunity strengthening function.

The present invention, in order to solve the above-mentioned problems,

In the functional coffee composition comprising a coffee bean extract,

The coffee bean extract includes a germinated coffee bean extract germinated using sulfuric water and a lactic acid bacteria fermented flower mushroom extract,

The composition provides a functional coffee composition comprising one or more of the following properties.

i) anticancer;

ii) anti-inflammatory;

iii) antibacterial;

iv) antioxidant; and

v) strengthening immunity.

In addition, the present invention, the sulfuric water is prepared by washing the mineral sulfur;

pulverizing the prepared mineral sulfur to 0.5 μm or less; and

It provides a functional coffee composition, characterized in that produced by a method for producing sulfuric water comprising the step of mixing the pulverized mineral sulfur with purified water.

In addition, the present invention, the germinated coffee bean extract,

1) preparing a mixture by mixing 99.9% by weight and 99.8% by weight of purified water with respect to 0.1% by weight or 0.2% by weight of pulverized mineral sulfur of 0.3 μm or 0.5 μm, respectively, in a state in which green coffee beans are evenly spread;

2) preparing germinated coffee beans by spraying the prepared mixture with sulfuric water 4 or 5 times a day;

3) washing, drying and roasting the prepared germinated coffee beans and then pulverizing them to prepare coffee beans; and

4) Providing a functional coffee composition, characterized in that it is prepared by a method for producing a germinated coffee bean extract comprising the step of extracting the prepared coffee beans by adding water at a temperature of 90 °C or 150 °C to prepare a coffee bean extract do.

In addition, the present invention, the lactic acid bacteria fermented chrysanthemum mushroom extract,

1) adding sterile distilled water to the oyster mushroom to absorb moisture required for fermentation;

2) performing dry fermentation by adding glucose to the flower mushroom obtained in step 1) and inoculating lactic acid bacteria;

3) washing, drying, roasting and pulverizing the fermented zinnia mushrooms; and

4) Extracting the flower mushroom powder obtained in step 3) with water to obtain a lactic acid bacterium fermented flower mushroom extract It provides a functional coffee composition, characterized in that it is prepared by the method for preparing the extract.

In addition, in the present invention, step 2) is Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus rhamnosus (Lactobacillus rhamnosus), Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus bulgaricus (Lactobacillus bulgaricus), Bacillus coagulans (Bacillus coagulans), and provides a functional coffee composition, characterized in that fermented with one or more lactic acid bacteria selected from the group consisting of Bifidobacterium Longum (Bifidobacterium Longum).

In addition, the present invention, the coffee bean extract, based on 100 parts by weight in total, 1 part by weight to 15 parts by weight of the germinated coffee bean extract and 5 parts by weight to 20 parts by weight of the lactic acid bacterium fermented blossom mushroom extract Functional A coffee composition is provided.

In addition, the present invention provides a method for producing coffee capsules comprising the step of filling a capsule (capsule) with the functional coffee composition, and sealing the capsule so that the internal components do not leak by covering the cap.

In addition, the present invention provides a method for producing coffee capsules, wherein the functional coffee composition is in liquid or powder form.

In addition, the present invention provides a method for producing coffee capsules, characterized in that the sealing is performed by putting an air pressure of 1 bar to 20 bar.

In addition, the present invention provides capsule coffee prepared by the method for producing capsule coffee according to the above.

The functional coffee composition according to the present invention includes a flower mushroom, but contains an extract for the flower mushroom, which is fermented with lactic acid bacteria for the flower mushroom, and additionally contains a germinated coffee bean extract germinated using jesulfite water. It is possible to provide coffee with higher efficacy, such as anti-cancer, anti-inflammatory, antibacterial, antioxidant, and immunity enhancement, while maintaining the flavor and aroma of the coffee. In addition, the functional coffee composition according to the present invention can maintain stability even in processing through high pressure, and thus can be easily provided as coffee capsules.

The accompanying drawings are intended to explain the contents of the present invention in more detail to those skilled in the art, but the technical spirit of the present invention is not limited thereto.

1 is a view showing the results for Experimental Example 3 of the present invention.

2 is a diagram showing another result of Experimental Example 3 of the present invention, and is a diagram showing an increase in tumor volume for murine B16F10 melanoma cell-related cancer compared to a control group and an experimental group.

3 is a diagram for comparing the secretion amount of TNF-α after the addition of β-glucan with respect to the secretion amount of TNF-α after adding the coffee bean extract in Experimental Example 4 of the present invention.

Hereinafter, the functional coffee composition according to the present invention, the production method of capsule coffee and the capsule coffee prepared from the capsule coffee production method will be described in detail, but the functional coffee composition, the production method of the capsule coffee and the production method of the capsule coffee The scope of rights of capsule coffee prepared from is not limited by the following description.

Throughout the specification, when a part "includes" a certain component, it means that other components may be further included, rather than excluding other components, unless otherwise stated.

In addition, as used herein, the term 'coffee bean extract' refers to coffee beans that have been pulverized after grinding with respect to germinated coffee beans obtained through fermentation or germination through the watering process for green coffee beans or coffee beans (coffee beans). and extracts obtained by hot water extraction of coffee beans.

The present invention relates to a coffee composition, and more particularly, the present invention relates to a functional coffee composition.

The functional coffee composition according to the present invention includes a coffee bean extract.

The coffee bean extract includes a germinated coffee bean extract germinated using sulfuric water and a lactic acid bacterium fermented Zinnia mushroom extract.

As used herein, the term 'sulfur water' may mean sulfur water from which toxicity is removed by diluting sulfur with purified water, and may have a similar meaning to 'admiral sulfur water' or 'legal sulfur water'.

Preparing the sulfuric water by washing the mineral sulfur; pulverizing the prepared mineral sulfur to 0.5 μm or less; and mixing the pulverized mineral sulfur with purified water.

In the case of the mineral sulfur, when it is first collected, since it contains a lot of foreign substances, it is possible to minimize the inclusion of foreign substances through the washing process.

After washing the mineral sulfur, a step of pulverizing the washed mineral sulfur into a fine powder of about 0.5 μm or less, about 0.25 μm or less, or about 0.1 μm or less may be performed.

In addition, the germinated coffee bean extract according to the present invention comprises: 1) 99.9 wt% and 99.8 wt% of purified water with respect to 0.1 wt% or 0.2 wt% of ground mineral sulfur of 0.3 μm or 0.5 μm, respectively, in a state where green coffee beans are evenly spread mixing to prepare a mixture; 2) preparing germinated coffee beans by spraying the prepared mixture with sulfuric water 4 or 5 times a day; 3) washing, drying and roasting the prepared germinated coffee beans and then pulverizing them to prepare coffee beans; and 4) extracting the prepared coffee beans by adding water at a temperature of 90°C or 150°C to prepare a coffee bean extract.

The size of 0.3 μm or 0.5 μm means the length of the diameter of one mineral sulfur particle, and the smaller the particle diameter, the easier it is to absorb coffee beans in the watering process to be described later, so more efficient coffee bean germination process can proceed.

In addition, the step of mixing the pulverized mineral sulfur with purified water may be mixed with an amount of purified water at a level necessary for the removal of toxicity of the pulverized mineral sulfur. Although the amount of the purified water is not particularly limited, for example, 99.5 wt% to 99.9 wt% of purified water and 0.1 wt% to 0.5 wt% of pulverized mineral sulfur; 99.75 wt% to 99.9 wt% purified water and 0.1 wt% to 0.25 wt% pulverized mineral sulfur; Or 99.8 wt% to 99.9 wt% of purified water and 0.1 wt% to 0.2 wt% of pulverized mineral sulfur.

When mixed in a mixing ratio of purified water and pulverized mineral sulfur within the above range, the prepared sulfuric water has no toxicity and can maximize the antioxidant and immune function of coffee beans.

The coffee bean extract may be performed through a coffee bean extraction method.

The method of extracting coffee beans may include a first step of spraying the prepared sulfuric water onto green coffee beans for germinating the coffee beans.

The process for the watering is not particularly limited, but for example, may include a process of supplying 4 to 5 times a day for 3 days and performing, and the coffee beans in a somewhat germinated state are 4 times a day It may additionally include a process of supplying for 2 days over 5 times.

The method for extracting coffee beans may include a second step of washing the germinated green coffee beans with purified water and then dehydrating them to obtain germinated coffee beans.

In the case of germinated green coffee beans, foreign substances may be additionally included by the watering process in the first step, so it is preferable to wash them with purified water.

After the washing, a separate dehydration process is required for a roasting process to be described later.

The coffee bean extraction method includes a third step of drying and roasting the obtained germinated coffee beans, followed by pulverization.

In the case of germinated coffee beans that have been removed from moisture to some extent through the dehydration process in the second step, the germinated coffee beans for roasting can be prepared by separately performing a drying process before performing the roasting process.

The drying conditions are not particularly limited, but for example, natural drying at about 70° C. to 80° C. for about 5 to 10 minutes or a drying process by applying heat may be performed.

The dried germinated coffee beans may be processed through a roasting process and a roasting process, and in this case, a roasting machine may be used.

The germinated coffee beans after the roasting can be ground to obtain ground coffee beans through the grinding process.

The coffee bean extraction method may include a fourth step of extracting the ground germinated coffee with hot water at 65 ° C. to 120 ° C., 70 ° C. to 110 ° C. or 80 ° C. to 100 ° C. to obtain a coffee bean extract.

When water having a temperature in the above-mentioned range is used, extraction of more pulverized coffee beans can be carried out easily and efficiently.

The coffee bean extract is not particularly limited, but for example, coffee using desulfurization water, characterized in that at least one selected from the group consisting of a fermentation extract, a germination extract, a ripening extract, a hot water extract, and a sulfur extract of the coffee beans. It may be a bean extract.

The lactic acid bacterium fermented flower mushroom extract according to the present invention comprises the steps of: 1) adding sterilized distilled water to the flower mushroom to absorb moisture required for fermentation; 2) performing dry fermentation by adding glucose to the flower mushroom obtained in step 1) and inoculating lactic acid bacteria; 3) washing, drying, and roasting and pulverizing the fermented Zinnia mushrooms; And 4) extracting the flower mushroom powder obtained in step 3) with water to obtain a lactic acid bacterium fermented flower mushroom extract.

To obtain the lactic acid bacterium fermented flower mushroom extract according to the present invention, first, sterile distilled water is added to the flower mushroom to absorb the moisture required for fermentation.

The flower mushroom of the present invention is applied to dry fermentation with lactic acid bacteria as it is collected. Specifically, after absorbing a certain level of moisture required for the fermentation of oyster mushroom before applying to dry fermentation, 1% to 10% (w/w) glucose, preferably 1, based on the weight of the oyster mushroom the moisture has been absorbed % to 5% (w/w) of glucose is added and lactic acid bacteria are inoculated and fermented.

As a more specific embodiment, in step 1), 10 ml of sterile distilled water is added per 100 g of oyster mushroom and immersed at 10° C. to 60° C. for 0.5 to 5 hours to absorb moisture into the cypress mushroom.

The lactic acid bacteria that can be used for the dry fermentation of the present invention are not particularly limited, but preferably Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus esidophilus ( Lactobacillus acidophilus), Lactobacillus bulgaricus (Lactobacillus bulgaricus), Bacillus coagulans (Bacillus coagulans), any one of Bifidobacterium Longum (Bifidobacterium Longum) can be used, preferably Lactobacillus plantarum can be used.

The present invention is followed by dry fermentation after inoculation of lactic acid bacteria in oyster mushroom. Dry fermentation made in a solid state is advantageous because the present invention uses the extracts of the flower mushrooms, and if the flower mushrooms are pulverized and cultured in liquid, the specific efficacy of the flower mushrooms may disappear.

Dry fermentation according to the present invention is performed by adding lactic acid bacteria, preferably Lactobacillus plantarum, to the collected cauliflower mushrooms 1×10 ⁵ to 1×10 ¹⁰ CFU/g, preferably 1×10 ⁸ to 1×10 ⁹ CFU/g After inoculation at a concentration of g, it can be carried out at 40°C to 60°C for 20 to 24 hours, preferably at 40°C to 50°C for 22 to 24 hours.

After the dry fermentation, the process of roasting and pulverizing is performed after the fermented oyster mushroom is washed and dried. In the present invention, the lactic acid bacteria fermented oyster mushroom is roasted at 100 ° C. to 200 ° C. for 3 to 5 minutes, and then pulverized with a grinder to prepare particles. Through the pulverization, the surface area of the blossom mushroom is increased to increase the contact area with water, thereby making it easy to extract the blossom mushroom.

Thereafter, the step of obtaining a lactic acid bacterium fermented flower mushroom extract is performed by extracting the obtained oyster mushroom powder with water.

The coffee bean extract according to the present invention may include 1 part by weight to 15 parts by weight of the germinated coffee bean extract and 5 parts by weight to 20 parts by weight of the lactic acid bacterium fermented blossom mushroom extract based on 100 parts by weight in total. Alternatively, the coffee bean extract may include 2.5 parts by weight to 12.5 parts by weight of the germinated coffee bean extract and 7.5 parts by weight to 17.5 parts by weight of the lactic acid bacterium fermented blossom mushroom extract based on 100 parts by weight of the total. The coffee bean extract is preferably contained in an amount of 15 to 25 parts by weight, more preferably 20 parts by weight, based on 100 parts by weight, of the mixed extract of the germinated coffee bean extract and the lactic acid bacterium fermented flower mushroom extract. While maintaining flavor and aroma, it is possible to maximize the anti-inflammatory and antioxidant effects of germinated coffee beans and the anti-cancer, anti-inflammatory, antioxidant, and immune-enhancing effects of the lactic acid bacteria-fermented chrysanthemum extract.

The functional coffee composition according to the present invention, as described above, may include one or more of the following properties.

i) anticancer;

ii) anti-inflammatory;

iii) antibacterial;

iv) antioxidant; and

v) strengthening immunity.

The content related to the functionality will be described in detail in Examples to be described later.

The present invention also relates to a method for preparing coffee capsules.

The method for producing coffee capsules may include filling a capsule with the functional coffee composition, and sealing the capsule so that internal components do not leak by covering the capsule with a cap.

In the method for producing coffee capsules according to the present invention, the sealing may be performed by putting an air pressure of 1 bar to 20 bar. Alternatively, the sealing may be performed at an atmospheric pressure of 1 bar to 15 bar, preferably 5 bar to 15 bar. Since the functional coffee composition according to the present invention has high stability even at high pressure, it may be packaged by sealing within the above atmospheric pressure range, so side effects such as bursting or cracking of the capsule while the coffee capsule product is extracted can be prevented.

Although not particularly limited, for example, the functional coffee composition may be used in the capsule coffee manufacturing method in liquid or powder form.

The present invention also relates to capsule coffee prepared by the method for producing capsule coffee described above.

The coffee capsule may mean that the raw material of coffee powder or liquid is included in the vitality container like a capsule made of a commonly known metal, plastic or synthetic resin, and may include a pod form.

The pod refers to the use of one cup of coffee beans in a natural paper non-woven pulp that is harmless to the human body. It refers to a form that can prevent the harmful effects of environmental hormones such as aluminum or plastic components that are not known. As the type of the pod, it can be divided into "hard pod" and "soft pod" based on the degree of tamping, and the "hard pod" hardens the ground coffee beans so that espresso can be extracted. It is extruded and wrapped in natural pulp, and the "soft pod" has the same content as the aforementioned 'hard pod', but it is not tamped hard and is soft, so it can be used to extract drip coffee.

In the case of the coffee capsules and pods, by including the functional coffee composition according to the present invention, the extract for the oyster mushroom, which was fermented with lactic acid bacteria for the oyster mushroom, and additionally germinated using sulfuric water By including the germinated coffee bean extract, it is possible to provide coffee with higher efficacy, such as anti-cancer, anti-inflammatory, antioxidant and immune enhancement, while maintaining the original flavor or aroma of coffee.

Hereinafter, the present invention will be described in more detail through specific experimental examples. However, these experimental examples are for illustrative purposes only and the scope of the present invention is not limited to these experimental examples.

[Preparation Example 1] Preparation of germinated coffee bean extract

Washing, drying, and roasting germinated coffee beans that were germinated by spraying green coffee beans evenly distributed with 0.5 μm of pulverized mineral sulfur and 0.2 wt% of purified water and 99.8 wt% of purified water 4 to 5 times a day. Then, pulverized germinated coffee beans were obtained. Thereafter, the ground germinated coffee beans were extracted by adding water at a temperature of 100° C. to obtain a germinated coffee bean extract.

[Preparation Example 2] Preparation of lactic acid bacteria fermented oyster mushroom extract

In this Preparation Example 2, the Lactobacillus plantarum KCTC3108 (hereinafter, L. plantarum) strain received from the Gene Bank (Korean Collection for Type Cultures, KCTC, Jeongeup, Korea) of the Korea Research Institute of Bioscience and Biotechnology was used.

For the initial culture of L. plantarum, MRS medium (Difco, Detroit, MI, USA) was used first. L. plantarum was inoculated into 5 ml of MRS medium and cultured for 24 hours in an incubator (SIF500, Jeiotech, Inc., Daejeon, Korea), washed with sterile physiological saline (NaCl 8.5 g/L), and 5 ml of the same solution diluted in Then, 2 ml of the dilution was inoculated into 200 ml of the edible medium, and then enriched and cultured for 24 hours at 37° C. and 100 rpm. The culture medium was centrifuged at 4℃, 3500rpm for 10 minutes (Combi 514R, Hanil scientific, Inc., Gimpo, Korea) to remove the medium, and then freeze-dried (FDV-7006, Operon-80, Gimpo, Korea) at -70℃ kept.

After that, using the oyster mushroom directly collected by the applicant, 30 ml of sterile distilled water was added to 300 g of oyster mushroom and immersed for 4 hours at 40° C. in order to absorb the moisture required for fermentation. Then, 2%, 5%, and 10% (w/w) of glucose were added based on the weight of the flower mushroom, respectively, and L. plantarum powder was inoculated at the same concentration of 1×10 ⁹ CFU/g at 40°C. Fermented for 24 hours. After the fermentation was completed, the oyster mushroom was washed twice with sterile distilled water, and then dried at 50° C. for 24 hours using an oven (OF-02G, Jeiotech, Inc., Daejeon, Korea).

Then, after drying, the dried oyster mushroom was put back into the oven and left for about 3 minutes from 220°C until the internal temperature dropped to 100°C, and then roasted for about 3 minutes while raising the temperature again to 200°C. Extraction of the roasted oyster mushroom was crushed to a size of 2 mm, and 250 g of the pulverized oyster mushroom was extracted using 2L of purified water for 10 hours. After the extraction was completed, the sample was refrigerated at -4°C and used to prepare the following coffee bean extract.

[Preparation Example 3] Preparation of coffee bean extract

The coffee bean extract was finally prepared by mixing the germinated coffee bean extract obtained in Preparation Example 1 and the lactic acid bacterium fermented flower mushroom extract at the mixing ratio as shown in Table 1 below.

Category (unit: g)	Example					comparative example
	One	2	3	4	5	One	2	3	4	5
Germinated coffee bean extract	2.5	5	7.5	10	12.5	0.1	-	15.5	16	23
Lactobacillus Fermented Blossom Mushroom Extract	17.5	15	12.5	10	7.5	21	23	3	3.5	-

[Experimental Example 1] Measurement of DPPH radical scavenging activity

In order to confirm whether the functional coffee composition according to the present invention has antioxidant efficacy, one of the antioxidant activity measurement experiments, DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity measurement method was used. The DPPH radical scavenging activity measurement method is a method of measuring antioxidant activity by using the degree of decolorization of purple due to the reduction by aromatic compounds and aromatic amines due to the electron donating ability of antioxidants as an index.

More specifically, each of the coffee bean extracts prepared in Examples 1 to 5 and Comparative Examples 1 to 5 was mixed with ethanol to prepare 2 ml of each concentration, 1 ml of a 200 μM DPPH solution was added, and then the reaction solution was completely mixed. After vortexing for 10 seconds, the reaction was carried out at room temperature for 30 minutes. In order to measure the amount of DPPH remaining, absorbance was measured at 517 nm using a UV-vis spectrophotometer (852A Diode Array Spectrophotometer, Hewlett Packard).

The measurement results are shown in Table 2 below.

division	Free radical scavenging ability by concentration (%)
Concentration (μM)	5	10	20	30	40	50
Example 1	25	43	61	81	93	98
Example 2	26	42	58	82	91	97
Example 3	24	44	55	80	90	96
Example 4	22	44	59	84	89	97
Example 5	26	41	64	85	89	95
Comparative Example 1	16	31	40	61	68	78
Comparative Example 2	14	33	40	63	67	81
Comparative Example 3	13	30	41	60	69	80
Comparative Example 4	15	33	44	63	69	84
Comparative Example 5	11	30	41	60	64	80

In the case of the functional coffee composition containing the coffee bean extract according to the present invention, the antioxidant activity showed a tendency to increase as the concentration increased in all experimental groups, and it was confirmed that the highest concentration of 50.0 μM exhibited a high DPPH scavenging ability of 95% or more. could

[Experimental Example 2] Measurement of nitric oxide (NO) production using macrophages

In order to determine whether the functional coffee composition according to the present invention has anti-inflammatory effects, the amount of nitric oxide (NO) production in macrophages was measured. NOS (nitric oxide synthase) in macrophages is influenced by various cytokines such as TNF-γ and TNF-α or bacterial endotoxins such as LPS (E. coli-derived lipopolysaccharide). The ability to generate NO was confirmed by administering the sample and LPS alone or together.

The cell line used for measurement of NO production was mouse-derived J774.1 macrophages, and the cells were cultured in 10% FBS and RPMI 1640 medium, put into a 24-well plate at a concentration of 4.5 × 10⁴cells/well, and then in Example Each of the two groups was divided into 1 to 5 and the coffee bean extracts of Comparative Examples 1 to 5 (sample, indicated as “Sam”), and all were cultured in an incubator at 37° C., 5% CO ₂ for 48 hours, followed by LPS (lipipilysaccharide) was treated at a concentration of 200 ng/well and the cells were cultured for 48 hours. Take 50 μg of the supernatant, add the same volume of Griess reagent (1% sulfanilamide/0.1% N-(1-naphthyl)-ethylenediamine dihtdrochloride/2.5% h3po4), and react at room temperature for 10 minutes. Then, using a microplate reader, absorbance at 540 nm was measured. Sodium nitrite was used as a standard material for nitrite, and the concentration was calculated by comparing with a standard curve obtained by diluting twice with RPMI 1640 medium from 32 μM to 0.25 μM.

The measurement and calculation results are shown in Table 3 below.

division	Nitric Oxide(㎛)
LPS	control	LPS	LPS+Sam	Sam
Example 1	3	9.5	6.5	3.1
Example 2			6.2	3.2
Example 3			6.7	3.3
Example 4			6.6	3.3
Example 5			6.6	3.2
Comparative Example 1			8.1	3.8
Comparative Example 2			8.3	3.9
Comparative Example 3			8.2	3.8
Comparative Example 4			8.6	3.7
Comparative Example 5			8.4	3.8

In the experimental group in which each of the functional coffee compositions containing the coffee bean extract according to the present invention was treated alone, the amount of NO production did not show a significant difference compared to the control group, but the functional coffee compositions containing the coffee bean extract according to the present invention were treated with LPS and In the experimental group treated together, it was confirmed that the NO production amount was lower than that of 9.5 μM treated with LPS alone.

[Experimental Example 3] Measurement of cancer cell proliferation inhibition

In order to measure the cancer cell proliferation inhibition of the functional coffee composition containing the coffee bean extract according to the present invention, MTT assay (methylthiazolyldiphenyl tetrazolium bromide assay) was performed.

Specifically, the colon cancer cell line HCT116 was inoculated at a concentration of 1 × 10 ⁵ cells/well, and the colorectal cancer cell line HCT116 was inoculated in DMEM/F-12 medium supplemented with 10% fatal bovine serum (FBS) in 96-well. After adding 100 μl of each to 96-well at a concentration of 1 × 10 ⁵ cells/well, culturing the cells for 24 hours, 5% CO ₂ , and culturing in an incubator at 37° C. for 24 hours, the above execution Samples dissolved in DMSO with 100 mg/ml of coffee bean extract prepared in Examples 1 to 5 and Comparative Examples 1 to 5 were diluted 1:1000 in DMEM/F-12 medium, and finally 100 μg/ml was injected into the cells. After additional incubation for 24 hours after addition, 50 μl of MTT (1 mg/ml) solution dissolved in phosphate buffered saline (PBS) was added to each well and incubated again for 2 hours. After that, the supernatant was removed so that the formazan formed on the bottom of the well was not dispersed, and 100 μl of dimethyl sulfoxide (DMSO) was added and dissolved slowly, followed by a UV/Visible spectrophotometer (UV/Visible spectrophotometer; The survival rate of HCT116 was measured by measuring absorbance at 540 nm using Human Cop., Xma-3000PC, Seoul, Korea).

The measurement results are shown in FIG. 1 .

As can be seen in FIG. 1 , it was confirmed that the functional coffee composition comprising the coffee bean extract according to the present invention has an effect in reducing the survival rate of colorectal cancer cells (HCT116), that is, cancer cell proliferation yesterday.

Additionally, murine B16F10 melanoma cells, which are known to cause cancer in vivo, were injected into the abdominal cavity of C57BL/6J mice to induce cancer. The tumor volume (the volume of the tumor) was measured for 21 days, and is shown in FIG. 2 . In this case, the coffee bean extract of Example 3 was fed, and the coffee bean extract of Comparative Example 5 was used as a control.

As can be seen in FIG. 2 , in the case of the mouse (control group) that ate the coffee bean extract of Comparative Example 5, the tumor volume increased rapidly from 15 days after injection of B16F10 cells. In the case of mice (experimental group) that ate the coffee bean extract of Example 3, it was found that the tumor volume did not significantly increase.

[Experimental Example 4] TNF-α assay

TNF-α is known as a representative cytokine secreted by macrophages for the defense of infected microorganisms. Therefore, in order to confirm the immunity-related efficacy of the functional coffee composition comprising the coffee bean extract according to the present invention, TNF-α analysis was performed using the TNF-α kit of Invitrogen (Carlsbad, CA, USA). Raw 264.7 cells were seeded in 96 well plates at 1 × 10 ⁴ cells/well, and then cultured for 24 hours. After treatment with a concentration of 1280㎍ / ㎖ and cultured for 24 hours, the supernatant was taken. 10 μl of the supernatant was transferred to a 96 well plate treated with TNF-α antibody-coated, 20 μl of standard diluents buffer and biotinylated antiTNF-α solution were added, respectively, and incubated at room temperature for 90 minutes. After incubation, after washing 4 times using wash buffer, 100 μl of steptavidin-HRP was added and reacted at room temperature for 30 minutes. After washing 4 times, 100 μl of stabilized chromogen was added to each well, and then reacted at room temperature for 30 minutes after blocking light.

After adding 100 μl of stop solution to each well, the amount of TNF-α secretion after adding the coffee bean extract of Example 1 is shown in FIG. 2 to compare with the amount of TNF-α secretion after adding β-glucan.

After adding 100 μl of a stop solution to each well, each of Examples 1 to 5 and Comparative Examples 1 to 5 was measured at 450 nm using an ELISA-reader (Bio-Rad Laboratories Inc.) within 30 minutes.

The measurement results are shown in Table 4 below.

division	absorbance (%)
Concentration (μg/ml)	Control	10	20	40	80	100	200
Example 1	80	68	75	80	82	88	94
Example 2		67	74	81	84	86	95
Example 3		66	76	82	85	89	97
Example 4		66	72	83	86	88	95
Example 5		60	75	81	85	87	96
Comparative Example 1		51	61	68	70	71	80
Comparative Example 2		55	66	67	73	74	82
Comparative Example 3		53	62	70	74	77	83
Comparative Example 4		51	61	71	71	76	84
Comparative Example 5		49	60	67	70	71	81

As can be seen in Table 4 above, when the functional coffee composition comprising the coffee bean extract according to the present invention is added, the secretion of TNF-α is significantly increased from the low concentration, indicating that the production of TNF-α is increased. By activating Raw 264.7 cells, which are macrophages, it is considered that there is sufficient immune-enhancing effect.

[Experimental Example 5] Sensory evaluation

The functional coffee composition comprising the coffee bean extract prepared in Examples 1 to 5 and Comparative Examples 1 to 5 was diluted 1:20 with hot water to perform a sensory evaluation test.

The sensory evaluation test was conducted on 20 ordinary people, and in particular, the upper, middle and lower grades were divided according to the preference according to the bitter taste.

The results of the sensory evaluation test are shown in Table 5.

division	Sensory evaluation results
Example 1	○○
Example 2	○
Example 3	○○
Example 4	○○
Example 5	○
Comparative Example 1	△
Comparative Example 2	×
Comparative Example 3	××
Comparative Example 4	△
Comparative Example 5	×
○○: When the preference “high” is 15 or more ○: When the preference “high” is 10 or more △: When the preference “high” is 5 or more, and the preference “medium” is 10 or more ×: Preference “low” In the case of 10 or more people ××: When the preference “lower” is 15 or more people

The above description of the present application is for illustration, and those of ordinary skill in the art to which the present application pertains will understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present application. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. For example, each component described as a single type may be implemented in a dispersed form, and likewise components described as distributed may also be implemented in a combined form.

The scope of the present application is indicated by the following claims rather than the above detailed description, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included in the scope of the present application.

Claims (10)

1. In the functional coffee composition comprising a coffee bean extract,

   The coffee bean extract includes a germinated coffee bean extract germinated using sulfuric water and a lactic acid bacterium fermented flower mushroom extract,

   The composition comprises a functional coffee composition comprising one or more of the following properties;

   i) anticancer;

   ii) anti-inflammatory;

   iii) antibacterial;

   iv) antioxidant; and

   v) strengthening immunity.
2. The method of claim 1, wherein the sulfurization water is prepared by washing mineral sulfur;

   pulverizing the prepared mineral sulfur to 0.5 μm or less; and

   Functional coffee composition, characterized in that it is prepared by a method for producing sulfuric water comprising the step of mixing the pulverized mineral sulfur with purified water.
3. According to claim 1, wherein the germinated coffee bean extract,

   1) preparing a mixture by mixing 99.9 wt% and 99.8 wt% of purified water with respect to 0.1 wt% or 0.2 wt% of ground mineral sulfur of 0.3 μm or 0.5 μm, respectively, in a state in which green coffee beans are evenly spread;

   2) preparing germinated coffee beans by sprinkling the prepared mixture with sulfuric water 4 or 5 times a day;

   3) washing, drying and roasting the prepared germinated coffee beans and then grinding them to prepare coffee beans; and

   4) A functional coffee composition, characterized in that it is prepared by a method for producing a germinated coffee bean extract comprising the step of extracting the prepared coffee beans by adding water at a temperature of 90°C or 150°C to prepare a coffee bean extract.
4. The method according to claim 1, wherein the lactic acid bacteria fermented chrysanthemum mushroom extract,

   1) adding sterile distilled water to the cypress mushroom to absorb moisture required for fermentation;

   2) performing dry fermentation by adding glucose to the flower mushroom obtained in step 1) and inoculating lactic acid bacteria;

   3) washing, drying, roasting and pulverizing the fermented Zinnia mushrooms; and

   4) Functional coffee composition, characterized in that it is prepared by a method for producing a lactic acid bacteria fermented cherry blossom mushroom extract comprising the step of extracting the flower mushroom powder obtained in step 3) with water to obtain a lactic acid bacteria fermented flower mushroom extract.
5. According to claim 4, wherein step 2) is Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus rhamnosus (Lactobacillus rhamnosus), Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus bulgaricus (Lactobacillus), Bacillus coagulans (Bacillus coagulans), and Bifidobacterium Longum (Bifidobacterium Longum) Functional coffee composition, characterized in that fermented with one or more lactic acid bacteria selected from the group consisting of.
6. The functional according to claim 1, wherein the coffee bean extract comprises 1 to 15 parts by weight of the germinated coffee bean extract and 5 to 20 parts by weight of the lactic acid bacterium fermented blossom mushroom extract based on 100 parts by weight of the total. coffee composition.
7. A method for producing coffee capsules, comprising the step of filling a capsule with the functional coffee composition according to any one of claims 1 to 6, and sealing the capsule so that the internal components do not leak by covering the capsule.
8. The method according to claim 7, wherein the functional coffee composition is in liquid or powder form.
9. The method of claim 7, wherein the sealing is performed by putting an air pressure of 1 bar to 20 bar.
10. [Claim 10] Coffee capsules prepared by the method for producing coffee capsules according to any one of claims 7 to 9.

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