KR20190001416A

KR20190001416A - A cosmetic composition comprising fermented coffee grounds

Info

Publication number: KR20190001416A
Application number: KR1020170081369A
Authority: KR
Inventors: 김미경; 류재덕
Original assignee: 주식회사 에이치앤비나인
Priority date: 2017-06-27
Filing date: 2017-06-27
Publication date: 2019-01-04

Abstract

The present invention relates to a novel functional cosmetic material utilizing a coffee bean as a natural material having a relatively large content of functional ingredients having easy raw material securing and physiological activity. The present invention is characterized in that the coffee bean is fermented with Bacillus subtilis or a complex microorganism including Bacillus subtilis, lactic acid bacteria, yeast bacteria and acetic acid bacterium, thereby significantly improving functionality such as antioxidation, whitening, and wrinkles. Further, the present invention relates to an extract of the coffee ground fermented product.

Description

A cosmetic composition comprising fermented coffee grounds,

The present invention relates to a cosmetic composition comprising a coffee fermented product.

Coffee is dried, roasted and processed into powder, which is harvested from coffee trees. Coffee contains various components such as caffeine, tannin, saturated fatty acids including palmitic acid and stearic acid, unsaturated fatty acids such as oleic acid, linolenic acid, organic acids and saccharides. The coffee grounds are called coffee grounds, and most have been treated as organic wastes. Despite the presence of various components in the coffee beans, they have been disposed of as simple wastes, but there have been various studies for recycling coffee beans. For example, coffee beans are used to make compost. It has been reported to produce biofuels or feed additives. In addition, there are examples of making coffee beans from various product materials such as tables using as a wood substitute material. Korean Patent Application No. 10-2015-0029239 describes a soil improvement agent using coffee beans and a method for producing the same, Korean Patent Application No. 10-2012-0103668 describes a low absorptive fuel powder using coffee beans and a method for producing the same And Korean Patent Application No. 10-2014-0107754 describes an invention of an antibacterial microbial fermentation agent for compost production using coffee beans and an antibacterial fermentation compost produced thereby. In addition, Korean Patent Application No. 10-2013-0069257 discloses a method for producing a composition for improving immunity by fermenting coffee beans with microorganisms.

On the other hand, the demand for highly functional natural materials is rapidly increasing in the cosmetics market, which is growing rapidly all over the world. In particular, in order to improve and prevent skin damage caused by various factors such as environmental pollution and stress, it is necessary to move away from the existing chemical synthetic material and to provide a safer and more functional material.

Accordingly, the present inventors have studied to develop a novel functional cosmetic material utilizing a coffee bean having a relatively high content of ingredients and a high content of functional ingredients as a natural material. As a result of analyzing various functions required as a cosmetic material after fermentation of a coffee bean into a specific microorganism, it has been confirmed that the coffee bean can be developed as a cosmetic material having superior functionality as compared with conventional chemical synthetic materials, Completed.

Korean Patent Application No. 10-2012-0103668 Korean Patent Application No. 10-2014-0107754 Korean Patent Application No. 10-2013-0069257

It is an object of the present invention to provide a fermented coffee bean fermented by a microorganism.

It is also an object of the present invention to provide a cosmetic composition comprising a coffee ground fermented product.

In addition, the present invention provides a method for fermenting coffee grounds having antioxidant, anti-inflammatory, whitening, skin moisturizing and skin wrinkle inhibitory effects.

In order to achieve the above object, the present invention provides a cosmetic composition comprising a coffee ground fermented product obtained by inoculating coffee grounds with a strain of Bacillus subtilis (KCTC 1027).

The present invention is lactic acid (in addition to the Bacillus subtilis Weissella coffee foil cibaria , KFCC11598P), Saccharomyces cerevisiae (ATCC9763) and Acetobacter Pasteurianus , ATCC 9432) is further inoculated and fermented to provide a cosmetic composition comprising the coffee fermented product. The cosmetic composition comprising the coffee bean fermented product provided in the present invention provides antioxidative, anti-inflammatory, whitening, skin moisturizing and skin wrinkle inhibiting effects.

(I) drying the coffee bean; And (ii) a, antioxidant, anti-inflammatory, bleaching, having a skin moisturizing and skin wrinkle formation inhibitory effects, coffee foil fermented by comprising inoculating Bacillus subtilis (Bacillus subtilis, KCTC 1027) to the dried coffee foil and fermentation The method comprising:

The present invention also relates to a method for the production of antioxidant, antiinflammatory, anti-inflammatory, anti-inflammatory and anti-inflammatory agents, which further comprises inoculating and fermenting lactic acid bacteria (Weissella cibaria and KFCC11598P), Saccharomyces cerevisiae (ATCC9763) and Acetobacter pasteurianus (ATCC 9432) , A whitening agent, a skin moisturizing agent and a skin wrinkle inhibiting effect.

The present invention also relates to a process for producing a coffee beverage having an antioxidant, anti-inflammatory, whitening, skin moisturizing and skin wrinkle formation inhibiting effect, which further comprises adding perchloric acid to the coffee ground fermented product after the step (ii) A method for producing a fermented product is provided.

The coffee ground fermented product according to the present invention maintains a high polyphenol content, has a collagenase inhibitory activity and a tyrosinase inhibitory activity. It has also been shown that it inhibits melanin formation and also has an anti-inflammatory function.

Fig. 1 shows the polyphenol content of the coffee grounds by Bacillus subtilis compared to the polyphenol content of the coffee grounds and Bacillus subtilis culture itself without Bacillus subtilis treatment.
Fig. 2 shows collagenase inhibitory activity of coffee fermented by Bacillus subtilis in comparison with collagenase inhibitory activity of coffee blot and Bacillus subtilis culture itself without Bacillus subtilis treatment.
Fig. 3 shows the tyrosinase inhibitory activity of coffee fermented by Bacillus subtilis compared to the tyrosinase inhibitory activity of the coffee bean without Bacillus subtilis and the culture of Bacillus subtilis itself.
Fig. 4 schematically shows a process for producing the fermented coffee bean of the present invention and the functional cosmetic comprising the same.

In one example, the present invention provides a comparative analysis of polyphenolic compound content and melanoidin content, including chlorogenic acid, which is a major physiologically active substance contained in coffee bean fermented product, Bacteria and lactobacillus strains were selected. Bacillus subtilis and lactic acid bacteria used in the present invention are Bacillus subtilis (KCTC 1027), Weissella cibaria (KFCC11598P). In the present invention, the fermented coffee bean fermented product comprises a fermentation broth fermented with a fermented coffee bean by inoculating lactic acid bacteria into a hot-air-dried coffee bean and the fermentation residue, and the fermented extract was prepared by extracting them.

In another embodiment, the present invention provides a method for establishing fermentation conditions for coffee beans using the selected Bacillus subtilis and lactic acid bacteria strains, and a method for controlling the fermentation conditions of the fermentation broth and the fermentation residue using physiological conditions (e.g., water, ethanol, An optimal extraction method was established by comparing and analyzing the active substance, for example, the content of polyphenol compounds and the content of melanoidins.

In another embodiment, the present invention relates to an effect of electron donating ability, skin cell regeneration, whitening, antioxidative function and wrinkle reducing function of coffee fermented product (hot air drying or freeze drying) and coffee ground extract (hot air drying or freeze drying) Respectively. The antioxidant function was confirmed by comparing the total amount of polyphenol compounds in the coffee grounds. The results showed that the electron donating effect was 87.34%, the collagen synthesis which is involved in skin cell regeneration and wrinkle improvement was similar to that of arbutin (38.09%), which is a positive control group of melanin synthesis inhibitor, by 36.72%.

Hereinafter, the present invention will be described in more detail with reference to Examples. The following examples are provided to illustrate the present invention and the scope of the present invention is not limited by the following examples.

Manufacturing example : Using Bacillus subtilis and complex microorganisms Coffee shop Preparation of fermentation broth and fermented extract

First, the coffee was obtained from Park Eun (Cheongju Melide Cafe, Cheonan Sunmoon University and Café Bene, Incheon Eiren and One Coffee Academy), and using a hot air dryer (Hanil Premium Dryer, Hanil Jeungco Co., Ltd.) After drying at ~ 60 ℃ for 10 ~ 14 hours, it was used for the next experiment. As a microorganism used in the present invention, Bacillus subtilis (KCTC 1027), which was purchased from the BRC, was used, and lactic acid bacteria ( Weissella cibaria , KFCC11598P), yeast ( Saccharomyces cerevisiae , ATCC9763), acetic acid bacteria ( Acetobacter pasteurianus , ATCC 9432) were purchased from the Korean Society of Bacteriocin and the International Center for Biological Resources. Next, for the fermentation of coffee beans, the dried coffee beans were inoculated at a concentration of 1 x 10 < ⁶ > cuf / g or higher and then fermented at 25 DEG C to 40 DEG C for 5 to 50 hours. For the fermentation by the complex microorganism, 10 to 30 (15%), 10 to 30 (20%), 10 to 30 (20%) and 10 to 30 15%) by weight, and 20 to 100 ppm of water-soluble silicon, which is inoculated at a concentration of 1 x 10 < ⁶ > cuf / Lt; / RTI > The coffee bean was fermented as described above to obtain a fermented product containing the fermentation broth and the fermentation residue, and the fermented product was extracted to prepare a fermented extract.

Example

Example One: Coffee shop (Antioxidant effect)

Most of the biologically active substances present in plants are secondary metabolites. Polyphenols, including flavonoids with two or more phenolic hyroxyl (OH) groups in one molecule, give the plant a special color and aroma , And acts as a substrate in the redox reaction. Polyphenol compounds vary in their properties and physiological activities according to their chemical structure. These polyphenols exhibit various physiological and pharmacological effects such as antioxidation, anti-aging, anti-inflammation and antibacterial activity as well as ultraviolet exposure and antiallergic. Which is a useful physiologically active ingredient.

(DF-100A, DOORI Scientific Co., Gyeonggi, Korea) was used for distilled water and distilled water for each of the coffee beans which were dried and stored according to the above Preparation Example, followed by high-pressure extraction at 121 ° C for 30 minutes . Ultrasonic extraction was performed using a 40 kHz ultrasonic bath (Daihan Science, Korea). The content of polyphenol compounds in the obtained extracts was measured by the Folin-Denis method. That is, 2 mL of the aqueous extract solution was mixed with 2 mL of 1 N Folin-Ciocalteu's phenol reagent, and the mixture was allowed to stand at room temperature for 3 minutes. Then, 2 mL of 10% Na ₂ CO ₃ was added, and the mixture was allowed to stand at room temperature for 1 hour. Absorbance was measured at 690 nm using an ELISA reader (BioTek, Italy). As a result, the polyphenol content of pressurized extract was higher than that of distilled water. Especially, the highest extractable amount of extract was 16.45 ± 0.18 mg / g. In case of extraction with ethanol solvent, extraction progress was impossible due to evaporation of solvent. Total amount of polyphenol was 12.00 ± 0.43 mg / g when sonicated with ethanol solvent. Table 1 below shows the total polyphenol content of the coffee bean according to the extraction solvent and extraction method.

Extraction solvent Extraction method Total polyphenol concentration (mg / g) Distilled water Pressure extraction method 12.58 ± 0.32 Ultrasonic extraction method 11.74 + 0.21 And arithmetic Pressure extraction method 16.45 ± 0.18 Ultrasonic extraction method 14.77 ± 0.22

* Total polyphenol content is in mg / g gallic acid equivalents (GAE)

Next, the total polyphenol content of the coffee bean fermentation broth by Bacillus subtilis according to the above Preparation Example was measured. The fermented coffee bean pulp with Bacillus subtilis showed 46.7% higher content than the control group using only coffee beans (blue in Fig. 1). In addition, it was confirmed that the content of total polyphenols increased by 1.3 times as compared to that of the initial culture and 1.5 times as compared with the control group using only Bacillus subtilis (green in FIG. 1) in the case of coffee bean fermentation using Bacillus subtilis. Therefore, it was found that the total polyphenol concentration was increased in the fermented coffee bean by the Bacillus subtilis, rather than the effect of polyphenol increase by the Bacillus subtilis own production material. The results are shown in Fig.

Example 2: Collagenase ( collagenase ) Inhibitory activity measurement (skin wrinkle improvement)

Collagen, a major constituent of the extracellular matrix, is a major substrate protein produced by dermal fibroblasts. It forms most of the organic substances in skin, bone and teeth and functions as skin firmness, connective tissue binding force, cell adhesion and cell differentiation induction. . Collagen is composed of more than 90% of the dermis layer and gives tension and strength to the skin to protect and maintain the skin against external stimuli. Representative symptoms of aged skin are the occurrence of fine lines and wrinkles, which can be attributed to the marked reduction of collagen, the main protein in the collagen of skin dermal tissue. Decrease due to collagen degradation is caused by destruction of connective tissue that maintains skin elasticity, which causes wrinkles and elasticity decrease and skin sagging. Collagenase inhibitory activity was measured by Wnsch E and Heindrich HG. In other words, the reaction mixture was incubated with a substrate solution (125 mg) containing 0.3 mg / mL of 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-DArg (Sigma, USA) in 0.1 M tris-HCl buffer (pH 7.5) uL and 50 μL of sample solution, 75 μL of collagenase (Sigma, USA) of 0.2 mg / mL was added and incubated at room temperature for 20 minutes. Then, 250 μL of 6% citric acid was added to stop the reaction. mL was added and the absorbance at 320 nm was measured. Collagenase inhibitory activity was expressed as the absorbance reduction ratio of the sample solution and the non - added sample.

The inhibition rate of collagenase was measured for the extracts obtained by varying the extraction solvent in coffee beans. As a result, when collagenase was used as an extraction solvent rather than distilled water, a higher collagenase inhibition rate appeared. In the case of the inhibition rate according to the extraction method, the inhibition rate was higher than that in the pressure extraction method rather than the ultrasonic extraction method. The extraction of ethanol by solvent evaporation was impossible due to the evaporation of solvent. The activity of collagenase inhibition by ethanol was 3.98 ± 0.96%, which was lower than that of hydrocolloid solvent extract. Table 2 below shows the inhibition rate of collagenase according to the extraction solvent and extraction method.

Extraction solvent Extraction method Inhibition rate (%) Distilled water Pressure extraction method 14.6 ± 0.95 Ultrasonic extraction method 1.26 + - 0.87 And arithmetic Pressure extraction method 26.32 ± 0.99 Ultrasonic extraction method 4.66 ± 0.96

* Inhibition ratio against retinol (%)

Next, the inhibition rate of collagenase using the coffee bean fermentation broth by Bacillus subtilis according to the above Preparation Example was confirmed. First, when the inhibition rate of collagenase was confirmed using only the culture of Bacillus subtilis, it was confirmed that the inhibitory effect was 36.6% (see FIG. 2Green). This suggests that Bacillus subtilis culture itself has collagenase inhibitory activity. Inhibition rate of the control group was 36.6% on average, whereas inhibition rate of coffee fermented product by Bacillus was 48.6% when control group was used with only Bacillus subtilis (FIG. 2 green) and only coffee beans (FIG. 2 blue) Which is 1.26 times higher than that of the previous year. The results are shown in Figure 2 in comparison with retinol.

Example 3: tyrosinase ( Tyrosinase ) Inhibition activity measurement (whitening effect)

Excessive production of active oxygen in the skin results in aging with wrinkles, skin sagging, and loss of elasticity, and exposure to ultraviolet rays for a long time is an external factor of aging, which causes pigmentation of spots and freckles. The most well-known pigment in relation to pigmentation is melanin, which is produced by polymerization of amino acids and proteins via derivatives such as dopaquinone, which is produced by the enzymatic action of tyrosine (tyrosine) . Therefore, the whitening effect can be expected by suppressing the melanin-producing enzyme tyrosine. Tyrosine is an enzyme involved in the initial crucial determination of the melanin biosynthesis pathway in the human body. It acts as a DOPA oxidase that oxidizes tyrosine to form L-3,4-dihydroxyphenyl-alanine (DOPA) and DOPA quinone, (5,6-dihydroxyindole) (DHI) is involved in the conversion of indole-5,6-quinone to the synthesis of melanin polymer . Tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein-2 (TRP-2) are also involved in the melanin biosynthesis pathway It is known that tyrosinase is an essential and important evaluation method for skin whitening and anti - aging cosmetics because it acts on various processes. To date, tyrosinase activity inhibitors isolated from natural products have been known to inhibit the activity of formononetin, glabrene, glabridin, glabrol, arbutin, oxyresveratrol, dihydromoriin, artocarbene, 4-prenyloxyresveratrol, kojic acid, a secondary metabolite of the fungus fungus, ), And arbutin and kojic acid are currently used as additives in functional cosmetics aiming at whitening. In addition, phenolic compounds, flavonoids, glycolic acid, ferulic acid, and oflavonoids are known as typical compounds.

The tyrosinase inhibitory activity was measured according to the method of Yagi et al. To the reaction mixture was added 200 U / mL mushroom tyrosinase (Sigma, USA) to a mixture of 40 μL of the substrate solution and 40 μL of the sample solution in 80 μL of 67 mM sodium phosphate buffer (pH 6.8) and 10 mM L-DOPA uL was added and reacted at 37 ° C for 10 minutes to measure the DOPA chrome produced in the reaction solution at 492 nm. The tyrosinase inhibitory activity was expressed as the absorbance reduction ratio of the sample solution and the non - added sample. Table 3 shows the inhibition rate of tyrosinase according to the extraction solvent and extraction method. As a result of measuring the tyrosinase inhibition rate of the extract obtained by extracting different kinds of coffee extracts from coffee beans, no inhibitory effect was observed in all the extracts

Extraction solvent Extraction method Inhibition rate (%) Distilled water Pressure extraction method N.D. Ultrasonic extraction method N.D. And arithmetic Pressure extraction method N.D. Ultrasonic extraction method N.D.

Next, inhibition rate of tyrosinase using coffee bran fermentation broth by Bacillus subtilis according to the above Preparation Example was confirmed. The fermentation of coffee pulp with Bacillus subtilis showed about 1.8-fold increase in tyrosinase inhibitory activity compared to the control without the strain. In addition, the longer the fermentation time of the Bacillus subtilis strain, the greater the whitening effect. This result supports the production of tyrosinase inhibitor continuously during the growth of Bacillus subtilis. In addition, it is presumed that the secondary metabolite - related substances produced in the stagnant phase of Bacillus subtilis through the growth phase directly inhibit tyrosinase or increase the inhibitory effect of tyrosinase in coffee bean.

Example 4: Using complex microorganisms Coffee shop Evaluation of efficacy of fermented extract

Example 4-1: Comparison of polyphenol contents

The total polyphenol contents of the fermented broth and the fermentation residue and the fermentation residue were determined and compared with that of the fermented broth using the complex microorganism according to the above Preparation Example. As a result, fermented residue was added with peroxide, (14.03 mg / g), which was the highest polyphenol content (20.39 mg / g). Table 4 shows the total polyphenol content of the coffee ground fermented product.

extract Total polyphenol (mg / g) Complex microorganism 5.80 + 0.017 Coffee fermentation residue and pressurized extract added with acid water 14.03 + 0.088 Coffee shop Fermentation liquid 20.39 ± 0.268

Example 4-2: Collagenase Inhibition activity comparison

The collagenase inhibitory activity of the fermented broth of fermented milk and fermentation residue was investigated by using the complex microorganism according to the above Preparation Example. At the concentration of 1,000 ppm, the coffee fermented liquid showed the highest inhibitory effect of 58.12%, which is about twice as high as that of the pressurized extract added with fermentation residue. Table 5 shows the collagenase inhibitory activity of the pressurized extract of the fermented coffee bean fermentation broth and fermentation residue added with peroxide.

extract Collagenase inhibition (%) Complex microorganism 35.65 + 0.147 Coffee fermentation residue and pressurized extract added with acid water 27.03 + - 0.351 Coffee ground fermentation broth 58.12 + - 0.139

Example 4-3: Comparison of tyrosine inhibitory activity

After the fermentation of the coffee bean using the complex microorganism according to the above Preparation Example, the tyrosine of the pressurized extract to which the fermentation broth and the fermentation residue were added with peroxide was measured in the same manner as in the above Example. At the concentration of 1,000 ppm, the fermented coffee had the highest inhibitory activity of 65.45%, which was about 2.9 times higher than that of the fermented residue. Table 6 below shows the effect of tyrosine inhibition of the coffee ground fermented product.

extract Inhibition rate of tyrosinase (%) Complex microorganism 15.77 ± 0.208 Coffee extract and pressed extract 22.48 ± 0.112 Coffee ground fermentation broth 65.45 + 0.127

Example 5: Coffee shop Efficacy evaluation by extraction method

Example 5-1: Electron donating ability effect

The electron donating ability of pressurized, steamed and mixed microbial fermentation broth of coffee grounds and pressurized extract (fermented extract) added with peroxide to fermentation residue was measured. The electron donating abilities (EDA) were measured by modifying the Blois method. 60 uL of DPPH solution (Sigma, U.S.A) and 120 uL of each extract were mixed and left for 15 minutes, and the absorbance was measured at 517 nm using a microplate reader. The electron donating ability was expressed by the absorbance reduction ratio of the sample solution and the non-additive solution. As a result, after extracting 78.28% of pressurized extract, 81.87% of hydrothermal extract, 81.87% of fermented extract, and fermented extract at 1,000 ppm of concentration, the residue was further extracted with distilled water and the fermented broth was extracted with 1: Mixed) showed an effect of 87.34%. Experimental results on electron donating ability and total polyphenol compound content suggest that the coffee ground fermented extract of the present invention can exhibit an excellent antioxidative function. Table 7 shows the effect of electron donating ability according to coffee bean extraction method.

Extraction method Electron donating ability (%) Pressure extraction 78.28 ± 1.34 And extraction of mountain water 81.87 ± 0.54 Fermentation extraction 87.34 + 0.89

Example 5-2: Inhibitory effect on melanin synthesis

Melanin synthesis inhibitory effect was measured using B16F10 cells for each concentration of 100 ppm of fermented extract extracted from pressurized, coffee, and mixed microbial fermentation broth and fermentation residues by pressurized extraction. The cells were cultured in B16F10 cells at 1 × 10 ⁵ for 24 hours in each well of a 6-well plate. 200 μg of the extracts obtained from the pressurized extraction method, the peroxide extraction method and the microbial fermentation method were dispensed into 2 mL of the culture, and 2 mM theophylline and 2 μM α-MSH were treated and cultured for one week. After the culture was completed, the medium was removed, washed twice with PBS, and 500 μL of lysis buffer (0.01 M sodium phosphate buffer, 1% Triton X-100, 5 mM EDTA) was added to dissolve the cells and centrifuged at 13,000 rpm for 20 minutes Respectively. The supernatant was discarded and the obtained cells were dried in a constant temperature oven at 60 DEG C for 24 hours. After drying, 300 uL of 1 N NaOH aqueous solution to which 10% DMSO was added was added, and the mixture was treated and dissolved at 90 DEG C for 1 hour. Absorbance was measured at a wavelength of 475 nm using a UV spectrometer. Based on the absorbance of the positive control group, the absorbance of the extracts extracted by each extraction method was expressed as a percentage. As a result, fermented extract of mixed microorganism (fermentation broth was separated and fermented broth was extracted with distilled water and mixed with 1: 1 by volume of fermented broth). The whitening effect of the fermented extract was confirmed to be comparable to that of the positive control. Table 8 shows the effect of inhibiting melanin synthesis by the coffee bean extraction method.

Extraction method Melarin synthesis inhibition (%) Pressure extraction 78.28 ± 1.34 And extraction of mountain water 81.87 ± 0.54 Fermentation extraction 87.34 + 0.89 Positive control (arbutin) 87.34 + 0.89

Example 5-3: Skin cell Playback effect

The skin consists of the epidermis, dermis, and subcutaneous tissue from the outside. The epidermis protects against external stimuli and pathogens, regulates body temperature, and maintains moisture and lipid components. The dermis consists of fiber component and lipid component. Collagen gives strength and tension to the skin, protects the skin, and accounts for 90% of the dermis. Collagen is mostly type-I collagen and is degraded by collagenase and elastinase. The dermis is closely related to skin aging, as well as its important role in determining the physical and chemical properties of the skin. In general, as the age increases, the action of fibroblasts and the number of cells decreases, the amount of collagen and elastin synthesis decreases, the moisture in the skin cells is lost, and the stratum corneum structure changes. Also, the action of collagenase is increased, and the cross-linked form of collagen is increased, so that smoothness, moisturizing and tightening are reduced.

According to the above preparation example. Extraction of coffee beans by pressurized extraction, extraction of hydrothermal solution, extraction of fermented extracts of mixed microbial fermentation broth and fermentation residues by pressurized extraction, and production of collagen for each of them were carried out. As a result, And the extract showed high synthesis in the fermentation extract (the extract as in 5-1 above). Table 9 shows the effect of collagen synthesis according to the coffee bean extraction method.

Extraction method density(%) Collagen synthesis (%) Jinseino Side (Rh2) - 22 Pressure extraction 0.5 21 One 26 5 31 And extraction of mountain water 0.5 23 One 28 5 33 Fermentation extraction 0.5 38 One 43 5 49

Example 5-4: Skin moisturizing effect

Skin aging is characterized by reduced skin elasticity, skin dryness and wrinkling. This phenomenon is closely related to the amount of collagen and elastin fibers present in the connective tissue of the skin. Collagen is synthesized in fibroblasts. Unlike other proteins, it consists of 10% of 4-hydroxyproline. Vitamin C is involved in the synthesis of 4-hydroxyproline, resulting in deficiency of vitamin C .

Coffee bean extracts and fermented liquids contain substances with antioxidant properties, thus affecting cells that synthesize collagen. First, amino acids digested by collagenase were reacted with ninhydrin reagent (manufacturer WACO, Deoksan Science Co., Ltd.) to measure indirectly CLP (collagen like polymer) . According to the above preparation example. The pressurized extract was prepared by adding perchloric acid to the coffee ground pressurized extract, the hydrothermal extract, and the mixed microbial fermentation residue, and each of them was added to NIH3T3 cells (KCLB 80051, Seoul National University) , 25 ㎍ / ㎖, 50 ㎍ / ㎖, and 100 ㎍ / ㎖, respectively, and the reaction was continued for 48 hours to determine type-I collagen. As a result, it was confirmed that the pressurized extract extracted by pressurized extraction, hydrothermal extraction, and fermentation extraction increased the type-I collagen production ratio in proportion to the concentration. In particular, the type-I collagen production rate was highest in the pressurized extract of mixed microbial fermentation broth and fermentation residue added with peroxide. Table 10 shows the amount of collagen type I produced according to the coffee bean extraction method.

Extraction method Sample concentration (ug / ml) Type-I collagen production rate (%) Remarks Pressure extraction Control group 100 After mixing with the extract, 1% DMSO was added 10 100 25 109 50 112 100 118 And extraction of mountain water Control group 100 After mixing with the extract, 1% DMSO was added 10 100 25 110 50 115 100 120 Fermentation extraction Control group 100 After mixing with the extract, 1% DMSO was added 10 105 25 119 50 123 100 131

Example 5-5: Anti-inflammatory effect

Inflammation is one of the natural biological responses to tissue damage that can be mediated by various stimuli such as infection, immune response. In the body inflammation process, inflammatory factors such as nitric oxide and prostaglandin E ₂ are formed by the inducible NO-producing enzyme and cyclooxygenase-2. Among these, NO has various physiological functions such as defense function in the body, signal transduction function, and vasodilation. However, overexpression of NO leads to inflammation, tissue damage, genetic mutations, and nerve damage.

The inhibitory effect of NO production on each of the fermented extracts (the extracts as described in 5-1) obtained by adding the fermented extracts of coffee bean pressurized extract, perennial extract, mixed microbial fermentation broth, Respectively. Nitric oxide (NO) measurement was carried out by measuring the amount of NO in the supernatant of the cells as nitrite and nitrate. When the confluence of RAW264.7 cells was 80% in a ^6- well plate (2 × 10 ⁶ cells / well), the cells were washed twice with PBS and incubated with serum-free After culturing for more than 12 hours using the medium, samples were treated for 1 hour, treated with 1 ug / mL of lipopolysacchride (LPS) and incubated for 24 hours. The supernatant of the culture was taken and reacted with the griess reagent. Absorbance was measured at 540 nm using an ELISA reader, and the NO production rate was expressed as a percentage. As a result, it was confirmed that NO production was decreased in a concentration dependent manner in the fermented extract. The lowest concentration was found at a concentration of 500 ppm to 3.7 μM. Table 11 shows the NO production amount according to the coffee bean extraction method.

Extraction method Extract concentration (ppm) NO production (uM) Pressure extraction blank 0 LPS 8 60 8.4 120 9 240 9.2 500 10.1 And extraction of mountain water blank 0 LPS 8 60 8.3 120 8.7 240 9 500 9.2 Fermentation extraction blank 0 LPS 8 60 6.3 120 5.4 240 4 500 3.7

Example 6: Coffee shop Analysis of physiologically active substance content of fermented extract

The content of caffeine was 0.71% in the sample prepared as the basic distilled water, 0.69% in the pressurized extract, 0.70% in the hydrothermal extract, and 0.21% in the fermented extract (the extract like 5-1). It is known that chlorogenic acid occupies the most part of the polyphenol compounds present in coffee beans and has physiological activities such as antioxidant activity, hepatoprotective activity and hypoglycemic activity . The content of chlorogenic acid was 1.78%, that of pressurized extract was 1.82% and the content of hydrolyzed extract was 1.98% and that of chloralic acid was 1.98% Fermented extract showed a content of 2.12%. Table 12 shows the contents of caffeine and chlorogenic acid according to the coffee bean extraction method.

Extraction method Caffeine content (%) Content of chlorogenic acid (%) 3rd distilled water extraction 0.73 ± 0.01 1.76 ± 0.01 Pressure extraction 0.71 ± 0.06 1.80 + 0.02 And extraction of mountain water 0.75 + 0.07 1.89 + 0.07 Fermentation extraction 0.18 + 0.03 2.03 ± 0.11

The present invention improves the utilization value of the microorganism by using a fermentation technique in a copy paper which is disposed of as waste, and it has been confirmed that the coffee ground fermented product according to the present invention has excellent effects on antioxidation, whitening and wrinkle improvement. Accordingly, the coffee bean composition of the present invention may be used in the cosmetics industry as a cosmetic composition.

Claims

A cosmetic composition comprising a coffee ground fermented product obtained by inoculating coffee grounds with a strain of Bacillus subtilis (KCTC 1027).

The method according to claim 1, wherein the lactic acid bacteria ( Weissella cibaria , KFCC11598P), Saccharomyces cerevisiae (ATCC9763) and Acetobacter pasteurianus , ATCC 9432). < / RTI >

The cosmetic composition according to claim 1 or 2, wherein the coffee ground fermented product has antioxidative, anti-inflammatory, whitening, skin moisturizing and skin wrinkle inhibiting effects.

(i) drying a coffee bean; And
(ii) including the inoculation and fermentation of the Bacillus subtilis (Bacillus subtilis, KCTC 1027) to the dried coffee foil,
Antioxidant, anti-inflammatory, whitening, skin moisturizing and skin wrinkle formation.

5. The method according to claim 4, wherein in (ii), the lactic acid bacteria ( Weissella cibaria , KFCC11598P), yeast ( Saccharomyces S. cerevisiae , ATCC9763) and acetic acid bacteria ( Acetobacter pasteurianus , ATCC 9432).
Antioxidant, anti-inflammatory, whitening, skin moisturizing and skin wrinkle formation.

The method according to claim 4 or 5, further comprising, after the step (ii), adding the ferric oxide to the coffee ground fermentation product followed by pressure extraction,
Antioxidant, anti-inflammatory, whitening, skin moisturizing and skin wrinkle formation.

A cosmetic composition having antioxidant, anti-inflammatory, whitening, skin moisturizing and skin wrinkle inhibiting effects, comprising a coffee ground fermented product produced according to claim 4 or 5.