WO2021261975A1 - Vegetable extract comprising vegetable collagen and vegetable mucin, and method for preparing same - Google Patents

Abstract

The present invention provides an extract comprising vegetable collagen and vegetable mucin, a food composition and a cosmetic composition comprising the extract, and a method for preparing the extract. The extract comprising vegetable collagen and vegetable mucin of the present invention contains low molecular weight collagen of 500 Da or less, and has an excellent skin moisturizing effect, a skin aging prevention effect, and a skin wrinkle improvement effect when administered orally and applied to the skin, and thus, can be helpfully used as a raw material of functional foods and cosmetics for improving skin condition.

Description

Plant extract containing plant collagen and plant mucin and method for preparing the same

This patent application is i) Korean Patent Application No. 10-2020-0078772, filed with the Korean Intellectual Property Office on June 26, 2020, ii) Korean Patent Application No. 10-2020, filed with the Korean Intellectual Property Office on December 23, 2020 -0182639, and iii) claim priority to Korean Patent Application No. 10-2021-0043663 filed with the Korean Intellectual Property Office on April 2, 2021, the disclosure of which is incorporated herein by reference. .

The present invention relates to a plant extract comprising plant collagen and plant mucin and a method for preparing the same.

Collagen is a fibrous structural protein constituting animal connective tissue, skin, bone, tendon, skin, cartilage, nails, toenails, teeth, blood vessels, and the like. Collagen accounts for about 30% of human protein, is the main component of connective tissue, and accounts for 70-90% of the dermal layer of the skin. The basic structural unit of collagen is tropocollagen, which is a very large macromolecular protein of 300,000 Daltons (Da) in which a peptide having a molecular weight of 100,000 Daltons (Da) has a triple helix structure. The amino acids constituting collagen are glycine, proline, hydroxyproline, and other amino acids combined with G-X-Y. Collagen contains relatively low hydroxyproline in other proteins, so the hydroxyproline content is also a measure of the collagen content in the organ. When collagen is treated with heat or chemicals, it denatures into gelatin. It has been expected that collagen can be ingested by ingesting pork skin or chicken feet, but collagen is a high molecular weight protein of 300,000 Daltons, so the absorption rate in the body is less than 10%. have. According to the 500 Dalton Rule, in order for collagen to be absorbed into the skin, it must be less than 500 Daltons. It is known that the absorption rate of 2,000 to 6,000 daltons of pharmaceutical grade collagen is 50%, and more than 90% of collagen hydrolyzed to 500 daltons is absorbed.

The collagen is a fibrous protein constituting skin, cartilage, hair, nails, toenails, teeth, and blood vessels, and serves to maintain the structure of the extracellular matrix (ECM) and maintain elasticity. When UV rays penetrate the dermis of the skin, collagen and elastin are decomposed, and oxidative stress and reactive oxygen species (ROS) increase, causing skin aging by UV rays. In addition, continuous exposure to smoking, alcohol, environmental pollution, etc. causes the body's antioxidant system to collapse, degeneration of collagen fibers in the dermis, and decrease in skin moisture, resulting in dry skin and loss of skin elasticity, resulting in wrinkles.

In the case of adolescents or pregnant women, the skin stretches rapidly due to excessive adrenal cortex hormones and rapid weight gain, and some of the bonds between collagen fibers are destroyed, leading to stretch marks. As we age, the cartilage in the joints wears away, making it difficult to walk. According to research results, enzymatically hydrolyzed collagen is well absorbed into the body, suppressing pain in the joints, anti-inflammatory, and increasing bone density. Collagen not only keeps skin, but also bones, joints, teeth, hair, eyes, nails and toenails healthy.

Plant collagen present in plants promotes collagen production in the body and inhibits collagen degradation by inhibiting MMP-1, a collagen-degrading enzyme. In addition, vegetable collagen is rich in antioxidants that animal collagen does not have. Most of the collagen products currently on the market are animal collagen extracted from fish or cattle, and there are concerns about consumers being uncomfortable to eat due to their unique smell or using mad cow disease or antibiotics. Most of the vegetable collagen of the present invention has a molecular weight of 100 to 400 Daltons and is hydrolyzed to an average of 500 Daltons or less, so the absorption rate is more than 90%. can be useful for

Numerous papers and patent documents are referenced throughout this specification and their citations are indicated. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to more clearly describe the level of the technical field to which the present invention pertains and the content of the present invention.

[Patent Literature]

PCT International Publication No. WO 2012-096760 A1 (published on July 19, 2012)

The present inventors made intensive research efforts to develop a method for producing a plant extract containing a large amount of vegetable collagen that can replace the existing animal collagen. As a result, viscous vegetable collagen and vegetable mucin are contained in large amounts in the extract extracted through enzyme and microbial fermentation after plant powder is immersed in a solvent. By identifying the excellent skin barrier improvement and skin moisturizing effect, the present invention was completed.

Accordingly, an object of the present invention is to provide an extract comprising vegetable collagen and vegetable mucin and a method for preparing the same.

Another object of the present invention is to provide a food composition or cosmetic composition for skin anti-aging, wrinkle improvement, skin barrier strengthening, or skin moisturizing comprising an extract containing vegetable collagen and vegetable mucin as an active ingredient.

According to one aspect of the present invention, the present invention provides a food composition for skin anti-aging, wrinkle improvement, skin barrier strengthening, or skin moisturizing comprising a plant extract as an active ingredient. The plant extract includes plant collagen and plant mucin.

The term "vegetable" in the present invention has the meaning of "derived from a plant". For example, vegetable collagen refers to collagen derived from plants.

In one embodiment of the present invention, the extract extracts at least one herbal material selected from the group consisting of spinach, Swiss chard, carrot, tomato, astragalus, chrysanthemum, mushrooms, round hemp, lotus root, rape flower, and rapeseed it was obtained by The one or more herbal ingredients may be one single herbal ingredient, or a mixture of two or more kinds of herbal ingredients. The mushrooms are not strictly classified as plants, but the plant extract of the present invention is a concept including the extract of mushrooms.

In one embodiment of the present invention, the food composition is pomegranate, tart cherry, blueberry, garlic, mango, orange, avocado, white lily, cashew nut, hibiscus, strawberry, goldfish, kiwi, guava, pineapple, soybean, An extract obtained by extracting a mixture of one or more herbal ingredients selected from the group consisting of bell pepper, raspberry, blackberry, goji berry, bokryeong, and sukjihwang is additionally included.

In a specific embodiment of the present invention, the mushrooms are white wood ear mushroom, wood ear mushroom, reishi mushroom, chima mushroom, matsutake mushroom, shiitake mushroom, truffle, situation mushroom, horseshoe mushroom, pine mushroom, chaga mushroom, neungi mushroom, roe deer At least one mushroom selected from the group consisting of oyster mushroom, oyster mushroom, and blackcurrant mushroom, but is not limited thereto.

In the present invention, the white woody mushroom belongs to the white woody mushroom and its scientific name is Tremella fuciformis. It is widely distributed in Korea, China, Japan, and tropical regions. The fruiting body (mushroom) of the white-necked ear is agaric, wrinkled and split, or has the shape of an earlobe or a crest, and the size is about 10 cm. When dried, it forms a thin film, but swells when water is absorbed. It is known to have various effects such as immunity enhancement, bone health enhancement, constipation improvement, anticancer action, skin beauty, prevention of vascular disease, and prevention of anemia. The present inventors have attempted to develop an extraction method according to the enzymatic treatment of the white wood ear mushroom in order to further improve the efficacy of the white wood ear mushroom and to improve the properties as a food composition.

In one embodiment of the present invention, the mushrooms refer to the fruiting bodies of each type of mushroom, but are not limited thereto.

In one embodiment of the present invention, the extract may be included in the range of 0.001 to 20% (w/w) based on the total weight of the food composition, but this is only exemplary and not limited thereto.

In one embodiment of the present invention, the extract contains hyaluronic acid. Accordingly, the extract of the present invention contains vegetable hyaluronic acid.

In one embodiment of the present invention, the plant extract, which is an active ingredient of the composition, contains vegetable collagen and plant mucin, and increases the expression of collagen in the skin.

In addition, the plant extract of the present invention contains collagen peptides and is prepared by fermentation. Therefore, the plant extract of the present invention can be called a fermented collagen peptide composition. When the raw material of the plant extract of the present invention is white wood ear mushroom, it may be called a white wood ear mushroom fermented collagen peptide composition.

Collagen, a major component of the extracellular matrix, is a protein expressed in fibroblasts of the skin and forms most of the organic materials of skin, bones, and teeth. It has various functions such as cell support (Brenneisen et al., 2002; Kim et al., 2015).

Collagen is known to be closely related to the formation of wrinkles in the skin, and a lack of collagen can cause wrinkles. Collagen is synthesized in the form of procollagen, a precursor, and is known to be cleaved and separated from collagen molecules during collagen polymerization (Lee et al., 2015). That is, by measuring the amount of propeptide, it is possible to determine the degree of collagen biosynthesis in the cell.

The food composition of the present invention may include ingredients commonly added during food production, as well as the extract. The additional ingredients include, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

According to another aspect of the present invention, the present invention provides a plant fermented food comprising the food composition.

In one embodiment of the present invention, the plant may be, for example, white wood ear mushroom, but is not limited thereto.

Accordingly, the present invention provides a fermented food of white wood ear mushroom. The fermented white wood ear mushroom food of the present invention may include not only the fermented collagen peptide composition prepared by the above manufacturing method, but also the ingredients commonly added during the preparation of the above-described food.

In another embodiment of the present invention, the food composition may be prepared in the form of a beverage. When the fermented food of the present invention is manufactured as a beverage, citric acid, high fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract, etc. may be additionally included in addition to the extract of the present invention.

Therefore, in a specific embodiment of the present invention, the fermented food may be a fermented white wood ear mushroom drink.

The food composition of the present invention includes processed forms of all natural materials such as food, functional food, nutritional supplement, health food, and food additives. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.

For example, as a health food, the extract (concentrate or powder) may be prepared in the form of tea, juice, and drink to be consumed, or granulated, encapsulated and powdered. In addition, as food, beverages (including alcoholic beverages), fruits and their processed foods (eg, canned fruit, bottled, jam, marmalade, etc.), fish, meat and their processed foods (eg, ham, sausage corn beef, etc.) ), breads and noodles (eg udon noodles, soba noodles, ramen, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, syrup, dairy products (eg yogurt, fermented milk, butter, cheese, etc.), edible vegetable oils and fats, margarine, Vegetable protein, retort food, frozen food, various seasonings (eg, soybean paste, soy sauce, sauce, etc.) may be prepared by adding the extract of the present invention. In addition, in order to use the extract of the present invention in the form of a food additive, it may be prepared and used in the form of a powder or a concentrate.

According to another aspect of the present invention, the present invention provides a cosmetic composition for skin anti-aging, wrinkle improvement, skin barrier strengthening or skin moisturizing comprising a plant extract as an active ingredient. The extract is also characterized in that it contains vegetable collagen and vegetable mucin.

The cosmetic composition of the present invention contains the same extract as the above-described food composition as an active ingredient, and descriptions of common contents between the two inventions are omitted in order to avoid the complexity of the present specification.

In the cosmetic, when the total weight of the cosmetic is based on 100% by weight, the extract containing the vegetable collagen and vegetable mucin may be included in an amount of 0.001 to 20% by weight.

The formulation of the cosmetic is not particularly limited, but preferably skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence , nutritional essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser.

The cosmetic of the present invention may also contain other ingredients that are formulated in conventional cosmetics. Other ingredients that may be added include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, colorants, fragrances, A blood circulation promoter, a cooling agent, a restrictive agent, purified water, etc. are mentioned.

Blending components that may be added other than the above are not limited thereto, and any of the above components can be blended within a range that does not impair the object and effect of the present invention, but preferably 0.01 to 5 wt%, more preferably, based on the total weight 0.01 - 3 wt%, 0.01 - 2 wt%, 0.01 - 1 wt%, 0.1 - 3 wt%, 0.5 - 3 wt%, 1 - 3 wt%, or 2 - 3 wt%, etc. may be blended.

The cosmetic of the present invention may take the form of a solution, an emulsion, a viscous mixture, or the like. The ingredients included in the cosmetic of the present invention may include ingredients commonly used in cosmetics in addition to the peptide composition as an active ingredient, for example, conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments and fragrances carrier. When the cosmetic formulation of the present invention is a paste, cream or gel, animal fiber, vegetable fiber, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, etc. can be used When the cosmetic formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propellants such as propane/butane or dimethyl ether.

When the cosmetic formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan.

When the cosmetic formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, and microcrystals Adult cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.

When the cosmetic formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, Fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linoline derivative or ethoxylated glycerol fatty acid ester and the like can be used.

When the formulation of the present invention is soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolysates, isethionate, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugar, etc. are used as carrier components may be, but is not limited thereto. These may be used alone or in combination of two or more.

According to another aspect of the present invention, the present invention provides a method for preparing a vegetable extract comprising the steps of:

(a) mixing a solvent in the herbal herbal material powder;

(b) adding an enzyme, lactic acid bacteria, or a combination thereof to the mixed solution and culturing;

(c) centrifuging or filtering the culture solution to remove sludge and starch and separating the viscous supernatant, or preparing a filtrate; and

(d) heating the viscous supernatant or the filtrate to inactivate and sterilize the enzyme, lactic acid bacteria, or a combination thereof.

As described above, the plant extract according to the present invention abundantly contains vegetable collagen, plant mucin, and the like.

In one embodiment of the present invention, the method for preparing the extract further comprises the step of (e) obtaining a powdery extract by drying the liquid extract or the same.

(a) step: mixing a solvent in the herbal herbal material powder;

In one embodiment of the present invention, the herbal material powder of step (a) is pulverized to a particle size in the range of 10 to 100 mesh (mesh). For the pulverization, a method of pulverizing and pulverizing food commonly used in the art may be used without limitation.

In one embodiment of the present invention, the herbal material is at least one selected from the group consisting of spinach, Swiss chard, carrot, tomato, astragalus, chrysanthemum, mushrooms, round hemp, lotus root, rape flower and rapeseed.

In one embodiment of the present invention, the herbal material is pomegranate, tart cherry, blueberry, garlic, mango, orange, avocado, white lily, cashew nut, hibiscus, strawberry, goldfish, kiwi, guava, pineapple, soybean, Bell pepper, raspberry, blackberry, goji berry, bokryeong, and additionally includes at least one selected from the group consisting of sukjihwang.

In a specific embodiment of the present invention, the mushrooms are white wood ear mushroom, wood ear mushroom, reishi mushroom, chima mushroom, matsutake mushroom, shiitake mushroom, truffle, situation mushroom, horseshoe mushroom, pine mushroom, chaga mushroom, neungi mushroom, roe deer At least one mushroom selected from the group consisting of oyster mushroom, oyster mushroom, and blackcurrant mushroom, but is not limited thereto.

In one embodiment of the present invention, the solvent is purified water or a C1 to C4 aqueous solution of lower alcohol, and more specifically, purified water or an aqueous ethanol solution.

In one embodiment of the present invention, in step (a), the solvent is added in an amount of 1 to 100 parts by weight based on 1 part by weight of the herbal herbal material powder. Specifically, the purified water may be added in an amount of 2 to 80 parts by weight, 2 to 50 parts by weight, 3 to 30 parts by weight, 5 to 20 parts by weight, 5 to 15 parts by weight, or 5 to 10 parts by weight, but is not limited thereto. .

In one embodiment of the present invention, the plant herbal material powder of step (a) is treated with an acid selected from citric acid, acetic acid, or dilute hydrochloric acid at a concentration of 1 to 80% (w/w) for 1 to 5 hours. can The acid is 1 to 60% (w/w), 1 to 50% (w/w), 1 to 40% (w/w), 1 to 30% (w/w), 1 to 20% (w/w) ), or 1 to 10% (w/w), but is not limited thereto.

In one embodiment of the present invention, the preparation method of the present invention may include neutralizing after treatment with an acid selected from citric acid, acetic acid, or dilute hydrochloric acid.

In one embodiment of the present invention, step (a) may include a step of adding an aqueous ethanol solution as a solvent to the plant herbal material powder.

The process of adding the aqueous ethanol solution as a solvent may be performed after acid treatment or acid treatment and neutralization of the above-described vegetable herbal material powder.

In addition to the extraction method using ethanol, the extract of the present invention may be extracted by a known natural product extraction method. For example, extraction may be performed by cold extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction, or heating extraction method, and specifically, extraction may be performed by hot water extraction or reflux cooling extraction method, 1 to 10 times, more specifically 2 Extraction can be repeated 7 to 7 times. In this case, the extraction solvent may be water (purified water).

In the present invention, the extract may be used in the form of a crude extract extracted with a solvent, and may be purified and used with high purity.

As used herein, the term 'extract' has the meaning commonly used as a crude extract in the art as described above, but in a broad sense also includes a fraction obtained by additionally fractionating the extract. That is, the extract of the herbal herbal material includes not only those obtained by using the above-described extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, dialysis, separation by various chromatography (prepared for separation according to size, charge, hydrophobicity or affinity), etc., additionally carried out The fractions obtained through various purification methods are also included in the extract of the herbal herbal material of the present invention.

In another embodiment of the present invention, in step (a), an aqueous ethanol solution is added to the vegetable herbal material powder as a solvent and extracted, followed by solid-liquid separation to obtain a liquid component, and the solid content is 60-95° C. after adding water After hot water extraction in the ethanol solution, it is mixed with the liquid component extracted with the aqueous ethanol solution.

In one embodiment of the present invention, in step (a), a solvent is added to the herbal herbal material powder, and the pressure treatment may be performed under 0.2 to 5 atmospheres. The pressurization treatment may be performed in parallel with the hot water extraction step.

The pressurization treatment is performed under conditions of 0.2 atm or more and less than 1 atm; pressure treatment under conditions of more than 1 atmosphere and less than 5 atmospheres; Alternatively, after being pressurized under conditions of 0.2 atm or more and less than 1 atm, it may be pressurized again under conditions of greater than 1 atm and below 5 atm.

The pressure treatment under the conditions of 0.2 to 5 atmospheres may be performed for 30 minutes to 2 days, and may be performed one or more times under different pressure and time conditions. The pressure treatment may be, for example, pressurized under conditions of 0.2 atm or more and less than 1 atm for 30 minutes to 1 day, and then pressurized under conditions of greater than 1 atm and below 5 atm, for 30 minutes to 1 day.

The pressurization process under the conditions of more than 0.2 atm and 1 atm or less expands and removes (de-foaming) fine air bubbles contained in the mixture of powder and solvent so that the herbal material powder can be mixed with the solvent better, and through this Increases extraction efficiency by allowing collagen, vegetable mucin, and vegetable polysaccharide to be better extracted into the solvent.

In addition, the pressurization process under the conditions of more than 1 atmosphere and less than 5 atmospheres increases the extraction efficiency by increasing the solubility of vegetable collagen and vegetable mucin contained in the herbal material powder by applying high pressure to the herbal powder and the solvent.

In one embodiment of the present invention, the heat treatment may be performed at a temperature condition of 50-130 °C during the pressure treatment under the 0.2 to 5 atmospheres pressure condition. The heat treatment process increases the solubility of useful components dissolved in the solvent, thereby increasing the extraction efficiency of vegetable collagen and vegetable mucin contained in the herbal material.

Therefore, in one embodiment of the present invention, in step (a), a solvent is added to the herbal herbal material powder, and the pressure and heat treatment are performed for 30 minutes to 2 days under conditions of 0.2 to 5 atmospheres and a temperature of 50-130 ℃. However, the present invention is not limited thereto.

Step (b): adding an enzyme, lactic acid bacteria, or a combination thereof to the mixture and culturing

This step is a step of adding an enzyme, lactic acid bacteria, or a combination thereof to the mixed solution prepared in step (a) and culturing. To the mixture, the enzyme or lactic acid bacteria may be added alone and cultured, or the enzyme and the lactic acid bacteria may be added together (sequentially or simultaneously) and cultured.

In one embodiment of the present invention, the enzyme of step (b) is selected from the group consisting of cellulase, hemicellulase, beta-glucosidase, betaglucanase, xylanase, arabinase, protease, and amylase One or more enzymes that become

In one embodiment of the present invention, the protease is at least one protease selected from the group consisting of pepsin, papain, trypsin, and elastase.

In one experimental example of the present invention, the enzyme may be used as a complex enzyme, such as Sumizyme, Ultimase, Viscozyme, Lyvarome A5, but is not limited thereto.

In one embodiment of the present invention, the enzyme may be added at a concentration of 0.01 to 3 wt%, but is not limited thereto.

In one embodiment of the present invention, the amount of the enzyme added is more specifically 0.1-4%, 0.1-3%, 0.1-2%, 0.1-1%, 1-4%, 1-3%, 1- 2%, 1%, 2%, or 3%, but not limited thereto. If the addition amount of the enzyme is less than 0.01%, the hydrolysis effect of the enzyme does not appear, and if it exceeds 5%, the manufacturing cost is excessive and not economical.

In one embodiment of the present invention, the temperature during the treatment of the enzyme is 40-70 ℃, more specifically 40-65 ℃, 45-65 ℃, or 50-65 ℃, but is not limited thereto. In the case of the enzyme treatment, it is preferable to perform the treatment while stirring in order to increase the contact frequency and reaction efficiency between the enzyme and the plant herbal material. The stirring speed is 50-250 rpm, but is not limited thereto.

In one embodiment of the present invention, when enzyme treatment is performed on the vegetable herbal material, the polysaccharide is reduced in molecular weight according to the hydrolysis of the polysaccharide and the taste is also improved. When the whole powder of vegetable herbal material is added here, the texture of the vegetable herbal material containing a large amount of vegetable collagen and the preference of the whole composition are greatly improved.

In one embodiment of the present invention, when the herbal material is white wood ear mushroom, the extract prepared by enzymatic treatment may be called a white wood ear mushroom enzyme-treated polysaccharide extract.

The enzyme-treated polysaccharide extract of the white oyster mushroom can be prepared as a food composition by adding the whole powder of the white rhododendron mushroom. In this case, the texture and taste of the food composition are improved.

In one embodiment of the present invention, the lactic acid bacteria of step (b) is Leuconostok genus, Lactobacillus genus, Bifidobacterium genus, Enterococcus sp. strain, Lactococcus genus, or a combination thereof.

In a specific embodiment of the present invention, the Leuconostok genus bacteria is a Leukonostok mecenteroides strain, and the Lactobacillus genus bacteria is Lactobacillus delbrooki spp. Bulgaricus, Lactobacillus helveticus, Lactobacillus permentum, Lactobacillus thermophilus, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus reuteri, or these is a combination of

In a specific embodiment of the present invention, the Lactobacillus genus fungus is Lactobacillus alementarius ( Lactobacillus alementarius ), and more specifically Lactobacillus alementarius Reuter strain ( Lactobacillus alementarius Reuter, ATCC 29643).

In a specific embodiment of the present invention, the Enterococcus sp. strain is Enterococcus faecium, Enterococcus faecalis, or a combination thereof.

In a specific embodiment of the present invention, the Lactobacillus genus bacteria are Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus reuteri, Lactobacillus permentum, Lactobacillus Helveticus, Lactobacillus brevis, Lactobacillus elementurius, Lactobacillus delbrooki bulgaricus, or a combination thereof.

In a specific embodiment of the present invention, the Bifidobacterium genus bacteria is Bifidobacterium bifidum, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium animalis lactis, or a combination thereof. to be.

In a specific embodiment of the present invention, the Lactococcus genus bacteria is Lactococcus lactis.

In one embodiment of the present invention, the culturing in step (b) is made for 1 to 50 days.

In a specific embodiment of the present invention, the culturing in step (b) is made for 1 day to 14 days, more specifically 1 day to 10 days, 1 day to 7 days, 1 day to 5 days, or 1 day to 3 days, but is not limited thereto.

In one embodiment of the present invention, the culture in step (b) is controlled within the range of pH 6 to 8, more specifically pH 6.5 to 7.5, and even more specifically 6.5 to 7, but limited thereto It can be adjusted appropriately depending on the enzyme used and the optimal culture conditions of the lactic acid bacteria.

Step (c): centrifuging or filtering the culture solution to remove sludge and starch and separating the viscous supernatant, or preparing a filtrate

The step is to remove the sludge and starch by centrifuging or filtering the culture solution of the enzyme, lactic acid bacteria, or a combination thereof prepared in step (b), and to separate the viscous supernatant, or to prepare the filtrate remaining after filtering. is a step

The centrifugation may be performed under suitable conditions that can be generally used for solid-liquid separation in the art, and it is sufficient as long as it can achieve the purpose of separating the supernatant from the precipitate.

Step (d): heating the viscous supernatant or the filtrate to inactivate and sterilize the enzyme, lactic acid bacteria, or a combination thereof

The step is a step of inactivating and sterilizing the enzyme, lactic acid bacteria, or a combination thereof contained therein by heating the viscous supernatant or the filtrate prepared in step (c).

In one embodiment of the present invention, the heating in step (d) is performed at 60-150° C. for 0.5 seconds to 3 hours.

In a more specific embodiment of the present invention, the heating in step (d) is performed at 61-65° C. for 30 minutes to 1 hour, or at 95-150° C. for 0.5 seconds to 30 seconds.

Step (e): obtaining a liquid extract or drying it to obtain a powdery extract

The step is a step of obtaining a liquid extract from step (d), or drying the liquid extract to obtain a powdery extract.

As the drying method, natural drying, drying under reduced pressure, and drying methods of the extract that can be used in the art, such as freeze-drying, may be used without limitation, but drying under reduced pressure or freeze-drying is preferable.

According to another aspect of the present invention, the present invention provides a method for producing a plant composition comprising a plant extract and whole powder of a plant herbal material, comprising:

Mixing the above-mentioned plant extract with a powder prepared by drying and pulverizing the precipitate remaining after centrifugation or filtration in step (c), or the filtrate.

In one embodiment of the present invention, the extract is prepared by a manufacturing method comprising steps (a) to (d), or (a) to (e) of the above-described method for preparing the extract. The plant extract may be prepared as a food composition by adding whole powder of plant raw materials. In this case, the texture and taste of the food composition are improved.

The present invention provides an extract comprising vegetable collagen and plant mucin, a food composition and a cosmetic composition comprising the extract, and a method for preparing the extract. The extract containing vegetable collagen and vegetable mucin of the present invention contains low molecular weight collagen of 500 Da or less, and has excellent skin moisturizing effect, skin aging prevention effect, and skin wrinkle improvement effect when orally administered and applied to the skin. It can be usefully used as a raw material of functional food and cosmetics for improvement.

1 is a diagram showing the results of a cytotoxicity test of the enzyme-treated extract of white wood ear mushroom of the present invention.

Figure 2 is a view showing the results of confirming the procollagen synthesis promoting effect of the enzyme-treated extract of white wood ear mushroom of the present invention.

3 is a schematic diagram of a method for producing a fermented composition for white wood ear mushroom according to the present invention.

4 is a diagram showing that the thickness of the epidermis of the UVB irradiation group (CON) was significantly increased as compared to the normal control group (NOR) as a result of H&E staining, and the epidermis thickness was decreased when the preparation of the present invention was administered to be.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Example

Throughout this specification, "%" used to indicate the concentration of a specific substance is (weight/weight) % for solid/solid, (weight/volume) % for solid/liquid, and Liquid/liquid is (volume/volume) %.

Example 1: Complex Enzyme Treatment

Example 1-1: Preparation of Enzyme-treated Polysaccharide Extract from White Wood Ear Mushroom

After quantifying 200 g of dried white oyster mushroom fruiting body, it was added to 2 kg of purified water, extracted under reflux at 121° C. for 2 hours, and the filtrate was separated. The filtered residual white oyster mushroom was dried and finely pulverized and used in Preparation Example 2. After cooling the filtrate at room temperature, a complex enzyme (SUMIZYME, Ultimase, Viscozyme, or Lyvarome A5) was added at a concentration of 1% (w/v) and treated at a temperature of 50-65° C. for 48 hours to inactivate the enzyme It was heated at 100° C. for 5 minutes. After filtration of the solution, the filtrate was dried under reduced pressure to obtain an enzyme-treated polysaccharide extract of white wood ear mushroom in powder form.

division SUMIZYME Ultimases Viscozyme Lyvarome A5 comparative example - - - - Preparation 1-1 1% (w/v) - - - Preparation 1-2 - 1% (w/v) - - Preparation 1-3 - - 1% (w/v) - Preparation Example 1-4 - - - 1% (w/v)

The SUMIZYME AC (Shin Nihon Chemical, Japan) is an enzyme complex produced by Aspergillus niger (A. niger), and is a complex enzyme including cellulase, hemicellulase and beta-glucosidase.

The Ultimase is a complex enzyme including beta-glucanase and xylanase.

The Viscozyme is a complex enzyme comprising arabanase, cellulase, beta-glucanase, hemicellulase, and xylanase.

The Lyvarome A5 is a pectinase extracted from Aspergillus niger.

Example 1-2: Cytotoxicity evaluation of polysaccharide extracts treated with Enzyme-treated White wood ear mushroom

Each of the HDFn cells was dispensed in a 96 well plate at 5×10 ⁴ cells/mL, and the concentration of each sample extract (Preparation Examples 1-1 to 1-4) prepared in Example 1-1 under Media-FBS 10% conditions. Each was treated for 72 hours. After that, after adding 1/10 times the volume of each cell culture solution, MTS lysate was added, followed by incubation at 37°C for 2 hours, and then absorbance was measured at 490 nm using an ELISA reader (Spectrostar-nano, BMG Labtech, Offenburg, Germany). did. Through this, it was tested whether each sample affects the cell viability.

The HDFn cells were purchased from Korea Cell Line Bank (KCLB, Seoul, Korea), and penicillin/streptomycin 100 unit/mL (10378, Invitrogen, Grand Island, NY, USA) and 10% FBS (16000, Invitrogen, Grand Island, NY) , USA) containing DMEM medium (11995, Invitrogen, Grand Island, NY, USA) at 37° C., 5% CO ₂ Incubated in an incubator (MCO-17A1, Sanyo, Osaka, Japan).

The results are shown in FIG. 1 .

As shown in FIG. 1 , the concentration of each polysaccharide extract treated with the enzymatic process of the white wood ear mushroom was treated in the range of 31.2 - 1,000 μg/mL. In Preparation Examples 1-1 to 1-4, all samples had an effect on cell viability from a concentration of 500 μg/mL, so the following experiments were performed at a concentration of 500 μg/mL or less showing a high viability.

Example 1-3: Measurement of Collagen Synthesis Promotion Effect of Enzyme-treated Polysaccharide Extract from White Wood Ear Mushroom

In order to see the collagen synthesis effect to test the skin wrinkle improvement effect, HDFn cells were cultured in a 96 well plate at 1 × 10 ⁵ cells/well for 24 hours, and then a sample diluted stepwise with a new serum-free medium was dispensed and added again. Incubated in a CO2 incubator for hours. After obtaining the culture medium, the procedure was performed according to the manual using the procollagen type I Cpepited EIA kit (MK101, Takara Bio inc, Shiga, Japan). After mixing 20 μl of cell culture solution and 100 μL of secondary antibody in a 96 well plate with primary collagen antibody, incubate at 37° C. for 3 hours, wash 4 times with PBS, and then 450 using an ELISA reader (BMG Labtech). Absorbance was measured in nm.

The results are shown in FIG. 2 .

As shown in FIG. 2, procollagen synthesis experiment was performed using polysaccharides treated with enzyme-treated white wood ear mushroom, and in the samples treated with Viscozyme, SUMIZYME, and Ultimase, procollagen synthesis was increased from 62.5 μg/mL concentration, and Lyvarome was treated. In one extract and in the sample not treated with the enzyme, procollagen synthesis was significantly increased from the concentration of 125 μg/mL. From the above results, it can be seen that the enzyme-treated extract of white wood ear mushroom of the present invention exhibits wrinkle improvement effect by promoting procollagen synthesis to make elastic skin.

Example 1-4: Preparation of a food composition comprising a white wood ear mushroom enzyme-treated polysaccharide extract and white wood ear mushroom whole powder

Among the enzyme-treated polysaccharide extracts prepared in Example 1-1, the Viscozyme-treated extract (Preparation Example 1-3), which had the best effect, and the white wood ear mushroom were hot water extracted and filtered, and the remaining white wood ear mushroom was dried and finely pulverized. The prepared powder was mixed in a weight ratio of 1:10 to prepare a food composition (Preparation Example 1-5) containing the white wood ear mushroom extract and the white wood ear mushroom whole powder. The extract that did not contain the white oyster mushroom powder was named Comparative Example 1-1 for the extract not treated with the enzyme, and Comparative Example 1-2 for the extract treated with the enzyme.

division SUMIZYME Ultimases Viscozyme Lyvarome A5 Whole White Wood Ear Mushroom Powder Comparative Example 1-1 - - - - unadded Comparative Example 1-2 - - 1% (w/v) - unadded Preparation 1-5 - - 1% (w/v) - adding

Example 1-5: Sensory test of a food composition comprising an enzyme-treated polysaccharide extract of a white wood ear mushroom and whole powder of a white wood ear mushroom

A beverage was prepared by taking a predetermined amount of each of the samples of Preparation Example 1-5, Comparative Example 1-1, and Comparative Example 1-2, and the odor, taste, and overall preference of the prepared beverage were evaluated. The sensory inspector finally selected 10 people through a preliminary experiment from among those who received identification training in advance, and the sensory evaluation items were sweetness, flavor, texture and overall preference. The degree of weakness was graded 1, and the degree of strength was graded 5.

The results are shown in Table 3.

division sweetness incense texture full symbol Comparative Example 1-1 2.8 3.1 2.88 2.93 Comparative Example 1-2 3.25 3.4 2.50 3.05 Preparation 1-5 3.19 3.5 4.13 3.61

In the case of sweetness, it was confirmed that the sweetness was strengthened in Comparative Examples 1-2 and Preparation Examples 1-5 treated with enzymes, and the flavor was improved in Comparative Examples 1-2 and Preparation Examples 1-5 treated with enzymes. confirmed that. However, in the case of texture, as a result of comparing Comparative Example 1-1 and Comparative Example 1-2 in which the enzyme was treated, it was confirmed that the texture was reduced in Comparative Example 1-2 in which the enzyme was treated. However, it was confirmed that the texture was greatly improved in Preparation Examples 1-5 in which the whole white wood ear mushroom powder was added. Therefore, as for the overall acceptability, Comparative Example 1-2 treated with the enzyme was evaluated better than Comparative Example 1-1, and it was found that the overall acceptability of Preparation Example 1-5 in which the enzyme treatment and whole white wood ear mushroom powder was added was the best.

As described above, by treating the enzyme, the enzyme hydrolyzes the components such as cellulose and hemicellulose contained in the mushroom, and the effect of improving the taste and flavor can be obtained. When added by recycling, it was possible to obtain the effect of improving the overall taste by greatly improving the texture.

Example 2: Lactic acid bacteria and complex enzyme treatment

Example 2-1: Preparation of Fermented Collagen Peptide Composition for White Wood Ear Mushroom

To obtain a fruiting body of the white wood ear mushroom grown in Deogyusan, it was ground finely and pulverized to a particle size of 10-100 mesh or less. 600 g of purified water was poured into 50 g of the pulverized dried oyster mushroom fruiting body powder, and the mixture was well mixed. A complex enzyme (SUMIZYME, Ultimase, Viscozyme, or Lyvarome A5) was added in a 2% portion (1 g) of dry oyster mushroom powder to the thoroughly mixed solution. In addition, Lactobacillus alimentarius Reuter (Lactobacillus alimentarius Reuter, ATCC 29643) was added as a lactic acid strain. The enzyme and lactic acid strain were added and cultured at 45° C. for 20 days. During the culture, the pH was adjusted so that the pH of the culture solution was 6 to 8. After the culture was completed, the culture solution was filtered to remove sludge. The filtered filtrate was collected and heated at 95° C. for 1 hour to sterilize the filtrate. After the sterilization was completed, the filtrate was concentrated under reduced pressure at 50-65° C. and then freeze-dried to obtain 8 g of a powdery white wood ear mushroom fermented collagen peptide composition.

Example 2-2: Peptide Amino Acid Content and Component Analysis of Fermented Collagen Peptide Composition of White Wood Ear Mushroom

The white wood ear mushroom fermented collagen peptide composition of the present invention prepared in Example 2-1 was diluted to a concentration of 1 wt%, and the content and components of peptide amino acids constituting it were analyzed. The results are shown in Table 4 below.

amino acid Molecular Weight Concentration (ppm) Aspartic acid 133.1 305.2 Glutamic acid 147.1 315.5 Serine 105.1 183.4 Histidine 155.2 0 Glycine 75.1 114.5 Threonine 119.1 182.1 Arginine 174.2 1372.0 Alanine 89.1 205.4 Tyrosine 181.2 82.3 Cystine 121.0 0 Valine 117.1 137.5 Methionine 149.2 0 Tryptophan 204.2 0 Phenylalanine 165.2 100.7 Isoleucine 130.0 113.8 Leucine 131.2 160.8 Lysine 145.2 177.7 Proline 115.0 336.7 Total amino acids - 3787.5

As shown in Table 4, it was confirmed that the amino acid content of the fermented white wood ear mushroom collagen peptide composition of the present invention was 3,787.5 mg/kg based on 1wt% concentration, and it was confirmed that it contained a very high content of amino acids. In particular, it contains 1372 mg/kg of Arginine, which is known to help improve blood flow by expanding blood vessels, and can be usefully used as a functional food for improving blood circulation. As a result, it was found that the potential for use as a composition for skin moisturizing and wrinkle improvement was high.

Example 2-3: Preparation of fermented Collagen Beverage with White Wood Ear Mushroom

3 g of the white wood ear mushroom fermented collagen peptide composition powder prepared in Example 2-1 and 200 ml of distilled water were mixed and stirred well. Even without heating after stirring, a viscous collagen beverage was prepared immediately, and a fermented beverage was prepared by adding syrup to adjust the sugar content to about 9 brix.

Example 2-4: sensory evaluation

Sensory evaluation was performed on the fermented collagen beverage prepared in Example 2-3. For sensory evaluation, 20 skilled agents (25-40 years old, female) were selected, the purpose of the experiment and evaluation items were explained, and the training process was conducted more than 10 times. The degree of preference for color, taste, and fragrance was evaluated using a five-point scale in which scores ranged from 1 (very bad) to 5 (very good). As a comparative example, 3 g of powder obtained by pulverizing the dried fruiting body of white wood ear mushroom into 10-100 mesh particles was put in water, boiled for 4 hours, and then syrup was added to adjust the sugar content to about 9 brix was used.

As a result, as shown in Table 5 below, the fermented white wood ear mushroom fermented beverage in which the enzyme and lactic acid strain of the present invention were cultured had the best overall preferences, such as sweetness and flavor. In particular, the fermented beverage of the present invention had more sweetness and superior color and flavor compared to the comparative example in which lactic acid bacteria and enzymes were not treated, despite the same sugar content.

strain used Enzyme used color sweetness Sour taste incense overall sign Lactobacillus alimentarius Reuter SUMIZYME 3.3 3.6 3.0 3.7 3.7 Ultimases 3.2 3.5 3.2 3.5 3.5 Viscozyme 3.3 3.3 3.2 3.6 3.4 Lyvarome A5 3.1 3.5 3.1 3.4 3.4 Comparative Example 2 (-) - 2.9 3.0 3.2 2.2 2.6

Example 2-5: Preparation of fermented collagen cosmetic composition for white oyster mushroom

A nutrient lotion containing 0.5% of the white wood ear mushroom fermented collagen peptide composition prepared in Example 2-1 based on the total weight was prepared at the component ratios shown in Table 6 below.

ingredient content (g) White Wood Ear Mushroom Fermented Collagen Peptide 0.5 glycerin 4.8 1,3-butylene glycol 3.0 PEG 1500 1.0 allantoin 0.1 DL-Panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Sodium Hyaluronate 5.0 ethanol 10.0 Octyldodeces-16 0.2 Polysorbate 20 0.2 Uniside-You 13 0.01 Distilled water remaining amount Sum 100

Example 2-6: Comparison of transdermal moisture loss, skin moisture content, skin elasticity, fine wrinkles improvement effect

The amount of transdermal moisture loss, the amount of skin moisture, the elasticity of the skin, and the effect of improving fine wrinkles were measured for the cosmetic composition containing the fermented collagen of the white wood ear mushroom of the present invention prepared in Examples 2-5.

Specifically, 50 women 19 years of age or older were selected as test subjects, and Examples 2-5 were typically applied to the skin of the test subject's face and forearm for 4 weeks, 8 weeks, and 12 weeks respectively. After application to the subjects, the applied skin condition was measured using anterior ocular imaging, Antera 3D imaging, Corneometer, Spectrophotometer CM2500d, and Skin-Visiometer. The results were averaged and shown in Table 7 below.

Experimental items before treatment after 4 weeks after 8 weeks after 12 weeks transdermal water loss 33.1±10.3 31.6±7.14 29.6±7.6 26.6±6.6 skin moisture content 64.1±4.9 65.1±5.1 67.2±5.1 68.5±4.6 skin elasticity 0.82±0.09 0.62±0.05 0.53±0.03 0.06±0.1 fine wrinkles 66.3±1.5 - - 65.9±1.2 skin texture 7.2±1.6 6.3±1.1

Referring to Table 7 above, when the cosmetic containing the fermented collagen of the white wood ear mushroom of the present invention prepared in Examples 2-5 according to the present invention is used, the amount of transdermal water loss is reduced, the amount of skin moisture is increased, the skin elasticity is increased, and the fine It can be seen that the effect of improving wrinkles and skin texture is excellent.

Example 2-7: Panel evaluation

The feeling of use was evaluated by using 30 women in their 30s and 40s as a panel. The cosmetic composition prepared in Example 2-5 was used as a test group, and a powder obtained by pulverizing the dried white wood ear mushroom fruiting body into 10-100 mesh particles was put in water, boiled for 4 hours, concentrated to dryness, and then powdered. , a cosmetic composition prepared at the same concentration as in Example 5 was used as a comparative example. Each sample was applied to the face 3 times a week for 2 weeks, and the skin cosmetic effect was measured on a 5-point scale for each item according to the items shown in Table 8 below, and the results are representatively shown.

evaluation item Example 5 comparative example overall sign 4.3 3.7 Skin feel after use 4.3 3.6 Spreadability when applied to the skin 4.2 3.6 Degree of skin irritation after use 0.5 1.1

As shown in Table 8 above, when the cosmetic containing the fermented collagen of the white wood ear mushroom of the present invention prepared in Examples 2-5 according to the present invention was used, skin irritation was low, overall preference and skin feel were excellent after use, In the case of Comparative Example 2 of the present invention, compared to Examples 2-5, it was confirmed that the skin irritation was large and the feeling of use was also poor.

Example 3

Example 3-1: Preparation of extracts (Preparation Examples 3-1 to 3-16) containing vegetable collagen and vegetable mucin using plant herbal materials

Spinach, Swiss chard, carrot, tomato, astragalus, chrysanthemum, wood ear mushroom, white wood ear mushroom, round hemp, lotus root, rape flower, rapeseed, pomegranate, tart cherry, blueberry, garlic, mango, orange, avocado, white lily, Powder of cashew nut, hibiscus, strawberry, goldfish, kiwi, guava, pineapple, soybean, green pepper, raspberry, blackberry, goji berry, bokryeong, and succulent yellow is immersed in 5-10% citric acid, acetic acid or dilute hydrochloric acid for 2 hours. After neutralization, 50 g of a solvent (purified water or 20% ethanol aqueous solution) as shown in Table 9 below was added and mixed well. The mixture was stirred at 70° C. for 1 hour so that the vegetable collagen and vegetable mucin contained in the herbal powder could be well extracted. The stirring process after the mixing was performed under high pressure conditions of 3 atmospheres.

0.1 g of a complex enzyme (SUMIZYME, Ultimase, Viscozyme, or Lyvarome A5; denoted as S, U, V, and L, respectively), lactic acid bacteria ( Lactobacillus alimentarius Reuter ATCC 29643, 1x10 ³ cfu/ml), or these was added and incubated at 37°C for 48 hours. The mixture was stirred once every 12 hours of incubation. During the culture, the pH was adjusted so that the pH of the culture solution was 6 to 8. After the culture was completed, the culture solution was centrifuged to remove the precipitated sludge and starch, and the viscous supernatant was separated. The separated viscous supernatant was heated at 130° C. for 2 seconds to inactivate the enzyme and sterilize the lactic acid bacteria. The viscous supernatant, which has been deactivated and sterilized, was filtered to obtain a liquid extract or a powdery extract by concentrating and freeze-drying it under reduced pressure.

production example Herbal medicine powder (g) Solvent (50 g) Enzyme (0.1 g) Lactobacillus strain (10 ³ cfu/ml) 3-1 5g Purified water SUMIZYME - 3-2 5g Purified water Ultimases - 3-3 5g Purified water Viscozyme - 3-4 5g Purified water Lyvarome A5 - 3-5 5g Purified water - O 3-6 5g Purified water - O 3-7 5g Purified water - O 3-8 5g Purified water - O 3-9 5g Purified water SUMIZYME O 3-10 5g Purified water Ultimases O 3-11 5g Purified water Viscozyme O 3-12 5g Purified water Lyvarome A5 O 3-13 5g 20% ethanol aqueous solution SUMIZYME - 3-14 5g 20% ethanol aqueous solution Ultimases - 3-15 5g 20% ethanol aqueous solution Viscozyme - 3-16 5g 20% ethanol aqueous solution Lyvarome A5 -

Of these, the extracts of Preparation Examples 3-1, 3-5, 3-9, and 3-13 were dried and powdery extracts were used in the following examples.

Examples 3-2 to 3-5 evaluated the efficacy as a food composition, and Experimental Examples 3-6 and 3-7 evaluated the efficacy as a cosmetic composition.

Example 3-2: Skin water retention, transepidermal water loss (TEWL) and erythema evaluation

Treatment of laboratory animals

The experimental group was a total of 6 groups (6 mice per group), a normal control group (normal, not UVB-induced), a negative control group (control, UVB-irradiated), Preparation Examples 3-1, 3-5, 3-9, and 3-13 of the treatment group.

As experimental animals, SKH-1 hairless mice (SkH: HR-1) were purchased from Orient Bio and were bred. The environment temperature of the animal breeding room was 22±2°C, the relative humidity was 50±10%, and the light and dark were adjusted in a 12-hour cycle. Drinking water and feed were ad libitum.

This experiment was conducted under the 'Guide for the Care and Use of Laboratory Animals (NRC)' after receiving approval from the Animal Experimental Ethics Committee of Korea University.

After the adaptation period for one week, UVB-ultraviolet lamp (T-8M; Vilber Lourmat, France) was attached to group 6 except for the normal control group (NOR) 3 times a week for 8 weeks (BLX-254; Vilber Lourmat, France) ) was used to induce photoaging by exposing the back skin to ultraviolet rays (Hong et al., 2015). The amount of UV irradiation was adjusted by measuring the dose inside the UV irradiator with a UV light meter (UV-340; Lutron, Taiwan), and then the UVB dose was adjusted as shown in Table 10 below.

Week One 2 3 4-8 UVB dosage 1 MED 2 MED 3 MED 4 MED

1 MED=75 mJ/cm ^{2 For} 8 weeks from the start date of UV irradiation, only drinking water was orally administered to the normal control group (NOR) and negative control group (CON). did Each mouse was sacrificed after 8 weeks of UV irradiation and the administration period of Preparation Example was completed.

<Drinking water intake and feed intake>

The present inventors measured the intake of water and feed once a week during the period of irradiating the mouse with UV light and administering the sample in order to confirm the effect of the composition of the present invention on the intake of water and feed of the mouse. There was no significant difference in dietary intake between groups (Tables 11 and 12).

division Feed intake (g/day) normal control group 14.6 negative control 17.2 Preparation Example 3-1 17.0 Preparation 3-5 16.5 Preparation 3-9 16.1 Preparation 3-13 18.3

division Water intake (mL/day) normal control group 14.5 negative control 16.3 Preparation Example 3-1 15.1 Preparation 3-5 12.8 Preparation 3-9 13.5 Preparation 3-13 17.6

<Evaluation of skin water retention, transepidermal water loss (TEWL) and erythema>

The present inventors used Multi Probe Adapter® MPA 6 (Courage und Khazaka, Germany) on the dorsal skin of rats irradiated with ultraviolet rays to confirm the effect of the composition of the present invention on the skin condition, the amount of moisture retention in the skin and the amount of transepidermal water loss ( TEWL) and erythema were measured. Skin measurements were carried out in all groups before the start of UV irradiation, and skin measurements were performed every week from 4 weeks after UV irradiation to confirm that there was a significant difference in skin measurement results between the NOR and CON groups. The skin measurement results are shown in Tables 13 to 15, showing the results of the last measurement on the day before sacrifice.

skin moisture retention

division skin moisture retention normal control group 43.1 negative control 27.5 Preparation Example 3-1 40.1 Preparation 3-5 39.8 Preparation 3-9 41.3 Preparation 3-13 38.6

Table 13 shows the amount of skin moisture retention for each experimental group of the present invention.

As shown in Table 13, water retention was significantly reduced in the negative control group (CON) compared to the normal control group (NOR) by about 36.2%, and water retention was increased in all sample administration groups compared to the negative control group.

Transepidermal water loss

division Transepidermal water loss (g/h/m ² ) normal control group 12.0 negative control 53.6 Preparation Example 3-1 23.3 Preparation 3-5 25.1 Preparation 3-9 19.8 Preparation 3-13 21.5

Table 14 shows the amount of transdermal water loss for each experimental group of the present invention.

As shown in Table 14, the amount of transepidermal water loss was significantly increased by about 4.47 times in the negative control group (CON) compared to the normal control group (NOR) (p<0.05). All sample administration groups significantly decreased transepithelial water loss compared to the negative control group (CON), and in particular, the greatest decrease in transepithelial water loss was achieved in Preparation Example 3-9 treated group, which was cultured by simultaneously treating enzymes and lactic acid bacteria. (p<0.05).

erythema level

division erythema level normal control group 142.1 negative control 224.5 Preparation Example 3-1 198.4 Preparation 3-5 192.7 Preparation 3-9 201.2 Preparation 3-13 195.2

As shown in Table 15, the erythema level in the negative control group (CON) showed a significant increase of about 1.58 times compared to the normal control group (NOR) (p<0.05), and the erythema level in the sample administration group was significant compared to the negative control group. decreased to

When the above results are taken together, it was confirmed that the extract containing vegetable collagen and vegetable mucin of the present invention exhibited positive activity in both water retention, transepidermal water loss, and erythema level. Therefore, it was found that the plant mixed extract of the present invention helps prevent skin moisture loss and damage to the skin barrier caused by UV rays, and helps the skin barrier function to function normally.

Example 3-3: Analysis of skin wrinkles

In order to confirm the anti-aging and anti-wrinkle effect of the food composition of the present invention, the present inventors photographed the dorsal skin of a rat and used a replica obtained using Visioline (VL650; CK electronic GmbH, Germany) to analyze the wrinkle pattern Thus, the area, number, length, and depth of wrinkles were analyzed. The results are shown in Table 16.

division Total crease area (mm ² ) Maximum wrinkle depth (μm) normal control group 4.7 221 negative control 23.8 728 Preparation Example 3-1 15.4 575 Preparation 3-5 13.7 560 Preparation 3-9 11.3 411 Preparation 3-13 12.4 432

As a result of analyzing the wrinkle pattern index through Table 16, the wrinkle area and depth of the negative control group (CON) were significantly increased compared to the normal control group (NOR) (p<0.05). In addition, when the extract containing vegetable collagen and vegetable mucin of the present invention was administered, it was confirmed that the index of the wrinkle pattern was significantly reduced compared to the negative control group (CON). In the group (Preparation Example 3-9), it was confirmed that the area and maximum depth of wrinkles were most significantly decreased (p<0.05).

Through this, it was confirmed that when the extract containing vegetable collagen and vegetable mucin of the present invention was ingested, there was an inhibitory effect on UVB-induced photoaging and wrinkle formation.

Example 3-4: Epidermal thickness and collagen measurement

In order to confirm the effect of the composition of the present invention on the collagen expression of the epidermis, the present inventors collected the dorsal skin of all mice after UV irradiation and the composition administration experiment was completed, put it in 10% formalin, and confirmed histopathological changes Hematoxylin and eosin (H&E) staining was performed, and immunohistochemistry (IHC) staining was performed to confirm collagen present in the dermis. After the stained tissue was imaged by taking a picture using an optical microscope, it was analyzed.

As shown in FIG. 4 , as a result of H&E staining, it was confirmed that the thickness of the epidermis of the UVB irradiation group was significantly increased (p<0.05) compared to the normal control group (NOR). In addition, in the total sample administration group, the epidermal thickness was significantly reduced compared to the negative control group (CON), and in particular, in the group cultured by treating enzymes and lactic acid bacteria at the same time (Preparation Example 3-9), the epidermal thickness was significantly higher than that of the negative control group was confirmed to show a decreasing trend (p<0.05).

division Epidermis thickness (μm) Collagen (%) normal control group 92 4.7 negative control 438 23.8 Preparation Example 3-1 210 15.4 Preparation 3-5 187 13.7 Preparation 3-9 143 11.3 Preparation 3-13 161 12.4

As a result of IHC staining for collagen, it was confirmed that the collagen ratio in the dermis of the negative control group (CON) was significantly decreased compared to the normal control group (NOR) (p<0.05). In addition, the collagen ratio in the dermis was significantly increased in the total sample administration group compared to the negative control group (CON) (p<0.05).

Experimental Example 3-5: Elastin measurement of the dermis

Elastin, a component of the skin dermis, is an elastic substance that affects the elasticity of the skin tissue. In order to confirm the effect of the composition of the present invention on skin elasticity, the present inventors extracted about 10 mg of skin tissue from each experimental group, and then used the method provided by Fastin Elastin assay (F2000; Biocolor, UK) to elastin in the skin tissue. The amount was measured.

division Elastin content (μg/mg) normal control group 1.4 negative control 0.9 Preparation Example 3-1 1.3 Preparation 3-5 1.5 Preparation 3-9 1.6 Preparation 3-13 1.3

As shown in Table 18, the composition administration group of the present invention showed a tendency to increase the elastin content in all compared to the negative control group (CON), in particular, the group cultured by treating the enzyme and lactic acid bacteria of the present invention at the same time (Preparation Example) 3-9) showed the most significant increase compared to the negative control group (CON) (p<0.05).

From the above results, when the plant extract containing vegetable collagen and vegetable mucin of the present invention is orally ingested, it is confirmed that the elasticity enhancing effect can be expected by enhancing the elastin content reduced by UV and improving the dermal layer network structure of the skin. did

Experimental Example 3-6: Effect of increasing total amount of collagen in human fibroblasts

The extracts of Preparation Examples 3-1, 3-5, 3-9, and 3-13 of the present invention were added to the culture solution of human-derived fibroblasts to measure the total amount of collagen at the cellular level. The total amount of collagen was quantified using the PICP EIA kit (Procollagen Type I C-Peptide Enzyme ImmunoAssay KIT). Before the experiment, human-derived fibroblasts were evaluated for cytotoxicity at concentrations of 10 ppm, 1 ppm, 0.1 ppm, 0.01 ppm, and 0.001 ppm of the test substance, and the total amount of collagen was measured by selecting a concentration without cytotoxicity. In the experiment, the concentrations of the preparation examples of the present invention were 1 ppm and 10 ppm, respectively, and each sample was added to the culture medium of human fibroblasts and cultured for 2 days, then the culture solution was taken and the total amount of collagen at each concentration with the PICP EIA kit. was measured at 450 nm using a spectrophotometer. For comparison of the effect, the total amount of collagen was measured in the same manner for the fibroblast culture medium (control group) to which nothing was added and the sample to which vitamin C was added to a final concentration of 52.8 ppm. The total amount of collagen was measured as UV absorbance, and the increase rate of the total amount of collagen was calculated as the ratio of the total amount of collagen relative to the control group, and the results are summarized in Table 19 below.

division Absorbance Average Collagen increase rate (%) normal control group 612 145.3 negative control 421 - Preparation Example 3-1 527 125.2 Preparation 3-5 510 121.1 Preparation 3-9 572 135.9 Preparation 3-13 533 126.6 vitamin C 586 139.2

As shown in Table 19, the extract of Preparation Example of the present invention generally exhibited an excellent collagen increase rate at a lower concentration than when vitamin C, which is known to have a collagen synthesis ability, was applied. Therefore, it was found that the extract of Preparation Example of the present invention can be used for skin regeneration and wrinkle improvement.

Experimental Example 3-7: Collagenase activity inhibitory effect

The effect of inhibiting collagenase activity in human-derived fibroblasts by the extracts of Preparation Examples 3-1, 3-5, 3-9, and 3-13 of the present invention was confirmed as follows.

Before the experiment, human-derived fibroblasts were evaluated for cytotoxicity at concentrations of 10 ppm, 1 ppm, 0.1 ppm, 0.01 ppm, and 0.001 ppm of the test substance, and a collagenase evaluation method was performed by selecting a concentration without cytotoxicity. Fibroblasts, which are normal human skin cells, ^{were inoculated in a 24-well microplate so as to become 2.5x10 4} cells per well, and cultured for 24 hours in 10% serum DMEM medium and 37° C., and then 10% serum DMEM medium was removed. After washing once with a phosphate buffer solution, it was further cultured for 30 minutes in serum-free DMEM medium to which the extract of Preparation Example of the present invention was added, and in serum-free DMEM medium as a normal control group and a negative control group. After 30 minutes of sample treatment, the cells were stimulated with 50 ng/mL of TNF-α (tumor necrosis factor-α), a substance known to produce MMP-1, a collagen degrading enzyme, and cultured for 24 hours. At this time, among the controls not containing the extract of the present invention, the group treated with TNF-α was defined as a negative control group, and the group not treated with TNF-α was defined as a normal control group. The supernatant of each well was collected and the amount of newly synthesized MMP-1 (ng/mL) was measured using an MMP-1 assay kit (Amersham, USA), and the collagenase activity inhibition rate was determined according to Equation 1 below. -1 production inhibition rate (%) was calculated, and the results are shown in Table 20 below.

[Equation 1]

MMP-1 production inhibition rate (%) = [1 - (Experimental group MMP-1 production amount - Negative control group MMP-1 production amount) / (Normal control group MMP-1 production amount - Negative control group MMP-1 production amount)]x100

division MMP-1 production Inhibition rate (%) Normal control group (no TNF-alpha treatment) 5.6 - Negative control group (TNF-alpha treatment) 20.4 - Preparation Example 3-1 15.8 125.2 Preparation 3-5 16.7 121.1 Preparation 3-9 8.5 135.9 Preparation 3-13 11.3 126.6

Formulation example

On the other hand, the formulation examples of the composition comprising the extract of Preparation Example of the present invention are described below. However, it is apparent to those skilled in the art that the following formulation examples are only intended to specifically describe examples of use of the present invention, and are not intended to limit the scope of the present invention to the following formulation examples.

Formulation Example 1: Preparation of nutritional lotion (lotion)

As shown in Table 21 below, a nutrient lotion (lotion) containing the extract obtained in Preparation Examples 3-1 to 3-16 was prepared according to a conventional method.

number Raw material content (wt%) One Preparation Examples 3-1 to 3-16 10.0 2 sitosterol 1.7 3 Polyglyceryl 2-oleate 1.5 4 Ceteares-4 1.2 5 cholesterol 1.5 6 dicetyl phosphate 0.4 7 glycerol 5.0 8 Carboxyvinyl Polymer 10.0 9 sunflower oil 0.2 10 Xanthan Gum 0.3 11 antiseptic a very small amount 12 Spices a very small amount 13 Purified water remaining amount

Formulation Example 2: Preparation of softening lotion (skin)

As shown in Table 22 below, a softening lotion (skin) containing the extract obtained in Preparation Examples 3-1 to 3-16 was prepared according to a conventional method.

number Raw material content (wt%) One Preparation Examples 3-1 to 3-16 5.0 2 glycerin 3.0 3 butylene glycol 2.0 4 propylene glycol 2.0 5 Polyoxyethylene (60) hydrogenated castor oil 1.0 6 ethanol 10.0 7 preethanolamine 0.1 8 antiseptic a very small amount 9 pigment a very small amount 10 Spices a very small amount 11 Purified water remaining amount

Formulation Example 3: Preparation of nourishing cream

As shown in Table 23 below, a nutritional cream containing the extract obtained in Preparation Examples 3-1 to 3-16 was prepared according to a conventional method.

number Raw material content (wt%) One Preparation Examples 3-1 to 3-16 10.0 2 sitosterol 4.0 3 Polyglyceryl 2-oleate 3.0 4 Ceteares-4 2.0 5 cholesterol 3.0 6 dicetyl phosphate 0.4 7 concentrated glycerol 5.0 8 sunflower oil 22.0 9 Carboxyvinyl Polymer 0.5 10 triethanolamine 0.5 11 antiseptic a very small amount 12 Spices a very small amount 13 Purified water remaining amount

Formulation Example 4: Preparation of Essence

As described in Table 24 below, an essence containing the extract obtained in Preparation Examples 3-1 to 3-16 was prepared according to a conventional method.

number Raw material content (wt%) One Preparation Examples 3-1 to 3-16 5.0 2 sitosterol 1.7 3 Polyglyceryl 2-oleate 1.5 4 Ceteares-4 2.0 5 cholesterol 3.0 6 dicetyl phosphate 0.4 7 concentrated glycerol 5.0 8 sunflower oil 22.0 9 Carboxyvinyl Polymer 0.5 10 triethanolamine 0.5 11 antiseptic a very small amount 12 Spices a very small amount 13 Purified water remaining amount

Formulation Example 5: Preparation of foundation

As shown in Table 25 below, a foundation containing the extract obtained in Preparation Examples 3-1 to 3-16 was prepared according to a conventional method.

number Raw material content (wt%) One Preparation Examples 3-1 to 3-16 1.0 2 beeswax 2.0 3 cyclomethicone 2.0 4 liquid paraffin 5.0 5 squalane 5.0 6 stearic acid 2.0 7 lipophilic monostearate glycerin 3.0 8 Caprylic/Capric Triglycerides 4.0 9 glycerin 4.0 10 propylene glycol 3.0 11 butylene glycol 3.0 12 triethanolamine 1.0 13 Aluminum Magnesium Silicate 0.5 14 pigment 12 15 antiseptic a very small amount 16 Spices a very small amount 17 Purified water remaining amount

Formulation Example 6: Preparation of hair conditioner

As shown in Table 26 below, hair conditioners containing the extracts obtained in Preparation Examples 3-1 to 3-16 were prepared according to a conventional method.

number Raw material content (wt%) One Preparation Examples 3-1 to 3-16 5.0 2 ethanol 3.5 3 glycerin 1.5 4 propylene glycol 2.5 5 Stearyl Triethine Ammonium Chloride 2.0 6 pigment a very small amount 7 Spices a very small amount 8 Purified water remaining amount

Claims (20)

A food composition for skin aging prevention, wrinkle improvement, skin barrier strengthening, or skin moisturizing comprising a plant extract containing vegetable collagen and plant mucin as an active ingredient. According to claim 1, wherein the plant is spinach, Swiss chard, carrot, tomato, astragalus, chrysanthemum, mushrooms, round hemp, lotus root, rape flower and rapeseed extract a mixture of one or more kinds of herbal ingredients selected from the group consisting of extract A food composition obtained by According to claim 2, wherein the food composition is pomegranate, tart cherry, blueberry, garlic, mango, orange, avocado, white lily, cashew nut, hibiscus, strawberry, goldfish, kiwi, guava, pineapple, soybean, green pepper, raspberry , blackberry, goji berry, bokryeong, and to further include an extract obtained by extracting a mixture of one or more herbal ingredients selected from the group consisting of sukjihwang, a food composition. According to claim 2, wherein the mushrooms are white wood ear mushroom, wood ear mushroom, reishi mushroom, chima mushroom, matsutake mushroom, shiitake mushroom, truffle, situation mushroom, horseshoe mushroom, pineal mushroom, chaga mushroom, neungi mushroom, enoki mushroom, Oyster mushroom, and at least one mushroom selected from the group consisting of blackcurrant, a food composition. The food composition according to claim 1, wherein the composition increases the expression of collagen and elastin in the skin. A cosmetic composition for skin anti-aging, wrinkle improvement, skin barrier strengthening, or skin moisturizing comprising a plant extract containing vegetable collagen and plant mucin as an active ingredient. A method for producing a plant extract comprising plant collagen and plant mucin comprising the steps of: (a) mixing a solvent in the herbal herbal material powder; (b) adding an enzyme, lactic acid bacteria, or a combination thereof to the mixed solution and culturing; (c) centrifuging or filtering the culture solution to remove sludge and starch and separating the viscous supernatant, or preparing a filtrate; and (d) heating the viscous supernatant or the filtrate to inactivate and sterilize the enzyme, lactic acid bacteria, or a combination thereof. The method according to claim 7, wherein the method for preparing the extract further comprises the step of (e) obtaining a powdery extract by drying the liquid extract or the same. The method according to claim 7, wherein the herbal material is at least one selected from the group consisting of spinach, Swiss chard, carrot, tomato, astragalus, chrysanthemum, mushrooms, round hemp, lotus root, rape flower and rapeseed. 10. The method of claim 9, wherein the herbal material is pomegranate, tart cherry, blueberry, garlic, mango, orange, avocado, white lily, cashew nut, hibiscus, strawberry, goldfish, kiwi, guava, pineapple, soybean, green pepper, raspberry , blackberry, goji berry, bokryeong, and the method for producing an extract further comprising at least one selected from the group consisting of sukhwanghwang. The method according to claim 7, wherein in step (a), the solvent is added in an amount of 1 to 100 parts by weight based on 1 part by weight of the plant herbal material powder. The method of claim 7, wherein in step (a), a solvent is added to the plant herbal material powder, and the process is pressurized at 0.2 to 5 atmospheres. The method of claim 7, wherein the enzyme in step (b) is cellulase, hemicellulase, beta-glucosidase, beta-glucanase, xylanase, arabinase, protease, and one selected from the group consisting of amylase The above enzyme, the manufacturing method. The method according to claim 7, wherein the lactic acid bacteria of step (b) are Leukonostok genus, Lactobacillus genus, Bifidobacterium genus, Enterococcus spp. strain, Lactococcus genus, or a combination thereof. The method according to claim 7, wherein the culturing in step (b) is made for 1 to 50 days. The method according to claim 7, wherein the heating in step (d) is performed at 60-150° C. for 0.5 seconds to 3 hours. The method according to claim 16, wherein the heating in step (d) is performed at 61-65°C for 30 minutes to 1 hour, or at 95-150°C for 0.5 seconds to 30 seconds. The method according to claim 7, wherein the plant herbal material powder of step (a) is treated with an acid selected from citric acid, acetic acid, or dilute hydrochloric acid at a concentration of 1 to 80% (w/w) for 1 to 5 hours, manufacturing Way. The method according to claim 18, comprising neutralizing after the acid treatment. (a) to (d) of claim 7, or to the extract prepared by the manufacturing method comprising the step (a) to (e) of claim 8, the precipitate remaining after centrifugation or filtration of step (c), Or drying and pulverizing the filtrate, comprising the step of mixing the powder prepared, a method for producing a plant composition comprising an extract comprising vegetable collagen and plant mucin and whole powder of plant herbal material.

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