Collagen peptide absorption-promoting composition and method for producing same

Abstract

To solve the problem in which: collagen peptides may not be fully absorbed when taken orally, requiring consumption in large quantities to achieve significant effects.SOLUTION: According to the present invention, a product derived from the fermentation of soy peptides with lactic acid bacteria can promote the absorption of collagen peptides, thus reducing the quantity of collagen peptides to be consumed for achieving significant effects.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a collagen peptide absorption-promoting composition and a method for producing the same. [Background technology]

[0002] Collagen is a protein found in large amounts in animal skin, bones, cartilage, etc. In the dermis layer of the skin, collagen forms a matrix structure together with hyaluronic acid and chondroitin sulfate. Therefore, collagen is one of the important components that play a role in supporting cells and skin tissue, retaining moisture in intercellular spaces, maintaining skin lubrication and flexibility, and protecting skin tissue from external factors such as ultraviolet rays, dry environments, mechanical irritation and damage, and microbial infection.

[0003] After oral ingestion, collagen is broken down into dipeptides and tripeptides by digestive enzymes in the body and absorbed into the bloodstream from the small intestine. Therefore, oral ingestion of collagen peptides, which are collagen hydrolysates, is expected to provide the beneficial effects of collagen.

[0004] Peptides containing hydroxyproline (Hyp), a characteristic amino acid that constitutes collagen, (peptide-type Hyp) have been shown to be an indicator for observing the dynamics of digestion and absorption of collagen peptides after oral ingestion (Non-Patent Documents 1-3). When observing collagen peptide absorption by measuring the transfer of peptide-type Hyp into the blood, no significant absorption was observed at 2 g of collagen peptide / 65 kg body weight, but transfer of peptide-type Hyp into the blood was observed at 10 g / 65 kg body weight (Non-Patent Document 2). This suggests that collagen peptides are poorly absorbed when ingested orally, and therefore that oral administration of collagen peptides will not achieve sufficient effects unless large amounts are ingested as is. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Journal of the Japanese Cosmetics Society Vol. 44, No. 4, pp.308-313(2020) [Non-patent document 2] Journal of the Japanese Society for Food and Nutrition Studies, Vol. 27, No. 3, pp. 147-152 (2016) [Non-patent document 3] Leather Science Vol.56, No. 2, pp.71-79(2010) Summary of the Invention [Problem to be solved by the invention]

[0006] Therefore, there is a need to enhance the absorption of collagen peptides, thereby reducing the amount of collagen peptide that needs to be ingested to achieve the desired effect. Furthermore, because collagen is used in various parts of the body, it is necessary to ensure that ingested collagen produces the desired effects. [Means for solving the problem]

[0007] The present inventors have found that the soybean peptide fermentation product of the present invention promotes the absorption of collagen peptides. Furthermore, the present inventors have found that the composition of the present invention, which contains the soybean peptide fermented product of the present invention and collagen peptide, improves skin condition.

[0008] The present invention includes the following aspects 1 to 5: [Aspect 1] A collagen peptide absorption-promoting composition containing a soybean peptide fermented by lactic acid bacteria; [Aspect 2] The collagen peptide absorption-enhancing composition according to Aspect 1, from which 70% or more of lactic acid bacteria have been removed; [Aspect 3] The collagen peptide absorption-enhancing composition according to Aspect 1 or 2, which is used by ingesting it together with, simultaneously with, before or after ingesting a collagen peptide or a collagen peptide-containing composition; [Aspect 4] A collagen peptide absorption-enhancing composition according to Aspect 1 or 2, used to improve skin condition; [Embodiment 5] A method for producing a collagen peptide absorption-promoting composition by mixing a soybean peptide fermented by lactic acid bacteria. [Embodiment 6] A skin condition improving composition containing a soybean peptide fermented by lactic acid bacteria and a collagen peptide. [Effects of the Invention]

[0009] The soybean peptide fermentation product of the present invention can promote the absorption of collagen peptides, thereby improving the moisture content and / or elasticity of the skin and improving skin condition. [Brief explanation of the drawings]

[0010] [Figure 1] 1 is a graph plotting the average amount of peptide-type Hyp in the plasma of subjects against the time elapsed after ingestion of the collagen drinks of Example 1, Example 2, and Comparative Example 1. [Figure 2] 1 is a graph plotting the average amount of Pro-Hyp in the plasma of subjects against the time elapsed after ingestion of the collagen drinks of Example 1, Example 2, and Comparative Example 1. [Figure 3] 1 is a graph plotting the average amount of Hyp-Gly in the plasma of subjects against the time elapsed after ingestion of the collagen drinks of Example 1, Example 2, and Comparative Example 1. [Figure 4] 1 is a graph plotting changes in the overall condition of the skin of subjects who took the collagen drinks of Example 3 and Comparative Example 2. [Figure 5] 1 is a graph plotting changes in the dryness of the skin of subjects who took the collagen drinks of Example 3 and Comparative Example 2. [Figure 6] 1 is a graph plotting changes in skin elasticity of subjects who took the collagen drinks of Example 3 and Comparative Example 2. [Figure 7]1 is a graph plotting changes in the overall condition of the skin of subjects who took the collagen drinks of Example 4, Comparative Example 3, and Comparative Example 4. [Figure 8] 1 is a graph plotting changes in the dryness of the skin of subjects who took the collagen drinks of Example 3, Comparative Example 3, and Comparative Example 4. [Figure 9] 1 is a graph plotting changes in skin elasticity of subjects who took collagen drinks of Example 3, Comparative Example 3, and Comparative Example 4. DETAILED DESCRIPTION OF THE INVENTION

[0011] Hereinafter, an embodiment of the present invention will be described in detail. In one embodiment of the present invention, a composition containing a soybean peptide lactic acid bacteria fermentation product of the present invention promotes collagen peptide absorption. Furthermore, a soybean peptide lactic acid bacteria fermentation product of the present invention from which 70% or more, for example 80% or more, or 90% or more of the lactic acid bacteria cells have been removed also promotes collagen peptide absorption in the same manner as before the lactic acid bacteria removal. Furthermore, a collagen peptide absorption-promoting composition of the present invention can be produced by mixing a soybean peptide lactic acid bacteria fermentation product or a soybean peptide lactic acid bacteria fermentation product from which lactic acid bacteria cells have been removed in any of the above proportions. The collagen peptide absorption-promoting composition of the present invention can promote the absorption of collagen peptide by taking it together with, at the same time as, before or after taking collagen peptide itself or a collagen peptide-containing composition. The soybean peptide fermentation product of the present invention is produced by fermenting soybean peptides with lactic acid bacteria.

[0012] The peptides used to produce the peptide lactic acid bacteria fermentation product of the present invention are obtained, for example, by hydrolyzing proteins of animal, plant, or microbial origin with acid, alkali, or protease, and include, for example, collagen peptides, soybean peptides, milk peptides, wheat peptides, or egg peptides, preferably soybean peptides. The soybean peptides used in the production of the soybean peptide lactic acid bacteria fermentation product of the present invention are, for example, those obtained by enzymatic hydrolysis of soybean protein, and are preferably those sold by Fuji Oil Co., Ltd. as the HINUTE series, and more preferably HINUTE AM (Fuji Oil Co., Ltd.). The amount of soybean peptide used in the production of the soybean peptide lactic acid bacteria fermentation product of the present invention is 0.1 to 20%, preferably 2 to 10%, and more preferably 3 to 5%.

[0013] Lactic acid bacteria used in the production of the soybean peptide lactic acid bacteria fermentation product of the present invention include, for example, Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus panis, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus acidophilus, Lactobacillus brevis, Lactobacillus paracasei subsp. paracasei, Lactobacillus gasseri, Lactobacillus plantarum subsp. plantarum, Lactobacillus fermentum, Lactobacillus kunkii, Lactococcus lactis subsp. lactis, Lactobacillus helveticus, or Streptococcus thermophilus, preferably Lactobacillus plantarum or Lactobacillus paracasei, and particularly preferably Lactobacillus plantarum. In addition, multiple lactic acid bacteria may be used, for example, one or more species may be selected from those listed above.

[0014] An example of a strain of Lactobacillus plantarum is Lactobacillus plantarum FL-664. An example of a strain of Lactobacillus paracasei is Lactobacillus paracasei NBRC15889. Examples of Lactobacillus reuteri strains include Lactobacillus reuteri JCM1112, Lactobacillus reuteri JCM1081, and Lactobacillus reuteri JCM1084. An example of a strain of Lactobacillus panis is Lactobacillus panis JCM11053. An example of a strain of Lactobacillus delbrueckii subsp. bulgaricus is Lactobacillus delbrueckii subsp. bulgaricus JCM1002. An example of a strain of Lactobacillus acidophilus is Lactobacillus acidophilus JCM1034. An example of a strain of Lactobacillus brevis is Lactobacillus brevis JCM1061.

[0015] An example of a strain of Lactobacillus paracasei subsp. paracasei is Lactobacillus paracasei subsp. paracasei JCM1109. An example of a strain of Lactobacillus gasseri is Lactobacillus gasseri JCM1131. An example of a strain of Lactobacillus plantarum subsp. plantarum is Lactobacillus plantarum subsp. plantarum JCM1149. An example of a strain of Lactobacillus fermentum is Lactobacillus fermentum JCM1173. An example of a strain of Lactobacillus kunkeii is Lactobacillus kunkeii JCM16173. An example of a strain of Lactococcus lactis subsp. lactis is Lactococcus lactis subsp. lactis JCM5805. An example of a strain of Lactobacillus helveticus is the Lactobacillus helveticus FL-65 strain. An example of a strain of Streptococcus thermophilus is the Streptococcus thermophilus FL-176 strain. The lactic acid bacteria strain used for producing the soybean peptide lactic acid bacteria fermentation product of the present invention is preferably Lactobacillus plantarum FL-664 strain.

[0016] The lactic acid bacteria strains used in the production of the soybean peptide lactic acid bacteria fermentation product of the present invention can be obtained, for example, from public microorganism collection institutions in Japan or abroad, such as the RIKEN BioResource Center or the Patent Microorganisms Deposit Center of the National Institute of Technology and Evaluation.

[0017] A pH adjuster may be added to the lactic acid bacteria fermentation medium used to produce the soybean peptide lactic acid bacteria fermentation product of the present invention. pH adjusters are inorganic bases (e.g., sodium hydroxide), inorganic acid salts (e.g., potassium carbonate), organic acid salts (e.g., trisodium citrate), or organic acids (e.g., lactic acid, citric acid, malic acid, or acetic acid). The amount of pH adjuster added to the medium is the amount necessary to adjust the pH of the medium to a pH range suitable for lactic acid bacteria fermentation (e.g., pH 3 to 9).

[0018] A flavoring may be added to the lactic acid bacteria fermentation medium used to produce the soybean peptide lactic acid bacteria fermentation product of the present invention. The flavoring may be, for example, a fruit flavor (e.g., apple, strawberry, grape, muscat, peach, or banana), a citrus flavor (lemon, orange, or lime), or a vanilla, cocoa, black tea, or coffee flavor. The type and amount of flavoring are appropriately selected depending on the desired flavor.

[0019] Sugars may be added to the lactic acid bacteria fermentation medium used to produce the soybean peptide lactic acid bacteria fermentation product of the present invention. Examples of the added sugars include glucose, sucrose, fructose, and maltose. The added sugars may be one type or a mixture of multiple types (e.g., a mixture of liquid sugar and starch syrup). Alternatively, sugars may be added in the form of a food product with a high sugar content (e.g., concentrated fruit juice). The amount of sugar added to the lactic acid bacteria fermentation medium used to produce the soybean peptide lactic acid bacteria fermentation product of the present invention is, for example, 0.1 to 6%, preferably 1 to 6%, and more preferably 2.5 to 4.5%, but is not limited to these amounts. The sugar added to the lactic acid bacteria fermentation medium used to produce the soybean peptide lactic acid bacteria fermentation product of the present invention is preferably GluFinal (Sanei Sugar Corporation) or HyMaltose M70-75C (Nippon Corn Starch Co., Ltd.). The amount of sugar added to the lactic acid bacteria fermentation medium used to produce the soybean peptide lactic acid bacteria fermentation product of the present invention is, for example, 0.1 to 6%, preferably 1 to 6%, and more preferably 2.5 to 4.5%, but is not limited to these amounts.

[0020] Amino acids may be added to the lactic acid bacteria fermentation medium used to produce the soybean peptide lactic acid bacteria fermentation product of the present invention. Examples of the amino acids added include alanine, arginine, aspartic acid, cysteine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, asparagine, and glutamine. The amino acids added may be one type or a mixture of multiple types. Alternatively, they may be added as an amino acid-containing food, such as yeast extract. The yeast extract is preferably Yeast Extract SL-W (Mitsubishi Corporation Life Sciences). The yeast extract may also be added in the form of a mixture with other nutrients. When yeast extract is added, the amount of yeast extract is, for example, 0.1 to 6%, preferably 0.1 to 3%, and more preferably 0.1 to 1%, but is not limited to these amounts.

[0021] The lactic acid bacteria fermentation medium used to produce the soybean peptide lactic acid bacteria fermentation product of the present invention may contain nutrients such as vitamins (e.g., biotin, folic acid, nicotinic acid, pantothenic acid, pyridoxal, riboflavin, thiamine, vitamin B12), minerals (e.g., manganese, magnesium, iron), and / or fatty acids (e.g., oleic acid). These nutrients may be added alone or as a food product containing these nutrients. An example of the nutrient to be added is Yeast Rich Manganese (Oriental Yeast Co., Ltd.).

[0022] In the production of the soybean peptide lactic acid bacteria fermentation product of the present invention, the fermentation time of the lactic acid bacteria is, for example, but not limited to, about 8 to about 108 hours, preferably about 12 to about 96 hours, and more preferably about 16 to about 72 hours. The fermentation time of the lactic acid bacteria is appropriately selected depending on the raw material, the microorganisms used, the fermentation temperature, the desired flavor, etc.

[0023] In the production of the soybean peptide lactic acid bacteria fermentation product of the present invention, the fermentation temperature of the lactic acid bacteria is, for example, but not limited to, 20 to 45° C., preferably 25 to 43° C., and more preferably 30 to 40° C. The fermentation temperature of the lactic acid bacteria is appropriately selected depending on the raw materials, the microorganisms used, the fermentation time, the desired flavor, etc.

[0024] The pH at the start of lactic acid bacteria fermentation in producing the soybean peptide lactic acid bacteria fermentation product of the present invention is, but is not limited to, for example, pH 3 to 9, preferably pH 4 to 8, and more preferably pH 5 to 7. The pH during lactic acid bacteria fermentation is appropriately selected depending on the raw materials, the microorganisms used, the fermentation temperature, the desired flavor, etc.

[0025] To obtain a product from which 70% or more, for example 80% or more, or 90% or more of the lactic acid bacteria cells have been removed from the soybean peptide lactic acid bacteria fermentation product of the present invention, methods such as decantation (decanting), centrifugation, centrifugation and decantation, filtration, vacuum filtration, membrane filtration, or ultrafiltration can be used. Preferably, the product can be obtained by decanting after centrifugation and removing the precipitated bacterial cells, thereby removing 70% or more of the lactic acid bacteria. The soybean peptide fermented product of the present invention promotes collagen peptide absorption whether or not the lactic acid bacteria cells are removed. For example, when the soybean peptide fermented product of the present invention is ingested as a beverage, it is preferable to use one in which the lactic acid bacteria cells have been removed to a certain extent, for example, 70% or more, from the viewpoint of mouthfeel and ease of drinking.

[0026] In another aspect of the present invention, the skin condition-improving composition of the present invention contains a soybean peptide lactic acid bacteria fermentation product and a collagen peptide. The soybean peptide lactic acid bacteria fermentation product contained in the skin condition-improving composition of the present invention is the soybean peptide lactic acid bacteria fermentation product contained in the collagen absorption-promoting composition of the present invention. Furthermore, the soybean peptide lactic acid bacteria fermentation product contained in the skin condition-improving composition of the present invention may be one before or after lactic acid bacteria removal.

[0027] The skin condition improving composition of the present invention may be a solid or a liquid, such as a powder, granules, pills, tablets, soft capsules, hard capsules, paste, solution, suspension, or emulsion. The skin condition improving composition of the present invention may also be a powder or granule that is reconstituted as a drink by mixing with a liquid such as water. The skin condition-improving composition of the present invention may also be in the form of a food or drink composition, such as a beverage, for example, juice, soup, or drink, or a food, for example, a nutritional supplement, a food for specified health uses, a functional food, or a health food.

[0028] The collagen peptide contained in the skin condition-improving composition of the present invention is a collagen peptide obtained by hydrolyzing collagen. The collagen used as a raw material for the collagen peptide is collagen obtained from mammals, such as the tails and limbs of cows or pigs; birds, such as bird combs; or fish, such as fins, skin, and scales of tuna, shark, cod, flounder, plaice, sea bream, tilapia, salmon, catfish, sea bass, or eel. The collagen peptide contained in the skin condition-improving composition of the present invention is made, for example, from porcine collagen.

[0029] The average molecular weight of the collagen peptide contained in the skin condition improving composition of the present invention is, for example, 8,000 or less, preferably 5,000 or less, more preferably 3,000 or less, and even more preferably 2,000 or less, for pig-derived collagen peptide. The amount of collagen peptide contained in the skin condition improving composition of the present invention is, for example, 5 to 40% by weight, preferably 5 to 30% by weight, and more preferably 10 to 25% by weight.

[0030] The skin condition-improving composition of the present invention may further contain other ingredients such as sweeteners (e.g., dextrin, glucose, lactose, or sugars (e.g., granulated sugar)), sugar alcohols, starch or processed products thereof, celluloses, colorants, flavorings, fruit juice, vitamins, minerals, protein, amino acids, antioxidants (e.g., vitamin C), preservatives, stabilizers, diluents, solubilizers, isotonic agents (e.g., salt), pH adjusters (e.g., citric acid or sodium citrate), acidulants, and various functional nutritional ingredients. The skin condition-improving composition of the present invention may further contain an ingredient that has the effect of promoting the absorption of collagens, such as a surfactant, sesamin, or phospholipids.

[0031] In this specification, the term "about" means a range of ±10%, preferably ±5% of the stated numerical value. Examples of the present disclosure will be described below, but the technical scope of the present disclosure is not limited to these descriptions. [Example]

[0032] (I) Production of the soybean peptide fermented product of the present invention (1) Production of lactic acid bacteria starter MRS (de Man Rogosa Sharpe) medium (Merck) was prepared according to the manufacturer's instructions and heated to 30°C. Next, 0.05% of Lactobacillus plantarum FL-664 strain was inoculated into the MRS medium, and the viable cell count was adjusted to approximately 1.0 × 10 5 ~1.0×10 7 After adjusting the concentration to cfu / ml, the mixture was fermented at 30°C for 20 hours. The fermented product was then centrifuged to collect the bacterial cells, which were then suspended in an equal volume of sterile saline and centrifuged again to wash the bacterial cells. The finally obtained bacterial cells were resuspended in sterile saline, and the viable cell count was adjusted to approximately 1.0 × 10 8 ~2.0×10 9 The culture was adjusted to cfu / ml and used as the FL-664 starter.

[0033] (2) Preparation of lactic acid bacteria fermentation medium Soybean peptide: HINUTE AM (Fuji Oil Co., Ltd.) 5%, Hymaltose M70-75C (Japan Cornstarch Co., Ltd.) 3%, Yeast extract SL-W (Mitsubishi Corporation Life Sciences Co., Ltd.) 0.3%, Yeast Rich Manganese (Oriental Yeast Co., Ltd.) 0.06%, Water 90.64% The prepared medium was sterilized at 90°C for 1 minute and then cooled to 30°C.

[0034] (3) Production of lactic acid bacteria fermentation products The FL-664 starter prepared in (1) was inoculated at 1% into the lactic acid bacteria fermentation medium prepared in (2), and fermentation was carried out at 30°C for 24 hours, after which the fermentation was stopped by heating at 85°C for 5 minutes. The resulting soybean peptide lactic acid bacteria fermentation product was designated as lactic acid bacteria fermentation product (A).

[0035] (4) Lactic acid bacteria removal The soybean peptide lactic acid bacteria fermentation product obtained in (3) was centrifuged at 3500 rpm for 15 minutes (using a tabletop centrifuge manufactured by Kokusan Co., Ltd., rotor RF-110). The supernatant was decanted, removing only the layer of solidly precipitated lactic acid bacteria, yielding a soybean peptide lactic acid bacteria fermentation product from which lactic acid bacteria had been removed. This was designated lactic acid bacteria-removed fermentation product (B). The number of lactic acid bacteria was measured using DAPI staining and compared with that in lactic acid bacteria fermentation product (A). It was confirmed that more than 90% of the lactic acid bacteria had been removed from lactic acid bacteria-removed fermentation product (B). Since the weight of the lactic acid bacteria removed here was approximately 2% of the total volume, the amount of fermentation product contained in lactic acid bacteria fermentation product (A) and lactic acid bacteria-removed fermentation product (B) was approximately the same.

[0036] (5) Collagen drink manufacturing To each of the collagen drinks (x3) containing 10 g of porcine collagen peptide, 6 g of granulated sugar, citric acid for adjusting pH and flavor, and flavoring, 10 g of the lactic acid bacteria fermentation medium before inoculation obtained in (2), lactic acid bacteria fermentation product (A), or lactic acid bacteria-removed fermentation product (B) was added, and the mixture was mixed with water to a total volume of 50 g. After mixing, the mixture was bottled, heated at 85°C for 10 minutes, and cooled. Here, the mixture mixed with the lactic acid bacteria fermentation medium before inoculation obtained in (2) is designated Comparative Example 1, the mixture mixed with the lactic acid bacteria fermentation product (A) is designated Example 1, and the mixture mixed with the lactic acid bacteria-removed fermentation product (B) is designated Example 2.

[0037] (6) Manufacturing collagen drink with reduced collagen peptides To each of the collagen drinks (x2) containing 5 g of porcine collagen peptide, 6 g of granulated sugar, citric acid for adjusting pH and flavor, and flavoring, 5 g of the lactic acid bacteria fermentation medium before inoculation obtained in (2) or the lactic acid bacteria fermentation product (A) was added, and water was added to bring the total volume to 50 g and mixed. After mixing, the mixture was filled into an aluminum pouch, heated at 85°C for 10 minutes, and cooled. Here, the mixture to which the lactic acid bacteria fermentation medium was added is Comparative Example 2, and the mixture to which the lactic acid bacteria fermentation product (A) was added is Example 3.

[0038] (7) Production of lactic acid bacteria fermentation product without peptide as a culture medium raw material (comparative example) Hymaltose M70-75C (Japan Cornstarch Co., Ltd.) 3%, Yeast extract SL-W (Mitsubishi Corporation Life Sciences Co., Ltd.) 0.6%, Yeast Rich Manganese (Oriental Yeast Co., Ltd.) 0.06%, L-monosodium glutamate (Mitsubishi Corporation Life Sciences Co., Ltd.) 2%, Dipotassium hydrogen phosphate (Yoneyama Chemical Co., Ltd.) 0.435%, Liquid citric acid 0.075% (Fuso Chemical Co., Ltd.), Water 92.83% A soybean peptide-free lactic acid bacteria fermentation medium was prepared by mixing the above ingredients. The prepared medium was sterilized at 90°C for 1 minute and then cooled to 30°C. This medium was inoculated with 1% of the FL-664 starter prepared in (1), fermented at 30°C for 24 hours, and then heated at 85°C for 5 minutes to stop the fermentation. To match the amount of soybean peptides in the lactic acid bacteria fermented product (A) used in collagen drinks, 5% soybean peptide: Hinute AM was added to the resulting lactic acid bacteria fermented product (C). The number of lactic acid bacteria was confirmed by DAPI staining, and it was confirmed that the number of lactic acid bacteria contained in the lactic acid bacteria fermented product (A) was similar to that in the lactic acid bacteria fermented product (C).

[0039] (8) Manufacturing of high-dose collagen drink made from lactic acid bacteria fermentation To each of the collagen drinks (x3) containing 5 g of porcine collagen peptide, 6 g of granulated sugar, citric acid to adjust the pH and flavor, and flavoring, 20 g of the lactic acid bacteria fermentation medium before inoculation obtained in (2), the lactic acid bacteria fermentation product (A), or the lactic acid bacteria fermentation product (C) obtained in (7) was added, and water was added to bring the total volume to 50 g and mixed. After mixing, the mixture was bottled, heated at 85°C for 10 minutes, and cooled. Here, the mixture to which the lactic acid bacteria fermentation medium was added is Comparative Example 3, the mixture to which the lactic acid bacteria fermentation product (A) was added is Example 4, and the mixture to which the lactic acid bacteria fermentation product (C) was added is Comparative Example 4.

[0040] The following table shows each example, the amount of collagen peptide in the collagen drink of each example, and the contents and amounts of fermented products, etc. [Table 1]

[0041] (II) Absorption enhancement test of collagen peptides in humans Five healthy men and women aged 20 to 40 years were used as subjects. The day before the test, the five subjects fasted for 12 hours before the start of the test and were only allowed to consume water. On the day of the test, blood was collected at time 0, and then the subjects were given the collagen drink of Comparative Example 1 prepared in (5), and blood samples were collected 1, 2, and 4 hours later.

[0042] The collected blood was centrifuged to obtain plasma. The plasma was then deproteinized with ethanol. The deproteinized plasma was divided into three portions: Sample 1, Sample 2, and Sample 3. Sample 1 was hydrolyzed with HCl at 110°C for 20 minutes, treated with PITC, and analyzed by HPLC to calculate the total amount of Hyp. Sample 2 was derivatized with PITC without hydrolysis, then analyzed by HPLC to calculate the amount of free Hyp. The amount of free Hyp was then subtracted from the total Hyp to calculate the amount of peptide-type Hyp transferred into the blood. Furthermore, Sample 3 was derivatized with AccQ and analyzed by LC-MS to measure Pro-Hyp and Hyp-Gly.

[0043] The collagen drinks of Examples 1 and 2 were tested in the same manner on the same subject. Blood was collected 1, 2, and 4 hours after ingestion of the collagen drink, and the total and free Hyp amounts were measured as described above, and the amount of peptide-type Hyp was calculated. Pro-Hyp and Hyp-Gly were also measured. After the test of Comparative Example 1 and the test of Example 1, a washout period of at least one week was allowed before the next test.

[0044] Figure 1 is a graph plotting the average amount of peptide-type Hyp in the plasma of five subjects against time (time 0 = time of collagen drink ingestion). Figure 2 is a graph plotting the average amount of Pro-Hyp in the plasma of five subjects against time (time 0 = time of collagen drink ingestion). Figure 3 is a graph plotting the average amount of Hyp-Gly in the plasma of five subjects against time (time 0 = time of collagen drink ingestion).

[0045] 1, within about two hours, the amounts of peptide-type Hyp in both Examples 1 and 2 were greater than the amount of peptide-type Hyp in Comparative Example 1. According to Fisher's least significant difference method, p<0.05 was found between Comparative Example 1 and Example 2 (indicated by *), confirming that the amount of collagen peptide absorbed in Example 2 was significantly higher than that in Comparative Example 1.

[0046] In the graph of FIG. 2, the amounts of Pro-Hyp in both Examples 1 and 2 were greater than the amount of Pro-Hyp in Comparative Example 1 within about two hours. In the graph of Figure 3, up to about 2 hours after the start of treatment, the amounts of Hyp-Gly in both Example 1 and Example 2 were higher than the amount of Hyp-Gly in Comparative Example 1. According to Fisher's least significant difference method, p<0.05 was found between Example 1 and Example 2 (indicated by *), and p<0.01 was found between Comparative Example 1 and Example 2 (indicated by *). In other words, it was confirmed that the amount of Hyp-Gly in Example 2 was significantly higher than that in Example 1, and that the amount of Hyp-Gly in Example 2 was significantly higher than that in Comparative Example 1.

[0047] 1 to 3, when comparing Comparative Example 1 and Example 1, the amounts of peptide-type Hyp, Pro-Hyp, and Hyp-Gly were greater in Example 1 than in Comparative Example 1. This suggests that the soybean peptide fermentation product or the lactic acid bacteria itself contained in the collagen peptide drink of Example 1 promotes the absorption of collagen peptides.

[0048] Furthermore, when comparing Comparative Example 1 with Example 2, peptide-type Hyp, Pro-Hyp, and Hyp-Gly were detected in large amounts, with peptide-type Hyp and Hyp-Gly being detected in significantly higher amounts. When comparing Example 1 with Example 2, peptide-type Hyp and Hyp-Gly were detected in large amounts in Example 2, with Hyp-Gly being detected in significantly higher amounts. This indicates that the collagen drink of Example 2, produced using the lactic acid bacteria-removed fermentation product (B), promoted collagen peptide absorption to a level equal to or greater than that of the lactic acid bacteria-free fermentation product (A). This suggests that the collagen peptide absorption-promoting effect of this fermentation liquid is primarily due to the product obtained by fermenting soy peptides with lactic acid bacteria, rather than the lactic acid bacteria themselves. This suggests that the effect of the product obtained by fermenting soy peptides with lactic acid bacteria is significant, regardless of the effectiveness of the lactic acid bacteria themselves.

[0049] (III) Sensory test of human skin condition (parallel group) Thirty-one healthy men and women in their 20s to 60s were divided into two groups, 16 people (Group 1) and 15 people (Group 2), to ensure equal age and gender distribution. The subjects in Group 1 were blindly provided with the collagen drink of Comparative Example 2, and the subjects in Group 2 were blindly provided with the collagen drink of Example 3. The test was conducted for a total of 10 days, including a 5-day pre-drinking observation period and a 5-day drinking period. During the 5-day drinking period, the subjects were required to drink one bottle of the collagen drink per day.

[0050] During the test period, the subjects looked in the mirror and palpated their skin daily, assessing the overall condition, dryness, and elasticity of their skin on a 5-point scale for 10 consecutive days, including the pre-drinking observation period and the drinking period. The evaluation was based on the following evaluation criteria: <Skin condition evaluation> Overall condition 1 point: I feel like my skin is in bad condition 3 points: Neither 5 points: My skin feels good The evaluation was done on a 5-point scale from 1 to 5 points. About drying 1 point: Feels dry 3 points: Neither 5 points: Feels moisturized The evaluation was done on a 5-point scale from 1 to 5 points. About elasticity 1 point: stiff and rigid 3 points: Neither 5 points: Soft and chewy The evaluation was done on a 5-point scale from 1 to 5 points.

[0051] [Table 2]

[0052] The evaluation results for skin condition are shown as mean values ​​± standard deviations in Table 2. In addition, p values ​​were calculated using the Wilcoxon signed rank test to confirm whether there was a significant difference between the results for the pre-drinking observation period and the results for the drinking and post-drinking observation periods.

[0053] Because both the Comparative Example 2 intake group and the Example 3 intake group consumed collagen peptides, their scores tended to be higher during the intake period compared to the observation period before intake. However, the results for the Comparative Example 2 intake group were all p>0.05, significantly greater than 0.05, and there was no significant difference. On the other hand, the Example 3 intake group showed a significant trend in overall skin condition and dryness with p=0.07, and particularly in elasticity with p<0.05, indicating a significant improvement. It was found that the Example 3 intake group showed more improvement than the Comparative Example 2 intake group in all of the overall skin condition, dryness, and elasticity during the intake period compared to the state before intake.

[0054] Furthermore, the average values ​​of the evaluations of the overall skin condition, dryness, and elasticity for each individual for the group taking Example 3 and the group taking Comparative Example 2 over the 5-day observation period before ingestion were calculated, and the differences between these values ​​and the evaluation values ​​for each day over the 5-day ingestion period were calculated. The average values ​​for each group were plotted and shown in Figures 4 to 6. From as early as the second day and as late as the third day of the 5-day ingestion period, the group taking Example 3 showed a clearer improvement in skin condition than the group taking Comparative Example 2.

[0055] From the above, it was found that even if the amount of collagen peptide was reduced to half the amount (5 g of collagen peptide) of the amount that had been confirmed to be absorbed in the past, the effect of collagen peptide, that is, the improvement of the skin condition of the subject, was observed.In addition, it was found that the improvement of the skin condition that the subject could feel was observed from the early stage of the drinking period.This is also commercially important in that it motivates consumers to continue drinking the skin condition improving composition of the present invention.

[0056] (IV) Sensory test of human skin condition (same group) A group-wide sensory test was conducted on three healthy men and women in their 30s and 40s on a high-dose lactic acid bacteria fermentation collagen drink. The test period consisted of a one-day pre-drinking observation period and a five-day drinking period, for a total of six days. Participants consumed one bottle of collagen drink per day during the five-day drinking period. During the test period, the subjects palpated their skin while looking in the mirror every day and self-evaluated their skin's overall condition, dryness, and elasticity on a five-point scale for six consecutive days during the pre-drinking observation period and drinking period. The evaluation was conducted according to the evaluation criteria described in (III).

[0057] The test was conducted on the same subject group, first using the collagen drinks of Comparative Example 3, then Example 4 and Comparative Example 4. After the tests of Comparative Example 3 and Example 4 were completed, a washout period of at least one week was allowed before the next test.

[0058] The differences between the individual values ​​for overall skin condition, dryness, and elasticity from the one-day pre-drinking observation period and the daily evaluation values ​​for the five-day drinking period were calculated for Comparative Example 3, Example 4, and the Comparative Example 4 intake group, and the average values ​​of these differences for each group were plotted and shown in Figures 7 to 9. During the five-day drinking period, the Example 4 intake group showed a clearer improvement in skin condition than the Comparative Examples 3 and 4 intake groups from the fourth day onwards. The results of Comparative Example 3 and Example 4 confirmed the skin condition-improving effect of the soy peptide lactic acid bacteria fermentation product, even in a high-dose collagen drink containing lactic acid bacteria fermentation product. Furthermore, the results of Comparative Example 4 and Example 4 showed that the lactic acid bacteria fermentation product in a medium not containing soy peptides was ineffective, and that unfermented soy peptides were ineffective, indicating that the improvement in skin condition was due to the effect of the "product obtained by fermenting soy peptides with lactic acid bacteria." Furthermore, although both Comparative Example 4 and Example 4 contain lactic acid bacteria cells, the effect of Example 4 was not observed in Comparative Example 4, suggesting that the effect of improving skin condition is due to the effect of the soybean peptide lactic acid bacteria fermentation product, rather than the lactic acid bacteria cells.

Claims

1. A cosmetic method for promoting collagen peptide absorption using a composition containing soybean peptide fermented by lactic acid bacteria (excluding medical procedures for humans (including diagnosis, treatment, prevention, or treatment to maintain health)).

2. A cosmetic method for promoting absorption of the collagen peptide described in claim 1, carried out by using the composition described in claim 1 to be ingested together with, at the same time as, or before or after ingestion of a collagen peptide or a collagen peptide-containing composition (excluding medical procedures on humans (including diagnosis, treatment, prevention, or procedures for maintaining health)).

3. A skin condition improving composition comprising 20% ​​to 40% by weight of a soybean peptide fermented by lactic acid bacteria and 10% to 40% by weight of a collagen peptide, wherein 70% or more of the lactic acid bacteria have been removed from the soybean peptide fermented by lactic acid bacteria.

4. A method for producing a skin condition improving composition containing 20% ​​to 40% by weight of soybean peptide lactic acid bacteria fermentation product and 10% to 40% by weight of collagen peptide, the composition being produced by mixing the soybean peptide lactic acid bacteria fermentation product and the collagen peptide.

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