A method for preparing oat polypeptides and its applications, and compositions.

Abstract

This invention provides a method for preparing oat polypeptides and their applications and compositions, relating to the field of cosmetics technology. The preparation method of this invention includes the following steps: (1) oat swells, is dried after steam explosion treatment, and then pectinase, cellulase, and amylase are added for enzymatic hydrolysis of carbohydrates to obtain enzymatically hydrolyzed oat residue; (2) a complex protease is added to the enzymatically hydrolyzed oat residue, and a first enzymatic hydrolysis is performed under the condition of 3-8% volume concentration of ethanol to obtain enzymatic hydrolysate A; the complex protease is papain, bromelain, and flavor protease in a mass ratio of 3-6:3-6:2; (3) trypsin is added to enzymatic hydrolysate A for a second enzymatic hydrolysis to obtain enzymatic hydrolysate B; (4) enzymatic hydrolysate B is ultrafiltered to obtain oat polypeptides. This application obtains oat polypeptides with small molecular weight and high content, which, when used in the preparation of cosmetics, have significant moisturizing, anti-wrinkle, and skin roughness-reducing effects.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a method for preparing oat polypeptides and their applications and compositions. Background Technology

[0002] Oats are an annual herbaceous plant belonging to the Poaceae family and are classified as a minor grain. Oats contain nutrients such as lipids, proteins, and antioxidants, making them a high-quality grain. The nutritional composition of oats is balanced and easily absorbed by the body. They also possess excellent health benefits, such as preventing and treating cardiovascular diseases and slowing down aging. The protein content of oats is significantly higher than that of other grains. Oat polypeptides are small-molecule peptides obtained from oat protein through enzymatic hydrolysis, fermentation, or other techniques, typically composed of 2-20 amino acids. Compared to whole oat protein, oat polypeptides have a smaller molecular weight, are more easily absorbed, and have enhanced activity.

[0003] Traditional protein extraction methods employ an "alkali-dissolution-acid-precipitation method," where the isolated protein is then hydrolyzed with enzymes to obtain hydrolyzed peptides. This method yields low peptide content and uses large amounts of acid and alkali, causing environmental pollution. To address this, Chinese invention patent CN115011660A discloses a method for preparing oat protein peptides. This method avoids the alkali-dissolution-acid-precipitation method. Instead, oats are pulverized and extracted with organic solvents to obtain oat protein. This is followed by primary enzymatic hydrolysis with pepsin and acidic protease, secondary enzymatic hydrolysis with neutral protease and papain, and tertiary enzymatic hydrolysis with alkaline protease and flavor protease, resulting in a tertiary enzymatic hydrolysate. This hydrolysate is then fermented with *Lactobacillus plantarum* and *Bacillus coagulans* to obtain the primary fermentation product of oat protein. Finally, it is fermented with selenium-enriched yeast to obtain metabolically beneficial and antioxidant oat protein peptides. However, this method uses at least six proteases for enzymatic hydrolysis, significantly increasing the cost; the fermentation process uses at least three types of bacteria, also increasing the complexity of the preparation process; and the patent uses organic components such as acetone, acetonitrile, and trichloroacetic acid for oat protein extraction, which have a certain degree of toxicity.

[0004] Chinese invention patent CN119498409A discloses an enzymatic hydrolysis method for oat milk, including the following steps: (1) regulating the activity of endogenous enzymes in oats and in-situ enzymatic hydrolysis, using hot water to blanch oat seeds to obtain heat-shocked oats; then, the heat-shocked oats are soaked in a solution containing gibberellin to obtain soaked oats; in-situ enzymatic hydrolysis is continued in the seeds while keeping them warm and moist to induce germination; (2) pulping and enzymatic hydrolysis by endogenous free enzymes in oats: water is added and pulped to obtain oat milk with residue, which is then kept at 40-45℃ for 1 hour and filtered; (3) the temperature is raised to 80-85℃, and high-temperature α-amylase is added to complete starch liquefaction; (4) saccharifying enzyme and papain are added, and the mixture is kept at 40-45℃ for 1-1.5 hours for enzymatic hydrolysis, homogenized, and sterilized to finally obtain enzymatically hydrolyzed oat milk. Using the method of this invention, the polypeptide content obtained is above 9 mg / mL.

[0005] Cui Bingqun (Study on the preparation process of oat polypeptides [J]. Modern Food Science and Technology, 2012, 28(08):1040-1042.DOI:10.13982 / j.mfst.1673-9078.2012.08.027.) Oat polypeptides were extracted using enzymatic technology from oat bran. The yield of oat polypeptides was used as the indicator. Single-factor experiments and orthogonal experiments were conducted to study the influencing factors of the oat polypeptide extraction process and determine the main parameters of the new extraction process: water addition of 18 times, reaction temperature of 50℃, alkaline protease as the hydrolytic enzyme, and enzyme addition of 1.0%. The process described in this article can achieve an oat polypeptide yield of 18.9%, and the purity of oat polypeptides after membrane filtration purification can reach 87.4%.

[0006] Currently, most existing technologies for preparing oat protein peptides utilize a single protease to achieve protein hydrolysis, without studying the synergistic effects of multiple proteases. Furthermore, the relationship between the types of proteases used in the hydrolysis process and the peptide content, moisturizing and anti-wrinkle effects remains unclear. In addition, oats contain a large amount of carbohydrates, which can negatively impact the hydrolysis, purity, and efficacy of oat peptides. Summary of the Invention

[0007] This invention addresses the problems existing in the prior art by providing a method for preparing oat polypeptides and their applications and compositions. The method involves sequentially subjecting oats to steam explosion, followed by enzymatic hydrolysis with pectinase, cellulase, and amylase, and then adding a complex protease and trypsin for further enzymatic hydrolysis to obtain oat polypeptides with smaller molecular weights. These oat polypeptides are used in the preparation of cosmetics, resulting in cosmetics with significant moisturizing, anti-wrinkle, and skin roughness-reducing effects.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0009] First, this invention provides a method for preparing oat polypeptides, comprising the following steps:

[0010] (1) The oats were moistened, dried after being steam-exploded, and then enzymatically hydrolyzed by adding pectinase, cellulase and amylase. After filtration, the enzymatically hydrolyzed oat residue was obtained.

[0011] (2) Add a compound protease to the enzymatically hydrolyzed oat residue and carry out the first enzymatic hydrolysis under the conditions of pH=5.5-7 and ethanol volume concentration of 3-8% to obtain enzymatic hydrolysate A;

[0012] The complex protease consists of papain, bromelain, and flavor protease in a mass ratio of 3-6:3-6:2.

[0013] (3) Adjust the pH of the enzymatic hydrolysate A to 8.5-9.5, add trypsin for a second enzymatic hydrolysis, and obtain enzymatic hydrolysate B;

[0014] (4) The enzyme inactivation of slurry B was carried out by enzyme hydrolysis, and the filtrate was subjected to ultrafiltration and concentration to obtain oat polypeptide.

[0015] Preferably, in step (1), before the oats are moistened, a crushing step is also included, with a crushing particle size of 50-100 mesh.

[0016] More preferably, in step (1), the particle size of the pulverized material is 80 mesh.

[0017] Preferably, in step (1), the oat soaking process specifically involves mixing oats with hot water at 50-70°C and soaking for 1-3 hours.

[0018] More preferably, in step (1), the oat soaking process specifically involves mixing oats with hot water at 60°C and soaking for 2 hours.

[0019] Preferably, in step (1), the steam explosion specifically involves: maintaining the swollen oats under a pressure of 0.8-2MPa for 60-200s, with a material-to-cavity ratio of 5:6-10, to obtain oats with a loose structure.

[0020] Preferably, in step (1), the steam explosion specifically involves: maintaining the swollen oats at a pressure of 1.0 MPa for 120 seconds with a material-to-cavity ratio of 5:8 to obtain loosely structured oats.

[0021] Preferably, in step (1), before drying, the oats after steam explosion treatment need to be washed to remove sugar, acidic substances, hemicellulose, etc. from the surface.

[0022] Preferably, in step (1), the mass ratio of pectinase, cellulase and amylase is 1:4-6:4-6.

[0023] More preferably, in step (1), the mass ratio of pectinase, cellulase and amylase is 1:5:5.

[0024] Preferably, in step (1), the enzymatic hydrolysis specifically involves: mixing dried steam-exploded oat flour with 15-30 times its mass of water and enzymatically hydrolyzing it at 45-52°C for 3-6 hours.

[0025] More preferably, in step (1), the enzymatic hydrolysis specifically involves: mixing dried steam-exploded oat flour with 20 times its mass of water and enzymatically hydrolyzing it at 48°C for 5 hours.

[0026] Preferably, in step (1), the total amount of pectinase, cellulase and amylase added during enzymatic hydrolysis is 80-130 mg of total enzyme mass / 1 g of oat mass.

[0027] Preferably, in step (1), the total amount of pectinase, cellulase and amylase added during enzymatic hydrolysis is: 100 mg of total enzyme mass / 1 g of oat mass.

[0028] Preferably, in step (1), after filtration, the oat residue needs to be washed with water to remove the starch, sugar generated after enzymatic hydrolysis, and by-products on the surface.

[0029] Preferably, in step (2), the complex protease is papain, bromelain, and flavor protease in a mass ratio of 4:5:2.

[0030] Preferably, in step (2), the amount of the complex protease added is: 30-50 mg of complex protease mass / 1 g of oat mass.

[0031] More preferably, in step (2), the amount of the complex protease added is: 40 mg of complex protease / 1 g of oat mass.

[0032] Preferably, in step (2), the first enzymatic hydrolysis is specifically performed at 42-50°C for 3-5 hours under the following conditions: solid-liquid ratio of 1:15-25 g / mL, pH=5.5-7, ethanol volume concentration of 3-8%.

[0033] More preferably, in step (2), the first enzymatic hydrolysis is specifically performed at 44°C for 4 hours under the conditions of a solid-liquid ratio of 1:20 g / mL, pH=6.0, and ethanol volume concentration of 6%.

[0034] In this invention, the ethanol volume concentration of 3-8% mentioned in step (2) means that the amount of ethanol added is equivalent to 3-8% of the total volume of the enzymatic hydrolysis liquid.

[0035] Preferably, in step (3), the amount of trypsin added is: 5-15 mg trypsin mass / 1 g oat mass.

[0036] More preferably, in step (3), the amount of trypsin added is: 10 mg trypsin mass / 1 g oat mass.

[0037] Preferably, in step (3), the second enzymatic hydrolysis specifically involves adjusting the pH of the enzymatic hydrolysate A to 8.5-9.2, adding trypsin, and enzymatically hydrolyzing at 37-44°C for 2-5 hours.

[0038] More preferably, in step (3), the second enzymatic hydrolysis specifically involves: adjusting the pH of the hydrolysate A to 8.6, adding trypsin, and performing enzymatic hydrolysis at 39°C for 3 hours.

[0039] Preferably, in step (4), the enzyme inactivation is performed at 121°C for 10-15 minutes.

[0040] Preferably, in step (4), the ultrafiltration is performed using an ultrafiltration membrane with a pore size of ≤8000Da.

[0041] Then, the present invention provides oat polypeptides prepared by the above preparation method.

[0042] Furthermore, the present invention provides the application of the above-mentioned oat polypeptide in the preparation of cosmetic raw materials or cosmetics, wherein the cosmetics used in the application are moisturizing and anti-wrinkle cosmetics.

[0043] Preferably, the cosmetic used in the application is a cosmetic that can moisturize, increase skin elasticity, increase collagen content in the skin, promote skin metabolism, and promote cell proliferation.

[0044] Furthermore, the present invention provides a cosmetic ingredient containing the above-mentioned oat polypeptide, comprising the following components: the above-mentioned oat polypeptide, a moisturizer, a skin conditioning agent, a preservative, and water.

[0045] Preferably, the cosmetic raw material comprises, by weight percentage: 0.1-0.4% of the above-mentioned oat peptides, 8-15% of moisturizer, 0.05-0.2% of skin conditioning agent, 0.5-1.5% of preservative and the balance being water.

[0046] More preferably, the cosmetic ingredient comprises, by weight percentage: 0.3% oat peptides, 10% moisturizer, 0.1% skin conditioning agent, 0.9% preservative and the balance being water.

[0047] Preferably, the moisturizer, skin conditioning agent, and preservative in the cosmetic ingredients are all conventionally used components in the art. In some embodiments of the present invention, the moisturizer is glycerin, the skin conditioning agent is ethylhexylglycerin, and the preservative is phenoxyethanol.

[0048] Finally, the present invention provides a moisturizing and anti-wrinkle composition comprising the following components: the above-mentioned oat polypeptide, hyaluronic acid, copper peptide and ceramide.

[0049] Preferably, in the moisturizing and anti-wrinkle composition, the mass ratio of oat peptides, hyaluronic acid, copper peptides and ceramides is 20:5-10:2-5:2-5.

[0050] More preferably, in the moisturizing and anti-wrinkle composition, the mass ratio of oat peptides, hyaluronic acid, copper peptides and ceramides is 20:8:4:4.

[0051] Compared with the prior art, the present invention has the following beneficial effects:

[0052] 1. The oat polypeptide prepared by the preparation method of the present invention has a structure similar to natural moisturizing factors and has excellent moisturizing properties. The oats are first soaked, steamed and exploded, and then subjected to enzymatic hydrolysis of sugars by pectinase, cellulase and amylase, which facilitates protein exposure and makes it easy to be hydrolyzed by proteases. The resulting oat residue is then subjected to a first enzymatic hydrolysis by a complex protease and a second endopeptidation by trypsin, so that the high molecular weight protein is efficiently hydrolyzed into small molecular weight oat polypeptides, thereby increasing the proportion of small molecular weight polypeptides.

[0053] 2. The oat polypeptide prepared by this invention has a high proportion of small molecule polypeptides, which is conducive to skin absorption and thus further improves moisturizing and anti-wrinkle properties.

[0054] 3. The composition prepared by using the oat polypeptide of the present invention in combination with hyaluronic acid, copper peptide and ceramide components has significant moisturizing, skin elasticity increase and roughness reduction effects. Detailed Implementation

[0055] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0056] When numerical ranges are given in the embodiments, it should be understood that, unless otherwise stated in the invention, both endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0057] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all chemical reagents used in the embodiments of the present invention are obtained through conventional commercial means.

[0058] In the following examples, the papain was purchased from Nanning Shanwan Biotechnology Co., Ltd., and the enzyme activity was 100,000 U / g;

[0059] The bromelain was purchased from Nanning Pangbo Biotechnology Co., Ltd., and the enzyme activity was 50,000 U / g.

[0060] The flavor protease was purchased from Nanning Shanwan Biotechnology Co., Ltd., and its enzyme activity was 20,000 U / g.

[0061] The trypsin was purchased from Nanning Pangbo Biotechnology Co., Ltd., and the enzyme activity was 4000 U / g.

[0062] The neutral protease was purchased from Shandong Longkete Enzyme Preparation Co., Ltd., and its enzyme activity was 100,000 U / g.

[0063] The alkaline protease was purchased from Shandong Longkete Enzyme Preparation Co., Ltd., and its enzyme activity was 50,000 U / g.

[0064] The above components do not have a significant impact on the effect depending on the manufacturer.

[0065] Example 1

[0066] A method for preparing oat polypeptides, comprising the following steps:

[0067] (1) Grind the oats to 80 mesh, soak them in water at 60℃ for 2 hours, and make sure the water covers the oats;

[0068] After the oats were swollen, they were steam-exploded under a pressure of 1.0 MPa for 120 seconds with a material-to-chamber ratio of 5:8. The oats were washed with water and dried at 60°C to obtain loosely structured oats (dried steam-exploded oat flour).

[0069] The dried steam-exploded oat flour was mixed with 20 times its weight of water, and pectinase, cellulase and amylase were added in a mass ratio of 1:5:5 (addition amount: 100mg total mass of enzymes / 1g mass of oat raw material). The mixture was enzymatically hydrolyzed at 48℃ for 5 hours, filtered, and the enzymatically hydrolyzed oat residue was obtained.

[0070] (2) Add compound protease to the enzymatically hydrolyzed oat residue, with the mass of 40 mg of compound protease / 1 g of oat raw material, to carry out the first enzymatic hydrolysis and obtain enzymatic hydrolysate A;

[0071] The complex protease consists of papain, bromelain, and flavor protease in a mass ratio of 4:5:2.

[0072] The conditions for the first enzymatic hydrolysis were: solid-liquid ratio 1:20 g / mL, pH=6.0, ethanol volume concentration 6%, and enzymatic hydrolysis at 44℃ for 4 h.

[0073] (3) Adjust the pH of the enzymatic hydrolysate A to 8.6, add trypsin, the amount of trypsin added is: 10 mg trypsin mass / 1 g oat raw material mass, enzymatic hydrolysis at 39℃ for 3 h, and perform a second enzymatic hydrolysis to obtain enzymatic hydrolysate B.

[0074] (4) The enzyme hydrolysate B was treated at 121℃ for 15 min to inactivate the enzyme, filtered, and then ultrafiltered using an ultrafiltration membrane with a pore size of ≤8000Da. The solution was then concentrated to obtain oat polypeptides.

[0075] Example 2

[0076] Unlike Example 1, the mass ratio of papain, bromelain, and flavor protease is different in step (2).

[0077] In this embodiment, the mass ratio of papain, bromelain, and flavor protease is 6:3:2.

[0078] Everything else is the same as in Example 1.

[0079] Example 3

[0080] Unlike Example 1, the mass ratio of papain, bromelain, and flavor protease is different in step (2).

[0081] In this embodiment, the mass ratio of papain, bromelain, and flavor protease is 3:6:2.

[0082] Everything else is the same as in Example 1.

[0083] Example 4

[0084] A method for preparing oat polypeptides, comprising the following steps:

[0085] (1) Grind the oats to 80 mesh, soak them in 50℃ water for 3 hours, and make sure the water covers the oats;

[0086] After the oats were soaked and expanded, they were subjected to steam explosion at a pressure of 0.8 MPa for 200 seconds with a material-to-cavity ratio of 5:10. After washing with water, the oats were freeze-dried to obtain loosely structured oats (dried steam-exploded oat flour).

[0087] The dried steam-exploded oat flour was mixed with 30 times its weight of water, and pectinase, cellulase and amylase were added in a mass ratio of 1:4:6 (addition amount: 80mg total mass of enzymes / 1g mass of oat raw material). The mixture was enzymatically hydrolyzed at 52℃ for 3 hours, filtered, and the enzymatically hydrolyzed oat residue was obtained.

[0088] (2) Add compound protease to the enzymatically hydrolyzed oat residue, with the mass of 50 mg of compound protease / 1 g of oat raw material, to carry out the first enzymatic hydrolysis and obtain enzymatic hydrolysate A;

[0089] The complex protease consists of papain, bromelain, and flavor protease in a mass ratio of 4:5:2.

[0090] The conditions for the first enzymatic hydrolysis were: solid-liquid ratio 1:25 g / mL, pH=7.0, ethanol volume concentration 3%, and enzymatic hydrolysis treatment at 42℃ for 5 h.

[0091] (3) Adjust the pH of the enzymatic hydrolysate A to 8.5, add trypsin, the amount of trypsin added is: 5 mg trypsin mass / 1 g oat raw material mass, enzymatic hydrolysis at 37℃ for 2 h, and perform a second enzymatic hydrolysis to obtain enzymatic hydrolysate B.

[0092] (4) The enzyme hydrolysate B was treated at 121℃ for 15 min to inactivate the enzyme, filtered, and then ultrafiltered using an ultrafiltration membrane with a pore size of ≤8000Da. The solution was then concentrated to obtain oat polypeptides.

[0093] Example 5

[0094] A method for preparing oat polypeptides, comprising the following steps:

[0095] (1) Grind the oats to 80 mesh and soak them in 70℃ water for 1 hour. The amount of water should cover the oats.

[0096] After the oats were soaked and expanded, they were subjected to steam explosion at a pressure of 2 MPa for 60 seconds with a material-to-chamber ratio of 5:6. The oats were then washed with water and freeze-dried to obtain loosely structured oats (dried steam-exploded oat flour).

[0097] The dried steam-exploded oat flour was mixed with 15 times its weight of water, and pectinase, cellulase and amylase were added in a mass ratio of 1:4:6 (addition amount: 130mg total mass of enzymes / 1g mass of oat raw material). The mixture was enzymatically hydrolyzed at 45℃ for 6 hours, filtered, and the enzymatically hydrolyzed oat residue was obtained.

[0098] (2) Add compound protease to the enzymatically hydrolyzed oat residue, with the mass of 30 mg of compound protease / 1 g of oat raw material, to carry out the first enzymatic hydrolysis and obtain enzymatic hydrolysate A;

[0099] The complex protease consists of papain, bromelain, and flavor protease in a mass ratio of 4:5:2.

[0100] The conditions for the first enzymatic hydrolysis were: solid-liquid ratio 1:15 g / mL, pH=5.5, ethanol volume concentration 8%, and enzymatic hydrolysis at 50℃ for 3 h.

[0101] (3) Adjust the pH of the enzymatic hydrolysate A to 9.2, add trypsin, the amount of trypsin added is: 15mg trypsin mass / 1g oat raw material mass, enzymatic hydrolysis at 44℃ for 5h, and perform a second enzymatic hydrolysis to obtain enzymatic hydrolysate B.

[0102] (4) The enzyme hydrolysate B was treated at 121℃ for 15 min to inactivate the enzyme, filtered, and then ultrafiltered using an ultrafiltration membrane with a pore size of ≤8000Da. The solution was then concentrated to obtain oat polypeptides.

[0103] Comparative Example 1

[0104] Unlike Example 1, in step (1), the steam explosion treatment is replaced with ultrasonic treatment.

[0105] (1) Grind the oats to 80 mesh, soak them in water at 60℃ for 2 hours, and make sure the water covers the oats;

[0106] After the oats were soaked and swollen, they were treated with ultrasonic waves at a frequency of 500W for 2 minutes, washed with water, and dried at 60℃ to obtain oats with a loose structure (dried ultrasonic oat powder).

[0107] The dried ultrasonic oat flour was mixed with 20 times its weight of water, and pectinase, cellulase and amylase were added in a mass ratio of 1:5:5 (addition amount: 100mg total mass of enzymes / 1g mass of oat raw material). The mixture was enzymatically hydrolyzed at 48℃ for 5 hours, filtered, and the enzymatically hydrolyzed oat residue was obtained.

[0108] Steps (2) to (4) are the same as in Example 1.

[0109] Comparative Example 2

[0110] Unlike Example 1, in step (1), pectinase is replaced with cellulase and amylase in a mass ratio of 1:1.

[0111] (1) Grind the oats to 80 mesh, soak them in water at 60℃ for 2 hours, and make sure the water covers the oats;

[0112] After the oats were swollen, they were steam-exploded under a pressure of 1.0 MPa for 120 seconds with a material-to-chamber ratio of 5:8. The oats were washed with water and dried at 60°C to obtain loosely structured oats (dried steam-exploded oat flour).

[0113] The dried steam-exploded oat flour was mixed with 20 times its weight of water, and cellulase and amylase were added in a mass ratio of 5.5:5.5 (addition amount: 100mg total mass of enzymes / 1g mass of oat raw material). The mixture was enzymatically hydrolyzed at 48℃ for 5 hours, filtered, and the enzymatically hydrolyzed oat residue was obtained.

[0114] Steps (2) to (4) are the same as in Example 1.

[0115] Comparative Example 3

[0116] Unlike Example 1, bromelain in step (2) was replaced with neutral protease. The complex protease consisted of papain, neutral protease, and flavor protease in a mass ratio of 4:5:2.

[0117] Everything else is the same as in Example 1.

[0118] Comparative Example 4

[0119] Unlike Example 1, in step (2), the mass ratio of papain, bromelain, and flavor protease is 8:1:2.

[0120] Everything else is the same as in Example 1.

[0121] Comparative Example 5

[0122] Unlike Example 1, in step (2), the ethanol volume concentration is 30% (relative to 30% of the total volume of the enzymatic hydrolysis liquid).

[0123] Everything else is the same as in Example 1.

[0124] Comparative Example 6

[0125] Unlike Example 1, in step (3), the trypsin is replaced with alkaline protease. Everything else is the same as in Example 1.

[0126] The oat peptides from Example 1 were freeze-dried and used in the preparation of the compositions in the following application examples and application comparative examples.

[0127] Application Example 1

[0128] A moisturizing and anti-wrinkle composition comprising, by weight parts: 20 parts oat polypeptide, 8 parts hyaluronic acid, 4 parts copper peptide and 4 parts ceramide.

[0129] Mix the above components evenly to obtain a moisturizing and anti-wrinkle composition.

[0130] Application Example 2

[0131] A moisturizing and anti-wrinkle composition comprising, by weight parts: 20 parts oat polypeptide, 5 parts hyaluronic acid, 5 parts copper peptide and 5 parts ceramide.

[0132] Mix the above components evenly to obtain a moisturizing and anti-wrinkle composition.

[0133] Application Example 3

[0134] A moisturizing and anti-wrinkle composition comprising, by weight parts: 20 parts oat peptides, 10 parts hyaluronic acid, 2 parts copper peptides and 2 parts ceramides.

[0135] Mix the above components evenly to obtain a moisturizing and anti-wrinkle composition.

[0136] Application Comparative Example 1

[0137] Unlike Application Example 1, oat peptides were replaced with commercially available soybean oligopeptides.

[0138] Application Comparative Example 2

[0139] Unlike Application Example 1, the copper peptide was replaced with acetyl hexapeptide-8.

[0140] Application Comparative Example 3

[0141] Unlike Application Example 1, the mass ratio of each component in the composition is different: 30 parts oat polypeptide, 1 part hyaluronic acid, 4 parts copper peptide and 1 part ceramide.

[0142] Experiment 1: Determination of polypeptide content and molecular weight

[0143] The oat peptides after ultrafiltration in each example and comparative example were concentrated and brought to a solid-liquid ratio of 1:10 g:mL (the solid mass refers to the mass of the raw oats). The resulting oat peptide products were then tested for peptide content and peptide molecular weight.

[0144] 1. Detection of peptide content:

[0145] Add 5 mL of concentrated oat peptides to 5 mL of 10% trichloroacetic acid solution, centrifuge at 8000 rpm for 3 min, and take the supernatant for testing (sample solution).

[0146] (1) Construction of the standard curve: Accurately pipette 0.16, 0.32, 0.48, 0.64, and 0.8 mL of 10 mg / mL bovine serum albumin standard solution into test tubes, add water to 0.8 mL, add 3.2 mL of biuret reagent, mix well, and react at 25℃ for 30 min. Use 1.0 mL of distilled water as a blank control, measure the absorbance value at 540 nm wavelength using a UV spectrophotometer, and take the average value after three repetitions. Plot the standard curve with protein content as the abscissa and OD 540 nm as the ordinate. It can be seen that the R2 of the linear regression equation of the standard curve is 0.9999, where y is the absorbance value and x is the protein content (mg / mL).

[0147] (2) Determination of peptides in the sample solution: Accurately pipette 0.8 mL of the sample solution to be tested, add 3.2 mL of biuret reagent, mix well, and react at 25 °C for 30 min. Using the reagent blank as a reference, measure the absorbance value at a wavelength of 540 nm using a UV spectrophotometer. Calculate the peptide content in the solution according to the standard curve. The peptide content detection results are shown in Table 1.

[0148] 2. Molecular weight determination was performed according to Appendix A of GB / T22492-2008, "Methods for Determination of Relative Molecular Mass Distribution of Peptides". The results of the molecular weight distribution of oat peptides are shown in Table 1.

[0149] Table 1

[0150]

[0151] In Table 1, # , ## This indicates that the effect of the comparative group is significantly different from that of the example group. # P < 0.05 ## P < 0.01.

[0152] As can be seen from Table 1, the oat polypeptide product prepared by this invention has a high polypeptide content; and the use of specific steam explosion and enzymatic hydrolysis conditions significantly increases the content of small molecule polypeptides, which is beneficial for skin absorption and thus enhances moisturizing and anti-wrinkle effects.

[0153] Experiment 2: Detection of the effect of improving skin roughness

[0154] Test method: Skin roughness was tested using the Frictiometer FR700 skin friction test probe from CK GmbH, Germany.

[0155] Test participants: A total of 66 people, all female, aged 25-45, were randomly divided into 11 groups of 6 people each.

[0156] Test sites and requirements: Select the inner sides of the left and right forearms of the test subject. Define four equal areas (3cm × 3cm) on the left forearm for the oat peptide test. The test is conducted in a room at 20-22℃ and 40%-60% relative humidity. The subject must acclimatize to the same environment for 20 minutes. During the test, the amount of oat peptide used is 1g, covering the test area.

[0157] The results were tested at two time points: before sample use (D0) and 56 days after sample use (D56). Skin friction was measured.

[0158] Skin roughness improvement effect (%) = (D0-D56) / D0×100%.

[0159] The mean values ​​of skin friction are shown in Table 2.

[0160] Table 2

[0161]

[0162] Table 2 shows a comparison of the effects of improving skin roughness. # , ## This indicates that the effect of the comparative group is significantly different from that of the example group. # P < 0.05 ## P < 0.01.

[0163] As shown in Table 2, the use of oat peptides improved skin friction and reduced roughness. The skin roughness in the example group decreased to a greater extent compared to the comparative example. This indicates that the oat peptides prepared using the examples of this application provide superior roughness improvement capabilities, and offer better skin improvement and smoothness enhancement compared to the comparative example and conventional oat peptides.

[0164] Experiment 3: Moisturizing Performance Test of the Composition

[0165] The moisturizing and anti-wrinkle compositions of Application Examples 1-3 and Comparative Examples 1-3 were formulated into a 1.0% (w / w) aqueous solution.

[0166] Experimental Methods: Twenty-four healthy men and women (12 men and 12 women) were randomly divided into six groups, with two men and two women in each group. Before the experiment, participants were required to wash their forearms and then sit quietly in a climate-controlled room (22±1℃, 50% relative humidity) for at least 20 minutes, maintaining a relaxed state. Their forearms were required to be free of scars, pigmentation, atrophy, port-wine stains, or other blemishes that could affect the interpretation of the experimental results.

[0167] Three 3cm x 3cm areas were randomly marked on the inner sides of the left and right forearms of each subject as test areas. The amount of skin applied to the inner sides of each subject's left and right hands was (2.0 ± 0.1) mg / cm². 2 Application Examples 1-3 and Comparative Examples 1-3 were used to test skin moisture content at time intervals of 2h and 6h. The skin moisture content Corneometer probe of the German CK multifunctional skin tester was used to test skin moisture content, calculate the change in skin moisture content and calculate the average value. The average skin moisture content results are shown in Table 3.

[0168] Table 3

[0169]

[0170] As can be seen from Table 3, the compositions of Application Examples 1-3 of this application have significantly better moisturizing effects than those of Comparative Examples 1-3. This indicates that the oat polypeptide prepared in this application, when combined with hyaluronic acid, copper peptide, and ceramide components, has a significant skin moisturizing effect. The components of the compositions in this application are not obtained through conventional selection or substitution, resulting in technical effects superior to conventional compositions.

[0171] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A method of making an oat polypeptide, comprising, Including the following steps: (1) Oat swells, is dried after steam explosion treatment, and is then hydrolyzed by adding pectinase, cellulase and amylase. After filtration, the hydrolyzed oat residue is obtained. The mass ratio of pectinase, cellulase and amylase is 1:4-6:4-6. The total amount of pectinase, cellulase and amylase added is 80-130 mg of total enzyme mass / 1 g of oat mass. (2) Add a compound protease to the enzymatically hydrolyzed oat residue and carry out the first enzymatic hydrolysis under the conditions of pH=5.5-7 and ethanol volume concentration of 3-8% to obtain enzymatic hydrolysate A; The compound protease is papain, bromelain, and flavor protease in a mass ratio of 3-6:3-6:2; the amount of the compound protease added is 30-50 mg of compound protease mass / 1 g of oat mass; (3) Adjust the pH of the enzymatic hydrolysate A to 8.5-9.5, add trypsin for a second enzymatic hydrolysis to obtain enzymatic hydrolysate B; the amount of trypsin added is: 5-15 mg trypsin mass / 1g oat mass; (4) The enzyme inactivation of slurry B was carried out by enzyme hydrolysis, and the filtrate was subjected to ultrafiltration and concentration to obtain oat polypeptide.

2. The production method according to claim 1, characterized by, In step (1), the oat soaking process specifically involves mixing oats with hot water at 50-70°C and soaking for 1-3 hours.

3. The preparation method according to claim 1, characterized in that, In step (1), the steam explosion specifically involves: maintaining the swollen oats under a pressure of 0.8-2MPa for 60-200s, with a material-to-cavity ratio of 5:6-10, to obtain loosely structured oats.

4. The preparation method according to claim 1, characterized in that, In step (1), the enzymatic hydrolysis specifically involves mixing dried steam-exploded oat flour with 15-30 times its mass of water and enzymatically hydrolyzing it at 45-52°C for 3-6 hours.

5. The preparation method according to claim 1, characterized in that, In step (2), the complex protease is papain, bromelain, and flavor protease in a mass ratio of 4:5:

2. In step (2), the first enzymatic hydrolysis is specifically performed at 42-50℃ for 3-5 hours under the following conditions: solid-liquid ratio of 1:15-25 g / mL, pH=5.5-7, ethanol volume concentration of 3-8%.

6. The preparation method according to claim 1, characterized in that, In step (3), the second enzymatic hydrolysis specifically involves adjusting the pH of the enzymatic hydrolysate A to 8.5-9.2, adding trypsin, and enzymatically hydrolyzing at 37-44℃ for 2-5 hours.

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