Preparation method, application and composition of oat polypeptide

Abstract

The invention provides a preparation method, application and composition of oat polypeptide, and relates to the technical field of cosmetics. The preparation method disclosed by the invention comprises the following steps: (1) moistening and swelling oat, drying after steam explosion treatment, and adding pectinase, cellulase and amylase to carry out carbohydrate enzymolysis, so as to obtain oat residues after enzymolysis; (2) adding compound protease into the oat residues subjected to enzymolysis, and carrying out primary proteolysis under the condition that the addition amount of ethanol with the volume concentration of 3-8%, so as to obtain enzymolysis slurry A; the compound protease comprises papain, bromelain and flavourzyme in a mass ratio of (3-6): (3-6): 2; (3) adding trypsin into the enzymolysis serous fluid A to carry out secondary proteolysis to obtain enzymolysis serous fluid B; and (4) carrying out ultrafiltration on the enzymolysis slurry B to obtain the oat polypeptide. The obtained oat polypeptide is small in molecular weight and high in content, is used for preparing cosmetics, and has remarkable effects of preserving moisture, resisting wrinkles and reducing skin roughness.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and in particular relates to a preparation method of an oat polypeptide and its application and composition. Background Art

[0002] Oats are an annual herbaceous plant in the Poaceae family and are classified as a minor grain. Oats contain nutrients such as lipids, protein, and antioxidants, making them a high-quality grain. Their balanced nutrient profile makes them easily absorbed by the human body. They also possess beneficial health benefits, such as preventing and treating cardiovascular and cerebrovascular diseases and delaying aging. The protein content in oats is far higher than that found in other grains. Oat peptides are small-molecule peptides derived from oat protein through enzymatic hydrolysis, fermentation, or other techniques. They are typically composed of 2-20 amino acids. Compared to complete oat protein, oat peptides have a smaller molecular weight, making them easier to absorb and enhancing their functional activity.

[0003] The traditional protein extraction method is to use the "alkali dissolution and acid precipitation method" to obtain separated protein and then hydrolyze it with an enzyme preparation to obtain hydrolyzed protein peptides. The polypeptide content obtained by this method is low, and a large amount of acid and alkali are used, which causes certain pollution to the environment. To this end, Chinese invention patent CN115011660A discloses a method for preparing oat protein peptides. This method does not use the alkali dissolution and acid precipitation method, but instead grinds the oats and extracts them with an organic solvent to obtain oat protein. After primary enzymatic hydrolysis with pepsin and acid protease, secondary enzymatic hydrolysis with neutral protease and papain, and tertiary enzymatic hydrolysis with alkaline protease and flavor protease, the oat protein tertiary enzymatic hydrolyzate is obtained. After fermentation with Lactobacillus plantarum and Bacillus coagulans, the oat protein primary fermentation product is obtained, and then fermented with selenium-enriched yeast to obtain metabolically promoting and antioxidant oat protein peptides. However, this method uses at least 6 proteases for enzymatic hydrolysis, which greatly increases the cost of enzymatic hydrolysis; the fermentation process uses at least 3 bacteria, which also increases the complexity of the preparation process; the patent uses organic components such as acetone, acetonitrile, and trichloroacetic acid to extract oat protein, which has certain toxicity.

[0004] Chinese invention patent CN119498409A discloses an enzymatic hydrolysis method for oat milk, comprising the following steps: (1) regulating the activity of endogenous enzymes in oats and performing in situ enzymatic hydrolysis, using hot water to scald oat seeds to obtain heat-shocked oats; then swell the heat-shocked oats in a soaking solution containing gibberellins to obtain soaked oats; and continuing in situ enzymatic hydrolysis in the seeds while maintaining heat and moisture, causing the seeds to germinate; (2) beating and enzymatic hydrolysis with endogenous free enzymes in oats: adding water to beat the oats to obtain oat milk with residue, which is then kept at 40-45°C for 1 hour and filtered; (3) heating to 80-85°C and adding high-temperature α-amylase to complete starch liquefaction; (4) adding saccharifying enzyme and papain, maintaining heat and enzymatic hydrolysis for 1-1.5 hours, homogenizing, and sterilizing to finally obtain enzymatic hydrolyzed oat milk. Using the method of the invention, the polypeptide content obtained is above 9 mg / mL.

[0005] Cui Bingqun (Study on the Preparation Technology of Oat Peptides [J]. Modern Food Science and Technology, 2012, 28(08):1040-1042. DOI:10.13982 / j.mfst.1673-9078.2012.08.027.) used oat bran as raw material and used enzymatic technology to extract oat peptides. The experiment used the yield of oat peptides as an indicator, and used single-factor and orthogonal experimental methods to study the factors affecting the oat peptide extraction process and determine the main parameters of the new extraction process: 18-fold water addition, reaction temperature of 50°C, alkaline protease as the hydrolysis enzyme, and enzyme addition of 1.0%. The process described in this article can achieve an oat peptide yield of 18.9%, and the purity of the oat peptide after membrane filtration purification can reach 87.4%.

[0006] Currently, existing technologies for preparing oat protein peptides mostly utilize a single protease to hydrolyze the protein, without studying the synergistic effects of multiple proteases. The relationship between the type of protease used in the hydrolysis process and the peptide content, moisturizing and anti-wrinkle efficacy, remains unclear. Furthermore, oats contain a large amount of carbohydrates, which can adversely affect the enzymatic hydrolysis, purity, and efficacy of oat peptides. Summary of the Invention

[0007] In response to the problems existing in the prior art, the present invention provides a preparation method of an oat polypeptide, an application thereof, and a composition thereof. The oats are sequentially steam-exploded, enzymatically hydrolyzed with pectinase, cellulase, and amylase, and then enzymatically hydrolyzed with a composite protease and trypsin to obtain an oat polypeptide with a relatively small molecular weight. The oat polypeptide is used in the preparation of cosmetics, and the cosmetics have significant moisturizing, anti-wrinkle, and skin roughness reducing effects.

[0008] To achieve the above object, the technical solution adopted by the present invention is as follows: First, the present invention provides a method for preparing oat polypeptide, comprising the steps of: (1) The oats are swollen, steam-exploded, and then dried. Pectinase, cellulase, and amylase are added for enzymatic hydrolysis, and the oat residue is obtained by filtration; (2) adding a composite protease to the oat residue after enzymatic hydrolysis, and performing the first enzymatic hydrolysis under the conditions of pH = 5.5-7 and ethanol volume concentration of 3-8% to obtain enzymatic hydrolysis slurry A; The composite protease comprises papain, bromelain and flavor protease in a mass ratio of 3-6:3-6:2; (3) The pH of the enzymatic hydrolyzed slurry A is adjusted to 8.5-9.5, and trypsin is added for a second proteolysis to obtain the enzymatic hydrolyzed slurry B; (4) The enzyme in the enzymatically hydrolyzed slurry B is inactivated and filtered. The filtrate is ultrafiltered and concentrated to obtain oat polypeptides.

[0009] Preferably, in step (1), before the oats are swollen, a crushing step is also included, and the crushed particle size is 50-100 mesh.

[0010] Further preferably, in step (1), the crushed particle size is 80 mesh.

[0011] Preferably, in step (1), the oats are allowed to swell, specifically by mixing the oats with hot water at 50-70° C. and swell for 1-3 hours.

[0012] Further preferably, in step (1), the oats are allowed to swell, specifically by mixing the oats with 60° C. hot water and allowing the oats to swell for 2 hours.

[0013] Preferably, in step (1), the steam explosion is specifically as follows: the swollen oats are maintained under a pressurized pressure of 0.8-2 MPa for 60-200 seconds, with a material-cavity ratio of 5:6-10, to obtain oats with a loose structure.

[0014] Preferably, in step (1), the steam explosion is specifically as follows: the swollen oats are maintained at a pressurized pressure of 1.0 MPa for 120 seconds, with a material-cavity ratio of 5:8, to obtain oats with a loose structure.

[0015] Preferably, in step (1), before drying, the oats after steam explosion treatment need to be washed to remove sugar, acidic substances, hemicellulose, etc. on the surface.

[0016] Preferably, in step (1), the mass ratio of pectinase, cellulase and amylase is 1:4-6:4-6.

[0017] Further preferably, in step (1), the mass ratio of pectinase, cellulase and amylase is 1:5:5.

[0018] Preferably, in step (1), the enzymatic hydrolysis is specifically as follows: the dried steam-exploded oat flour is mixed with 15-30 times the mass of water, and enzymatically hydrolyzed at 45-52° C. for 3-6 hours.

[0019] Further preferably, in step (1), the enzymatic hydrolysis is specifically as follows: the dried steam-exploded oat flour is mixed with 20 times the mass of water and subjected to enzymatic hydrolysis at 48° C. for 5 h.

[0020] Preferably, in step (1), the total amount of enzymatic hydrolysis, pectinase, cellulase and amylase added is: 80-130 mg total mass of enzymes per 1 g mass of oats.

[0021] Preferably, in step (1), the total amount of the enzymatic hydrolysis, pectinase, cellulase and amylase added is: 100 mg of the total mass of the enzyme / 1 g of the mass of oats.

[0022] Preferably, in step (1), after filtration, the oat residue needs to be washed with water to remove surface starch, sugars generated after enzymatic hydrolysis, and by-products.

[0023] Preferably, in step (2), the composite protease is papain, bromelain, and flavor protease in a mass ratio of 4:5:2.

[0024] Preferably, in step (2), the added amount of the composite protease is: 30-50 mg composite protease mass / 1 g oat mass.

[0025] Further preferably, in step (2), the added amount of the composite protease is: 40 mg composite protease mass / 1 g oat mass.

[0026] Preferably, in step (2), the first enzymatic hydrolysis is specifically performed at a solid-liquid ratio of 1:15-25 g / mL, pH = 5.5-7, and an ethanol volume concentration of 3-8%, and at 42-50°C for 3-5 hours.

[0027] Further preferably, in step (2), the first enzymatic hydrolysis is specifically performed at a solid-liquid ratio of 1:20 g / mL, pH = 6.0, and an ethanol volume concentration of 6%, and the enzymatic hydrolysis is performed at 44°C for 4 hours.

[0028] In the present invention, the ethanol volume concentration of 3-8% in step (2) means that the amount of ethanol added is equivalent to 3-8% of the total volume of the enzymatic hydrolysis liquid.

[0029] Preferably, in step (3), the amount of trypsin added is: 5-15 mg trypsin mass / 1 g oat mass.

[0030] Further preferably, in step (3), the amount of trypsin added is: 10 mg trypsin mass / 1 g oat mass.

[0031] Preferably, in step (3), the second enzymatic hydrolysis is specifically as follows: the pH of the enzymatic hydrolysis slurry A is adjusted to 8.5-9.2, trypsin is added, and the enzymatic hydrolysis treatment is carried out at 37-44°C for 2-5 hours.

[0032] Further preferably, in step (3), the second enzymatic hydrolysis is specifically as follows: the pH of the enzymatic hydrolysis slurry A is adjusted to 8.6, trypsin is added, and the enzymatic hydrolysis is carried out at 39° C. for 3 h.

[0033] Preferably, in step (4), the enzyme is inactivated by treating at 121°C for 10-15 minutes.

[0034] Preferably, in step (4), the ultrafiltration is performed using an ultrafiltration membrane with a pore size of ≤8000Da.

[0035] Then, the present invention provides the oat polypeptide prepared by the above preparation method.

[0036] Furthermore, the present invention provides the use of the above-mentioned oat polypeptide in the preparation of cosmetic raw materials or cosmetics, wherein the cosmetics in the application are moisturizing and anti-wrinkle cosmetics.

[0037] Preferably, the cosmetics used in the application are cosmetics that can moisturize, increase skin elasticity, increase collagen content in the skin, promote skin metabolism, and promote cell proliferation.

[0038] Furthermore, the present invention provides a cosmetic raw material containing the above oat polypeptide, comprising the components: the above oat polypeptide, a moisturizer, a skin conditioner, a preservative and water.

[0039] Preferably, the cosmetic raw material comprises, by mass percentage, 0.1-0.4% of the above-mentioned oat polypeptide, 8-15% of moisturizer, 0.05-0.2% of skin conditioner, 0.5-1.5% of preservative and the balance of water.

[0040] Further preferably, the cosmetic raw material comprises, by weight percentage, 0.3% oat polypeptide, 10% moisturizer, 0.1% skin conditioner, 0.9% preservative and the balance water.

[0041] Preferably, the moisturizer, skin conditioner, and preservative in the cosmetic raw materials are all ingredients commonly used in the art. In some embodiments of the present invention, the moisturizer is glycerin, the skin conditioner is ethylhexylglycerin, and the preservative is phenoxyethanol.

[0042] Finally, the present invention provides a moisturizing and anti-wrinkle composition, comprising the oat polypeptide, hyaluronic acid, copper peptide and ceramide.

[0043] Preferably, in the moisturizing and anti-wrinkle composition, the mass ratio of oat polypeptide, hyaluronic acid, copper peptide and ceramide is 20:5-10:2-5:2-5.

[0044] Further preferably, in the moisturizing and anti-wrinkle composition, the mass ratio of oat polypeptide, hyaluronic acid, copper peptide and ceramide is 20:8:4:4.

[0045] Compared with the prior art, the present invention has the following beneficial effects: 1. The oat polypeptide prepared by the preparation method of the present invention has a structure similar to that of a natural moisturizing factor and has excellent moisturizing properties. The oats are first subjected to swelling, steam explosion, and carbohydrate enzymatic hydrolysis by pectinase, cellulase, and amylase, which is conducive to protein exposure and easy hydrolysis by proteases. The obtained oat residue is then subjected to a first enzymatic hydrolysis by a composite protease and a second endoproteolytic hydrolysis by trypsin, so that the high-molecular-weight protein is efficiently enzymatically hydrolyzed into small-molecule oat polypeptides, thereby increasing the proportion of small-molecule polypeptides.

[0046] 2. The oat polypeptide prepared by the present invention has a high proportion of small molecule polypeptides, which is conducive to being absorbed by the skin, thereby further improving the moisturizing and anti-wrinkle properties.

[0047] 3. The oat polypeptide of the present invention works synergistically with hyaluronic acid, copper peptide and ceramide components to prepare a composition that significantly moisturizes, increases skin elasticity and reduces roughness. DETAILED DESCRIPTION

[0048] The following non-limiting examples are provided to enable those skilled in the art to more fully understand the present invention, but are not intended to limit the present invention in any way. The following are merely illustrative of the scope of the present invention, and those skilled in the art may make various changes and modifications to the present invention based on the disclosed content, which should also fall within the scope of the present invention.

[0049] When numerical ranges are given in the examples, it should be understood that, unless otherwise specified herein, 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 herein have the same meaning as commonly understood by those of ordinary skill in the art to which the invention belongs.

[0050] The present invention is further described below by way of specific examples. Unless otherwise specified, the various chemical reagents used in the examples of the present invention were obtained through conventional commercial channels.

[0051] In the following examples, the papain was purchased from Nanning Shanwan Biotechnology Co., Ltd., with an enzyme activity of 100,000 U / g; Bromelain was purchased from Nanning Pangbo Bioengineering Co., Ltd., with an enzyme activity of 50,000 U / g; The flavor protease was purchased from Nanning Shanwan Biotechnology Co., Ltd., with an enzyme activity of 20,000 U / g; The trypsin was purchased from Nanning Pangbo Bioengineering Co., Ltd., with an enzyme activity of 4000 U / g; The neutral protease was purchased from Shandong Longkote Enzyme Preparation Co., Ltd., with an enzyme activity of 100,000 U / g; The alkaline protease was purchased from Shandong Longkote Enzyme Preparation Co., Ltd., and the enzyme activity was 50,000 U / g.

[0052] For the above components, products from different manufacturers have no significant impact on the effects.

[0053] Example 1 A method for preparing oat polypeptide comprises the steps of: (1) Grind the oats into 80 mesh and soak them in 60℃ water for 2 hours, with the amount of water covering the oats; The swollen oats were steam exploded at a pressure of 1.0 MPa for 120 seconds, with a material-to-cavity ratio of 5:8, washed with water, and dried at 60°C to obtain loose oats (dried steam-exploded oat flour). The dried steam-exploded oat flour was mixed with 20 times the mass of water, and pectinase, cellulase, and amylase were added in a mass ratio of 1:5:5 (addition amount: 100 mg of total enzyme mass / 1 g of oat raw material mass). The mixture was enzymatically hydrolyzed at 48°C for 5 h and filtered to obtain the enzymatically hydrolyzed oat residue.

[0054] (2) Adding a composite protease to the oat residue after enzymatic hydrolysis, with the weight of 40 mg composite protease per 1 g of oat raw material, to perform the first enzymatic hydrolysis to obtain enzymatic slurry A; The composite protease is papain, bromelain, and flavor protease in a mass ratio of 4:5:2; The first enzymatic hydrolysis conditions were: solid-liquid ratio 1:20 g / mL, pH = 6.0, ethanol volume concentration 6%, and enzymatic hydrolysis at 44 °C for 4 h.

[0055] (3) The pH of the enzymatic hydrolysis slurry A was adjusted to 8.6, and trypsin was added at a rate of 10 mg trypsin / 1 g oat raw material. The enzymatic hydrolysis was carried out at 39°C for 3 h, and a second enzymatic hydrolysis was performed to obtain the enzymatic hydrolysis slurry B.

[0056] (4) The enzymatic hydrolysis slurry B was treated at 121°C for 15 min to inactivate the enzyme, filtered, and ultrafiltered using an ultrafiltration membrane with a pore size of ≤8000 Da. The concentrated solution was then concentrated to obtain oat polypeptides.

[0057] Example 2 The difference from Example 1 is that in step (2), the mass ratios of papain, bromelain and flavor protease are different.

[0058] In this embodiment, the mass ratio of papain, bromelain and flavor protease is 6:3:2.

[0059] The rest are the same as in Example 1.

[0060] Example 3 The difference from Example 1 is that in step (2), the mass ratios of papain, bromelain and flavor protease are different.

[0061] In this embodiment, the mass ratio of papain, bromelain and flavor protease is 3:6:2.

[0062] The rest are the same as in Example 1.

[0063] Example 4 A method for preparing oat polypeptide comprises the steps of: (1) Grind the oats into 80 mesh and soak them in 50℃ water for 3 hours, with the amount of water covering the oats; The swollen oats were steam exploded at a pressure of 0.8 MPa for 200 s with a material-to-cavity ratio of 5:10, washed with water, and freeze-dried to obtain loosely structured oats (dried steam-exploded oat flour). The dried steam-exploded oat flour was mixed with 30 times the mass of water, and pectinase, cellulase, and amylase were added in a mass ratio of 1:4:6 (added amount: 80 mg of total enzyme mass / 1 g of oat raw material mass). The oat flour was enzymatically hydrolyzed at 52°C for 3 h and filtered to obtain the enzymatically hydrolyzed oat residue.

[0064] (2) Adding a composite protease to the oat residue after enzymatic hydrolysis, with the mass of 50 mg composite protease per mass of 1 g oat raw material, to perform the first enzymatic hydrolysis to obtain enzymatic slurry A; The composite protease is papain, bromelain, and flavor protease in a mass ratio of 4:5:2; The first enzymatic hydrolysis conditions were: solid-liquid ratio 1:25 g / mL, pH = 7.0, ethanol volume concentration 3%, and enzymatic hydrolysis at 42 °C for 5 h.

[0065] (3) The pH of the enzymatic hydrolysis slurry A was adjusted to 8.5, and trypsin was added at a rate of 5 mg trypsin / 1 g oat raw material. The enzymatic hydrolysis was carried out at 37°C for 2 h, and a second enzymatic hydrolysis was performed to obtain the enzymatic hydrolysis slurry B.

[0066] (4) The enzymatic hydrolysis slurry B was treated at 121°C for 15 min to inactivate the enzyme, filtered, and ultrafiltered using an ultrafiltration membrane with a pore size of ≤8000 Da. The concentrated solution was then concentrated to obtain oat polypeptides.

[0067] Example 5 A method for preparing oat polypeptide comprises the steps of: (1) Grind the oats into 80 mesh and soak them in 70℃ water for 1 hour, with the amount of water covering the oats; The swollen oats were steam exploded at a pressure of 2 MPa for 60 seconds, with a material-to-cavity ratio of 5:6, washed with water, and freeze-dried to obtain loosely structured oats (dried steam-exploded oat flour). The dried steam-exploded oat flour was mixed with 15 times the mass of water, and pectinase, cellulase, and amylase were added in a mass ratio of 1:4:6 (added amount: 130 mg total mass of enzyme / 1 g mass of oat raw material). The oat flour was enzymatically hydrolyzed at 45°C for 6 h and filtered to obtain the enzymatically hydrolyzed oat residue.

[0068] (2) Adding a composite protease to the oat residue after enzymatic hydrolysis, with the mass of 30 mg composite protease per mass of 1 g oat raw material, to perform the first enzymatic hydrolysis to obtain enzymatic slurry A; The composite protease is papain, bromelain, and flavor protease in a mass ratio of 4:5:2; The first enzymatic hydrolysis conditions were: solid-liquid ratio 1:15 g / mL, pH = 5.5, ethanol volume concentration 8%, and enzymatic hydrolysis at 50 °C for 3 h.

[0069] (3) The pH of the enzymatic hydrolysis slurry A was adjusted to 9.2, and trypsin was added at a rate of 15 mg trypsin per 1 g of oat raw material. The enzymatic hydrolysis was carried out at 44°C for 5 h, and a second enzymatic hydrolysis was performed to obtain the enzymatic hydrolysis slurry B.

[0070] (4) The enzymatic hydrolysis slurry B was treated at 121°C for 15 min to inactivate the enzyme, filtered, and ultrafiltered using an ultrafiltration membrane with a pore size of ≤8000 Da. The concentrated solution was then concentrated to obtain oat polypeptides.

[0071] Comparative Example 1 The difference from Example 1 is that in step (1), the steam explosion treatment is replaced by ultrasonic treatment.

[0072] (1) Grind the oats into 80 mesh and soak them in 60℃ water for 2 hours, with the amount of water covering the oats; The swollen oats were treated at an ultrasonic frequency of 500W for 2 minutes, washed with water, and dried at 60°C to obtain oats with a loose structure (dried ultrasonic oatmeal powder). The dried ultrasonic oatmeal powder was mixed with 20 times the mass of water, and pectinase, cellulase and amylase were added in a mass ratio of 1:5:5 (addition amount: 100 mg total mass of enzyme / 1 g mass of oat raw material). The oatmeal powder was enzymatically hydrolyzed at 48°C for 5 h and filtered to obtain the oatmeal residue after enzymatic hydrolysis.

[0073] Steps (2) to (4) are the same as in Example 1.

[0074] Comparative Example 2 Different from Example 1, in step (1), pectinase is replaced by cellulase and amylase in a mass ratio of 1:1.

[0075] (1) Grind the oats into 80 mesh and soak them in 60℃ water for 2 hours, with the amount of water covering the oats; The swollen oats were steam exploded at a pressure of 1.0 MPa for 120 seconds, with a material-to-cavity ratio of 5:8, washed with water, and dried at 60°C to obtain loose oats (dried steam-exploded oat flour). The dried steam-exploded oat flour was mixed with 20 times the mass of water, and cellulase and amylase were added in a mass ratio of 5.5:5.5 (added amount: 100 mg total mass of enzyme / 1 g mass of oat raw material). The mixture was enzymatically hydrolyzed at 48°C for 5 h and filtered to obtain the enzymatically hydrolyzed oat residue.

[0076] Steps (2) to (4) are the same as in Example 1.

[0077] Comparative Example 3 The difference from Example 1 is that bromelain in step (2) is replaced by neutral protease. The composite protease is papain, neutral protease, and flavor protease in a mass ratio of 4:5:2.

[0078] The rest are the same as in Example 1.

[0079] Comparative Example 4 The difference from Example 1 is that in step (2), the mass ratio of papain, bromelain and flavor protease is 8:1:2.

[0080] The rest are the same as in Example 1.

[0081] Comparative Example 5 The difference from Example 1 is that in step (2), the volume concentration of ethanol is 30% (relative to 30% of the total volume of the enzymatic hydrolysis liquid).

[0082] The rest are the same as in Example 1.

[0083] Comparative Example 6 The difference from Example 1 is that in step (3), the trypsin is replaced by alkaline protease. The rest is the same as Example 1.

[0084] The oat polypeptide of Example 1 was freeze-dried and used in the preparation of the compositions of the following application examples and comparative application examples.

[0085] Application Example 1 A moisturizing and anti-wrinkle composition comprises the following components in parts by mass: 20 parts of oat polypeptide, 8 parts of hyaluronic acid, 4 parts of copper peptide and 4 parts of ceramide.

[0086] The above components are mixed evenly to obtain a moisturizing and anti-wrinkle composition.

[0087] Application Example 2 A moisturizing and anti-wrinkle composition comprises the following components in parts by mass: 20 parts of oat polypeptide, 5 parts of hyaluronic acid, 5 parts of copper peptide and 5 parts of ceramide.

[0088] The above components are mixed evenly to obtain a moisturizing and anti-wrinkle composition.

[0089] Application Example 3 A moisturizing and anti-wrinkle composition comprises the following components in parts by mass: 20 parts of oat polypeptide, 10 parts of hyaluronic acid, 2 parts of copper peptide and 2 parts of ceramide.

[0090] The above components are mixed evenly to obtain a moisturizing and anti-wrinkle composition.

[0091] Comparative Application Example 1 The difference from Application Example 1 is that the oat polypeptide is replaced by commercially available soybean oligopeptide.

[0092] Application Comparative Example 2 The difference from Application Example 1 is that the copper peptide is replaced by acetyl hexapeptide-8.

[0093] Application Comparative Example 3 The difference from Application Example 1 is that the mass ratio of the components in the composition is different: 30 parts of oat polypeptide, 1 part of hyaluronic acid, 4 parts of copper peptide and 1 part of ceramide.

[0094] Experiment 1 Determination of polypeptide content and polypeptide molecular weight The oat polypeptides obtained after ultrafiltration in each example and comparative example were concentrated and fixed to a solid-liquid ratio of 1:10 g:mL (the solid mass refers to the mass of the raw oats). The obtained oat polypeptide products were tested for polypeptide content and polypeptide molecular weight.

[0095] 1. Detection of polypeptide content: 5 mL of concentrated oat peptide was added with 5 mL of 10% trichloroacetic acid solution, centrifuged at 8000 rpm for 3 min, and the supernatant was collected for testing (test sample solution).

[0096] (1) Drawing 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 a test tube, add water to 0.8 mL, add 3.2 mL of biuret reagent, mix well, and react at 25°C for 30 min. Use 1.0 mL of distilled water as a blank control, measure the absorbance at a wavelength of 540 nm using a UV spectrophotometer, and average the three replicates. Draw a standard curve with protein content as the horizontal axis and OD 540 nm as the vertical axis. The linear regression equation of the standard curve has an R2 of 0.9999, where y is the absorbance and x is the protein content (mg / mL).

[0097] (2) Determination of peptides in 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 at a wavelength of 540 nm using a UV spectrophotometer. Calculate the peptide content in the solution based on the standard curve. The peptide content test results are shown in Table 1.

[0098] 2. Molecular weight determination was performed in accordance with Appendix A of GB / T 22492-2008, "Determination of Relative Molecular Mass Distribution of Peptides." The results of the molecular weight distribution of oat peptides are shown in Table 1.

[0099] Table 1

[0100] In Table 1, # 、 ## The results of the comparative example group were significantly different from those of the example group. # P<0.05, ## P<0.01.

[0101] As can be seen from Table 1, the oat polypeptide product prepared by the present 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 conducive to skin absorption and thus enhances moisturizing and anti-wrinkle effects.

[0102] Experiment 2: Testing the effect of improving skin roughness Test method: The skin roughness was tested using the skin friction test probe Frictiometer FR700 produced by German CK Company.

[0103] Test population: A total of 66 people, all female, aged 25-45 years old, were randomly divided into 11 groups, with 6 people in each group.

[0104] Test Site and Requirements: The inner sides of the left and right forearms were selected. Four identical areas (3cm x 3cm) were defined on the left forearm for oat peptide testing. Testing was conducted in a room at 20-22°C and 40%-60% relative humidity. Subjects were required to acclimate to the same environment for 20 minutes. During testing, 1g of oat peptide was applied to cover the test area.

[0105] The test results were collected at the following two time periods: before using the sample (D0) and 56 days after using the sample (D56).

[0106] Skin roughness improvement effect (%) = (D0-D56) / D0×100%.

[0107] The mean skin friction test results are shown in Table 2.

[0108] Table 2

[0109] In Table 2, the comparison of skin roughness improvement effect is shown. # 、 ## The results of the comparative example group were significantly different from those of the example group. # P<0.05, ## P<0.01.

[0110] As can be seen in Table 2, after using oat peptides, skin friction was improved and roughness decreased. The skin roughness of the Example group decreased to a greater extent than that of the Control Group. This indicates that the oat peptides prepared in the Examples of this application have a superior ability to improve roughness, and provide a more effective effect on improving skin and enhancing smoothness than the Control Group and conventional oat peptides.

[0111] Experiment 3: Moisturizing performance test of the composition The moisturizing and anti-wrinkle compositions of Application Examples 1-3 and Comparative Application Examples 1-3 were prepared into 1.0% (w / w) aqueous solutions.

[0112] Test Methods: 24 healthy men and women were randomly divided into 6 groups, 12 men and 12 women in each group. Before the test, the subjects were asked to sit quietly in a climate-controlled room (22±1°C, 50% relative humidity) for at least 20 minutes and remain relaxed 2 hours after washing their arms. The subjects were required to have no scars, pigmentation, atrophy, port-wine stains, or other blemishes on their forearms that could affect the test results.

[0113] Three 3 cm × 3 cm skin areas were randomly marked on the inner side of the left and right forearms of the subjects as test areas. The application amount on the inner side of the left and right hands of each subject was (2.0 ± 0.1) mg / cm 2 In Application Examples 1-3 and Comparative Examples 1-3, the skin moisture content was tested at time intervals of 2h and 6h. The skin moisture content was tested using the Corneometer test probe of the German CK multifunctional skin tester. The change in skin moisture content was calculated and the average value was calculated. The average skin moisture content results are shown in Table 3.

[0114] Table 3

[0115] As can be seen from Table 3, the moisturizing effects of the compositions of Application Examples 1-3 of the present application are significantly better than those of Comparative Examples 1-3. This indicates that the oat peptides prepared in the present application, when compounded with hyaluronic acid, copper peptide, and ceramide components, have a significant moisturizing effect on the skin. The components of the compositions of the present application are not conventionally selected or replaced, resulting in a technical effect that is superior to conventional compositions.

[0116] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A method for preparing oat polypeptide, characterized in that: Including steps: (1) The oats are swollen, steam-exploded, and then dried. Pectinase, cellulase, and amylase are added for enzymatic hydrolysis, and the oat residue is obtained by filtration; (2) adding a composite protease to the oat residue after enzymatic hydrolysis, and performing the first enzymatic hydrolysis under the conditions of pH = 5.5-7 and ethanol volume concentration of 3-8% to obtain enzymatic hydrolysis slurry A; The composite protease comprises papain, bromelain and flavor protease in a mass ratio of 3-6:3-6:2; (3) The pH of the enzymatic hydrolyzed slurry A is adjusted to 8.5-9.5, and trypsin is added for a second proteolysis to obtain the enzymatic hydrolyzed slurry B; (4) The enzyme in the enzymatically hydrolyzed slurry B is inactivated and filtered. The filtrate is ultrafiltered and concentrated to obtain oat polypeptides.

2. The preparation method according to claim 1, characterized in that In step (1), the oats are allowed to swell, specifically by mixing the oats with hot water at 50-70° C. and allowing the mixture to swell for 1-3 hours.

3. The preparation method according to claim 1, characterized in that In step (1), the steam explosion is specifically as follows: the swollen oats are maintained under a pressurized pressure of 0.8-2 MPa for 60-200 seconds, with a material-cavity ratio of 5:6-10, to obtain oats with a loose structure.

4. The preparation method according to claim 1, characterized in that In step (1), 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; In step (1), the enzymatic hydrolysis of the carbohydrates is specifically as follows: the dried steam-exploded oat flour is mixed with 15-30 times the mass of water, and the mixture is enzymatically hydrolyzed at 45-52° C. for 3-6 hours.

5. The preparation method according to claim 1, characterized in that In step (2), the composite protease is papain, bromelain, and flavor protease in a mass ratio of 4:5:2; the added amount of the composite protease is: 30-50 mg composite protease mass / 1 g oat mass; In step (2), the first enzymatic hydrolysis is specifically carried out at a solid-liquid ratio of 1:15-25 g / mL, pH = 5.5-7, and an ethanol volume concentration of 3-8%, and at 42-50°C for 3-5 hours.

6. The preparation method according to claim 1, characterized in that In step (3), the second enzymatic hydrolysis is specifically as follows: the pH of the enzymatic hydrolysis slurry A is adjusted to 8.5-9.2, trypsin is added, and the enzymatic hydrolysis is carried out at 37-44°C for 2-5 hours; In step (3), the amount of trypsin added is: 5-15 mg trypsin mass / 1 g oat mass.

7. The oat polypeptide prepared by the preparation method according to any one of claims 1 to 6.

8. Use of the oat polypeptide according to claim 7 in the preparation of cosmetic raw materials or cosmetics.

9. A cosmetic raw material, characterized in that: The invention comprises the following components: the oat polypeptide prepared by the preparation method according to any one of claims 1 to 6 or the oat polypeptide according to claim 7, a moisturizer, a skin conditioner, a preservative and water.

10. A moisturizing and anti-wrinkle composition, characterized in that: The invention comprises the following components: the oat polypeptide prepared by the preparation method according to any one of claims 1 to 6 or the oat polypeptide according to claim 7, hyaluronic acid, copper peptide and ceramide.

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