Oat bran fermentation product and methods of making and using same

Abstract

The application discloses an oat bran fermentation product and a preparation method and application thereof, and combines an enzymolysis method and a fermentation method to take oat bran as a main raw material, and obtains the oat bran fermentation product through gelatinization, primary enzymolysis, fermentation and secondary enzymolysis, wherein the oat bran fermentation product contains active substances such as beta-glucan, oat protein, oat polypeptide, polysaccharide, polyphenol, ferulic acid and feruloyl oligosaccharide.The oat bran fermentation product has high content of active substances, is easy to be absorbed by skin, has good anti-aging, antioxidant, anti-inflammatory and bacteriostatic effects and moisturizing effect, can be applied to the field of cosmetics, and is used for preparing emulsions, water agents, creams, gels, lotions and masks, and greatly improves the bioavailability of the oat bran, and has low production cost and high enzymolysis and fermentation efficiency.

Description

Technical Field

[0001] This invention relates to the field of bio-fermentation technology, and in particular to an oat bran fermentation product, its preparation method, and its application. Background Technology

[0002] Oats, as a functional ingredient, have wide applications in food, feed, medicine, cosmetics, and industrial raw materials. Oat bran, a major byproduct of oat processing, is generally treated cheaply as livestock feed or discarded as waste, failing to be effectively utilized and its value fully realized, resulting in significant resource waste. However, research shows that oat bran contains various functional active ingredients, such as proteins, polysaccharides, fats, dietary fiber, lignin, phenolic substances, and vitamins.

[0003] Currently, beta-glucan has become a research hotspot as a skincare ingredient due to its remarkable effects, such as promoting collagen synthesis, increasing skin elasticity, and providing high-efficiency moisturizing. However, other active ingredients in oat bran also have significant research value. For example, oat skin-firming protein, a unique component of oat bran, has excellent skin-firming, moisturizing, and anti-aging effects. In addition, oat bran also contains polyphenols with antioxidant activity and oat peptides with anti-inflammatory, antibacterial, and anti-aging properties. Therefore, oat bran has broad application prospects in the cosmetics industry and possesses great development value.

[0004] In recent years, the cosmetics industry has developed rapidly, with people paying more attention to functional cosmetics such as moisturizing, whitening, anti-aging, and anti-allergy products, no longer limited to basic skincare products aimed at cleansing and moisturizing. As living standards improve, people prefer natural cosmetics to chemically synthesized moisturizers. Cosmetics with natural active ingredients are favored by more and more people due to their low irritation, good penetration, and significant effects. Therefore, there is an urgent need to develop an oat fermentation extract for use in cosmetics, which would both increase the added value of oats and meet consumer demand. Summary of the Invention

[0005] The main objective of this invention is to provide a method for preparing oat bran fermentation products and their applications, aiming to solve the technical problem of low utilization rate of oat bran.

[0006] To achieve the above objectives, the present invention provides a method for preparing oat bran fermentation products, comprising the following steps:

[0007] Gelatinization: Oat bran and water are mixed and gelatinized to obtain oat gelatinized liquid;

[0008] One-step enzymatic hydrolysis: α-amylase and saccharifying enzyme are added to the oat gelatinized liquid for enzymatic hydrolysis and enzyme inactivation treatment to obtain oat bran fermentation medium;

[0009] Fermentation: Yellow wine yeast was inoculated into the oat bran fermentation medium and fermentation was carried out to obtain fermentation broth;

[0010] Secondary enzymatic hydrolysis: The pH value of the oat bran fermentation broth was adjusted, and then neutral protease and ferulic acid esterase were added for secondary enzymatic hydrolysis. The supernatant was sterilized to obtain the oat bran fermentation product.

[0011] In some embodiments of this application, the oat bran is further subjected to micronization and sieving before the gelatinization process.

[0012] In some embodiments of this application, the oat bran is sieved through a mesh size of 100-200.

[0013] In some embodiments of this application, in the gelatinization step, the ratio of oat bran to water is 1:10-25 (g / ml);

[0014] And / or, in the gelatinization step, the gelatinization temperature is 55℃-95℃;

[0015] And / or, in the gelatinization step, the gelatinization time is 35 min to 60 min.

[0016] In some embodiments of this application, in the single enzymatic hydrolysis step, based on 100% of the total mass of the oat gelatinized liquid, the amount of α-amylase added is 0.05%-0.15%;

[0017] And / or, in the first enzymatic hydrolysis step, based on 100% of the total mass of the oat gelatinized liquid, the amount of saccharifying enzyme added is 0.05%-0.15%;

[0018] And / or, in the first enzymatic hydrolysis step, the temperature of the first enzymatic hydrolysis is 55℃-95℃;

[0019] And / or, in the single enzymatic hydrolysis step, the time for the single enzymatic hydrolysis is 35 min-60 min;

[0020] And / or, in the single enzymatic hydrolysis step, the α-amylase is a high-temperature α-amylase.

[0021] In the fermentation step, based on 100% of the mass of the oat bran fermentation medium, the amount of rice wine yeast added is 1%-5%.

[0022] And / or, in the fermentation step, the fermentation temperature is 25°C-30°C;

[0023] And / or, in the fermentation step, the fermentation time is 3-4 days.

[0024] In some embodiments of this application, during the secondary enzymatic hydrolysis step, the pH of the oat bran fermentation broth is adjusted to neutral.

[0025] And / or, based on 100% of the total mass of the oat bran fermentation broth, the amount of neutral protease added is 0.001%-0.015%;

[0026] And / or, based on 100% of the total mass of the oat bran fermentation product, the amount of ferulic acid esterase added is 0.01%-0.15%;

[0027] And / or, the temperature of the secondary enzymatic hydrolysis is 45℃-55℃;

[0028] And / or, the secondary enzymatic hydrolysis time is 3-5 hours;

[0029] And / or, in the secondary enzymatic hydrolysis step, the supernatant is obtained by centrifugation;

[0030] And / or, after completing the sterilization operation of the supernatant, the method further includes adding a preservative.

[0031] In some embodiments of this application, after the supernatant is taken in the secondary enzymatic hydrolysis step, there is residual filter residue. The filter residue is subjected to cell wall breaking, water extraction and centrifugation to obtain the supernatant to obtain oat bran fermentation lysate.

[0032] In addition, to achieve the above objectives, the present invention also provides an oat bran fermentation product, which is prepared by the preparation method described above.

[0033] In some embodiments of this application, the oat bran fermentation product contains β-glucan, oat protein, oat polypeptide, polysaccharide, polyphenols, ferulic acid, and feruloyl oligosaccharide.

[0034] In addition, to achieve the above objectives, the present invention also provides an application of oat bran fermentation product in the cosmetics field.

[0035] The beneficial effects that this invention can achieve are:

[0036] This invention combines enzymatic hydrolysis and fermentation, resulting in low production costs and high efficiency in both processes. This significantly improves the extraction rate of active substances from oat bran. The resulting fermented oat bran product contains active substances such as β-glucan, oat protein, oat polypeptides, polysaccharides, polyphenols, ferulic acid, and feruloyl oligosaccharides, enhancing the utilization rate of active ingredients in oat bran. Furthermore, the fermented small-molecule active substances are more easily absorbed by the skin, exhibiting excellent anti-aging, antioxidant, anti-inflammatory, antibacterial, and moisturizing effects. It can be applied in the cosmetics field for preparing emulsions, liquids, creams, gels, lotions, and masks, greatly improving the bioavailability of oat bran.

[0037] Furthermore, the present invention can also break down the cell walls, extract water, and centrifuge the fermented filter residue of oat bran to obtain fermented oat bran lysate. This fermented oat bran lysate also contains active substances such as β-glucan, oat protein, oat polypeptide, polyphenols, ferulic acid, and feruloyl oligosaccharides, which can also be applied in the cosmetics field. Thus, the full utilization of oat bran can be achieved, and its bioavailability can be further improved. Attached Figure Description

[0038] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art are briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the structures shown in these drawings without creative effort.

[0039] Figure 1 This is a schematic flowchart of a method for preparing an oat bran fermentation product according to the present invention.

[0040] Figure 2 This is a graph showing the results of the stratum corneum moisture content test in an application example of the present invention.

[0041] Figure 3 This is a graph showing the results of the transdermal moisture loss test in an application example of the present invention.

[0042] Figure 4 This is a graph showing the results of the skin elasticity R2 test in an application example of the present invention.

[0043] Figure 5 This is a graph showing the results of the F4 skin elasticity test in an application example of the present invention.

[0044] The realization of the objective, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0045] It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0046] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0047] In this invention, the terms "first," "second," etc., are used for descriptive purposes only and should not be construed as indicating or implying their relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined with "first" or "second" may explicitly or implicitly include at least one of that feature. Furthermore, the technical solutions of the various embodiments can be combined with each other, but only on the basis of being achievable by those skilled in the art. If the combination of technical solutions is contradictory or impossible to implement, such a combination of technical solutions should be considered non-existent and not within the scope of protection claimed by this invention.

[0048] This invention provides a method for preparing fermented oat bran products, referring to... Figure 1 The preparation method of oat bran fermentation products includes the following steps:

[0049] Gelatinization: Oat bran and water are mixed and gelatinized to obtain oat gelatinized liquid;

[0050] One-step enzymatic hydrolysis: α-amylase and saccharifying enzyme are added to the oat gelatinized liquid for enzymatic hydrolysis and enzyme inactivation treatment to obtain oat bran fermentation medium;

[0051] Fermentation: Yellow wine yeast was inoculated into the oat bran fermentation medium to carry out fermentation and obtain oat bran fermentation liquid;

[0052] Secondary enzymatic hydrolysis: The pH value of the oat bran fermentation broth was adjusted, and then neutral protease and ferulic acid esterase were added for secondary enzymatic hydrolysis. The supernatant was sterilized to obtain the oat bran fermentation product.

[0053] This invention combines enzymatic hydrolysis and fermentation, resulting in low production costs and high efficiency in both processes. The fermented oat bran product contains active substances such as β-glucan, oat protein, oat polypeptides, polyphenols, ferulic acid, and feruloyl oligosaccharides, which are easily absorbed by the skin and have excellent anti-aging, antioxidant, anti-inflammatory, antibacterial, and moisturizing effects. It can be applied in the cosmetics field to prepare emulsions, lotions, creams, gels, lotions, and masks, greatly improving the bioavailability of oat bran.

[0054] In some embodiments, oat bran can be pre-treated before gelatinization, for example, by pulverizing it with a micro grinder and then sieving it. This promotes gelatinization of the oat bran and improves its bioactivity. The mesh size of the sieve can be 100-200 mesh, resulting in smaller oat bran particles, which facilitates the release of active substances from the oat bran.

[0055] In this invention, the purpose of the gelatinization step is to remove some of the starch and impurities from the oat bran. At the same time, it can also remove the endogenous β-glucanase in the oat bran, preventing the endogenous β-glucanase from enzymatically hydrolyzing the active substance β-glucan and increasing the content of active substances in the oat bran fermentation product.

[0056] In some embodiments of the gelatinization step, the ratio of oat bran to water is 1:10 to 25 (g / ml). For example, it can be any ratio within the range of 1:10 (g / ml), 1:11 (g / ml), 1:14 (g / ml), 1:15 (g / ml), 1:17 (g / ml), 1:18 (g / ml), 1:20 (g / ml), 1:21 (g / ml), 1:23 (g / ml), 1:25 (g / ml), etc. This can yield oat gelatinized liquid with better gelatinization effect, thereby improving the efficiency of subsequent enzymatic hydrolysis and fermentation.

[0057] In some embodiments of the gelatinization step, the gelatinization temperature is 55℃-95℃, and the gelatinization time is 35min-60min. For example, the gelatinization temperature can be any temperature within the range of 55℃-95℃, such as 55℃, 60℃, 65℃, 70℃, 75℃, 80℃, 85℃, 90℃, or 95℃, and the gelatinization time can be any time value within the range of 35min-60min, such as 35min, 40min, 45min, 50min, 55min, or 60min. Under the above gelatinization temperature and time conditions, it is beneficial to obtain an oat gelatinized liquid with better gelatinization effect, while removing its endogenous β-glucanase, which is also beneficial to improve the efficiency of subsequent enzymatic hydrolysis and fermentation.

[0058] It is understandable that the restrictions on the ratio of oat bran to water, the gelatinization time, and the gelatinization temperature in the above gelatinization process can be met simultaneously, or only one of them can be met. Meeting all of them can improve the gelatinization effect of oat bran, remove a large amount of starch and impurities, improve the efficiency of subsequent enzymatic hydrolysis and fermentation, and obtain oat bran fermentation products with high content of functional substances.

[0059] In this invention, the purpose of adding α-amylase in the first enzymatic hydrolysis is to hydrolyze the glycogen in oat bran, the α-1,4 glucosinolate bonds within the gelatinized degradation products, and some residual starch to obtain soluble dextrin and a small amount of oligosaccharides. This also rapidly reduces the viscosity of the originally gelatinous oat gelatinized liquid, making the enzymatic hydrolysis more favorable. The addition of saccharifying enzymes allows for the production of glucose, removing starch impurities while providing energy for subsequent fermentation by the rice wine yeast.

[0060] In some examples of a single enzymatic hydrolysis, based on 100% of the total mass of the oat gelatinized liquid, the amount of α-amylase added is 0.05%-0.15%, for example, it can be any value in the range of 0.05%-0.15%, such as 0.05%, 0.08%, 0.1%, 0.12%, 0.13%, 0.14%, 0.15%, etc.

[0061] In some examples of enzymatic hydrolysis, based on 100% of the total mass of oat gelatinized liquid, the amount of saccharifying enzyme added is 0.05%-0.15%, for example, it can be any value in the range of 0.05%-0.15%, such as 0.05%, 0.08%, 0.1%, 0.12%, 0.13%, 0.14%, 0.15%, etc.

[0062] In some embodiments of a single enzymatic hydrolysis, the temperature of the single enzymatic hydrolysis is 55℃-95℃, for example, it can be any value in the range of 55℃-95℃, such as 55℃, 60℃, 65℃, 70℃, 75℃, 80℃, 85℃, 90℃, 95℃, etc.

[0063] In some embodiments of a single enzymatic hydrolysis, the hydrolysis time is 35-60 minutes. For example, it can be any time value within the range of 35-60 minutes, such as 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, or 60 minutes.

[0064] In some examples of enzymatic hydrolysis, the α-amylase is a high-temperature α-amylase, which can hydrolyze the α-1,4-glucosidic bonds inside starch, glycogen and their degradation products at a relatively high temperature of 90℃-95℃, producing soluble dextrin and a small amount of oligosaccharides, which causes the viscosity of the colloidal starch solution to decrease rapidly.

[0065] It is understandable that in the above enzymatic hydrolysis process, the restrictions on the temperature, time, α-amylase and saccharifying enzyme, etc., can be met simultaneously, or only one of them can be met. Meeting all of them can improve the efficiency and effect of the enzymatic hydrolysis, remove a large amount of residual starch, obtain more active substances, and provide more energy for the subsequent fermentation of rice wine yeast.

[0066] In the fermentation step of this invention, the rice wine yeast has a strong and complete enzyme system. It uses undigested macromolecules such as starch, cellulose, and protein in oat bran, as well as glucose, as nutrients for fermentation and hydrolysis, which is beneficial to improving the extraction rate of active substances in oat bran.

[0067] Meanwhile, oat bran contains phenolic compounds, primarily ferulic acid. These phenolic compounds typically exist in three forms: free, bound, and conjugated. Conjugated polyphenols are linked to carbohydrates or other small molecules via ester bonds, while bound polyphenols are usually covalently bound to cell wall skeletal polysaccharides and proteins, making them difficult to release and utilize. However, *Saccharomyces cerevisiae* produces specific enzyme systems that can hydrolyze and disrupt the cell walls of oat bran and the crystal structure of oat starch, weakening the bonds between phenolic substances and large cell wall molecules, thus releasing a large amount of free phenolic substances. Simultaneously, it also weakens the ether bonds between conjugated and bound polyphenolic compounds and cell wall components, making them easier to extract.

[0068] In some embodiments of the fermentation process, the rice wine yeast can be activated before fermentation, which is beneficial to improving the fermentation effect and efficiency. Methods well known to those skilled in the art can be used to activate the rice wine yeast.

[0069] In some embodiments of the fermentation step, based on 100% of the mass of the oat bran fermentation medium, the inoculation amount of rice wine yeast is 1%-5%, for example, any value in the range of 1%-5%, such as 1%, 2%, 3%, 4%, 5%, etc. This can enhance the hydrolysis of macromolecules such as starch, cellulose, and protein in oat bran that have not been enzymatically hydrolyzed, which is beneficial to improving the extraction rate of active substances in oat bran and is less likely to cause waste.

[0070] In some embodiments of the fermentation process, fermentation is carried out at 25°C-30°C. For example, any temperature within the 25°C-30°C range, such as 25°C, 26°C, 27°C, 28°C, 29°C, or 30°C. These fermentation temperatures are beneficial for promoting fermentation efficiency and prevent the enzyme activity from being destroyed due to excessively high fermentation temperatures.

[0071] In some embodiments of the fermentation process, fermentation can also be carried out at a rotation speed of 100-300 r / min, which is beneficial to increase the fermentation rate and make the unhydrolyzed macromolecules such as starch, cellulose, and protein in oat bran more complete, which is beneficial to improve the extraction rate of active substances in oat bran.

[0072] In some embodiments of the fermentation step, the fermentation time is 3-4 days.

[0073] It is understandable that the various fermentation conditions mentioned above can be met individually or simultaneously. Meeting all conditions simultaneously can achieve better fermentation results and efficiency, allowing the undigested starch, cellulose, protein and other macromolecules in oat bran to be hydrolyzed more completely, which is more conducive to improving the extraction rate of active ingredients in oat bran.

[0074] This invention also includes a secondary enzymatic hydrolysis process. The added neutral protease breaks down the large, difficult-to-utilize proteins in oat bran into smaller oat polypeptides and amino acids. The resulting oat polypeptides are mainly short peptide chains, possessing protein functions and high activity. Furthermore, due to their small molecular weight, they diffuse rapidly and are more easily absorbed by the human body.

[0075] In some embodiments, neutral protease can be added in multiple batches, which helps to increase the content of polyphenols and β-glucan in the fermentation product.

[0076] During fermentation, the degradative enzymes metabolized by the rice wine yeast may release a large amount of free polyphenols by hydrolyzing the oat cell wall structure. Ferulic acid is mainly found in the cell wall of oat bran, where it is cross-linked with compounds such as xylan by ester bonds. During the fermentation of oat bran, the enzymes secreted by the rice wine yeast may weaken these ester bond structures, making ferulic acid easier to extract. The addition of ferulic acid esterase during the secondary enzymatic hydrolysis allows for further release of ferulic acid from the oat bran. In addition, some unhydrolyzed feruloyl oligosaccharides were also found to be produced during the entire fermentation process.

[0077] Feruloyl oligosaccharides are an important class of functional oligosaccharides formed by the linkage of ferulic acid to arabinose residues on the xylan side chain via ester bonds. They are widely found in the cell walls of oat bran, have water solubility and thermal stability, and are ideal raw materials for the food, pharmaceutical and cosmetic industries. Because feruloyl oligosaccharides contain both hydrophobic ferulic acid moieties and hydrophilic oligosaccharide moieties, they have a number of physiological functions, including antioxidant activity, probiotic effects and inhibition of glycation.

[0078] In some embodiments of secondary enzymatic hydrolysis, the pH of the oat bran fermentation product is adjusted to neutral, for example, by adjusting the pH to 6.8-7.0 with an acid-base regulator, which is more conducive to promoting the enzymatic hydrolysis of neutral proteases.

[0079] In some embodiments of secondary enzymatic hydrolysis, based on 100% of the total mass of oat bran fermentation product, the amount of neutral protease added is 0.001%-0.015%. For example, the amount added can be any value within the range of 0.001%-0.015%, such as 0.001%, 0.005%, 0.008%, 0.010%, 0.012%, 0.013%, 0.014%, or 0.015%. Within the above range, the enzymatic hydrolysis of large protein molecules can be efficiently promoted, while minimizing resource waste.

[0080] In some embodiments of secondary enzymatic hydrolysis, the enzyme can be added in multiple stages, which is beneficial for promoting the enzymatic hydrolysis of large protein molecules.

[0081] In some examples of secondary enzymatic hydrolysis, based on 100% of the total mass of oat bran fermentation products, the amount of ferulic acid esterase added is 0.01%-0.15%, for example, any value within the range of 0.01%-0.15%, such as 0.01%, 0.03%, 0.05%, 0.08%, 0.10%, 0.13%, 0.15%, etc. Within the above addition range, the hydrolysis of the polyphenol ferulic acid can be efficiently promoted, while minimizing resource waste.

[0082] In some embodiments of secondary enzymatic hydrolysis, the temperature of secondary enzymatic hydrolysis is 45℃-55℃, for example, it can be a temperature in the range of 45℃-55℃ such as 45℃, 48℃, 50℃, 52℃, 55℃, etc., which can exert the best enzymatic hydrolytic activity of neutral protease and ferulic acid esterase.

[0083] In some embodiments of secondary enzymatic hydrolysis, the secondary enzymatic hydrolysis time is 3-5 hours. For example, it can be any time value within the range of 3-5 hours, such as 3 hours, 3.5 hours, 4 hours, 4.5 hours, or 5 hours, which can make the enzymatic hydrolysis of polyphenols and large protein molecules more complete.

[0084] In some embodiments, after the secondary enzymatic hydrolysis is completed, the supernatant can be obtained by centrifugation. Centrifugation can reduce impurities and improve the purity of active substances in oat bran.

[0085] In some embodiments, the centrifugation speed can be 8000-10000 rpm, and the centrifugation time can be 20-30 min.

[0086] In some embodiments, sterilizing the supernatant by autoclaving at 90-120°C for 15-30 minutes can inactivate the rice wine yeast.

[0087] In some embodiments, after sterilization of the supernatant, preservatives may be added to extend the shelf life of the oat bran fermentation product.

[0088] The preservative may be selected from those commonly used in the art, and in some embodiments, includes one or more of 1,2-pentanediol, 1,2-hexanediol, 1,3-propanediol, methylpropanediol, 1,2-octanediol, 1,2-decanediol, p-hydroxyacetophenone, ethylhexylglycerin, capryloyl hydroxamic acid, glyceryl caprylate, chlorphenesin, sorbitan caprylate, phenoxyethanol, and capryloyl glycine.

[0089] In some embodiments, the preservative content is 0.1-1% of the total mass of the oat bran fermentation product.

[0090] In some embodiments of the secondary enzymatic hydrolysis step, the filter residue separated from the supernatant still contains a certain amount of active substances. The filter residue can be processed by cell wall breaking, water extraction, and centrifugation to obtain oat bran fermentation lysate. This oat bran fermentation lysate also contains active substances such as β-glucan, oat protein, oat polypeptide, polyphenols, ferulic acid, and feruloyl oligosaccharides, and can also be applied in the cosmetics field.

[0091] By processing the filter residue, oat bran can be fully utilized, further improving its bioavailability.

[0092] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following specific embodiments are only used to explain the present invention and are not intended to limit the present invention.

[0093] Example 1

[0094] The preparation method of the oat bran fermentation product in this embodiment includes the following steps:

[0095] Step 1, Pre-treatment: Use a micro grinder to grind the oat bran, and then pass it through a 100-200 mesh sieve to obtain the ground oat bran raw material.

[0096] Step 2, gelatinization: Mix oat bran raw material and water at a material-to-liquid ratio of 1:10 (g / ml) and add to the fermentation bottle. Gelatinize in a water bath at 95℃ for 60 minutes to obtain oat gelatinized liquid.

[0097] Step 3, one-time enzymatic hydrolysis: Based on 100% of the oat gelatinized liquid, add 0.05% high-temperature α-amylase and 0.05% saccharifying enzyme to the oat gelatinized liquid, and enzymatically hydrolyze in a water bath at 55℃ for 60 minutes, and then perform enzyme inactivation treatment to obtain oat bran fermentation medium.

[0098] Step 4: Fermentation: Based on 100% of the oat bran fermentation medium, inoculate with 5% rice wine yeast and ferment at 28℃ and 200r / min for 4 days to obtain oat bran fermentation liquid.

[0099] Step 5, Secondary Enzymatic Hydrolysis: Adjust the pH of the oat bran fermentation broth to 6.8 using an acid-base regulator. Based on 100% of the total mass of the oat bran fermentation broth, add 0.010% neutral protease and 0.10% ferulic acid esterase. Perform a secondary enzymatic hydrolysis at 45℃ for 4 hours, then add 0.005% neutral protease and incubate at 50℃ for 1 hour. Next, centrifuge at 10,000 rpm for 20 minutes, collect the supernatant, and autoclave at 90℃ for 30 minutes to obtain the oat bran fermentation product.

[0100] Example 2

[0101] The preparation method of the oat bran fermentation product in this embodiment includes the following steps:

[0102] Step 1, Pre-treatment: Use a micro grinder to grind the oat bran, and then pass it through a 100-200 mesh sieve to obtain the ground oat bran raw material.

[0103] Step 2, gelatinization: Mix oat bran raw material and water at a material-to-liquid ratio of 1:15 (g / ml) and add to the fermentation bottle. Gelatinize in a water bath at 85℃ for 50 minutes to obtain oat gelatinized liquid.

[0104] Step 3, one-time enzymatic hydrolysis: Based on 100% of the oat gelatinized liquid, add 0.15% high-temperature α-amylase and 0.15% saccharifying enzyme to the oat gelatinized liquid, and carry out enzymatic hydrolysis at 60℃ for 50 min, and then perform enzyme inactivation treatment to obtain oat bran fermentation medium.

[0105] Step 4: Fermentation: Based on 100% of the oat bran fermentation medium, inoculate with 1% rice wine yeast and ferment at 28℃ and 100r / min for 3 days to obtain oat bran fermentation liquid.

[0106] Step 5, Secondary Enzymatic Hydrolysis: Adjust the pH of the oat bran fermentation broth to 6.8 using an acid-base regulator. Based on 100% of the total mass of the oat bran fermentation broth, add 0.015% neutral protease and 0.15% ferulic acid esterase. Perform secondary enzymatic hydrolysis at 50℃ for 4.5 hours. Then, centrifuge at 8000 rpm for 25 minutes, collect the supernatant, and autoclave at 120℃ for 15 minutes to obtain the oat bran fermentation product.

[0107] Example 3

[0108] Step 1, Pre-treatment: Use a micro grinder to grind the oat bran, and then pass it through a 100-200 mesh sieve to obtain the ground oat bran raw material.

[0109] Step 2, gelatinization: Mix oat bran raw material and water at a material-to-liquid ratio of 1:20 (g / ml) and add to the fermentation bottle. Gelatinize in a water bath at 80℃ for 40 minutes to obtain oat gelatinized liquid.

[0110] Step 3, one-time enzymatic hydrolysis: Based on 100% of the oat gelatinized liquid, add 0.05% high-temperature α-amylase and 0.05% saccharifying enzyme to the oat gelatinized liquid, and carry out enzymatic hydrolysis at 70℃ for 55 minutes, and then perform enzyme inactivation treatment to obtain oat bran fermentation medium.

[0111] Step 4: Fermentation: Based on 100% of the oat bran fermentation medium, inoculate with 3% rice wine yeast and ferment at 25℃ and 300r / min for 4 days to obtain oat bran fermentation liquid.

[0112] Step 5, Secondary Enzymatic Hydrolysis: Adjust the pH of the oat bran fermentation broth to 7.0 using an acid-base regulator. Based on 100% of the total mass of the oat bran fermentation broth, add 0.010% neutral protease and 0.15% ferulic acid esterase. Perform secondary enzymatic hydrolysis at 45℃ for 5 hours. Then, centrifuge at 9000 rpm for 20 minutes, collect the supernatant, and autoclave at 100℃ for 25 minutes to obtain the oat bran fermentation product.

[0113] Example 4

[0114] The preparation method of the oat bran fermentation product in this embodiment includes the following steps:

[0115] Step 1, Pre-treatment: Use a micro grinder to grind the oat bran, and then pass it through a 100-200 mesh sieve to obtain the ground oat bran raw material.

[0116] Step 2, gelatinization: Mix oat bran raw material and water at a material-to-liquid ratio of 1:25 (g / ml) and add to the fermentation bottle. Gelatinize in a water bath at 70℃ for 40 minutes to obtain oat gelatinized liquid.

[0117] Step 3, one-time enzymatic hydrolysis: Based on 100% of the oat gelatinized liquid, add 0.10% high-temperature α-amylase and 0.10% saccharifying enzyme to the oat gelatinized liquid, and carry out enzymatic hydrolysis at 65℃ for 40 minutes. Then, perform enzyme inactivation treatment to obtain oat bran fermentation medium.

[0118] Step 4: Fermentation: Based on 100% of the oat bran fermentation medium, inoculate with 4% rice wine yeast and ferment at 30℃ and 100r / min for 3 days to obtain oat bran fermentation liquid.

[0119] Step 5, Secondary Enzymatic Hydrolysis: Adjust the pH of the oat bran fermentation broth to 6.8 using an acid-base regulator. Based on 100% of the total mass of the oat bran fermentation broth, add 0.001% neutral protease and 0.01% ferulic acid esterase. Perform secondary enzymatic hydrolysis at 45℃ for 3 hours. Then, centrifuge at 10,000 rpm for 20 minutes, collect the supernatant, and autoclave at 110℃ for 30 minutes to obtain the oat bran fermentation product.

[0120] Example 5

[0121] The preparation method of the oat bran fermentation product in this embodiment includes the following steps:

[0122] Step 1, Pre-treatment: Use a micro grinder to grind the oat bran, and then pass it through a 100-200 mesh sieve to obtain the ground oat bran raw material.

[0123] Step 2, gelatinization: Mix oat bran raw material and water at a material-to-liquid ratio of 1:20 (g / ml) and add to the fermentation bottle. Gelatinize in a water bath at 65℃ for 50 minutes to obtain oat gelatinized liquid.

[0124] Step 3, one-time enzymatic hydrolysis: Based on 100% of the oat gelatinized liquid, add 0.15% high-temperature α-amylase and 0.15% saccharifying enzyme to the oat gelatinized liquid, and carry out enzymatic hydrolysis at 95℃ for 35 minutes. Then, perform enzyme inactivation treatment to obtain oat bran fermentation medium.

[0125] Step 4: Fermentation: Based on 100% of the oat bran fermentation medium, inoculate with 2% rice wine yeast and ferment at 25℃ and 300r / min for 3 days to obtain oat bran fermentation liquid.

[0126] Step 5, Secondary Enzymatic Hydrolysis: Adjust the pH of the oat bran fermentation broth to 6.8 using an acid-base regulator. Based on 100% of the total mass of the oat bran fermentation broth, add 0.001% neutral protease and 0.01% ferulic acid esterase. Perform secondary enzymatic hydrolysis at 45℃ for 3.5 hours. Then, centrifuge at 10,000 rpm for 20 minutes, collect the supernatant, and autoclave at 90℃ for 30 minutes to obtain the oat bran fermentation product.

[0127] Example 6

[0128] Example 6 Preparation method of oat bran fermentation product is the same as that of Example 1, except that in the secondary enzymatic hydrolysis, after adjusting the pH of the oat bran fermentation product to 6.8 with an acid-base regulator, 0.015% of neutral protease is added at once.

[0129] Comparative Example 1

[0130] The preparation method of the oat bran fermentation product in Comparative Example 1 is the same as that in Example 1, except that gelatinization is not performed.

[0131] Comparative Example 2

[0132] The preparation method of the oat bran fermentation product in Comparative Example 2 is the same as that in Example 1, except that instead of using rice wine yeast for fermentation, the same amount of Bifidobacterium longum was selected for fermentation.

[0133] Comparative Example 3

[0134] The preparation method of the oat bran fermentation product in Comparative Example 3 is the same as that in Example 1, except that no secondary enzymatic hydrolysis is performed.

[0135] Performance Test 1

[0136] The content of active substances in the products obtained in the examples and comparative examples was determined, and the results are shown in Table 1.

[0137] Table 1. Content of active substances in the oat bran fermentation products obtained in the examples and comparative examples.

[0138]

[0139] Performance Test 2

[0140] The antioxidant effects of the products obtained in the examples and comparative examples were determined, and the results are shown in Table 2.

[0141] Table 2. Antioxidant effects of the products obtained from the examples and comparative examples.

[0142] Active substances Total reducing power Scavenging DPPH free radicals Example 1 0.508 53.8％ Example 2 0.493 50.57％ Example 3 0.348 52.14％ Example 4 0.301 49.5％ Example 5 0.447 50.7％ Example 6 0.486 52.1％ Comparative Example 1 0.378 38.25％ Comparative Example 2 0.452 37.94％ Comparative Example 3 0.411 37.22％

[0143] As shown in Table 1, the oat bran fermentation product prepared by this invention contains abundant polysaccharides, β-glucan, oat protein, oat polypeptides, polyphenols, ferulic acid, and feruloyl oligosaccharides.

[0144] As shown in Table 2, the oat bran fermentation product prepared by this invention has a high scavenging rate of DPPH free radicals and has a good antioxidant effect.

[0145] Furthermore, comparing Example 1 and Example 6, in the secondary enzymatic hydrolysis step, the addition of neutral protease in multiple stages is beneficial to increasing the content of polyphenols and β-glucan in the fermentation product.

[0146] Compared to Example 1, which did not undergo gelatinization, the resulting product contained excessive impurities. The contents of polysaccharides, β-glucan, oat protein, oat polypeptides, polyphenols, ferulic acid, and feruloyl oligosaccharides were significantly reduced. Furthermore, it may have retained a large amount of endogenous β-glucanase from the oat bran, leading to enzymatic hydrolysis of β-glucan and consequently a substantial decrease in β-glucan content. Ultimately, this also affected the product's antioxidant properties.

[0147] Comparative Example 2 did not use rice wine yeast for fermentation, but instead used Bifidobacterium for fermentation, which reduced the extraction rate of active substances in oat bran.

[0148] Comparative Example 3 did not undergo secondary enzymatic hydrolysis, and the large molecular proteins in oat bran that are difficult to utilize were not further enzymatically hydrolyzed into small molecular oat peptides. As a result, the active substances in the product were reduced, and the utilization rate of oat bran was not significantly improved.

[0149] Application Example 1: Moisturizing, Repairing, and Firming Efficacy Test

[0150] The oat bran fermentation product obtained in Example 1 was made into a basic essence formulation as a test sample, wherein the concentration of the oat bran fermentation product was 10%.

[0151] Twenty-six healthy Chinese subjects aged 18 to 55 years (4 males and 22 females) were selected and used the test sample continuously for 28 days under normal conditions.

[0152] The cheeks of the subjects were selected as the test site. Skin stratum corneum moisture content, transepidermal water loss (TEWL) value, skin elasticity (R2) value, and F4 value were measured using the instrument probe before use of the test sample and on days 14 and 28 of continuous use. The results are shown in the table below. Figures 2-5 , Figures 2-5 The mean data are for 26 subjects (P < 0.001).

[0153] in, Figure 2 The results show the moisture content of the skin's stratum corneum after use. Figure 3 This represents the amount of transepidermal moisture lost from the skin after use. Figures 4 to 5 These represent the changes in skin elasticity R2 and F4 after use, respectively.

[0154] Depend on Figure 2 It can be seen that after 28 days of use, the moisture content of the stratum corneum on the face of the subjects increased significantly by 76.40% (P<0.001); the above indicates that continuous use for 28 days can significantly increase the moisture content of the stratum corneum and has a moisturizing effect.

[0155] Depend on Figure 3It was found that after 28 days of use, the transepidermal water loss of the subjects' faces was significantly reduced by 13.27% (P<0.001); the above indicates that continuous use for 28 days can significantly reduce transepidermal water loss and has the effect of enhancing the skin barrier and repairing.

[0156] Depend on Figure 4 , Figure 5 It can be seen that after 28 days of product use, the R2 value of facial elasticity of the subjects increased significantly by 5.22% (P=0.003); after 28 days of product use, the F4 value of facial elasticity of the subjects decreased significantly by 11.25% (P=0.003).

[0157] The above indicates that continuous use of the test product for 28 days can significantly improve skin elasticity and has a skin-tightening effect.

[0158] In summary, the oat bran fermentation product obtained by fermenting with rice wine yeast in this invention has excellent moisturizing, repairing and firming effects.

[0159] The above are merely preferred embodiments of the present invention and do not limit the scope of the patent. Any equivalent structural or procedural transformations made based on the description and drawings of the present invention, or direct or indirect applications in other related technical fields, are similarly included within the scope of patent protection of the present invention.

Claims

1. A method of making a fermented oat bran product, characterized by, Includes the following steps: Gelatinization: Oat bran and water are mixed and gelatinized to obtain oat gelatinized liquid. The gelatinization temperature is 55℃-95℃ and the gelatinization time is 35min-60min. One-time enzymatic hydrolysis: α-amylase and saccharifying enzyme are added to the oat gelatinized liquid for enzymatic hydrolysis and enzyme inactivation treatment to obtain oat bran fermentation medium. Based on 100% of the total mass of the oat gelatinized liquid, the amount of α-amylase added is 0.05%-0.15%, the amount of saccharifying enzyme added is 0.05%-0.15%, the temperature of the first enzymatic hydrolysis is 55℃-95℃, and the time of the first enzymatic hydrolysis is 35min-60min. The α-amylase is a high-temperature α-amylase. Fermentation: Yellow wine yeast is inoculated into the oat bran fermentation medium and fermented to obtain oat bran fermentation liquid. Based on the total mass of the oat bran fermentation medium of 100%, the amount of yellow wine yeast added is 1%-5%, the fermentation temperature is 25℃-30℃, and the fermentation time is 3-4 days. Secondary enzymatic hydrolysis: The pH value of the oat bran fermentation broth is adjusted, and then neutral protease and ferulic acid esterase are added for secondary enzymatic hydrolysis. The supernatant is sterilized to obtain the oat bran fermentation product. Based on 100% of the total mass of the oat bran fermentation broth, the amount of neutral protease added is 0.001%-0.015%, the amount of ferulic acid esterase added is 0.01%-0.15%, the temperature of the secondary enzymatic hydrolysis is 45℃-55℃, and the time of the secondary enzymatic hydrolysis is 3-5 hours.

2. The method of claim 1, wherein the oat bran fermentation product is prepared by, Prior to the gelatinization process, the oat bran was also subjected to micronization and sieving.

3. The method of claim 1, wherein the oat bran fermentation product is prepared by, In the gelatinization step, the ratio of oat bran to water is 1:10~25 (g / ml).

4. The method of claim 1, wherein the oat bran fermentation product is prepared by, In the secondary enzymatic hydrolysis step... The supernatant was obtained by centrifugation; And / or, after completing the sterilization operation of the supernatant, the method further includes adding a preservative.

5. The method of producing a fermented oat bran product according to claim 1, wherein, In the secondary enzymatic hydrolysis step, after taking the supernatant, there is a filter residue. The filter residue is subjected to cell wall breaking, water extraction, and centrifugation to obtain the supernatant to obtain oat bran fermentation lysate.

6. An oat bran fermentation product prepared by the preparation method according to any one of claims 1 to 5.

7. The oat bran fermentation product of claim 6, characterized in that, The fermented oat bran product contains β-glucan, oat protein, oat polypeptide, polysaccharides, polyphenols, ferulic acid, and feruloyl oligosaccharides.

8. The use of the oat bran fermentation product as described in claim 6 or 7 in the preparation of cosmetics.

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