Oat preparation, oat product containing oat preparation, preparation method therefor and use thereof

Abstract

An oat preparation, an oat product comprising the oat preparation, a preparation method therefor and the use thereof. The oat preparation is obtained by fermenting a fermentation broth; the fermentation broth comprises a fermentation substrate and a starter culture; the fermentation substrate mainly consists of hydrolyzed oat bran; the starter culture comprises lactobacillus reuteri, lactobacillus plantarum, lactobacillus acidophilus, bifidobacterium lactis and lactobacillus rhamnosus, and comprises streptococcus salivarius subsp. thermophilus and lactobacillus delbrueckii subsp. bulgaricus.

Description

Oat products, oat products containing oat products, their preparation methods and uses Technical Field

[0001] This invention belongs to the field of fermentation technology, and specifically relates to fermentation products and post-fermentation products derived from oat bran, as well as oat products containing such fermentation products or post-fermentation products, their preparation methods and uses. Background Technology

[0002] The gut is a vital digestive organ, responsible not only for nutrient absorption but also for preventing bacterial invasion. Gut health involves the normal presence and balance of beneficial bacteria in the gut, as well as its interaction with other bodily systems. Gut health is crucial for overall health, as it is related not only to food digestion and nutrient absorption but also closely linked to immune function.

[0003] Fermented foods are generally considered to promote gut health; however, some commercially available fermented foods have not only failed to show positive effects on gut health and other functions, but have even led to gut dysfunction. Therefore, there remains a demand for fermented foods that can improve gut health and related functions. Summary of the Invention

[0004] In a first aspect of the invention, an oat product is provided, which is obtained by fermenting a fermentation solution comprising a fermentation substrate and a starter culture, wherein the fermentation substrate is mainly composed of hydrolyzed oat bran, and the starter culture comprises a concentration of 1*10⁻⁶ per gram of fermentation solution. 4 CFU to 1*10 11 The CFU contains *Lactobacillus reuteri*, *Lactobacillus plantarum*, *Lactobacillus acidophilus*, *Bifidobacterium lactis*, and *Lactobacillus rhamnosus*, and contains 1 μg to 100 μg of *Streptococcus thermophilus* and *Lactobacillus delbrueckii* subsp. bulgaricus per 1 gram of fermentation solution, with a weight ratio of 50 to 150:1.

[0005] In a second aspect of the invention, a method for preparing oat products is provided, the method comprising: enzymatically hydrolyzing oat bran to prepare hydrolyzed oat bran; and mixing a fermentation substrate mainly composed of hydrolyzed oat bran with water, and adding a starter culture for fermentation to obtain a fermentation product, wherein the starter culture comprises a concentration of 1*10⁻⁶ per gram of fermentation solution. 4 CFU to 1*10 11The CFU contains *Lactobacillus reuteri*, *Lactobacillus plantarum*, *Lactobacillus acidophilus*, *Bifidobacterium lactis*, and *Lactobacillus rhamnosus*, and contains 1 μg to 100 μg of *Streptococcus thermophilus* and *Lactobacillus delbrueckii* subsp. bulgaricus per 1 gram of fermentation solution, with a weight ratio of 50 to 150:1.

[0006] In a third aspect of the invention, an oat product for enhancing gut health or improving immune function is provided, comprising the oat product of the first aspect of the invention.

[0007] In a fourth aspect of the invention, a method for enhancing gut health or improving immunity is provided, wherein the method comprises giving an oat product of the first aspect of the invention or an oat product of the third aspect of the invention to a person in need. Attached Figure Description

[0008] Figure 1 shows the effect of different doses of the oat product of the present invention on the immune age of the subjects: control group (15 g / d unfermented oat bran powder), low-dose treatment group (5 g / d oat bran postbiotic), and high-dose treatment group (15 g / d oat bran postbiotic), where * indicates a statistically significant difference relative to the control group (P<0.05), and ** indicates a highly statistically significant difference relative to the control group (P<0.01). Detailed Implementation

[0009] The present invention will now be described in further detail to enable those skilled in the art to readily put it into practice. However, it should be understood that the present invention can be implemented in various forms and is not limited to the specific embodiments described herein, and the embodiments of the present invention are merely examples.

[0010] As described in this article, the term “about” or “approximately” applied to numerical values ​​covers both precise values ​​and reasonable variances.

[0011] Unless otherwise stated, the terms “include” and “contain” and their grammatical variations are intended to indicate an “open” or “inclusive” language that includes the listed elements but also allows for the inclusion of additional, unlisted elements.

[0012] Unless otherwise defined, all terms have and should be given the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0013] This invention uses oat bran as a substrate and selects specific beneficial bacteria for fermentation. During fermentation, the number of various bacteria is significantly increased. At the same time, macromolecular nutrients are broken down into smaller molecules (such as proteins being broken down into small amino acids). It also produces a variety of metabolites, such as phenols, flavonoids, short-chain fatty acids, γ-aminobutyric acid (GABA), and catechin compounds. This makes the fermented product easier to absorb and utilize when it enters the body, promoting intestinal health and enhancing immunity.

[0014] In a first aspect, the present invention provides an oat product obtained by fermenting a fermentation solution comprising a fermentation substrate and a starter culture. The fermentation substrate refers to the substrate utilized by the starter culture during fermentation, which is decomposed by the starter culture to produce metabolites. The fermentation substrate of the present invention mainly consists of hydrolyzed oat bran. The starter culture includes *Streptococcus thermophilus* subsp. *salivarius*, *Lactobacillus delbrueckii* subsp. *bulgaricus*, *Lactobacillus reuteri*, *Lactobacillus plantarum*, *Lactobacillus acidophilus*, *Bifidobacterium lactis*, and *Lactobacillus rhamnosus*.

[0015] Oat bran is the outer layer of the oat endosperm after the outer husk has been removed. Oat bran is rich in protein, especially essential amino acids. In addition, it contains a large amount of insoluble dietary fiber, which helps promote intestinal motility and the elimination of toxins from the body. Despite its many health benefits, in industrial applications, oat bran tends to become too viscous during fermentation, making it difficult to ferment fully and thus preventing the complete dissolution of active ingredients.

[0016] In one embodiment, the *Lactobacillus reuteri* in the starter culture of the present invention can be *Lactobacillus reuteri* GL104. In one embodiment, the *Lactobacillus plantarum* in the starter culture of the present invention can be *Lactobacillus plantarum* LPL28. In one embodiment, the *Lactobacillus acidophilus* in the starter culture of the present invention can be *Lactobacillus acidophilus* NCFM. In one embodiment, the *Bifidobacterium lactis* in the starter culture of the present invention can be *Bifidobacterium lactis* HN019. In one embodiment, the *Lactobacillus rhamnosus* in the starter culture of the present invention can be *Lactobacillus rhamnosus* GG.

[0017] In one embodiment, the total content of *Streptococcus thermophilus* subsp. *salivarius* and *Lactobacillus delbrueckii* subsp. *bulgaricus* in the fermentation agent can be approximately 1 μg to 100 μg per 1 gram of fermentation solution, for example, approximately 2 μg to 90 μg, 3 μg to 80 μg, 4 μg to 70 μg, 5 μg to 60 μg, 6 μg to 50 μg, 7 μg to 40 μg, approximately 8 μg to 30 μg, or approximately 9 μg to 20 μg per 1 gram of fermentation solution. In one embodiment, the content of *Streptococcus thermophilus* subsp. *salivarius* and *Lactobacillus delbrueckii* subsp. *bulgaricus* may be unequal. In one embodiment, the weight ratio of *Streptococcus thermophilus* subsp. *salivarius* and *Lactobacillus delbrueckii* subsp. *bulgaricus* can be approximately 50 to 150:1, for example, approximately 60 to 140:1, approximately 70 to 130:1, approximately 80 to 120:1, or approximately 90 to 110:1, for example, approximately 95 to 105:1. In one embodiment, the starter culture can comprise a concentration of approximately 1*10⁻⁶ per gram of fermentation solution. 4 CFU to 1*10 11 The CFU contains *Lactobacillus reuteri*, *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bifidobacterium lactis*, with each species having a concentration of approximately 1*10⁻⁶ per gram of fermentation solution. 5 CFU to 1*10 10 CFU, approximately 1*10 6 CFU to 1*10 9 CFU, or approximately 1*10 6 CFU to 1*10 8 CFU.

[0018] Unhydrolyzed oat bran contains a large amount of macromolecules such as starch, which easily form a high-viscosity matrix when hydrated. This high-viscosity matrix is ​​difficult to further process commercially because it does not flow easily through pipes, and as noted herein, its high viscosity acts as a barrier to the starter culture, making it difficult to ferment oat bran to a commercial scale (greater than 1 L). Therefore, this invention uses hydrolyzed oat bran. During hydrolysis, amylase breaks down the macromolecules of starch in the oat bran powder into smaller polysaccharides, disaccharides, and monosaccharides, reducing the viscosity of the matrix and making it more suitable for subsequent industrial production.

[0019] The oat products of this invention are mainly made from hydrolyzed oat bran, which contains numerous beneficial components that promote gut health and enhance immunity. In one embodiment, the hydrolyzed oat bran constitutes approximately 80 wt% or more of the fermentation substrate, for example, approximately 85 wt% or more, approximately 90 wt% or more, approximately 95 wt% or more, approximately 96 wt% or more, approximately 97 wt% or more, approximately 98 wt% or more, approximately 99 wt% or more, or even 100 wt%.

[0020] In one embodiment, the oat product of the present invention is a fermentation product obtained by fermenting the fermentation solution of the present invention. The fermentation product may be liquid, semi-solid, or solid. In one embodiment, the oat product of the present invention is an epigenetic agent prepared by sterilization of the fermentation product. The epigenetic agent of the present invention may be liquid, semi-solid, or solid. "Sterilization" or "sterilization" as used herein refers to a method of killing all pathogenic and non-pathogenic microorganisms, as well as their spores and fungal spores, in an object such as the fermentation product of the present invention, so that the object is in a state free of living microorganisms. The epigenetic agent of the present invention can be obtained by killing the microorganisms in the fermentation product of the present invention through heat sterilization. For example, the fermentation product can be sterilized by heating at approximately 70 to 140°C, such as approximately 80 to 135°C, approximately 90 to 130°C, or approximately 100 to 125°C, for approximately 10 to 30 seconds, such as approximately 10 to 20 seconds, approximately 10 to 10 seconds, approximately 10 to 5 seconds, approximately 10 to 1 minute, approximately 10 to 45 seconds, or approximately 10 to 30 seconds, to remove viable microorganisms, thereby preparing a postbiotic. For example, sterilization methods commonly used in the food industry, such as pasteurization or ultra-high temperature sterilization, can be used to prepare the postbiotic. In one embodiment, the fermentation product can be sterilized by heating at approximately 70 to 140°C, such as approximately 80 to 135°C, approximately 90 to 130°C, or approximately 100 to 125°C, to remove viable microorganisms, thereby preparing a postbiotic. In one embodiment, the fermentation product can be pasteurized, for example, by heating at approximately 70 to 90°C for approximately 10 to 120 seconds, at approximately 75 to 85°C for approximately 15 to 60 seconds, or at approximately 77 to 80°C for approximately 20 to 45 seconds, to remove any live microorganisms, thereby preparing a metabiotic. The metabiotic may include inactivated bacteria, bacterial components (cytoplasmic polypeptides, phosphoglycolic acid, intracellular and extracellular polysaccharides (EPSs), and surface proteins), bacterial metabolites (short-chain fatty acids (SCFAs), organic acids, bacteriocins, and enzymes), etc. Because the metabiotic does not contain live microorganisms, it minimizes the risk of ingesting probiotics, while the presence of probiotic components allows it to exhibit beneficial health effects through mechanisms similar to those of probiotics.

[0021] In one embodiment, the oat product comprises at least one of the following: amino acids or amino acid metabolites, phenols, and flavonoids.

[0022] In one embodiment, the amino acid includes L-tryptophan. Tryptophan is one of the essential amino acids for the human body. Tryptophan derived from gut microbiota metabolism can regulate the development of inflammation and related diseases in the intestinal and central nervous systems. It is metabolized by microorganisms in the intestine, which can affect intestinal health, and its metabolites can also regulate the body's immune response. In one embodiment, based on the total dry weight of the oat product, L-tryptophan is, for example, more than about 25 μg / g, more than about 30 μg / g, more than about 40 μg / g, more than about 50 μg / g, or even more. In one embodiment, the amino acid includes γ-aminobutyric acid (GABA). GABA is an important functional non-protein amino acid with functions such as enhancing brain activity, regulating hormone secretion, improving lipid metabolism, and lowering blood pressure. In one embodiment, based on the total dry weight of the oat product, GABA is more than about 100 mg / kg, more than about 200 mg / kg, more than about 300 mg / kg, more than about 400 mg / kg, more than about 500 mg / kg, more than about 600 mg / kg, or even higher.

[0023] In one embodiment, the amino acid metabolites include indole metabolites. Under the action of the fermentation spawn of the present invention, tryptophan can be metabolized to generate indole derivatives, which can, for example, activate aryl hydrocarbon receptors (AhR), which mediate inflammation control and are effective immunomodulatory factors, promote the production of IL-22, and prevent colonic inflammation. In one embodiment, the indole metabolites include indoleacetic acid, indoleacetaldehyde, indoleacrylic acid, or combinations thereof. Using the fermentation spawn of the present invention, tryptophan in the fermentation product of the present invention can be converted into the intermediate metabolite indoleacetaldehyde, and further converted into indoleacetic acid. Regarding intestinal and immune function, indoleacetic acid can maintain the structural integrity of intestinal epithelial cells, reduce intestinal inflammation, and help maintain intestinal homeostasis.

[0024] In one embodiment, the oat product contains phenols. In another embodiment, the phenols include catechin compounds. Catechins have antioxidant, anti-inflammatory, glucose and lipid metabolism-improving, and tumor-preventing effects. By fermenting oat bran with the fermenting agent of the present invention, the catechin compounds in the oat bran can be dissolved. In the oat product of this application, the catechin compounds may include gallic acid, protocatechuic acid, protocatechuic aldehyde, epicatechin, or combinations thereof. In one embodiment, based on the total dry weight of the oat product, the gallic acid content is about 40 ng / g or more, about 50 ng / g or more, about 60 ng / g or more, or even higher, for example, from about 40 ng / g to about 200 ng / g, from about 50 ng / g to about 150 ng / g, from about 60 ng / g to about 100 ng / g. In one embodiment, based on the total dry weight of the oat product, the protocatechuic acid content is approximately 110 ng / g or more, approximately 120 ng / g or more, approximately 130 ng / g or more, or even higher, for example, approximately 110 ng / g to approximately 300 ng / g, approximately 120 ng / g to approximately 250 ng / g, or approximately 130 ng / g to approximately 200 ng / g. In one embodiment, based on the total dry weight of the oat product, the protocatechuic aldehyde content is approximately 210 ng / g or more, approximately 230 ng / g or more, approximately 260 ng / g or even higher, for example, approximately 210 ng / g to approximately 400 ng / g, approximately 230 ng / g to approximately 350 ng / g, or approximately 250 ng / g to approximately 300 ng / g. In one embodiment, based on the total dry weight of the oat product, the content of epicatechin can be more than about 1.40 g / g, more than about 1.50 ng / g, or more than about 1.60 ng / g, for example, from about 1.40 ng / g to about 100 ng / g, from about 1.50 ng / g to about 80 ng / g, or from about 1.60 ng / g to about 60 ng / g.

[0025] In one embodiment, the oat product comprises flavonoids. In one embodiment, the flavonoids include quercetin. In one embodiment, based on the total dry weight of the oat product, the quercetin content is approximately 110 μg / g or higher, approximately 130 μg / g or higher, approximately 160 μg / g or higher, approximately 190 μg / g or higher, for example, approximately 110 to approximately 400 μg / g, approximately 150 to approximately 300 μg / g, or approximately 200 to approximately 250 μg / g. Quercetin can regulate the diversity and abundance of gut microbiota, promote gut microecological balance, and restore the structure and function of the intestinal barrier.

[0026] In one embodiment, the flavonoids may further include dihydromyricetin, dihydroquercetin, or a combination thereof. In one embodiment, based on the total dry weight of the oat product, the content of dihydromyricetin may be more than about 1 μg / g, more than about 2 μg / g, more than about 3 μg / g, for example, more than 1 to 20 μg / g, more than 2 to 15 μg / g, or more than 3 to 10 μg / g. In one embodiment, based on the total dry weight of the oat product, the content of dihydroquercetin may be more than about 7 μg / g, more than about 8 μg / g, more than about 9 μg / g, or even more, for example, more than 7 to about 20 μg / g.

[0027] In one embodiment, the hydrolyzed oat bran is obtained by hydrolyzing the oat bran with an amylase. In one embodiment, the amylase is at least one selected from α-amylase, β-amylase, saccharifying enzyme, γ-amylase, isoamylase, and debranching amylase. In one embodiment, the hydrolyzed oat bran is obtained by mixing the amylase and oat bran with water to form a hydrolysis solution, and hydrolyzing the oat bran at 50°C to 70°C for 30 minutes to 3 hours. The amylase accounts for 0.005 wt% to 0.05 wt% of the hydrolysis solution by weight, and the oat bran accounts for 1 wt% to 40 wt% of the hydrolysis solution by weight. In one embodiment, the hydrolyzed oat bran is obtained by hydrolyzing oat bran at, for example, about 5 wt% to 35 wt%, about 10 wt% to 30 wt%, or about 12 wt% to 25 wt% of the hydrolysis solution at, for example, about 55°C to 68°C, about 58°C to 66°C, or about 60°C to 65°C, with an amylase at, for example, about 0.008 wt% to 0.03 wt%, about 0.001 wt% to 0.02 wt% of the weight of the hydrolysis solution, for example, for 1 to 2 hours, or for example, 1 hour. The hydrolysis time can be adjusted according to the amount of enzyme added, the amount of fermentation substrate, or the temperature.

[0028] In one embodiment, the hydrolyzed oat bran accounts for approximately 1 wt% to 40 wt% of the fermentation solution, for example, approximately 5 wt% to 35 wt%, approximately 10 wt% to 30 wt%, or approximately 12 wt% to 25 wt%. In one embodiment, the oat bran is fermented at 25°C to 40°C, for example, 28°C to 38°C, 29°C to 37°C, such as 37°C, for approximately 8 to 48 hours, for example, approximately 12 to 36 hours, approximately 16 to 32 hours, approximately 18 to 28 hours, or approximately 20 to 24 hours. The fermentation time can be adjusted according to the amount of starter culture added, the amount of fermentation substrate, or the fermentation temperature. In one embodiment, the fermentation substrate of the present invention comprises hydrolyzed oat bran and may also contain other carbon sources, other nitrogen sources, or combinations thereof. In one embodiment, based on the total weight of the fermentation solution, the content of the other carbon sources is approximately 0.5-3 wt%, and the content of the other nitrogen sources is approximately 0.5-3 wt%. In one embodiment, the other carbon source may be selected from glucose, maltose, lactose, sucrose, or combinations thereof. In one embodiment, the other nitrogen source includes proteins, such as whey protein.

[0029] In one embodiment, the oat product of the present invention can be obtained by drying the fermentation product or post-fermentation agent using various drying methods known in the art, such as vacuum drying, airflow drying, freeze drying, etc. In one embodiment, the fermentation product or post-fermentation agent can be dispersed into tiny droplets using a mechanical device such as an atomizer and exchanged with a hot air stream at approximately 180 to 200°C for 5 to 10 seconds, thereby evaporating the moisture in the fermentation product and obtaining the powdered or granular oat product of the present invention. In one embodiment, the fermentation product or post-fermentation agent of the present invention can be dried by drum drying at 40°C to 110°C to obtain the oat product of the present invention. In one embodiment, the fermentation product or post-fermentation agent of the present invention can be dried by freezing at -20°C to -35°C to obtain the oat product of the present invention.

[0030] In a second aspect, the present invention provides a method for preparing oat products. The method includes: enzymatically hydrolyzing oat bran to prepare hydrolyzed oat bran; and mixing a fermentation substrate, mainly composed of hydrolyzed oat bran, with water and adding a starter culture for fermentation to obtain a fermentation product. The starter culture includes *Streptococcus thermophilus* subsp. *salivarius*, *Lactobacillus delbrueckii* subsp. *bulgaricus*, *Lactobacillus reuteri*, *Lactobacillus plantarum*, *Lactobacillus acidophilus*, *Bifidobacterium lactis*, and *Lactobacillus rhamnosus*.

[0031] In one embodiment, the *Lactobacillus reuteri* in the starter culture of the present invention can be *Lactobacillus reuteri* GL104. In one embodiment, the *Lactobacillus plantarum* in the starter culture of the present invention can be *Lactobacillus plantarum* LPL28. In one embodiment, the *Lactobacillus acidophilus* in the starter culture of the present invention can be *Lactobacillus acidophilus* NCFM. In one embodiment, the *Bifidobacterium lactis* in the starter culture of the present invention can be *Bifidobacterium lactis* HN019. In one embodiment, the *Lactobacillus rhamnosus* in the starter culture of the present invention can be *Lactobacillus rhamnosus* GG.

[0032] In one embodiment, the total content of *Streptococcus thermophilus* subsp. *salivarius* and *Lactobacillus delbrueckii* subsp. *bulgaricus* in the fermentation agent can be approximately 1 μg to 100 μg per 1 gram of fermentation solution, for example, approximately 2 μg to 90 μg, 3 μg to 80 μg, 4 μg to 70 μg, 5 μg to 60 μg, 6 μg to 50 μg, 7 μg to 40 μg, approximately 8 μg to 30 μg, or approximately 9 μg to 20 μg per 1 gram of fermentation solution. In one embodiment, the content of *Streptococcus thermophilus* subsp. *salivarius* and *Lactobacillus delbrueckii* subsp. *bulgaricus* may be unequal. In one embodiment, the content ratio of *Streptococcus thermophilus* subsp. *salivarius* and *Lactobacillus delbrueckii* subsp. *bulgaricus* can be approximately 50 to 150:1, for example, approximately 60 to 140:1, approximately 70 to 130:1, approximately 80 to 120:1, or approximately 90 to 110:1, for example, approximately 95 to 105:1. In one embodiment, the starter culture can comprise a concentration of approximately 1*10⁻⁶ per gram of fermentation solution. 4 CFU to 1*10 11 The CFU contains *Lactobacillus reuteri*, *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bifidobacterium lactis*, with each species having a concentration of approximately 1*10⁻⁶ per gram of fermentation solution. 5 CFU to 1*10 10 CFU, approximately 1*10 6 CFU to 1*10 9 CFU, or approximately 1*10 6 CFU to 1*10 8 CFU.

[0033] In one embodiment, the hydrolysis comprises hydrolyzing oat bran with an amylase. In one embodiment, the hydrolysis comprises hydrolyzing unprocessed or physically processed oat bran, such as milled oat bran, with an amylase. In one embodiment, the amylase is at least one selected from α-amylase, β-amylase, saccharifying enzyme, γ-amylase, isoamylase, and debranching amylase. In one embodiment, the method comprises mixing the amylase and oat bran with water to form a hydrolysis solution, and hydrolyzing the oat bran at 50°C to 70°C for 30 minutes to 3 hours, wherein the amylase comprises 0.005 wt% to 0.05 wt% of the hydrolysis solution by weight, and the oat bran comprises 1 wt% to 40 wt% of the hydrolysis solution by weight. In one embodiment, the method includes adjusting the viscosity of the hydrolysate prior to hydrolysis so that its viscosity, as measured at 40°C using a Brookfield viscometer at a shear rate of 30 rpm / 30 s, is below approximately 1000 cp, approximately 900 cp, approximately 800 cp, approximately 700 cp, approximately 600 cp, or even lower. The viscosity of the hydrolysate can be controlled by controlling the concentration of the fermentation substrate in the hydrolysate. In one embodiment, the method includes hydrolyzing oat bran, for example, at approximately 55°C to 68°C, approximately 58°C to 66°C, or approximately 60°C to 65°C, with an amylase at a weight ratio of, for example, approximately 0.008 wt% to 0.03 wt%, approximately 0.001 wt% to 0.02 wt% of the hydrolysate, for example, approximately 5 wt% to 35 wt%, approximately 10 wt% to 30 wt%, or approximately 12 wt% to 25 wt% of the hydrolysate, for example, for approximately 1 hour. The hydrolysis time and temperature can be adjusted according to the enzyme concentration and the concentration of the hydrolysate.

[0034] In one embodiment, the method includes heating the hydrolysate after enzymatic hydrolysis to inactivate the enzyme. The heating can be performed using methods known in the art, as long as they can terminate the enzymatic hydrolysis process.

[0035] In one embodiment, oat bran is hydrolyzed until the viscosity of the hydrolysate is less than approximately 500 cp, less than approximately 400 cp, less than approximately 300 cp, less than approximately 200 cp, less than approximately 100 cp, less than approximately 50 cp, or even lower, as measured by a Brookfield viscometer at a shear rate of 10 rpm / 30 s at 30°C, so as to facilitate passage through pipes, for example, into subsequent processes, such as into a sterilization tank or fermenter.

[0036] In one embodiment, the method includes performing a first sterilization on the hydrolysate after enzymatic hydrolysis and before adding the fermentation agent of the present invention. The first sterilization can be performed using methods commonly used in the food industry, such as pasteurization or ultra-high temperature (UHT) sterilization. In one embodiment, the method includes subjecting the hydrolysate to UHT sterilization, for example, heating at approximately 90 to 140°C for approximately 5 to 400 seconds, for example, heating at approximately 95 to 135°C for approximately 10 to 350 seconds, or heating at approximately 100 to 130°C for approximately 10 to 100 seconds. In one embodiment, heating at approximately 95 to 100°C for approximately 200 to 350 seconds, for example, heating at approximately 95 to 135°C for approximately 10 to 300 seconds. In one embodiment, after the first sterilization, the viscosity of the fermentation solution is approximately 200 cp or less, approximately 100 cp or less, approximately 50 cp or less, approximately 40 cp or less, approximately 30 cp or less, approximately 20 cp or less, or even lower, as measured by a Brookfield viscometer at a shear rate of 10 rpm / 30 s at 23°C.

[0037] In one embodiment, the method includes mixing the hydrolyzed oat bran, a starter culture, and water to form a fermentation solution for fermentation. The hydrolyzed oat bran accounts for approximately 1 wt% to 40 wt% of the fermentation solution by weight, for example, approximately 5 wt% to 35 wt%, approximately 10 wt% to 30 wt%, or approximately 12 wt% to 25 wt%. In one embodiment, the method includes fermenting the oat bran at 25°C to 40°C, for example, 28°C to 38°C, 29°C to 37°C, or 37°C, for approximately 8 to 48 hours, for example, approximately 12 to 36 hours, approximately 16 to 32 hours, approximately 18 to 28 hours, or approximately 20 to 24 hours. The fermentation time can be adjusted according to the amount of starter culture added, the amount of substrate, or the fermentation temperature. In one embodiment, the fermentation substrate of the present invention may also contain other carbon sources, other nitrogen sources, or combinations thereof. In one embodiment, the content of the other carbon source is approximately 0.5-3 wt% and the content of the other nitrogen source is approximately 0.5-3 wt% based on the total weight of the fermentation solution. In one embodiment, the other carbon source may be selected from glucose, maltose, lactose, sucrose, or combinations thereof. In one embodiment, the other nitrogen source includes proteins, such as whey protein.

[0038] In one embodiment, the method includes fermenting with a starter culture until the viscosity of the fermentation product is less than approximately 200 cp, less than approximately 100 cp, less than approximately 50 cp, less than approximately 40 cp, less than approximately 30 cp, less than approximately 20 cp, less than approximately 10 cp, or even lower, as measured by a Brookfield viscometer at a shear rate of 10 rpm / 30 s at 23°C.

[0039] In one embodiment, the method of the present invention includes a second sterilization after fermentation, which can be performed using methods commonly used in the food industry, such as pasteurization or ultra-high temperature (UHT) sterilization. In one embodiment, the fermentation product can be sterilized by heating at approximately 70 to 140°C, for example, approximately 80 to 135°C, approximately 90 to 130°C, or approximately 100 to 125°C, for approximately 10 seconds to 30 minutes, for example, approximately 10 seconds to 20 minutes, approximately 10 seconds to 10 minutes, approximately 10 seconds to 5 minutes, approximately 10 seconds to 1 minute, approximately 10 seconds to 45 seconds, or approximately 10 seconds to 30 seconds, to remove any viable microorganisms, thereby preparing a post-biotic. In one embodiment, the method includes pasteurizing the fermentation product, for example, by heating at approximately 70 to 90°C for approximately 10 seconds to 120 seconds, for example, by heating at approximately 75 to 85°C for approximately 15 seconds to 60 seconds, or by heating at approximately 77 to 80°C for approximately 20 seconds to 45 seconds. In one embodiment, after the second sterilization, the viscosity of the fermentation product is approximately 200 cp or less, approximately 100 cp or less, approximately 50 cp or less, approximately 40 cp or less, approximately 30 cp or less, approximately 20 cp or less, or even lower, as measured by a Brookfield viscometer at a shear rate of 10 rpm / 30 s at 23°C.

[0040] In one embodiment, the oat product of the present invention can be obtained by drying the fermentation product or post-biotic using various drying methods known in the art, such as vacuum drying, airflow drying, freeze drying, etc. In one embodiment, the fermentation product or post-biotic can be dispersed into tiny droplets using a mechanical device such as an atomizer and exchanged with a hot air stream of approximately 180 to 200°C for 5 to 10 seconds to evaporate the moisture in the fermentation product, thereby obtaining the powdered or granular oat product of the present invention. In one embodiment, the fermentation product or post-biotic of the present invention can be obtained by drum drying at 40°C to 110°C to obtain the oat product of the present invention. In one embodiment, the fermentation product or post-biotic of the present invention can be obtained by freezing at -20°C to -35°C to obtain the oat product of the present invention. In one embodiment, the oat product of the present invention can be obtained by inactivating and drying the fermentation product in the same step, for example, by heating and evaporating the moisture at temperatures of approximately 70 to 140°C, such as approximately 80 to 135°C, approximately 90 to 130°C, or approximately 100 to 125°C, thereby obtaining the dried post-biotic of the present invention.

[0041] In a third aspect, the present invention provides an oat product for enhancing gut health or improving immunity, comprising the oat products of the first aspect.

[0042] As used in this article, "enhancing gut health" or "improving gut health" refers to improving the functional state of the gut, such as improvements in excretion, microbial diversity, and / or an increase in the amount of beneficial bacteria in the gut. Whether gut health is enhanced or improved can be determined by, but is not limited to, measuring gut microbiota (e.g., measuring Bifidobacteria, Lactobacillus, Mycoprotein Actinomycetes, Clostridium plasmidonum, and other probiotics), analyzing bowel habits, and assessing the absence of significant discomfort symptoms. Improved gut health promotes efficient nutrient absorption, waste elimination, and maintains the normal functioning of the immune system. The inventors have demonstrated, through gut microbiota measurement and the Bristol stool score, that the application of the oat product of this invention can enhance the gut health of subjects.

[0043] As used in this article, "immunity" or "immune function" refers to the body's resistance to disease through the immune system, and these terms can be used interchangeably. Immunity can be assessed through methods including cellular immune function, humoral immune function, monocyte-macrophage function, and NK cell activity. In this application, the inventors, following the methods and experimental procedures outlined in the paper "Immune-Ageing Evaluation of Peripheral T and NK Lymphocyte Subsets in Chinese Healthy Adults" published by Zhenghu Jia et al. in Phenomics on May 23, 2023, quantified cellular immune function and NK cell activity by measuring and analyzing the immune age of the subjects. Immune age is obtained based on peripheral blood immune cell function parameters of the subjects, including T cell and NK cell subsets, phenotype, cell differentiation state, and quantity, applied to an immune aging model formed by artificial intelligence deep learning. The smaller the value, the stronger the immune function. The inventors found that the immune age of the subjects significantly decreased after administering the oat product of this invention, thus demonstrating that the oat product of this invention can improve the body's immunity.

[0044] In the body's immune system, immune-related cytokines secreted by immune cells also play a crucial role in fighting disease. The inventors assessed the immunity of subjects administering the oat product of this invention by analyzing the levels of four important immune-related cytokines: interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10), and tumor necrosis factor-α (TNF-α). Interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) are typical pro-inflammatory cytokines that play an important role in acute inflammatory responses, but overactivation can lead to chronic inflammation and related diseases. Interleukin-10 (IL-10) is an important anti-inflammatory cytokine that plays a key role in preventing excessive inflammation. The inventors discovered that administering the oat product of this invention effectively reduced the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in subjects, while increasing the level of interleukin-10 (IL-10), thus demonstrating that the oat product of this invention helps improve the body's immunity. In one embodiment, the oat product is selected from beverages, instant beverages, solid ready-to-eat foods, solid non-ready-to-eat foods, or semi-solid foods. For example, the oat product can be solid granules and can be eaten directly. The oat product can also be a solid non-ready-to-eat food that requires slight heating before consumption. The oat product can be a flake-like solid that, after being added to warm water, can become a paste or solution, thus facilitating consumption. In one embodiment, the oat product can be a dietary food, health food, or functional preparation, etc. The oat product can improve the intestinal microenvironment, promote the digestion and absorption of nutrients, relieve constipation, or improve immunity, etc., but is not limited to these.

[0045] In one embodiment, the oat product may further comprise a sweetener, a thickener, etc. In one embodiment, the sweetener may include at least one selected from granulated sugar, crystalline fructose, erythritol, sucralose, aspartame, acesulfame potassium, steviol glycosides, honey, etc. In one embodiment, the sweetener comprises from about 0.001 wt% to about 20 wt% of the oat product by weight, for example, from about 0.01 wt% to about 10 wt%, or from about 0.1 wt% to about 1 wt%. In one embodiment, the thickener may include at least one selected from guar gum, xanthan gum, starch, etc. In one embodiment, the sweetener comprises from about 0.001 wt% to about 10 wt% of the oat product by weight, for example, from about 0.01 wt% to about 5 wt%, or from about 0.1 wt% to about 1 wt%.

[0046] In one embodiment, the oat product of the present invention is formulated as a dosage form selected from the group consisting of: oral solutions, syrups, granules, capsules, powders, pills, tablets, etc. In one embodiment, the oat product of the present invention is formulated as a unit dosage form.

[0047] In a fourth aspect, the present invention provides a method for enhancing gut health or improving immunity, wherein the method comprises giving an oat product as described in the first aspect or the oat product as described in the third aspect of the present invention to a person in need.

[0048] In one implementation, the object is a human being, including adults, children, newborns, and humans in the prenatal stage.

[0049] In one embodiment, the method includes administering the oat product described in the first aspect or the oat product described in the third aspect of the present invention to a desired subject in the form of a liquid dosage form, a solid dosage form, or a semi-solid dosage form. In one embodiment, the method includes administering an effective amount of the active ingredient, i.e., the oat product present in the oat product described in the first aspect or the oat product described in the third aspect of the present invention, to a desired subject. In this invention, "effective amount" refers to the content at which a predetermined amount of the oat product present in the oat product described in the first aspect or the oat product described in the third aspect of the present invention, when administered to a desired subject, produces the desired effect (e.g., enhanced immunity, regulation of intestinal flora, and bowel movement). The actual content level of the active ingredient in the product of the present invention (e.g., fermentation product or post-generic dry weight) can be modified to effectively achieve the expected response for a specific subject. In one embodiment, the method includes administering the oat product described in the first aspect or the oat product described in the third aspect of the present invention to a recipient at a daily dose of approximately 1g or more, approximately 2g or more, approximately 3g or more, approximately 4g or more, approximately 6g or more, approximately 7g or more, approximately 8g or more, approximately 9g or more, approximately 10g or more, approximately 15g or more, approximately 20g or more, or even higher, and said daily dose may be administered to the recipient once, twice, three times, or more times. In one embodiment, the method includes administering the oat product described in the first aspect or the oat product described in the third aspect of the present invention to the recipient daily, every other day, every three days, or even for a longer period, such as for more than 15 days, more than 20 days, more than one month, more than two months, or even longer. In one embodiment, the oat product described in the first aspect or the oat product described in the third aspect of the present invention may be administered to the recipient in unit dose form. In one embodiment, the method includes using the oat products described in the first aspect or the third aspect of the present invention separately, in combination, or sequentially with drugs or preparations for enhancing gut health or boosting immunity to further improve gut health or enhance immunity.

[0050] The following will describe in detail the oat bran fermentation product, post-biotic or product, its preparation method and uses, with reference to specific embodiments, but the present invention is not limited thereto.

[0051] Example

[0052] Materials and strains

[0053] Oat bran flour (AQ 6 mesh) was supplied by PepsiCo Inc. *Lactobacillus reuteri* GL104 and *Lactobacillus plantarum* LPL28 were purchased from Bioflag (Taiwan). *Lactobacillus rhamnosus* GG, *Bifidobacterium lactis* HN019, *Lactobacillus acidophilus* NCFM, and *Streptococcus salivarius* subsp. *thermophilus* and *Lactobacillus delbrueckii* subsp. *bulgaricus* (catalog number Yo-Mix 485LYO 200DCU; the weight ratio of *Streptococcus salivarius* subsp. *thermophilus* and *Lactobacillus delbrueckii* subsp. *bulgaricus* was approximately 100:1) were purchased from IFF (USA). Lactose and whey protein (food grade) were commercially available products. Unless otherwise specified, all materials, reagents, and instruments used in this application are conventional materials, reagents, and instruments in the art, and can be obtained commercially or prepared by conventional methods.

[0054] The content or concentration of each strain in the fermentation agent of the present invention used in the examples is shown in Table 1 below:

[0055] Table 1. Content or concentration of each strain in the fermentation antagonist of this invention.

[0056] Example 1: Preparation of Fermentation Products and Metabiotics

[0057] Experimental group 1

[0058] Unhydrolyzed oat bran powder and hydrolyzed oat flour (Shanghai Jinshan Dele Food Ingredients Co., Ltd.; Product No.: 810274) were mixed as fermentation substrates at a weight ratio of 3:1. The fermentation substrates were mixed with lactose and whey protein and added to water at 50±5℃, and subjected to high-speed shear hydration for 10-15 minutes. The solution was sieved for the first homogenization. The filtrate was subjected to ultra-high temperature instantaneous sterilization (125±2℃, 15s), and then cooled to 37±1℃. The starter culture of the present invention was added to a total of 50 liters of the cooled solution, stirred for 1 to 2 minutes, and then allowed to ferment at 37±1℃ for 24 hours. The fermentation product was demulsified, stirred, and sieved for the second homogenization. The fermentation product was then subjected to high-temperature sterilization (75±2℃, 25s) to prepare post-biotics. The sterilized fermentation products are mechanically dispersed into very fine droplets. By increasing the surface area for water evaporation, they are brought into contact with hot air at 195 degrees Celsius, accelerating the drying process and removing most of the moisture instantly, thus drying the substances in the fermentation products into post-fermentation powder.

[0059] Experimental group 2

[0060] While stirring, oat bran powder, whey protein powder, lactose, and α-amylase were added to hot water at 60±5℃, so that the weight ratios of oat bran powder, whey protein powder, lactose, and α-amylase in the solution were 16.665wt%, 0.914%, 1.83%, and 0.01675wt%, respectively. The mixture was dissolved and homogenized for hydrolysis. After hydrolysis for approximately 30 minutes to 2 hours, the hydrolyzed solution was sieved for the first homogenization. The filtrate was subjected to ultra-high temperature instantaneous sterilization (97±2℃, 300s), and then cooled to 37±1℃. The starter culture of this invention was added to 2 tons of the cooled solution, and the mixture was stirred for 5 minutes to 1 hour according to the overall volume of the fermentation system, followed by static fermentation at 37±1℃ for 24 hours. After the acidity of the fermentation product reached the standard range (120-140°T), the fermented solution was demulsified, stirred, and sieved for the second homogenization. The fermentation products were subjected to high-temperature sterilization (77±2℃, 30s) to prepare post-biotics. The sterilized fermentation products were mechanically dispersed into very fine droplets. By increasing the surface area for water evaporation, the products were exposed to hot air at 195℃ to accelerate the drying process, removing most of the moisture instantly and drying the substances in the fermentation products into post-biotic powder.

[0061] Control group 1

[0062] Unhydrolyzed oat bran powder, whey protein powder, lactose, and water are mixed, with the weight ratios of oat bran powder, whey protein powder, and lactose in the hydrolysis system being 16.665 wt%, 0.914 wt%, and 1.83 wt%, respectively. The starter culture of the present invention is then added, and the mixture is immediately inactivated.

[0063] Control group 2

[0064] Fermentation was carried out in the same manner as in Example 2, except that unhydrolyzed oat bran powder was used. This experiment was difficult to conduct on an industrial scale for the following reasons: 1) Industrial-scale production requires a large amount of oat bran powder, and the time required for mixing oat bran powder with water is much longer (e.g., for industrial-scale production with a 5-ton fermentation solution, the time required for full hydration is up to 3 hours). During the long hydration process, the viscosity of the fermentation solution will continuously increase, which will greatly affect the efficiency of industrial production; 2) Using a fermentation substrate mainly composed of unhydrolyzed oat bran powder will create a highly viscous fermentation medium, which will prevent the strain from growing fully, resulting in incomplete fermentation and inconsistencies between batches; 3) The excessive viscosity of the fermented solution will gelatinize and clog the pipes, thus preventing it from passing smoothly through the pipes to the subsequent sterilization tank.

[0065] Control group 3

[0066] Unhydrolyzed oat bran powder, whey protein powder, and lactose were mixed in a weight ratio of 16.665:0.914:1.83 to obtain an unfermented mixed powder.

[0067] Control group 4

[0068] Unhydrolyzed oat bran powder, whey protein powder, and lactose were mixed with water in weight ratios of 16.665 wt%, 0.914 wt%, and 1.83 wt%, respectively, to prepare an unfermented mixed solution.

[0069] Example 2: Viscosity of fermentation products

[0070] The solid content of the product obtained in experimental group 1 of Example 1 at each stage was measured and calculated using a moisture analyzer; the viscosity of the product obtained in experimental group 1 of Example 1 at each stage was measured using Brookfield, and the results are shown in Table 2 below:

[0071] Table 2

[0072] The solid content of the products obtained in experimental group 2 of Example 1 at each stage was measured and calculated using a moisture analyzer; the viscosity of the products obtained in experimental group 2 of Example 1 at each stage was measured using Brookfield, and the results are shown in Table 3 below:

[0073] Table 3

[0074] Both experimental groups 1 and 2 can prepare fermentation products by fermenting the fermentation substrate. However, when using a fermentation substrate containing unhydrolyzed oat bran, the fermentation solution after ultra-high temperature instantaneous sterilization is too viscous due to the presence of a large number of macromolecules, which easily clogs the feed pipes during the feeding process. Hydrolyzing the oat bran breaks down the macromolecules in it into smaller molecules, thereby reducing the viscosity. This avoids clogging the pipelines when adding the hydrolyzed product to the fermenter. As shown in Tables 2 and 3, the viscosity of the product obtained in experimental group 2 at each stage is significantly lower than that obtained in experimental group 1 at each stage, thus ensuring that the process of this invention can be used industrially to prepare fermentation products.

[0075] Example 3: Growth of the strain

[0076] Two samples (Sample 1 and Sample 2) were taken from the liquid before and after fermentation in both Experimental Group 1 and Experimental Group 2. The lactic acid bacteria (including *Lactobacillus reuteri* GL104, *Lactobacillus plantarum* LPL28, *Lactobacillus acidophilus* NCFM, and *Rhamnosus rhamnosus* KPGG) and bifidobacteria (*Bifidobacterium lactis* HN019) in the samples were counted according to the National Food Safety Standard of the People's Republic of China GB4786.35-2016, "Examination of Lactic Acid Bacteria". The test was performed in triplicate, and the average value was calculated.

[0077] By plate counting, the growth factor of lactic acid bacteria in experimental groups 1 and 2 after 24 hours of fermentation (fermentation products) relative to the time of addition (0 hours) is shown in Table 4 below:

[0078] Table 4

[0079] By plate counting, the growth fold of Bifidobacteria in experimental groups 1 and 2 after 24 hours of fermentation (fermentation product) relative to the time of addition (0 hours) is shown in Table 5 below:

[0080] Table 5

[0081] As can be seen from the table above, lactic acid bacteria can grow in both experimental groups 1 and 2. Bifidobacteria still showed significant proliferation after fermentation in experimental group 2, while proliferation was not obvious or even non-existent in experimental group 1.

[0082] Example 4: Absolute Quantitative Method for Determining the Gene Copy Number of Each Strain in Post-Shengyuan Powder

[0083] The copy numbers of the bacterial strains in the post-fermentation powder of experimental groups 1 and 2, and the copy number of the bacterial strains in control group 1 (i.e., the amount of bacterial strains introduced) were detected by real-time quantitative PCR, and then the data were analyzed. Control group 1 was a liquid with a water content of 80.59 wt% and a solid content of 19.41 wt% of the fermentation product. The post-fermentation powder of experimental groups 1 and 2 had a water content of 4 wt% and a solid content of 96 wt% of the post-fermentation powder.

[0084] In the multiple comparison, the bacterial copy number of control group 1 at a moisture content of 80.59% was converted to the bacterial copy number at a moisture content of 4%, i.e., according to...

[0085] We get x = a·4.95

[0086] In the control group 1, the average copy number of the strains was a / g sample, and the average copy number of the strains after conversion into powder was x / g sample. The copy numbers of the corresponding strains after conversion were compared with the copy numbers of the strains in the post-biotic powder of experimental groups 1 and 2, and the fold increase of the strains is shown in Table 6 below.

[0087] Table 6. The ratio of the number of bacterial strains in the post-biotic powder of experimental groups 1 and 2 to the number of bacterial strains in control group 1:

[0088] As can be seen from Table 6, in experimental group 2, which used hydrolyzed oat bran as the fermentation substrate, the copy number of all strains increased, especially Lactobacillus acidophilus and Streptococcus salivarius thermophilus subsp.

[0089] Example 5: Analysis of changes in components during oat fermentation

[0090] 5.1 Analysis of Microbial Metabolites

[0091] Experimental group 3 was prepared in the same way as experimental group 1, but the starter culture used 6 strains, namely Streptococcus salivarius subsp. thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus reuteri GL104, Lactobacillus plantarum LPL28, Lactobacillus acidophilus NCFM, and Bifidobacterium lactis HN019.

[0092] Experimental group 4 was prepared in the same way as experimental group 1, but the starter culture used 6 strains, namely Streptococcus salivarius subsp. thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus reuteri GL104, Lactobacillus plantarum LPL28, Lactobacillus acidophilus NCFM, and Lactobacillus rhamnosus LGG.

[0093] Microbial metabolite analysis was performed on the fermentation products from experimental groups 1-4 and the unfermented mixed solution from control group 4 to confirm the potential of these fermentation products to enhance gut health and improve immunity. First, the microbial metabolites in the fermentation products of Examples 1-4 were compared with the unfermented mixed solution from control group 4. Attention was paid to metabolites that were 0% abundant in control group 4 but not 0% abundant in the fermentation products of Examples 1-4; these metabolites were likely products of microbial fermentation. Data showed that the number of metabolites in experimental groups 1-4 was significantly increased compared to control group 4 in the fermentation products.

[0094] The fold increase in the content of metabolites in the fermentation products and post-fermentation powder of experimental groups 1-4 relative to the content of metabolites in the unfermented mixed powder of control group 3 and the unfermented mixed solution of control group 4 was calculated. Metabolites with a fold increase greater than 2 in the content of fermentation products and post-fermentation powder of experimental groups 1-4 were screened. The results showed significant changes in amino acid metabolism after fermentation. Experimental groups 1 and 2 contained more amino acids or amino acid metabolites compared to the other two experimental groups. Specifically, in the fermentation products, 3-indoleacrylic acid in experimental groups 1, 2, and 4 showed significant changes compared to control group 4. The fermentation products of experimental group 2 also contained 3-indoleacetic acid. The post-fermentation powder of experimental group 1 contained only one indole metabolite: indoleacetaldehyde. In the post-fermentation powder of experimental group 2, three indole metabolites showed significant changes: indoleacetic acid, indoleacetaldehyde, and 3-indoleacrylic acid. Indole metabolites are products of microbial metabolism of tryptophan and offer numerous benefits.

[0095] 5.2 Component Detection in Oat Products

[0096] The compounds in the test samples were separated chromatographically using an Agilent 1290 Infinity II series (Agilent Technologies) ultra-high performance liquid chromatograph (UHPLC) with a Waters ACQUITY UPLC BEH Amide (100×2.1mm, 1.7μm, Waters) column according to the instrument manufacturer's instructions. The L-tryptophan content was determined by mass spectrometry analysis using an Agilent 6460 triple quadrupole mass spectrometer equipped with an AJS-ESI ion source in multiple reaction monitoring (MRM) mode, according to the instrument manufacturer's instructions. The phenolic and flavonoid components and contents in the test samples were identified and analyzed using UHPLC (Vanquish, UPLC, Thermo, USA) and high-resolution mass spectrometry (Q Exactive, Thermo, USA) equipped with a Waters HSS T3 (50*2.1mm, 1.8μm) column, according to the instrument manufacturer's instructions. The results of L-tryptophan, phenolic, and flavonoid content detection in the unfermented mixed powder of control group 3, and the post-fermentation powders of experimental groups 1 and 2 are shown in Table 7 below:

[0097] Table 7

[0098] Compared to control group 3, experimental group 2 showed a significant increase in L-tryptophan. Tryptophan is one of the essential amino acids for the human body. Tryptophan derived from gut microbiota metabolism can regulate inflammation in the intestines and central nervous system. Metabolites of tryptophan metabolized by gut microbes, especially indoles, can regulate the body's immune response. Compared to control group 1, experimental groups 1 and 2 showed significant increases in phenolic and flavonoid substances. Furthermore, compared to experimental group 1, experimental group 2 showed a significant increase in phenolic and flavonoid substances. Phenolic compounds, such as gallic acid, protocatechuic acid, and protocatechuic aldehyde, inhibit the production of pro-inflammatory factors, thus exerting an anti-inflammatory effect. In addition, flavonoid substances, such as quercetin, can regulate the distribution of gut microbiota, promote gut microecological balance, and restore the intestinal barrier's results and function. This also explains why, in Example 6 below, the immune and intestinal indicators of subjects treated with oat bran post-fermentation powder improved compared to those treated with unfermented powder.

[0099] γ-aminobutyric acid (GABA) was separated from the unfermented mixed powder of control group 3 and the post-fermentation powder of experimental group 2 (three copies and the average value was taken) by ion exchange chromatography, and post-column derivatization was performed using ninhydrin. The GABA was then detected by an automated amino acid analyzer. The results are shown in Table 8 below.

[0100] Table 8

[0101] *Below the instrument's detection limit

[0102] The results showed that γ-aminobutyric acid was significantly increased in the post-natal powder of experimental group 2.

[0103] 5.3 Determination of antioxidant activity

[0104] DPPH (1,1-diphenyl-2-trinitrophenylhydrazine) radical is a very stable nitrogen-centered free radical with a single electron. Its alcoholic solution is purple and exhibits strong absorption at 515 nm. In the presence of antioxidants, DPPH radicals are scavenged, resulting in a lighter solution color and a decrease in absorbance at 515 nm. Within a certain range, the change in absorbance is directly proportional to the degree of free radical scavenging. This experiment measured the DPPH scavenging ability of the post-fermentation powder in experimental group 2 and the unfermented mixed powder in control group 3 using an enzyme-linked immunosorbent assay (ELISA) reader. The DPPH scavenging ability of the post-fermentation powder was quantified using the equivalent antioxidant capacity of vitamin C. The results are shown in Table 9 below.

[0105] Table 9

[0106] The results showed that the antioxidant capacity of the post-natal powder in experimental group 2 was significantly improved.

[0107] Example 6: Effects of Oat Bran Postbiotic Powder on Gut Health and Immune Indicators

[0108] 6.1. Experimental Design

[0109] This experiment is a clinical trial testing the effects of post-natal vitamin powder (Experimental Group 2) on gut health and immune indicators. It includes a control group, a low-dose treatment group, and a high-dose treatment group. The experimental design employs randomization, includes a placebo control group, and is a double-blind, parallel design.

[0110] In addition, to enhance the comparability of data between groups, this study also conducted rigorous balance checks on several key factors and major efficacy indicators that may interfere with the final results, including but not limited to gender distribution, age group, body mass index (BMI) and fasting blood glucose level, striving to maintain balance among groups in these dimensions, thereby ensuring the accuracy and reliability of the trial results.

[0111] 6.2. Experimental Subjects

[0112] A total of 75 participants met the criteria for participation in this study. They voluntarily signed informed consent forms and promised to maintain their original diet and lifestyle during the study period, not to consume other probiotics, postbiotics, fermented oats and related foods or products, and to take the experimental products daily. They also agreed to allow us to collect their blood and stool samples.

[0113] Inclusion criteria for the study participants included: healthy adults (male or female), aged 50 to 65 years, with a body mass index (BMI) below 28 kg / m². 2 Participants should have normal or slightly elevated blood pressure, blood lipid levels, and blood sugar levels, requiring no medication; and should not have undergone major surgery or hospitalization in the past five years. Participants with the following conditions will be excluded: chronic diseases such as diabetes, hypertension, hyperlipidemia, or atherosclerosis; gastrointestinal diseases treated surgically in the past five years; antibiotic use in the past six months; allergic reactions to any component of the experimental product; frequent consumption of fermented products; participation in any oat-related research in the past six months; or inability to complete the study as expected due to personal reasons.

[0114] 6.3. Experimental Materials

[0115] The experimental sample in the control group was unfermented oat bran powder; the experimental sample in the low-dose treatment group was a 1:2 mixture of oat bran post-biotic powder and maltodextrin from experimental group 2; and the experimental sample in the high-dose treatment group was post-biotic powder from experimental group 2. A trace amount of potassium acesulfame potassium was added to all three groups to balance the overall flavor.

[0116] 6.4. Experimental Methods

[0117] Seventy-five participants were randomly divided into three groups: a control group, a low-dose treatment group, and a high-dose treatment group, with 25 participants in each group. Each participant took 15g of the experimental product corresponding to their group's daily intake; the corresponding daily intake of oat bran post-biotic powder for the three groups was 0g, 5g, and 15g, respectively. Each participant was instructed to take the product once daily, between meals, mixing it with 150ml of warm water (40-50°C), shaking well, and ensuring complete dissolution before consumption, for 30 consecutive days. During this period, participants were required to maintain their original dietary and exercise habits.

[0118] Data, blood, and / or stool samples were collected at corresponding time points during the 30-day experiment to detect the indicators shown in Table 10 below:

[0119] Table 10

[0120] In addition, the immune age of the subjects was calculated. The methods and experimental procedures used in the paper "Immune-Ageing Evaluation of Peripheral T and NK Lymphocyte Subsets in Chinese Healthy Adults" published by Zhenghu Jia et al. in Phenomics on May 23, 2023, were employed to assess the immune age of the subjects before and after treatment. Peripheral blood was collected from the three groups of subjects on day 0 and day 30, and flow cytometry was used to analyze the functions of different immune cells (including overall T cell function, CD4+ T cells, CD8+ T cells, GDT cells, and NK cells) under different treatments. Random forest machine learning was applied to the data to model immune aging, and neural network analysis was used for confirmation.

[0121] 6.5. Data Analysis

[0122] Normality was tested using the Shapiro-Wilk method (IBM SPSS Statistics 27.0). After the test, normally distributed data were compared between groups using one-way ANOVA (P < 0.05); non-normal data were described using 95% confidence intervals (Mean, 95% CI) of the mean, and compared between groups using nonparametric tests (P < 0.05).

[0123] 6.6. Experimental Results

[0124] A total of 72 subjects were ultimately included as valid samples in the statistical analysis. The control group, low-dose treatment group, and high-dose treatment group had 25, 23, and 24 subjects, respectively.

[0125] There were no significant differences in age, sex, and various biochemical indicators of blood glucose and blood lipids among the three groups at the start of the experiment. Furthermore, according to the intake questionnaire, there were no statistically significant differences in energy and energy-producing nutrient intake among the three groups before and after the experiment.

[0126] Regarding immune indicators (Table 11), there were no significant differences in IL-10 levels among the three groups on day 0, but the high-dose group showed a significantly higher level than the control group on day 30 (P<0.05). The high-dose treatment group showed a significantly greater decrease in IL-1β, IL-6, and TNF-α levels than the control group (P<0.05), and the decrease in IL-1β and IL-6 levels was significantly greater than that in the low-dose treatment group (P<0.05). Therefore, the treated experimental product can significantly increase the level of the anti-inflammatory cytokine IL-10, while significantly decreasing the levels of the pro-inflammatory cytokines IL-1β, IL-6, and TNF-α.

[0127] Table 11

[0128] a indicates a statistically significant difference compared to the control group (P<0.05), and b indicates a statistically significant difference compared to the low-dose treatment group (P<0.05).

[0129] Regarding intestinal indicators, the levels of various bacterial communities were comparable across the three groups on day 0, with no significant differences. On day 30, Bifidobacterium gene expression was significantly higher in the low-dose treatment group than in the control group (P<0.05), and the high-dose treatment group also showed an increasing trend in Bifidobacterium gene expression. On day 15, Lactobacillus gene expression was significantly higher in both the low-dose and high-dose treatment groups than in the control group (P<0.05). On day 30, Akkermansia myxophilus gene expression showed an increasing trend relative to the control group in both the low-dose and high-dose treatment groups. On day 15, Clostridium plasmidonum gene expression showed an increasing trend relative to the control group in both the low-dose and high-dose treatment groups. Overall, the treated samples had a regulatory effect on the expression of specific genes in the intestinal flora.

[0130] Regarding the defecation questionnaire (Table 12), the Bristol stool characteristics score in the high-dose treatment group showed a decreasing trend, indicating a significant improvement in stool characteristics. Both the high-dose and low-dose treatment groups showed an increasing trend in defecation habit satisfaction and maintained a satisfactory level. Therefore, the treatment group's experimental product improved the subjects' stool characteristics, which means it improves intestinal comfort and increases defecation habit satisfaction in a relatively short period of time.

[0131] Table 12

[0132] Between-group comparisons of defecation questionnaire results (mean, 95% CI)

[0133] * indicates a statistically significant difference relative to baseline (P<0.05)

[0134] Regarding immune age (Figure 1), the immune age of the high-dose treatment group was significantly lower than that of the control group after 30 days of administration.

Claims

1. An oat product obtained by fermenting a fermentation solution comprising a fermentation substrate and a starter culture, wherein the fermentation substrate is primarily composed of hydrolyzed oat bran, and the starter culture comprises a concentration of 1*10⁻⁶ per gram of fermentation solution. 4 CFU to 1*10 11 The CFU contains *Lactobacillus reuteri*, *Lactobacillus plantarum*, *Lactobacillus acidophilus*, *Bifidobacterium lactis*, and *Lactobacillus rhamnosus*, and contains 1 μg to 100 μg of *Streptococcus thermophilus* and *Lactobacillus delbrueckii* subsp. bulgaricus per 1 gram of fermentation solution, with a weight ratio of 50 to 150:

1.

2. The oat product according to claim 1, characterized in that, The hydrolyzed oat bran constitutes more than 80 wt% of the fermentation substrate.

3. The oat product according to claim 1, characterized in that, The oat product is a fermentation product obtained by fermenting a fermentation solution or by sterilizing the fermentation product.

4. The oat product according to any one of claims 1 to 3, characterized in that, The oat product contains at least one of the following: amino acids or amino acid metabolites, phenols, and flavonoids.

5. The oat product according to claim 4, characterized in that, The amino acid or amino acid metabolite includes at least one of tryptophan and indole metabolites.

6. The oat product according to claim 5, characterized in that, Indole metabolites include at least one of indoleacetic acid, indoleacetaldehyde, and indoleacrylic acid.

7. The oat product according to claim 4, characterized in that... The phenols include at least one of the following: gallic acid, protocatechuic acid, protocatechuic aldehyde, and epicatechin.

8. The oat product according to claim 4, characterized in that, The flavonoids include at least one of the following: dihydromyricetin, dihydroquercetin, and quercetin.

9. The oat product according to any one of claims 1 to 3, characterized in that, The hydrolyzed oat bran is obtained by mixing amylase and oat bran with water to form a hydrolysis solution, and hydrolyzing the oat bran at 50°C to 70°C for 30 minutes to 3 hours. The amylase accounts for 0.005 wt% to 0.05 wt% of the hydrolysis solution by weight, and the oat bran accounts for 1 wt% to 40 wt% of the hydrolysis solution by weight.

10. The oat product according to any one of claims 1 to 3, characterized in that, The oat product is made by fermenting hydrolyzed oat bran at a concentration of 1 wt% to 40 wt% of the fermentation solution at 25 to 40°C for 8 to 48 hours.

11. A method for preparing oat products, the method comprising: Hydrolyzing oat bran to prepare hydrolyzed oat bran; The fermentation substrate and the starter culture are mixed to obtain a fermentation solution. The fermentation substrate mainly consists of the hydrolyzed oat bran, and the starter culture contains 1*10⁻⁶ components per gram of fermentation solution. 4 CFU to 1*10 11 The CFU contains *Lactobacillus reuteri*, *Lactobacillus plantarum*, *Lactobacillus acidophilus*, *Bifidobacterium lactis*, and *Lactobacillus rhamnosus*, and includes *Streptococcus thermophilus* subsp. *salivarius* and *Lactobacillus delbrueckii* subsp. *bulgaricus* per 1 gram of fermentation solution, with a total weight ratio of 50 to 150:

1. The fermentation solution is then fermented.

12. The method according to claim 11, characterized in that, The hydrolyzed oat bran accounts for more than 80 wt% of the fermentation substrate.

13. The method according to claim 11, characterized in that, The method includes mixing amylase and oat bran with water to form a hydrolysate, and hydrolyzing the oat bran at 50°C to 70°C for 30 minutes to 3 hours, wherein the amylase accounts for 0.005 wt% to 0.05 wt% of the hydrolysate and the oat bran accounts for 1 wt% to 40 wt% of the hydrolysate.

14. The method according to claim 11, characterized in that, The method comprises fermenting the hydrolyzed oat bran, in an amount of 1 wt% to 40 wt% of the fermentation solution, at 25 to 40 degrees Celsius for 8 to 48 hours.

15. The method according to claim 11, characterized in that, The method includes adding other carbon sources and other nitrogen sources to a fermentation solution for fermentation, wherein the other carbon sources are selected from at least one of glucose, maltose, lactose and sucrose and constitute 0.5 wt% to 3 wt% of the weight of the fermentation solution, and the other nitrogen sources include whey protein and constitute 0.5 wt% to 3 wt% of the weight of the fermentation solution.

16. The method according to claim 11, characterized in that, The method includes sterilizing the fermentation product at a temperature of 70°C to 140°C to prepare the oat product.

17. An oat product for enhancing gut health or boosting immunity, comprising the oat products as described in any one of claims 1 to 10.

18. The oat product according to claim 17, characterized in that, The oat products are selected from beverages, instant beverages, solid ready-to-eat foods, solid non-instant foods, or semi-solid foods.

19. The oat product according to claim 17, characterized in that, The oat product is obtained by drying the oat products.

20. Use of the oat product according to any one of claims 1 to 10 in the preparation of functional formulations or dietary products for enhancing gut health or improving immunity.

21. A method for enhancing gut health or boosting immunity, wherein the method comprises giving an oat product as described in any one of claims 1 to 10 or any oat product as described in any one of claims 17 to 19 to a subject in need.

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