Grain dietary fiber, composite dietary fiber and preparation method and application thereof

Abstract

The invention provides a cereal dietary fiber, a composite dietary fiber and a preparation method and application thereof, and the cereal dietary fiber comprises the following components based on the total weight of the cereal dietary fiber: less than or equal to 2% of oil content, less than or equal to 5% of starch content, more than or equal to 50% of total dietary fiber content, and more than or equal to 2% of soluble pentosan content. According to the cereal dietary fiber provided by the invention, osteoblasts can be remarkably increased, and the sparse degree of bone trabecula can be improved, so that the cereal dietary fiber has the effect of improving bone metabolism, and is particularly suitable for improving bone metabolism of old people. Furthermore, by compounding the cereal dietary fiber and the inulin provided by the invention, the cereal dietary fiber and the inulin have a synergistic effect, and as a food raw material, the food texture can be improved, and the nutrition can be improved; and one or more effects of improving the anti-aging and anti-oxidation effects of the body, regulating intestinal flora, relieving chronic inflammation, improving the athletic ability and cognitive ability, improving the immunity and the like are also facilitated.

Description

Technical Field

[0001] The present invention relates to a cereal dietary fiber, and more particularly to a cereal dietary fiber, a composite dietary fiber, a preparation method and an application thereof. Background Art

[0002] Human bone metabolism refers to the processes of growth, regeneration, destruction, and repair that bone tissue undergoes during physiological processes, involving the regulation and activities of bone cells. Bone metabolism is an important process for maintaining bone structure and function, and it is regulated by various factors, including hormones, cytokines, nutrients, and mechanical stimuli. The human skeleton is a complex tissue composed of bone cells, bone matrix, and bone marrow. Bone cells include osteoblasts and osteoclasts. The former is responsible for synthesizing and depositing bone matrix, while the latter is responsible for absorbing and degrading bone matrix. Bone matrix is the main component of bone tissue, consisting of collagen and inorganic salts (mainly calcium phosphates), which endow bones with strength and hardness. Bone growth is a dynamic process in which osteoblasts are responsible for synthesizing new bone matrix to increase bone length, while osteoclasts are responsible for removing excessive or aged bone tissue. Bone metabolism is affected by multiple regulatory factors. For example, hormones play an important role in this process. Estrogen and androgen have important effects on bone health. In addition, the skeleton is also regulated by cytokines and growth factors, which all play important roles in the life cycle of bone cells. The aging of the human body is a natural law and one of the main factors leading to osteoporosis. As people age, the human skeleton gradually loses calcium and bone density, making the bone tissue become fragile and prone to fractures. The mechanism by which aging leads to osteoporosis involves multiple physiological and biochemical processes, such as: (1) Increased bone resorption: As people age, the body's ability to absorb calcium decreases, while bone resorption (the degradation of bone proteins) increases. This leads to a decrease in the quality of bone tissue and bone density, thereby increasing the risk of fractures; (2) Decreased bone formation: During the aging process, the activity and function of osteoblasts gradually decline, resulting in a decrease in the formation of new bone tissue and a slowdown in the bone formation rate; (3) Changes in hormone levels: Hormones play a key role in bone health. After menopause, estrogen levels decrease in women, and androgen also gradually decreases in men during the aging process. The decrease in hormones leads to bone imbalance and an increased risk of osteoporosis; (4) Changes in bone microstructure: Aging leads to changes in bone microstructure, including the loosening of trabeculae and the reduction of the fine structure of bones, which will affect the mechanical properties of bones; (5) Inflammation and oxidative stress: Chronic inflammation and oxidative stress may play a certain role in the development of osteoporosis. These physiological processes may affect the function of bone cells, accelerating bone resorption and bone damage; (6) Accumulation of bone marrow fat: As people age, the fat content in bone marrow may increase, which will replace part of the bone tissue, further reducing the strength and quality of bones. In summary, the mechanism by which aging leads to osteoporosis is multi-faceted, involving multiple biological processes such as bone resorption, bone formation, hormone levels, bone microstructure, and inflammation. It is currently believed that a balanced diet, exercise, supplementation of necessary nutrients, and medical treatment can help slow down the process of osteoporosis.

[0003] Rice bran is a high - value - added by - product in the rice processing process. It is the substance attached to the surface layer of rice after the paddy is husked, mainly composed of the hull, seed coat, cross - linked layer, and aleurone layer, accounting for about 6 - 8% of the total paddy. Because it contains rich dietary fiber, amino acids, and essential physiological active ingredients and nutrients for the human body, it has been increasingly favored and valued by people. However, currently, rice bran is basically used as feed or raw material for extracting oil, resulting in the waste of some of its nutritional components. According to previous research reports, rice bran contains a high level of dietary fiber, which has certain effects on defecation and reducing blood lipid.

[0004] However, there are few reports on the effect of rice bran on bone metabolism. Patent CN202310061156.X mentions the application of a short - peptide derived from rice bran in drugs for preventing and treating bone - related diseases. The reported substance is a short - peptide compound isolated from rice bran protein after protease hydrolysis and then artificially synthesized by chemical synthesis and other methods. It can be seen that this substance is not the original substance in rice bran, but a new substance processed specifically, and it is difficult to expect that other sequences of short - peptides can also play a role in preventing and treating bone - related diseases.

[0005] An article in the journal Nutrients in 2021 mentioned that using rice bran extract can have a certain impact on bone metabolism. The rice bran used in the article comes from a self - cultivated dark rice with a high content of anthocyanins. The main components of its extract are small - molecule substances such as anthocyanins, oryzanol, and phenolic acids, which are significantly different from the main components in rice bran, such as dietary fiber and protein, which are macromolecules. At the same time, the article believes that the antioxidant effect of the above - mentioned small - molecule substances mainly affects bone metabolism.

[0006] Patent KR20120126533A also discloses that using the alcohol extract of rice bran can prevent and treat osteoporosis. However, the alcohol extract of rice bran is mainly small - molecule active substances, and its effect is to inhibit the expression of osteoclasts and does not involve the improvement of osteoblasts. Summary of the Invention

[0007] An object of the present invention is to overcome the defects existing in the prior art and provide a cereal dietary fiber and its preparation method. This cereal dietary fiber can significantly increase the activity of osteoblasts, effectively improve the sparsity of trabecular bone, and has the effect of improving bone metabolism.

[0008] In the first aspect of the present invention, a cereal dietary fiber is provided, and its characteristics include: based on the total mass of the cereal dietary fiber, the oil content in the cereal dietary fiber is ≤2%, preferably ≤1.7 wt%, more preferably 0.4 - 1.7 wt%, and most preferably 0.9 - 1.7 wt%; the starch content is ≤%, preferably 2.0 - 5.0 wt%; the total dietary fiber content is ≥50%, preferably 53.0 - 60.0 wt%; the protein content is 20 - 30 wt%, preferably 22 - 25 wt%; the soluble pentosan content is ≥2%, preferably 2.0 - 4.0 wt%.

[0009] In the second aspect of the present invention, a preparation method of a cereal dietary fiber is provided. The preparation method includes degreasing, de-starching, and hemicellulose enzymolysis treatment of cereal bran. Optionally, extrusion treatment / steam explosion treatment is also included before hemicellulose enzymolysis.

[0010] In one or more embodiments, the cereal bran is selected from one or more of rice bran, wheat bran, barley bran, oat bran, and corn bran.

[0011] In one or more embodiments, the degreasing includes treating the cereal bran with a solvent capable of dissolving lipids for degreasing treatment.

[0012] In one or more embodiments, the oil content in the cereal bran obtained by degreasing treatment is ≤2 wt%. In one or more embodiments, the solvent is n-hexane and / or petroleum ether.

[0013] In one or more embodiments, the mass ratio of the cereal bran to the degreasing solvent is 1:2 - 10, preferably 1:5 - 8.

[0014] In one or more embodiments, the cereal bran is treated with the degreasing solvent 1 - 5 times, preferably 2 - 5 times.

[0015] In one or more embodiments, precipitation is obtained, and the solvent in the precipitation is removed to obtain degreased cereal bran.

[0016] In one or more embodiments, the de-starching is carried out using amylase.

[0017] In one or more embodiments, the starch content in the cereal bran obtained by de-starching is ≤5 wt%.

[0018] In one or more embodiments, the amylase is a heat-resistant (such as resistant to above 70 °C, such as resistant to 70 - 100 °C) α-amylase. Preferably, the optimal action pH range of this heat-resistant α-amylase is 5.8 - 7.8, and the optimal action temperature is 90 - 95 °C.

[0019] In one or more embodiments, the mass ratio of cereal bran to amylase is 100 to 2000:1, preferably 200 to 1500:1, 200 to 1000:1, or 250 to 500:1.

[0020] In one or more embodiments, the de-starching enzymatic reaction system further contains water, and the mass ratio of water to cereal bran is 2 to 10:1, more preferably 3 to 8:1 or 4 to 6:1.

[0021] In one or more embodiments, the de-starching enzymatic hydrolysis time is from 30 minutes to 5 hours, more preferably from 30 minutes to 2 hours, and even more preferably from 30 minutes to 1 hour.

[0022] In one or more embodiments, after the de-starching enzymatic hydrolysis is completed, the precipitate part of the enzymatic hydrolysate is separated, and this precipitate part is dried.

[0023] In one or more embodiments, the steam pressure for the steam explosion treatment is 0.9 to 1.5 MPa, and the pressure holding time is 90 to 150 seconds.

[0024] In one or more embodiments, the cereal bran is added into a steam explosion tank, saturated steam is introduced to make the pressure in the container 0.9 to 1.5 MPa, and this pressure is maintained in the container for 90 to 150 seconds, thereby completing the steam explosion treatment.

[0025] In one or more embodiments, the temperature for extrusion is 110 - 130 °C.

[0026] In one or more embodiments, the hemicellulase hydrolysis treatment is carried out using hemicellulase.

[0027] In one or more embodiments, the mass ratio of hemicellulase to cereal bran is 1:100 to 1500, preferably 1:200 to 1000 or 1:200 to 500.

[0028] In one or more embodiments, the enzymatic reaction system for hemicellulase treatment further contains water, and the mass ratio of water to cereal bran is 2 to 10:1, more preferably 3 to 8:1 or 4 to 6:1.

[0029] In one or more embodiments, the enzymatic hydrolysis for hemicellulase treatment is carried out at a temperature of 40 - 60 °C. In one or more embodiments, the enzymatic hydrolysis time for hemicellulase treatment is 4 - 6 hours, and the pH is 4 - 6.

[0030] In one or more embodiments, after the enzymatic hydrolysis for hemicellulase treatment is completed, the pH is adjusted to neutral, the enzyme is inactivated, the enzymatic hydrolysate is concentrated and dried to obtain the cereal dietary fiber.

[0031] In one or more embodiments, the cereal dietary fiber is rice bran dietary fiber.

[0032] The present invention also protects a cereal dietary fiber prepared by the method described in the present invention.

[0033] The third aspect of the present invention provides an application of a cereal dietary fiber.

[0034] In one or more embodiments, there is provided an application of a cereal dietary fiber in a product for improving bone metabolism, increasing osteoblasts, and improving the degree of trabecular bone rarefaction.

[0035] In one or more embodiments, there is provided an application of a cereal dietary fiber in a product for increasing the level of bone-specific alkaline phosphatase in serum.

[0036] The fourth aspect of the present invention provides a composite dietary fiber, which contains the cereal dietary fiber and inulin described in the present invention.

[0037] In one or more embodiments, the inulin is a natural polysaccharide derived from chicory or Jerusalem artichoke, and the average chain length is 8-13 monomers.

[0038] In one or more embodiments, the ratio of the cereal dietary fiber to inulin in the composite dietary fiber is 5-1:1, preferably 3-2:1.

[0039] In one or more embodiments, based on the total weight of the composite dietary fiber, the content of total dietary fiber (TDF) in the composite dietary fiber is ≥70%, preferably 70-80%, and the proportion of soluble dietary fiber (SDF) is 20-50%.

[0040] The fifth aspect of the present invention provides a preparation method of a composite dietary fiber, which comprises compounding the cereal dietary fiber and inulin in a ratio of 5-1:1, preferably 3-2:1, to obtain the composite dietary fiber.

[0041] The sixth aspect of the present invention provides an application of a composite dietary fiber in a product beneficial to human health. Preferably, the benefits to human health include one or more of anti-aging, antioxidant, regulating intestinal flora, reducing chronic inflammation, improving exercise ability and cognitive ability, and enhancing immunity.

[0042] In one or more embodiments, there is provided an application of a composite dietary fiber in a product for increasing the content of short-chain fatty acids in the intestine. More preferably, it is to increase the content of total acid, acetic acid, propionic acid, and butyric acid.

[0043] In one or more embodiments, there is provided an application of a composite dietary fiber in a product for improving the abundance of Bifidobacterium and Faecalibacterium prausnitzii in the intestine, and in a product for improving the diversity of intestinal flora.

[0044] In one or more embodiments, a composite dietary fiber is provided for use in a product for increasing the levels of SOD, T-AOC and GSH in the blood, and for use in a product for reducing the content of inflammatory markers produced in the intestine, such as calprotectin and LCN-2.

[0045] In one or more embodiments, a composite dietary fiber is provided for use in a product for improving the immunity of an aging mammal. Preferably, it is to increase the number of immune primitive cells (T Naive ) levels, reducing T cell CD4 + 、CD8 + T CM The proportion of subtypes can enhance the immunity of the elderly population.

[0046] A seventh aspect of the present invention provides a food comprising the cereal dietary fiber and / or composite dietary fiber described in the present invention.

[0047] In one or more embodiments, the food is: a pasta product, a meat product, a rice product, a baked food, a snack food, a nutritional food, a meal replacement product, a filling or a solid beverage.

[0048] Other aspects of the present invention will be apparent to those skilled in the art in view of the disclosure herein.

[0049] Technical effects of the present invention:

[0050] 1. The present invention provides a cereal dietary fiber that can improve bone metabolism and a preparation method thereof. The product has been shown through mouse experiments to significantly increase osteoblasts and improve the sparseness of trabeculae, thereby having the effect of improving bone metabolism, and is particularly suitable for improving bone metabolism in the elderly.

[0051] 2. The main components of the product of the present invention are still the macromolecular nutrients such as dietary fiber and protein naturally present in rice bran, but they are modified through certain processes to have the functional characteristics of improving bone metabolism. Different from the currently reported rice bran active ingredient extracts or rice bran peptide synthetic compounds, the product has greater safety and a higher degree of utilization of raw materials.

[0052] The rice bran product prepared in the present invention has dietary fiber and other macromolecular substances as main components, the protein components contained in the raw materials are not enzymatically hydrolyzed, and the rice bran raw materials used are common rice varieties, which do not contain anthocyanins, and oryzanol has been removed during the defatting process.

[0053] 3. Data in previous Nutrients literature showed that, compared with mice in the sham operation group, the content of alkaline phosphatase (ALP) in the blood of female ovariectomized mice was significantly increased, while after using rice bran extract, this index could be significantly decreased. However, in the examples of the present invention, after feeding the rice bran prepared by the present invention, the ALP index in the blood of aged mice increased significantly. This index is an indicator of osteoblasts in bone metabolism, and our experimental results are contrary to the previous literature reports. This also shows that the improvement of bone metabolism in the present invention is achieved through the increase of osteoblasts, and it also proves that there are obvious differences between the modified rice bran product of the present invention and the previous literature reports in terms of the product itself and the mechanism of action.

[0054] 4. The raw materials used in the present invention are derived from food ingredients. After certain processing, the raw materials are easily available and can be used in various foods to improve bone metabolism through food intake. Compared with the extracts or synthetic compounds reported in previous literature and patents, the usage scenarios are more diverse. Especially for the elderly population, more choices bring more convenience.

[0055] 5. The composite dietary fiber provided by the present invention is composed of cereal dietary fiber and inulin, wherein the cereal dietary fiber is derived from the by-products after cereal processing and has improved nutritional value after modification.

[0056] 6. The composite dietary fiber provided by the present invention contains both soluble dietary fiber (SDF) and insoluble dietary fiber (IDF).

[0057] 7. The present invention combines two different sources of dietary fiber and, through scientific proportioning, enables them to have a synergistic effect. As a food raw material, it can not only improve the texture of food, enhance nutrition, but also help to improve one or more functions of the body such as anti-aging, antioxidant, regulating intestinal flora, reducing chronic inflammation, improving exercise ability and cognitive ability, and enhancing immunity. Description of the Drawings

[0058] Figure 1 It is a CT observation diagram of trabecular bone. The small diagram below is an enlarged image of a partial field of view in the cross-section observed from top to bottom. Detailed Description of the Invention

[0059] The following details each aspect of the present invention. Without specific description, various raw materials of the present invention can be obtained commercially or prepared according to conventional methods in the art. Unless otherwise defined or described, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. Any methods and materials similar or equivalent to the described content can be applied to the method of the present invention.

[0060] As used in the present invention, all features defined in the form of numerical ranges or percentage ranges, such as numerical values, quantities, contents, and concentrations, are only for the sake of brevity and convenience. Accordingly, the description of a numerical range or percentage range should be regarded as having covered and specifically disclosed all possible sub-ranges and individual numerical values (including integers and fractions) within the range.

[0061] As used in the present invention, the term "above" or "below" means including the recited number.

[0062] As used in the present invention, for the sake of brevity, not all possible combinations of all technical features in each embodiment or example are described. Therefore, as long as there is no contradiction in the combination of these technical features, the technical features in each embodiment or example can be combined arbitrarily, and all possible combinations should be considered as falling within the scope described in this specification.

[0063] Cereal dietary fiber

[0064] The cereal dietary fiber provided by the present invention, based on the total weight of the cereal dietary fiber, has an oil content of ≤2% in the cereal dietary fiber, a starch content of ≤5%, a total dietary fiber content of ≥50%, a protein content of 20 - 30 wt%, and a soluble pentosan content of ≥2%.

[0065] Preferably, the oil content is ≤2 wt%, preferably ≤1.7 wt%. In some embodiments, the oil content of the cereal product is 0.4 - 1.7 wt%, more preferably 0.7 - 1.7 wt%, and most preferably 0.9 - 1.7 wt%.

[0066] Preferably, the starch content is ≤5 wt%. In some embodiments, the starch content of the cereal product is 2.0 - 5.0 wt%.

[0067] Preferably, the protein content is 20 - 30 wt%, more preferably 20 - 25 wt%, and most preferably 22 - 25 wt%.

[0068] Preferably, the total dietary fiber content is 53.0 - 60.0 wt%. Herein, the total dietary fiber includes soluble dietary fiber and insoluble dietary fiber.

[0069] Preferably, in the cereal product, the content of soluble pentosan is ≥2.5 wt%. In some embodiments, the content of soluble pentosan in the cereal product is 2.0 - 4.0 wt%.

[0070] Preparation method of cereal dietary fiber

[0071] The present invention provides a method for preparing cereal dietary fiber, which includes degreasing, de-starching and hemicellulose enzymolysis treatment of cereal bran. Optionally, extrusion treatment / steam explosion treatment is also included before hemicellulose enzymolysis.

[0072] In this article, the cereal bran is selected from one or more of rice bran, wheat bran, barley bran, oat bran and corn bran.

[0073] In this article, the degreasing of cereal bran can be carried out by the conventional methods for degreasing oil crop seeds in the art. Specifically, the cereal can be treated with a solvent capable of dissolving lipids for degreasing. The solvents include n-hexane, petroleum ether, etc. Generally, the mass ratio of cereal bran to degreasing solvent can be 1:2 - 10, such as 1:4 - 8, and more preferably 1:5 - 8. The cereal bran can be treated with the degreasing solvent several times, such as 1 - 5 times, preferably 2 - 5 times, and then a precipitate is obtained, and the solvent in the precipitate is removed to obtain degreased cereal bran. The oil content in the degreased cereal bran obtained by degreasing treatment is ≤2wt%. In one or more embodiments, the solvent is n-hexane and / or petroleum ether.

[0074] In this article, amylases can be used for de-starching treatment. Any amylases well-known in the art can be used in the present invention. In some embodiments, the amylase is a thermostable (such as resistant to high temperatures above 70°C, such as resistant to 70 - 100°C) α-amylase, and the optimal pH range for the action of this thermostable α-amylase is 5.8 - 7.8, and the optimal action temperature is 90 - 95°C. The mass ratio of degreased cereal bran to the enzyme can be 100 - 2000:1, such as 200 - 1500:1, 200 - 1000:1 or 250 - 500:1. The enzymolysis reaction system also contains water, and the mass ratio of water to degreased cereal bran can be 2 - 10:1, such as 3 - 8:1 or 4 - 6:1. The enzymolysis can be carried out within the optimal pH range and optimal action temperature range of the used enzyme. For example, the enzymolysis can be carried out at 90 - 95°C. The enzymolysis time is determined according to factors such as the amount of materials and the performance of the used enzyme, and generally can be from 30 minutes to 5 hours. After the enzymolysis is completed, the precipitate part of the enzymolysis solution can be separated and then dried. The precipitate can be separated by, for example, centrifugation.

[0075] In this article, steam explosion, also known as "Steam Explosion", is a technology for pretreating biomass by applying the principle of steam ejection. The essence of this technology is to instantaneously release all the steam molecules that have penetrated into the plant tissue, converting the internal energy of the steam into mechanical energy and acting on the intercellular layer of the biomass tissue, so as to decompose the raw material according to the purpose with less energy. In this article, the steam pressure for steam explosion treatment can be 0.8 - 1.5 MPa, and the pressure holding time can be 60 - 150 seconds. For example, a cereal bran sample can be added to a suitable container, such as a steam explosion tank, and then saturated steam is introduced to make the pressure in the container 0.8 - 1.5 MPa and maintain this pressure in the container for 60 - 150 seconds, thereby completing the steam explosion treatment. In some embodiments, the pressure for steam explosion treatment is 0.9 - 1.2 MPa, and the time is 90 - 150 s. The steam explosion equipment well-known in the art can be used to implement the present invention, and these equipment can be commercially available products. After the steam explosion treatment is completed, the cereal sample is taken out and dried.

[0076] In this article, the temperature for extrusion is 110 - 130 °C.

[0077] In this article, the hemicellulase treatment includes the step of enzymatic hydrolysis using hemicellulase. Various hemicellulases well-known in the art can be used to implement this enzymatic hydrolysis, such as commercially available HC-90 hemicellulase. The mass ratio of the enzyme to the cereal bran can be 1:100 - 1500, such as 1:200 - 1000 or 1:200 - 500. The enzymatic hydrolysis reaction system also contains water, and the mass ratio of water to the cereal can be 2 - 10:1, such as 3 - 8:1 or 4 - 6:1. The enzymatic hydrolysis can be carried out within the optimal pH range and the optimal action temperature range of the enzyme used. For example, the enzymatic hydrolysis can be carried out at 40 - 60 °C. The pH value of the enzymatic hydrolysis system can be adjusted in a manner well-known in the art, such as adding acid or base to adjust the pH value of the enzymatic hydrolysis system to 4 - 6. The enzymatic hydrolysis time is determined according to factors such as the amount of the material and the performance of the enzyme used, and it can generally be 2 - 8 hours, preferably 4 - 6 hours. After the enzymatic hydrolysis is completed, the pH is adjusted to neutral, the enzymatic hydrolysis solution is concentrated and dried, and the cereal dietary fiber of the present invention can be obtained. The dried product can be pulverized to 50 - 150 mesh, such as 80 - 100 mesh.

[0078] Preferably, the cereal dietary fiber is rice bran dietary fiber.

[0079] In some embodiments, the cereal dietary fiber of the present invention can be prepared by a method including the following steps: degreasing, de-starching, steam explosion, and hemicellulase hydrolysis of cereal bran, and the order of de-starching and steam explosion can be interchanged.

[0080] In some embodiments, the method for preparing the cereal dietary fiber of the present invention is as follows:

[0081] (1) The by-product rice bran obtained in rice processing is added with n-hexane with a volume 5-8 times that of the rice bran for defatting treatment, so that the residual oil content in the sample is below 2%.

[0082] (2) The defatted rice bran obtained in step (2) is subjected to de-starching treatment. The method is to take a certain amount of the rice bran sample obtained in step (1), add distilled water with a volume 4-6 times that of the rice bran sample, add thermostable amylase for enzymatic hydrolysis reaction. The enzymatic hydrolysis conditions are: the enzyme dosage is 0.1-0.5%, the temperature is 90-95 °C, and the enzymatic hydrolysis time is 0.5-1 h. After enzymatic hydrolysis, centrifuge and discard the supernatant, and dry the precipitate so that the residual starch content in the sample is below 5%.

[0083] (3) The product obtained in step 2 is subjected to a combined treatment of physical modification and enzymatic modification. The method is to first subject the material to steam explosion under the conditions of 0.9-1.5 MPa and a time of 90-150 s. After the treatment, add distilled water with a volume 6 times that of the material, add hemicellulase for enzymatic hydrolysis, the enzyme dosage is 0.1-0.5%, the temperature is 40-60 °C, pH is 4-6, and the enzymatic hydrolysis time is 4-6 h. After enzymatic hydrolysis is completed, adjust the pH to neutral with sodium hydroxide, boil to inactivate the enzyme for 15 min, concentrate the enzymatic hydrolysate and dry it at 105 °C, and pulverize it after drying to obtain the final sample.

[0084] The amylase is a thermostable α-amylase (such as resistant to high temperatures above 70 °C, such as resistant to 70-100 °C), and any amylase well-known in the art can be used in the present invention. For this enzymatic hydrolysis, various hemicellulases well-known in the art can be used for the hemicellulase, for example, commercially available HC-90 hemicellulase or hemicellulose complex enzyme type 606 can be used.

[0085] Furthermore, the present invention also protects a cereal dietary fiber prepared by the foregoing method for preparing cereal dietary fiber.

[0086] Application of cereal dietary fiber

[0087] Through in-depth research, the present inventor has found that by subjecting cereal bran to defatting, de-starching, steam explosion treatment and hemicellulase hydrolysis treatment, a cereal product can be obtained, which can significantly increase the activity of osteoblasts, effectively improve the sparsity of bone trabeculae, and has the effect of improving bone metabolism.

[0088] The present invention provides an application of a cereal dietary fiber in a product for improving bone metabolism, increasing osteoblasts, and improving the sparsity of bone trabeculae. The method includes administering an effective amount of the cereal product described in any embodiment herein to the subject. The cereal product can be consumed as three meals a day for the subject, and / or can be consumed as a snack.

[0089] In one or more embodiments, there is provided an application of cereal dietary fiber in a product for increasing the level of bone-specific alkaline phosphatase in serum.

[0090] Compound dietary fiber

[0091] The inventors of the present application found that the compounding of the cereal dietary fiber and inulin of the present invention has a synergistic effect. As a food raw material, it can not only improve the texture of food and enhance nutrition, but also contribute to one or more effects such as anti-aging, antioxidant, regulation of intestinal flora, reduction of chronic inflammation, improvement of exercise ability and cognitive ability, and enhancement of immunity, thereby completing the present invention.

[0092] In this article, the inulin is a natural polysaccharide derived from chicory or Jerusalem artichoke, and the average chain length is 8-13 monomers. The ratio of the composite dietary fiber to inulin in the composite dietary fiber is 5-1:1, preferably 3-2:1. Based on the total weight of the composite dietary fiber, the content of total dietary fiber (TDF) in the composite dietary fiber is ≥70%, preferably 70-80%, and the proportion of soluble dietary fiber (SDF) is 20-50%.

[0093] Preparation method of compound dietary fiber

[0094] The present invention provides a preparation method of a composite dietary fiber, in which the cereal dietary fiber and inulin are compounded in a ratio of 5-1:1, preferably 3-2:1, to obtain the composite dietary fiber.

[0095] Application of compound dietary fiber

[0096] The present invention provides an application of a composite dietary fiber in a product beneficial to human health. Preferably, the benefits to human health include one or more of anti-aging, antioxidant, regulation of intestinal flora, reduction of chronic inflammation, improvement of exercise ability and cognitive ability, and enhancement of immunity. The method includes administering an effective amount of the composite dietary fiber product described in any embodiment herein to the subject. The composite dietary fiber product can be consumed as the three meals a day of the subject and / or can be consumed as a snack.

[0097] Preferably, there is provided an application of a composite dietary fiber in a product for increasing the content of short-chain fatty acids in the intestine. More preferably, it is to increase the content of total acid / acetic acid / propionic acid / butyric acid.

[0098] Preferably, there is provided an application of a composite dietary fiber in a product for improving the abundance of Bifidobacterium and Faecalibacterium prausnitzii in the intestine, and in a product for improving the diversity of intestinal flora.

[0099] Preferably, provided is the use of a composite dietary fiber in a product for increasing the levels of SOD, T-AOC and GSH in the blood, and in a product for reducing the levels of inflammatory markers produced in the intestine, such as calprotectin and LCN-2.

[0100] Preferably, provided is the use of a composite dietary fiber in a product for improving the immunity of aging mammals. Preferably, it is to increase the level of immune progenitor cells (T Naive ), and reduce the proportion of T cell CD4 + , CD8 + of the T CM subtype, and enhance the body immunity of the elderly population.

[0101] Food

[0102] The present invention provides a food product, which contains the cereal dietary fiber and / or composite dietary fiber described in the present invention. Preferably, the food product is: flour products, meat products, rice products, baked foods, snack foods, nutritional foods, meal replacement products, fillings or solid beverages.

[0103] The present invention will be described below by way of specific examples. It should be understood that these examples are merely illustrative and are not intended to limit the scope of the present invention. In the present invention, the prepared cereal dietary fiber sample can be detected as follows:

[0104] 1. Detection method for fat content: Detection is carried out with reference to the method of GB 5009.6-2016;

[0105] 2. Detection of starch content: Detection is carried out with reference to the acid hydrolysis method of GB 5009.9-2016;

[0106] 3. Detection of dietary fiber content: Detection is carried out with reference to the method of GB 5009.88-2014;

[0107] 4. Detection of protein content: GB 5009.5-2016 National Food Safety Standard Determination of Protein in Foods;

[0108] 5. Detection of soluble pentosan content: Weigh 2.0 g of the sample precisely into an Erlenmeyer flask, add 100 mL of water, and shake and extract at 30 °C for 120 min. After shaking well, transfer it to a centrifuge tube, centrifuge at a centrifugal force of 4000×g for 15 min. Measure 10 mL of the supernatant, add 10 mL of 4 mol / L hydrochloric acid solution, place it in a 25-mL stoppered test tube, stopper it, and hydrolyze in a boiling water bath for 120 min. After cooling, filter with filter paper, collect the filtrate, dilute it, and reserve for use. Pipette 3 mL of the diluted sample solution into a 10-mL stoppered test tube, successively add 0.3 mL of 1% orcinol-anhydrous ethanol solution and 3 mL of 0.1% ferric chloride-hydrochloric acid solution, and mix well by vortex. Heat in a boiling water bath for 30 min, cool to room temperature, transfer it to a 10-mL volumetric flask, and make up the volume with distilled water. Measure the absorbance values at wavelengths of 670 nm and 580 nm respectively, and calculate the difference between the two. Use xylose as a reference substance to make a standard curve, and calculate the soluble pentosan content in the sample through the standard curve.

[0109] 6. Experiment on acute aging animal model (bone metabolism): Six-month-old male C57BL / 6J mice were randomly divided into different groups according to their body weights, with 8 mice in each group, including a normal control group (NC group), an acute aging model control group injected with D-gal for modeling (M-AG group. Inject a physiological saline solution of D-gal subcutaneously at the back of the neck at a dose of 300 mg / kg / d for 8 consecutive weeks), and each intervention group fed with customized rice bran feed. Among them, the normal control group and the acute aging model control group were fed with ordinary feed, and the customized rice bran feed was used as the intervention group to feed the acute aging model mice. The mice were all placed under standard experimental conditions, at a room temperature of 25±2 °C, a relative humidity of 50±10%, and a light-dark cycle of 12 h. During the whole adaptation and experimental period, the mice could freely obtain food and water. The intervention time was 8 weeks.

[0110] 7. Experiment on natural aging animal model (compound): Take 8-month-old C57BL / 6J mice, with half males and half females, and randomly divide them into different groups according to their body weights, with 10 mice in each group, including a natural aging model control group (N-AG group, fed with ordinary feed), the example group of the present invention, and each intervention group fed with the customized rice bran feed prepared in the comparative example group. Place the mice under standard experimental conditions, at a room temperature of 25±2 °C, a relative humidity of 50±10%, and a light-dark cycle of 12 h. During the whole adaptation and experimental period, the mice could freely obtain food and water. After two months of adaptive feeding with ordinary maintenance feed, change to customized rice bran feed and feed for 3 months.

[0111] 8. Detection of various indicators: During the experiment, the mice were weighed and the food intake was recorded every week; various relevant indicators of the mice were detected during and at the end of the experiment.

[0112] Serum bone metabolism indicators: After feeding the acute aging model mice for 8 weeks, an Elisa kit method was used to detect tartrate-resistant acid phosphatase (TRACP) and bone-specific alkaline phosphatase (BALP) in the serum of each mouse to evaluate its bone metabolism indicators.

[0113] Detection of bone microstructure: After feeding the acute aging model mice for 8 weeks, the femurs of the mice were taken for Micro-CT scanning, and the bone mineral density (BMD), trabecular separation (Tb.Sp), and bone volume fraction (BV / TV) were analyzed, and the sparsity of the trabecular structure was observed.

[0114] Short-chain fatty acids (SCFAs) and beneficial bacteria: Gas chromatography was used to determine the contents of acetic acid, propionic acid, and butyric acid in the feces of mice. qPCR was used to detect the proliferation numbers of Bifidobacterium and Faecalibacterium prausnitzii.

[0115] Antioxidant experiment: The contents of SOD (WST-1 method), T-AOC (ABTS method), and GSH (microplate method) in the blood of mice were detected to evaluate the oxidative stress level of mice.

[0116] Inflammatory factor detection: ELISA method was used to analyze the contents of calprotectin (CALP) and lipocalin-2 (LCN-2) in the feces of mice.

[0117] Immune T cell detection: Flow cytometry was used to analyze CD4 + , CD8 + , and the proportion of its T CM subtypes.

[0118] Mouse aging score: The appearance aging degree of mice was evaluated every month during the experiment. The observation indicators included behavioral activity categories considered related to the aging process and the general appearance of the skin, eyes, and spine. According to the degree of change, the score for each category was 0 to 4 points, and the aging score of each mouse was the sum of the scores for each category.

[0119] Motor and cognitive ability tests: Include: Pole test, Y-maze, New object recognition, Open field test. The test methods are as follows.

[0120] Mouse pole test: A test widely used to evaluate movement disorders related to the basal ganglia in mice. The mice were trained to complete the pole test in two training trials. The mice were placed head-up at the top of the pole, and the animals would usually orient themselves downward naturally and descend along the length of the pole to return to their cages. In five trials, the total time required for the animals to descend to the bottom of the pole was recorded.

[0121] Open field test: The open field test was used to evaluate the spontaneous activity ability of mice. The mice were placed in a black test chamber of 50 cm × 50 cm × 40 cm and allowed to freely explore for 5 minutes. A camera was placed at the top of the open area to record and calculate the proportion of time the mice spent in the central area.

[0122] Y maze: The Y maze is a test of the working memory of mice, which depends on the animal's preference for the new arms of the maze. The Y maze (35 cm × 16 cm × 16 cm) was used for the test, where the three opaque arms were at 120° angles to each other. The mice were introduced to the same initial position in the maze and allowed to freely explore the arms for 5 minutes. Entering three different arms was counted as one alternation. The number of arm entries and the number of alternations were recorded to calculate the percentage of alternation behavior. Spontaneous alternation percentage (%) = [Number of successful alternations / (Total number of arm entries - 2)] × 100.

[0123] Novel object recognition test: It was used to evaluate the short-term memory ability of mice. The experimental process was divided into an adaptation stage, a training stage, and a test stage. In the adaptation stage, each mouse was placed in an open field without any objects and allowed to freely explore for 5 minutes. The next day, there were two odorless objects with the same shape and color in the open field, and each mouse was placed in the box for 10 minutes of exploration, which was called the training process stage. The third day was the test stage, using the same process as the training stage, but one of the objects was replaced with a new object with different shape and color. The exploration behavior was defined as follows: When the mouse licked, sniffed, and touched the object with its paw or head within a range of 2 cm from the object, the time of the mouse's exploration behavior was recorded. The novel object recognition index (NOI) was used to define the novel object recognition ability of mice: NOI = Exploration time of novel object / (Exploration time of novel object + Exploration time of old object).

[0124] 9. In vitro fermentation experimental method for middle-aged and elderly groups: Using the samples obtained from the examples and comparative examples as substrates, the INFOGEST method was used for in vitro simulated digestion experiments. After simulating the absorption of small molecules by human intestinal epithelial cells, the remaining substances (simulating the substances after reaching the human colon) were prepared into a culture medium for inoculation and fermentation. The steps were as follows:

[0125] (1) Preparation of simulated digestive fluid: 0.5 mL of CaCl2(H2O)2 (0.3 M), 30 mL of KCl (0.5 M), 6 mL of KH2PO4 (0.5 M), 65 mL of NaHCO3 (1 M), 25 mL of NaCl (2 M), 2 mL of MgCl2(H2O)6 (0.15 M), and 2 mL of (NH4)2CO3 (0.5 M) were taken respectively, and water was added to prepare a 400 mL solution. The required pH value was adjusted with 1 M NaOH or 1 M HCl.

[0126] (2) Mix each sample with water at a ratio of 1:6.25 and incubate in a water bath at 100 °C for 5 min. Take 1 g of the mixture, add simulated digestive fluid, mix well, and then add α - salivary amylase to make the enzyme activity in the final mixture 75 U / mL. Incubate in a water bath at 37 °C for 2 min;

[0127] (3) Add simulated gastric phase digestive fluid (adjust the pH of the simulated digestive fluid to 1.5 - 2 with HCl) to the sample that has completed oral simulated digestion above, mix well, and then add pepsin to make the enzyme activity in the final mixture 2000 U / mL. Incubate in a water bath at 37 °C for 2 hours;

[0128] (4) Add simulated digestive fluid to the sample that has completed gastric phase digestion above, mix well, and then add bile solution to make the bile concentration in the final mixture 10 mM, and add trypsin to make the enzyme activity of trypsin in the final mixture 100 U / mL. Incubate in a water bath at 37 °C for 2 hours to complete intestinal simulated digestion;

[0129] (5) After the above reaction is completed, heat in a water bath at 100 °C for 5 min to terminate the reaction;

[0130] (6) Cool the reaction solution and then perform dialysis. Add an appropriate amount of dialysis fluid (3.75 g / L sodium bicarbonate solution) to a beaker and stir it on a magnetic stirrer. Dialysis lasts for 3 hours and the dialysis fluid is changed every hour. Obtain the substrate for preparing the culture medium.

[0131] (7) Mix the obtained substrates with the basal medium and boil. The basal medium contains the following components: 2.5 g / L yeast extract; 10 g / L tryptone; 0.8 g / L L - cysteine hydrochloride; 0.05 g / L hemin chloride; 0.9 g / L NaCl; 0.09 g / L MgSO4·7H2O; 0.09 g / L CaCl2·6H2O; 0.45 g / L KH2PO4; 0.45 g / L K2HPO4; a mixture of trace vitamins and 0.1 mg / L Resazurin. Prepare the anaerobic culture medium by the N2 flushing method, dispense it into 5 - mL vials and perform autoclaving to obtain a culture medium with a substrate concentration of 4‰ (W / V) for in vitro fermentation culture.

[0132] Detection methods for SCFA production, microbial diversity, and beneficial bacteria in in vitro fermentation: Collect feces from the subjects, requiring that all subjects are non - vegetarians and have not taken antibiotics and probiotics for at least 2 months before sample collection. After processing the feces, inoculate it into the culture medium for in vitro fermentation experiments, detect SCFA, and perform qPCR to detect Bifidobacterium and Faecalibacterium prausnitzii.

[0133] Raw material source:

[0134] α - high temperature amylase: purchased from Novozymes Biotechnology Company;

[0135] Hemicellulase HC - 90: purchased from Tianye Enzyme Preparation Co., Ltd.;

[0136] Hemicellulase complex enzyme type 606: purchased from Baiyin Sinogene Biotechnology Co., Ltd.;

[0137] Other raw materials and reagents used in the examples and comparative examples are conventional raw materials and reagents in this field and can be obtained through commercial channels.

[0138] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions in the following examples are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight.

[0139] Example 1:

[0140] Take 1.5 kg of rice bran, add 7.5 L of n - hexane for degreasing treatment, perform the treatment twice, after filtration, evaporate the solvent in the precipitate to dryness, and obtain degreased rice bran. Take 1 kg of the degreased rice bran sample, add 6 kg of distilled water, mix well and heat to 95 °C, add 5 g of α - high temperature amylase, and stir and enzymatically hydrolyze at 95 °C for 1 h. After the enzymatic hydrolysis is completed, centrifuge, discard the supernatant, and dry the precipitate part.

[0141] Load the above - mentioned dried precipitate part into a steam explosion device for steam explosion treatment, with a pressure of 0.9 MPa and a time of 90 seconds. After the treatment, dry the material. Take 500 g of the material after steam explosion, add 3000 mL of distilled water, adjust the pH to 4 with hydrochloric acid, add 1 g of hemicellulase HC - 90 for enzymatic hydrolysis, the enzymatic hydrolysis temperature is 50 °C, and the enzymatic hydrolysis time is 6 h. After the enzymatic hydrolysis is completed, adjust the pH to neutral with sodium hydroxide, boil to inactivate the enzyme for 15 min, concentrate the enzymatic hydrolysate and dry it at 105 °C, and then pulverize it to 80 - 100 mesh.

[0142] Example 2:

[0143] Take 1.5 kg of rice bran, add 9 L of n - hexane for degreasing treatment, perform the treatment twice, after filtration, evaporate the solvent in the precipitate to dryness, and obtain degreased rice bran. Take 1 kg of the degreased rice bran sample, add 4 kg of distilled water, mix well and heat to 90 °C, add 1 g of α - high temperature amylase, and stir and enzymatically hydrolyze at 90 °C for 1 h. After the enzymatic hydrolysis is completed, centrifuge, discard the supernatant, and dry the precipitate part.

[0144] Load a portion of the dried precipitate described above into a steam explosion device for steam explosion treatment at a pressure of 1.5 MPa for 120 seconds. After the treatment, dry the material. Take 500 g of the material after steam explosion, add 3000 mL of distilled water, adjust the pH to 5 with hydrochloric acid, add 2.5 g of hemicellulase HC-90 for enzymatic hydrolysis at an enzymatic hydrolysis temperature of 60 °C for 4 h. After the enzymatic hydrolysis is completed, adjust the pH to neutral with sodium hydroxide, boil to inactivate the enzyme for 15 min, concentrate the enzymatic hydrolysate, dry it at 105 °C, and pulverize it to 80-100 mesh after drying.

[0145] Example 3:

[0146] Take 1.5 kg of rice bran, add 12 L of n-hexane for degreasing treatment twice. After filtration, evaporate the solvent in the precipitate to obtain degreased rice bran. Take 1 kg of the degreased rice bran sample, add 5 kg of distilled water, mix well and heat to 95 °C, add 3 g of α-thermostable amylase, and stir and enzymatically hydrolyze at 95 °C for 0.5 h. After the enzymatic hydrolysis is completed, centrifuge, discard the supernatant, and dry the precipitate part.

[0147] Load a portion of the dried precipitate described above into a steam explosion device for steam explosion treatment at a pressure of 1.2 MPa for 150 seconds. After the treatment, dry the material. Take 500 g of the material after steam explosion, add 3000 mL of distilled water, adjust the pH to 6 with hydrochloric acid, add 0.5 g of hemicellulase HC-90 for enzymatic hydrolysis at an enzymatic hydrolysis temperature of 40 °C for 5 h. After the enzymatic hydrolysis is completed, adjust the pH to neutral with sodium hydroxide, boil to inactivate the enzyme for 15 min, concentrate the enzymatic hydrolysate, dry it at 105 °C, and pulverize it to 80-100 mesh after drying.

[0148] Example 4:

[0149] Take 1.5 kg of rice bran, add 7.5 L of n-hexane for degreasing treatment twice. After filtration, evaporate the solvent in the precipitate to obtain degreased rice bran. Take 1 kg of the degreased rice bran sample, add 6 kg of distilled water, mix well and heat to 90 °C, add 1 g of α-thermostable amylase, and stir and enzymatically hydrolyze at 90 °C for 1 h. After the enzymatic hydrolysis is completed, centrifuge, discard the supernatant, and dry the precipitate part.

[0150] Load a portion of the dried precipitate described above into a steam explosion device for steam explosion treatment at a pressure of 0.9 MPa for 150 seconds. After the treatment, dry the material. Take 500 g of the material after steam explosion, add 3000 mL of distilled water, adjust the pH to 5 with hydrochloric acid, add 1.5 g of hemicellulose complex enzyme 606 for enzymatic hydrolysis at an enzymatic hydrolysis temperature of 45 °C for 5 h. After the enzymatic hydrolysis is completed, adjust the pH to neutral with sodium hydroxide, boil to inactivate the enzyme for 15 min, concentrate the enzymatic hydrolysate, dry it at 105 °C, and pulverize it to 80-100 mesh after drying.

[0151] Example 5:

[0152] Take 1.5 kg of rice bran, add 9 L of n-hexane for degreasing treatment, conduct the treatment twice, evaporate the solvent in the precipitate after filtration, and obtain degreased rice bran. Take 1 kg of the degreased rice bran sample, add 5 kg of distilled water, mix well and heat to 95 °C, add 5 g of α-thermostable amylase, and stir and enzymatically hydrolyze at 95 °C for 0.5 h. After the enzymatic hydrolysis is completed, centrifuge, discard the supernatant, and dry the precipitate part.

[0153] Load the above-mentioned dried precipitate part into a steam explosion device for steam explosion treatment, with a pressure of 1.1 MPa and a time of 120 seconds. After the treatment, dry the material. Take 500 g of the material after steam explosion, add 3000 mL of distilled water, adjust the pH to 4 with hydrochloric acid, add 0.5 g of hemicellulose complex enzyme 606 for enzymatic hydrolysis, with an enzymatic hydrolysis temperature of 55 °C and an enzymatic hydrolysis time of 6 h. After the enzymatic hydrolysis is completed, adjust the pH to neutral with sodium hydroxide, boil to inactivate the enzyme for 15 min, concentrate the enzymatic hydrolysate and dry it at 105 °C, and pulverize it to 80 - 100 mesh after drying.

[0154] Example 6:

[0155] Take 1.5 kg of rice bran, add 12 L of n-hexane for degreasing treatment, conduct the treatment twice, evaporate the solvent in the precipitate after filtration, and obtain degreased rice bran. Take 1 kg of the degreased rice bran sample, add 4 kg of distilled water, mix well and heat to 90 °C, add 4 g of α-thermostable amylase, and stir and enzymatically hydrolyze at 90 °C for 0.5 h. After the enzymatic hydrolysis is completed, centrifuge, discard the supernatant, and dry the precipitate part.

[0156] Load the above-mentioned dried precipitate part into a steam explosion device for steam explosion treatment, with a pressure of 1.5 MPa and a time of 90 seconds. After the treatment, dry the material. Take 500 g of the material after steam explosion, add 3000 mL of distilled water, adjust the pH to 6 with hydrochloric acid, add 2.5 g of hemicellulose complex enzyme 606 for enzymatic hydrolysis, with an enzymatic hydrolysis temperature of 50 °C and an enzymatic hydrolysis time of 4 h. After the enzymatic hydrolysis is completed, adjust the pH to neutral with sodium hydroxide, boil to inactivate the enzyme for 15 min, concentrate the enzymatic hydrolysate and dry it at 105 °C, and pulverize it to 80 - 100 mesh after drying.

[0157] Example 7

[0158] Take 500 g of rice bran, add 2.5 L of n-hexane for degreasing treatment, evaporate the solvent in the precipitate after filtration, and obtain defatted rice bran. Take 100 g of the defatted rice bran sample, add 600 mL of distilled water, mix well and heat to 95 °C, add 0.2 g of α-thermostable amylase, and stir and enzymatically hydrolyze at 95 °C for 1 h. After the enzymatic hydrolysis is completed, centrifuge, discard the supernatant, and dry the precipitate part.

[0159] Add a portion of the dried precipitate mentioned above to 600 mL of distilled water, mix well and heat to 60 °C. Adjust the pH of the system to 5 with food-grade citric acid, add 0.1 g of hemicellulase type 606, and enzymatically hydrolyze for 6 h. After the enzymatic hydrolysis is completed, adjust the pH to neutral with edible alkali, boil to inactivate the enzyme for 15 min, concentrate the enzymatic hydrolysate and dry it at 105 °C. After drying, pulverize it to 80 - 100 mesh. Compound the modified rice bran and inulin in a ratio of 2:1 to obtain the composite dietary fiber.

[0160] Example 8

[0161] Take 500 g of rice bran, add 3.5 L of n-hexane for defatting treatment. After filtration, evaporate the solvent in the precipitate to obtain defatted rice bran. Take 100 g of the defatted rice bran sample, add 500 mL of distilled water, mix well and heat to 95 °C, add 0.3 g of α-thermostable amylase, and stir and enzymatically hydrolyze at 95 °C for 0.6 h. After the enzymatic hydrolysis is completed, centrifuge, discard the supernatant, and dry the precipitate part.

[0162] Add a portion of the dried precipitate mentioned above to 500 mL of distilled water, mix well and heat to 60 °C. Adjust the pH of the system to 4.5 with food-grade citric acid, add 0.5 g of hemicellulase HC90, and enzymatically hydrolyze for 4 h. After the enzymatic hydrolysis is completed, adjust the pH to neutral with edible alkali, boil to inactivate the enzyme for 15 min, concentrate the enzymatic hydrolysate and dry it at 105 °C. After drying, pulverize it to 80 - 100 mesh. Compound the modified rice bran and inulin in a ratio of 1:1 to obtain the composite dietary fiber.

[0163] Example 9

[0164] Take 500 g of rice bran, add 3 L of n-hexane for defatting treatment. After filtration, evaporate the solvent in the precipitate to obtain defatted rice bran. Load the above defatted rice bran into a steam explosion device for steam explosion treatment at a pressure of 1.5 MPa for 90 s. After the treatment, dry the material. Take 100 g of the rice bran sample after steam explosion, add 500 mL of distilled water, mix well and heat to 95 °C, add 0.5 g of α-thermostable amylase, and stir and enzymatically hydrolyze at 95 °C for 0.5 h. After the enzymatic hydrolysis is completed, centrifuge, discard the supernatant, and dry the precipitate part.

[0165] Add a portion of the dried precipitate mentioned above to 500 mL of distilled water, mix well and heat to 50 °C. Adjust the pH of the system to 5 with food-grade citric acid, add 0.4 g of hemicellulase HC-90, and enzymatically hydrolyze for 6 h. After the enzymatic hydrolysis is completed, adjust the pH to neutral with edible alkali, boil to inactivate the enzyme for 15 min, concentrate the enzymatic hydrolysate and dry it at 105 °C. After drying, pulverize it to 80 - 100 mesh. Compound the modified rice bran and inulin in a ratio of 3:1 to obtain the composite dietary fiber.

[0166] Example 10

[0167] Take 500 g of rice bran, add 4 L of n-hexane for degreasing treatment. After filtration, evaporate the solvent in the precipitate to dryness to obtain defatted rice bran. Load the above defatted rice bran into an extrusion equipment for extrusion treatment at a temperature of 110 °C. After the treatment, dry the material. Take 100 g of the extruded rice bran sample, add 600 mL of distilled water, mix well and heat to 95 °C. Add 0.3 g of α-thermostable amylase and stir for enzymatic hydrolysis at 95 °C for 1 h. After the enzymatic hydrolysis is completed, centrifuge and discard the supernatant. Dry the precipitate part.

[0168] Add the above dried precipitate part to 600 mL of distilled water, mix well and heat to 60 °C. Adjust the pH of the system to 4.5 with food-grade citric acid, add 0.3 g of cellulase type 606, and carry out enzymatic hydrolysis for 5 h. After the enzymatic hydrolysis is completed, adjust the pH to neutral with edible alkali, boil to inactivate the enzyme for 15 min. Concentrate the enzymatic hydrolysate and dry it at 105 °C. After drying, pulverize it to 80 - 100 mesh. Compound the modified rice bran and inulin in a ratio of 5:1 to obtain composite dietary fiber.

[0169] Control 1:

[0170] Take 1 kg of rice bran, add 6 L of n-hexane for degreasing treatment, carry out the treatment twice. After filtration, evaporate the solvent in the precipitate to dryness to obtain defatted rice bran, and pulverize it to 80 - 100 mesh.

[0171] Control 2:

[0172] Take 1.5 kg of rice bran, add 9 L of n-hexane for degreasing treatment, carry out the treatment twice. After filtration, evaporate the solvent in the precipitate to dryness to obtain defatted rice bran. Take 1 kg of the defatted rice bran sample, add 5 kg of distilled water, mix well and heat to 95 °C. Add 5 g of α-thermostable amylase and stir for enzymatic hydrolysis at 95 °C for 0.5 h. After the enzymatic hydrolysis is completed, centrifuge and discard the supernatant. Dry the precipitate part and pulverize it to 80 - 100 mesh.

[0173] Control 3

[0174] Take 500 g of rice bran, add 2.5 L of n-hexane for degreasing treatment, carry out the treatment twice. After filtration, evaporate the solvent in the precipitate to dryness to obtain defatted rice bran, and pulverize it to 80 - 100 mesh. Take 100 g of the defatted rice bran sample, add 600 mL of distilled water, mix well and heat to 95 °C. Add 0.4 g of α-thermostable amylase and stir for enzymatic hydrolysis at 95 °C for 0.5 h. After the enzymatic hydrolysis is completed, centrifuge and discard the supernatant. Dry the precipitate part.

[0175] Add a portion of the dried precipitate mentioned above to 600 mL of distilled water, mix well and heat to 60 °C. Adjust the pH of the system to 5 with food-grade citric acid, add 0.2 g of cellulase type 606, and enzymatically hydrolyze for 6 h. After the enzymatic hydrolysis is completed, adjust the pH to neutral with edible alkali, boil to inactivate the enzyme for 15 min, concentrate the enzymatic hydrolysate and dry it at 105 °C, and then pulverize it to 80 - 100 mesh to obtain a dietary fiber sample.

[0176] Control Example 4 Inulin

[0177] Control Example 5

[0178] Take 500 g of rice bran, add 3.5 L of n-hexane for degreasing treatment twice. After filtration, evaporate the solvent in the precipitate to obtain defatted rice bran, and pulverize it to 80 - 100 mesh. Compound the defatted rice bran and inulin in a ratio of 1:1 to obtain a composite dietary fiber.

[0179] Detection Example 1: Detection of physical and chemical indexes:

[0180] Perform physical and chemical index detection on the rice bran fiber samples prepared in the above Examples 1 - 10 and Control Examples 1 - 5. The detection indexes include fat content, starch content, dietary fiber content, and soluble pentosan content. The results are shown in Tables 1 and 2.

[0181] Table 1. Physical and chemical detection of samples (for acute aging model experiment)

[0182]

[0183] Table 2. Physical and chemical detection of samples (for natural aging model experiment)

[0184]

[0185]

[0186] Detection Example 2: Feeding and intervention of acute aging mice

[0187] Fifty-six 6-month-old C57BL / 6J male mice were purchased from Jiangsu Jicui Yakang and randomly divided into 7 groups according to body weight, with 8 mice in each group, namely the normal control group (NC), the senescence model control group (M-AG), and the rice bran customized feed intervention groups (Example 1, Example 2, Example 6, Comparative Example 1, Comparative Example 2). Mice in the senescence model control group and the intervention groups were subcutaneously injected with a physiological saline solution of D-gal at a dose of 300 mg / kg / d on the nape of the neck for 8 consecutive weeks. Mice in the normal control group were subcutaneously injected with the same amount of physiological saline as the senescent mice. Mice in the normal control group and the M-AG group were fed with ordinary feed, while mice in the intervention groups were fed with rice bran customized feed. The rice bran customized feed was a sample obtained from an example or a comparative example, replacing a part of the cellulose raw materials in the ordinary feed, but ensuring that the energy and nutritional components of each group of feed were equivalent. During the experiment, the body weight and food intake of the mice were recorded weekly; at the end of the experiment, serum bone metabolism indicators, oxidative stress indicators, and short-chain fatty acids in feces were detected in the mice, and Micro-CT scanning analysis was performed on the femurs of the mice.

[0188] Detection Example 3: Detection of Bone Metabolism Indicators

[0189] After 8 weeks of feeding the acute senescence model mice, the bone metabolism indicators in the blood of each mouse were detected, and the results are shown in Table 2. Tartrate-resistant acid phosphatase (TRACP) is a marker of bone resorption and osteoclast activity. Measuring the concentration of TRACP, especially TRACP-5b, in serum helps to understand the bone metabolism status under physiological conditions and various pathological conditions. Bone-specific alkaline phosphatase (BALP) is involved in the bone formation process, is stable in serum, and is a marker of osteoblast maturation and activity; quantitative determination and dynamic observation of serum BALP provide effective basis for the early diagnosis of bone metabolism diseases, monitoring of treatment effects, and judgment of disease prognosis. The detection results are shown in Table 2. There was no significant difference in TRACP-5b among the groups; while in BALP, the normal group and the rice bran example groups were significantly higher than the AG group and the comparative example groups, and the Example 2 group was significantly higher than the NC group. This indicates that the rice bran samples prepared in the present invention can significantly increase the activity of osteoblasts. Since the type of osteoporosis caused by aging is low-turnover osteoporosis, its main feature is the decrease in osteoblast activity. At the same time, there was no obvious change in the osteoclast index in this experiment, which also indicates that the rice bran products prepared in the present invention mainly improve bone metabolism by increasing the activity of osteoblasts rather than inhibiting the activity of osteoclasts.

[0190] Table 3. Detection of Bone Metabolism Indicators in Acute Senescent Mice (n = 8, after 8 weeks of feeding)

[0191]

[0192]

[0193] Note: There were significant differences compared with the M-AG group (*p < 0.05, **p < 0.01)

[0194] Detection Example 4: Detection of bone microstructure

[0195] After feeding the acute aging model mice for 8 weeks, the femurs of the mice were taken for Micro-CT scanning and the obtained data were analyzed. Bone mineral density (BMD) represents the amount of bone mineral density in the bone tissue of the region of interest. The lower the bone density, the higher the risk of fracture. The results showed that the BMD of the femurs of the mice in the NC group and the rice bran sample example group was significantly higher than that in the AG group and the comparative example group. Trabecular separation (Tb.Sp) is the main index for evaluating the spatial morphological structure of trabeculae. The higher the trabecular separation, the higher the risk of fracture. The results showed that the Tb.Sp in the AG group was significantly higher than that in the NC group and the example group. Bone volume fraction (BV / TV) represents the ratio of bone tissue volume to tissue volume. The lower the bone volume fraction, the higher the risk of fracture. The results showed that the BV / TV values in the NC group and the example group were significantly higher than that in the AG group and the comparative example group. The detection results of bone microstructure indexes are shown in Table 2, and the CT results of femur scanning are shown in Figure 1 shown. The CT results of femur scanning showed that the trabecular structure of the mice in the normal group (NC group) was dense, while the trabecular structure of the mice in the aging group (M-AG group) was sparse. After intervention with rice bran, the trabecular structure density of the mice in each of Comparative Example 1, Comparative Example 2, and Comparative Example 6 was significantly improved, while the improvement in the trabecular density of Comparative Example 1 and Comparative Example 2 groups compared with the AG group was not significant.

[0196] Detection Example 5: Experimental results of in vitro fermentation to produce SCFA in middle-aged and elderly groups

[0197] Collect 10 feces samples each from middle-aged people aged 45 - 60 years and elderly people aged 60 - 80 years. Weigh 1 g ± 0.02 g of the feces sample, put it into a feces sample box, seal it and then put it into a feces treatment instrument. Add 10 mL of feces dilution solution (thiamine and riboflavin at 5 mg / L were added to the PBS solution) to prepare a 10% feces suspension, and shake it evenly. Use a syringe to inoculate the sample into the required fermentation medium. Put the inoculated medium into a 37°C incubator. Draw 500 mL of the fermentation broth and add 100 mL of crotonic acid after culturing for 24 hours. Draw 500 mL of the fermentation broth and add 100 mL of crotonic acid after culturing for 48 hours to stop the fermentation. Acidify at -30°C for 24 hours, and aliquot the remaining fermentation broth.

[0198] SCFA detection: The SCFA concentration in the fermentation broth was determined by gas chromatography (GC) (Shimadzu, GC-2010Plus, Japan) equipped with a DB-FFAP column (0.32 mm × 30 m × 0.5 μm) (Agilent Technologies, United States) using a H2 flame ionization detector. Crotonic acid was used as the internal standard, and the internal standard method was adopted to determine the contents of acetic acid, propionic acid, and butyric acid in the fermentation broth filtrate, and calculate the total acid content. The results are shown in Tables 2 - 5 below. The results show that: whether it is the middle-aged group or the elderly group, the ability of the experimental group of the present invention to produce acetic acid, propionic acid, and butyric acid in vitro fermentation is significantly improved compared with the comparative group, especially in terms of propionic acid, and with the increase of fermentation time, the advantage becomes more obvious. The middle-aged group and the elderly group show the same trend, and the effect on the elderly group is more obvious than that on the middle-aged group. Since short-chain fatty acids have a protective effect on the human intestinal environment, it shows that the rice bran compound product prepared by the present invention has better physiological effects than using it alone.

[0199] Table 4. SCFA production in the in vitro fermentation of the middle-aged group for 24 h (n = 10)

[0200] Group Total acid (μM) Acetic acid (μM) Propionic acid (μM) Butyric acid (μM) Control Example 1 24.13±17.55 19.60±15.83 2.68±1.27 1.37±0.94 Control Example 3 25.91±13.71 19.61±12.66 2.89±1.12 1.25±0.85 Example 3 28.82±19.29 19.36±10.79 2.75±1.22 1.47±0.80 Control Example 4 20.32±7.47 13.67±7.01 2.85±0.89 1.06±0.91 Control Example 5 29.84±15.31 22.69±14.32 3.56±1.42 1.79±0.89 Example 7 30.82±14.50 24.59±12.79 3.85±0.62 2.06±0.79 Example 9 29.72±12.61 22.16±10.43 3.92±0.86 2.39±0.98

[0201] Table 5. SCFA production in the in vitro fermentation of the middle-aged group for 48 h (n = 10)

[0202] Group Total acid (μM) Acetic acid (μM) Propionic acid (μM) Butyric acid (μM) Control Example 1 30.46±12.44 25.06±12.07 2.70±0.68 1.85±0.82 Control Example 3 31.19±10.52 24.87±9.18 2.63±0.57 1.98±0.52 Example 3 32.45±16.08 26.87±10.85 2.62±0.61 1.86±0.98 Control Example 4 24.36±11.18 15.23±9.52 2.62±0.64 1.35±0.63 Control Example 5 36.37±10.70 27.39±9.99 3.68±0.12 2.05±0.92 Example 7 41.29±10.16 32.73±13.09 5.23±0.63* 2.25±0.48 Example 9 38.22±13.42 30.06±10.16 4.05±0.28* 2.53±0.32

[0203] Note: *Significant difference compared with the comparative group (*p < 0.05, **p < 0.01)

[0204] Table 6. SCFA production in the in vitro fermentation of the elderly group for 24 h (n = 10)

[0205] Group Total acid (μM) Acetic acid (μM) Propionic acid (μM) Butyric acid (μM) Control Example 1 28.60±12.00 21.61±9.62 3.19±0.61 1.08±0.74 Control Example 3 29.31±11.20 22.04±5.68 3.64±0.59 1.10±0.37 Example 3 27.83±13.31 25.04±8.04 3.56±0.56 1.27±0.27 Control Example 4 27.85±8.26 19.18±5.97 3.21±0.42 1.26±0.68 Control Example 5 34.68±10.87 27.79±8.82 4.02±0.54 1.32±0.87 Example 7 38.84±10.75 31.64±9.84 4.86±0.52* 1.45±0.54 Example 9 36.30±7.78 27.14±5.80 5.37±0.39* 1.68±0.36

[0206] Note: *Significant difference compared with the comparative group (*p < 0.05, **p < 0.01)

[0207] Table 7. SCFA production in the in vitro fermentation of the elderly group for 48 h (n = 10)

[0208] Group Total acid (μM) Acetic acid (μM) Propionic acid (μM) Butyric acid (μM) Control Example 1 35.47±13.26 28.02±7.22 4.42±0.41 1.72±0.36 Control Example 3 36.04±10.67 28.30±5.71 4.22±0.63 1.89±0.50 Example 3 36.17±8.60 29.96±7.55 4.33±0.58 1.78±0.44 Control Example 4 32.79±10.23 23.51±5.28 3.40±0.60 1.73±0.26 Control Example 5 43.46±9.51 30.7±5.84 5.36±0.37 1.92±0.39 Example 7 52.24±6.24 40.45±5.92 6.52±0.26* 2.37±0.41 Example 9 55.50±5.43* 43.37±4.15* 6.80±0.34** 2.64±0.28*

[0209] Note: *Significant difference compared with the comparative group (*p < 0.05, **p < 0.01)

[0210] Detection Example Six: Experimental results of the diversity of in vitro fermentation flora and the detection of beneficial bacteria in the middle-aged and elderly groups:

[0211] The samples after in vitro fermentation in Example 5 were detected. The bacterial genomic DNA of the fermentation broth precipitate was extracted by the TIANamp Stool DNA Kit and quantified by NanoDrop ND-2000 (NanoDrop Technologies, United States). The CFX96TM Real-Time PCR Detection System (Bio-Rad, United States) was used, and the qPCR technique was adopted to detect the quantities of Bifidobacteria spp. and F. prausnitzii spp. in the fermentation broth precipitate. The specific primers Bififi601F: 5’-GGGTGGTAATGCCGGATG and Bififi601R: 5’-TAAGCCATGGACTTTCACACC-3’ were used to measure the total Bifidobacteria species. The specific primers Fae-F: 5’-CACCGCTACACATGGAG-3’ and Fae-R: 5’-AGCAGTAGGGAATCTTCCA-3’ were used to measure the total F. prausnitzii species. The total DNA of the intestinal flora was extracted from the fecal samples, 16S rDNA was amplified, and the flora diversity was analyzed. The results are shown in Table 6. The results show that compared with the control group, the Bifidobacteria and F. prausnitzii in the intestines of the middle-aged and elderly populations in the example group of the present invention have been improved to a certain extent, especially the increase in F. prausnitzii is more obvious. Both Bifidobacteria and F. prausnitzii are beneficial bacteria in the human intestine. Therefore, it can be shown that the rice bran compound sample prepared in the present invention has a role in proliferating beneficial bacteria in the human intestine. At the same time, the experimental results also show that the group adopting the example group has also been improved in terms of flora diversity compared with the control group.

[0212] Table 8. Changes in beneficial bacteria in the middle-aged and elderly group after 48 h of in vitro fermentation (log CFU / g, n = 10)

[0213]

[0214] Note: *Significant difference compared with the control group (*p < 0.05, **p < 0.01)

[0215] Detection Example 7: Feeding and intervention of naturally aging mice

[0216] 80 eight - month - old C57BL / 6J male mice were purchased from Jiangsu Jicui Yakang, with an equal number of males and females. They were randomly divided into 8 groups of 10 mice each according to body weight, including a natural aging model control group (N - AG group, fed with normal feed) and rice bran customized feed intervention groups (Example 7, Example 9, Comparative Example 1, Comparative Example 3, Example 3, Comparative Example 4, Comparative Example 5). After two months of adaptive feeding with normal maintenance feed, the intervention groups were changed to rice bran customized feed for 3 months. The rice bran customized feed was prepared by using the samples obtained in the examples or comparative examples to replace part of the cellulose raw materials in the normal feed, while ensuring that the energy and nutritional components of each group of feed were equivalent. During the experiment, the mice were weighed and their food intake was recorded weekly; the corresponding indicators of the mice were detected during and at the end of the experiment.

[0217] Detection Example VIII: Experimental results of measuring SCFA and beneficial bacteria proliferation in animal experiments of natural aging mice:

[0218] After the intervention experiment of natural aging mice, the SCFA in the feces of the mice was detected by GC method, including acetic acid, propionic acid, butyric acid, etc., and the total acid production was calculated. And q - PCR was used for quantitative analysis of whether Bifidobacterium spp. and F. prausnitzi proliferated. The results are shown in Table 7. The experimental results showed that compared with the old - age mouse control group and the comparative example groups, after feeding the mice with the compound rice bran samples prepared in the examples for 3 months, more short - chain fatty acids were produced in the feces, and the contents of total acid, acetic acid, propionic acid, and butyric acid were all significantly increased, showing statistical differences; at the same time, obvious proliferation effects were also shown on Bifidobacterium and F. prausnitzi in the intestinal flora. These results indicate that the compound rice bran samples prepared according to the present invention have an obvious improvement effect on the intestinal flora of old - age mice, and there is a synergistic effect after the modified rice bran and inulin are compounded in a certain proportion, and the effect is significantly better than the single use of the two.

[0219] Table 9. Production of SCFA and proliferation of beneficial bacteria in natural aging mice after 3 - month intervention (n = 10, fed for 3 months)

[0220]

[0221] Note: *Significant difference compared with the comparative example group (*p < 0.05, **p < 0.01)

[0222] Detection Example IX: Experimental results of the effects on antioxidant and inflammatory factors in animal experiments of natural aging mice:

[0223] After the intervention experiment on naturally aging mice was completed, the antioxidant and inflammatory indexes of mice in each group were detected. The antioxidant indexes included superoxide dismutase (SOD), total antioxidant capacity (T-AOC), and glutathione (GSH) content in the blood. The inflammatory indexes were analyzed by ELISA method to detect the content of calprotectin and lipocalin-2 (LCN-2) in the feces of mice. The results are shown in Table 8. The experimental results showed that after 3 months of feeding, compared with the old-age mouse control group (AG) and the comparative example group, the three antioxidant indexes of the example group prepared by the present invention all increased significantly. Among them, there were statistical differences in SOD and T-AOC, and GSH also increased significantly, indicating that the sample of the example of the present invention can significantly enhance the antioxidant capacity of the mouse body. At the same time, the content of calprotectin and LCN-2 in the feces of the mice in the example group decreased significantly, indicating that the inflammation of the mouse intestine was alleviated.

[0224] Table 10. Detection of antioxidant and inflammatory factors in naturally aging mice after 3 months of intervention (n = 10, fed for 3 months)

[0225]

[0226]

[0227] Note: *Significant difference compared with the comparative example group (*p < 0.05, **p < 0.01)

[0228] Detection Example Ten: Experimental Results of the Influence of the Animal Experiment on Naturally Aging Mice on Immune T Cells

[0229] After the intervention experiment on naturally aging mice, the related indexes of immune T cells in the mice were detected: Antibodies and 100 μL of blood were added to a fresh 15 ml test tube and incubated in the dark at room temperature for 15 minutes. Then, 2 ml of 1x lysis buffer was added to each test tube and incubated in the dark at 37 °C for 3 min. The supernatant was discarded by centrifugation. 2 ml of PBS was added to each test tube and the test tube was centrifuged at 300 g for 5 minutes at room temperature, and the blood was resuspended in 350 μL of PBS. Flow cytometry was used to analyze the proportions of CD4+, CD8+ in the peripheral blood T cells of the mice, the proportion of their TCM subtypes, and the proportion of immune naive cells (T Naive). The results are shown in Table 9. Generally speaking, as the degree of aging deepens, the immunity of the body will decline, manifested in the decrease of the CD4+ / CD8+ ratio, and the immune naive cells will gradually differentiate into memory cells (TCM), resulting in an increase in the number of CD4+ TCM and CD8+ TCM subtypes, while the number of immune naive cells (T Naive) decreases. In this experiment, the sample prepared in the experimental group of the present invention can increase CD4+ / CD8+ in aging mice, while reducing the values of CD4+ TCM and CD8+ TCM. At the same time, it can also increase the proportion of immune naive cells (T Naive). Among them, the results of CD8+ TCM and CD4+ T Naive have statistical differences, indicating that the compound sample prepared by the present invention has a certain anti-aging effect on the immune T cells of aging mice and can enhance the body immunity of the elderly population.

[0230] Table 11. Detection of immune T cells in naturally aging mice after 3 months of intervention (n = 10, fed for 3 months)

[0231]

[0232] Note: *Significant difference compared with the control group (*p < 0.05, **p < 0.01)

[0233] Detection Example XI: Measurement results of exercise ability and cognitive ability indexes of naturally aging mice

[0234] After 3 months of intervention in naturally aging mice, pole climbing tests, Y-maze tests, novel object recognition tests, and open field experiments were conducted on the mice, and the results are shown in Table 10 below. The results showed that there was a certain degree of improvement in the maze data in the example group, indicating an improvement in the memory ability of aging mice; in the pole climbing experiment, the movement time of the mice in the example group was significantly shortened, indicating an enhancement in the motor ability of aging mice; in the open field experiment, the time of the mice in the example group in the central area increased significantly, indicating an enhanced desire of aging mice to explore the new environment and a reduced anxiety level; in the novel object recognition experiment, the novel object recognition index NOI of the example group increased, indicating an improvement in the short-term memory ability of the mice. The above results indicate that the compound rice bran sample prepared according to the present invention has different degrees of improvement effects on the motor ability and cognitive ability of aging mice.

[0235] Table 12. Detection of motor ability and cognitive ability of naturally aging mice after 3 months of intervention (n = 10, fed for 3 months)

[0236]

[0237] Note: *Significant difference compared with the control group (*p < 0.05, **p < 0.01)

[0238] Detection Example Twelve: Experiment on the aging score index of natural mice:

[0239] During the intervention of naturally aging mice, the aging degree of the mice was scored every month. The aging score included multiple indicators such as reactivity, passive avoidance response, fur luster, fur roughness, hair loss degree, skin ulcer, periorbital damage, corneal opacity, corneal ulcer, cataract, kyphosis, etc. The score for each category was from 0 to 4, and the aging degree score of each mouse was the sum of the scores of each category. The higher the score, the higher the aging degree. The test results are shown in Table 11. The results showed that the compound rice bran obtained by using the example had an obvious reducing effect on the aging degree score value of aging mice, especially after the 2nd and 3rd months, and there was a statistical difference in the aging score. It shows that the compound rice bran product prepared by the present invention has an obvious improvement and delaying effect on the aging of mice.

[0240] Table 13. Aging degree score of naturally aging mice (n = 10)

[0241]

[0242]

[0243] Note: *Significant difference compared with the control group (*p < 0.05, **p < 0.01)

[0244] Application Example One

[0245] The composite dietary fibers obtained from each example and comparative example were applied to the preparation of baked bread as follows:

[0246] (1) Bread formula

[0247] 500 g of high-gluten flour or (450 g of high-gluten flour + 50 g of composite dietary fiber), 305 g of water, 75 g of granulated sugar, 30 g of butter, 7.5 g of yeast, 5 g of salt, and 2 g of improver.

[0248] (2) Bread making process

[0249] Pour all the dry ingredients into a dough mixer, add an appropriate amount of water and stir at a low speed for about 4 min, then stir at a high speed for about 3 min. Add the butter and stir at a low speed for about 4 min until the butter is absorbed by the dough, and then stir at a high speed for about 3 min until the dough can be stretched into a film.

[0250] Take out the dough, smooth it, and let it rest for 30 min. Divide the dough into pieces of 450 g each, smooth them, and let them rest for 30 min.

[0251] After the dough has finished resting, shape it into a spindle, roll it out into a long strip with a rolling pin, roll it up from one side, gather the two sides, and shape it into a loaf pan.

[0252] Place the dough in the proofing box and proof for 90 min at a proofing temperature of 35 °C and a humidity of 85%.

[0253] After proofing is completed, place it in the oven for baking at an upper heating temperature of 180 °C and a lower heating temperature of 170 °C for 25 min.

[0254] (3) Baked bread height

[0255] After the bread is taken out of the oven, directly measure the height of the baked bread with a scale in mm, and the results are shown in Table 12.

[0256] (4) Bread volume

[0257] 5 min after the bread is taken out of the oven, measure the volume of the bread with a volume measuring instrument in cm3, and the results are shown in Table 12.

[0258] (5) Evaluation of bread baking quality

[0259] After the bread has cooled at room temperature, it is placed in a self-sealing bag for storage. After 18 h, the external and internal characteristics of the bread are subject to sensory evaluation, mainly including the following: the appearance of the bread, the color of the bread crumb, the texture of the bread crumb, and the texture structure of the bread crumb, etc. (Table 13). When evaluating, first score the appearance, cut the bread, and evaluate it in the order of the texture, color, and texture structure of the bread crumb. Twelve trained personnel in the sensory evaluation group score the baking quality of the bread, and the score is the average value of the twelve people. The results are shown in Table 2.

[0260] (6) Dietary fiber content of bread

[0261] After the evaluation of the bread quality is completed, an appropriate amount of bread is cut into pieces, dried, and ground into powder for standby. Refer to the method of GB5009.88-2014 for the detection of dietary fiber content.

[0262] The application test results show that the compound fiber obtained by adding the examples has a weaker deteriorating effect on the baking quality of the bread compared with the fiber in the comparative example, and has a higher sensory acceptance, indicating that the compound rice bran product prepared by the present invention can be used in baking products to improve the nutritional value of the products.

[0263] Table 14. Bread height and volume

[0264] Height mm <![CDATA[Volume cm 3 > Sensory score Dietary fiber % Example 7 149.16 2043.34 92 6.37 Example 8 147.38 2028.96 91 6.78 Example 9 146.32 2020.11 90 6.45 Example 10 144.21 2005.57 90 6.22 Control Example 1 138.01 1897.11 82 3.68 Control Example 3 135.30 1864.98 80 5.25 Example 3 134.07 1849.33 78 5.80 Control Example 4 143.24 1977.45 87 7.69 Control Example 5 140.98 1945.83 85 5.63 Blank 158.81 2195.34 96 0.48

[0265] Table 15. Baking quality score table of bread

[0266]

[0267]

[0268] Finally, it should be understood that although the various aspects of this specification describe specific embodiments, those skilled in the art will easily understand that these disclosed embodiments are only illustrations of the principles of the subject matter disclosed herein. Therefore, it should be understood that unless expressly stated otherwise, the disclosed subject matter is not limited to the specific combinations, methods, and / or formulations, etc. described herein. In addition, those of ordinary skill in the art will recognize that certain changes, modifications, substitutions, alterations, additions, deletions, and sub-combinations can be made in accordance with the teachings herein without departing from the spirit of this specification. Therefore, the appended claims of this application are intended to be construed as including all such changes, modifications, substitutions, alterations, additions, deletions, and sub-combinations within their true spirit and scope.

Claims

1. A cereal dietary fiber, characterized in that, Based on the total weight of the cereal dietary fiber, the oil content in the cereal dietary fiber is ≤2%, preferably ≤1.7 wt%, more preferably 0.4 - 1.7 wt%, and most preferably 0.9 - 1.7 wt%; the starch content is ≤5%, preferably 2.0 - 5.0 wt%; the total dietary fiber content is ≥50%, preferably 53.0 - 60.0 wt%; the protein content is 20 - 30 wt%, preferably 22 - 25 wt%; the soluble pentosan content is ≥2%, preferably 2.0 - 4.0 wt%.

2. The preparation method of the cereal dietary fiber according to claim 1, characterized in that, The preparation method includes degreasing, de-starching and hemicellulose enzymolysis treatment of cereal bran. Optionally, extrusion treatment / steam explosion treatment is also included before hemicellulose enzymolysis.

3. The preparation method of the cereal dietary fiber according to claim 2, wherein, The preparation method further includes one or more of the following features: (1) The cereal bran is selected from one or more of rice bran, wheat bran, barley bran, oat bran and corn bran; (2) The degreasing includes treating the cereal bran with a solvent capable of dissolving lipids for degreasing treatment; (3) The oil content in the cereal bran obtained by degreasing treatment is ≤2 wt%. Preferably, the solvent is n-hexane and / or petroleum ether; (4) The mass ratio of cereal bran to degreasing solvent is 1:2 - 10, preferably 1:5 - 8; (5) The cereal bran is treated with the degreasing solvent 1 - 5 times, preferably 2 - 5 times; (6) Precipitation is obtained, and the solvent in the precipitation is removed to obtain degreased cereal bran; (7) The de-starching is carried out using amylase; (8) The starch content in the cereal bran obtained by de-starching is ≤5 wt%; (9) The amylase is a high-temperature resistant α-amylase, preferably resistant to above 70°C, more preferably resistant to 70 - 100°C α-amylase. Preferably, the optimal action pH range of this high-temperature resistant α-amylase is 5.8 - 7.8, and the optimal action temperature is 90 - 95°C; (10) The mass ratio of cereal bran to amylase is 100 - 2000:1, preferably 200 - 1500:1, 200 - 1000:1 or 250 - 500:1; (11) The enzymatic hydrolysis reaction system for de-starching also contains water, and the mass ratio of water to cereal bran is 2 - 10:1, more preferably 3 - 8:1 or 4 - 6:1; (12) The enzymatic hydrolysis time for de-starching is 30 minutes to 5 hours, more preferably 30 minutes to 2 hours, and even more preferably 30 minutes to 1 hour; (13) After the enzymatic hydrolysis for de-starching is completed, the precipitated part of the enzymatic hydrolysate is separated and dried; (14) The steam pressure of the steam explosion treatment is 0.9 - 1.5 MPa, and the pressure holding time is 90 - 150 seconds; (15) The cereal bran is added into the steam explosion tank, saturated steam is introduced to make the pressure in the container 0.9 - 1.5 MPa, and this pressure is maintained in the container for 90 - 150 seconds, thereby completing the steam explosion treatment; (16) The temperature of the extrusion is 110 - 130°C; (17) The hemicellulose enzymolysis treatment is carried out using hemicellulase; (18) The mass ratio of hemicellulase to cereal bran is 1:100 - 1500, preferably 1:200 - 1000 or 1:200 - 500; (19) The enzymatic hydrolysis reaction system treated with hemicellulase also contains water, and the mass ratio of water to cereal bran is 2-10:1, more preferably 3-8:1 or 4-6:1; (20) The enzymatic hydrolysis with hemicellulase is carried out at a temperature of 40-60 °C; (21) The enzymatic hydrolysis time with hemicellulase is 4-6 hours, and the pH is 4-6; (22) After the enzymatic hydrolysis with hemicellulase is completed, the pH is adjusted to neutral, the enzyme is inactivated, the enzymatic hydrolysate is concentrated and dried to obtain the cereal dietary fiber; (22) The cereal dietary fiber is rice bran dietary fiber.

4. Use of a cereal dietary fiber, characterized in that, It includes one or more of the following applications, (1) The application of cereal dietary fiber in products for improving bone metabolism, increasing osteoblasts, and improving the degree of trabecular bone rarefaction; (2) The application of cereal dietary fiber in products for increasing the level of bone-specific alkaline phosphatase in serum.

5. A composite dietary fiber, characterized in that, It includes the cereal dietary fiber described in any one of claims 1-3 and inulin.

6. The composite dietary fiber according to claim 5, wherein The composite dietary fiber meets one or more of the following limitations: (1) The inulin is a natural polysaccharide derived from chicory or Jerusalem artichoke, and the average chain length is 8-13 monomers; (2) The ratio of cereal dietary fiber to inulin in the composite dietary fiber is 5-1:1, preferably 3-2:1; (3) Based on the total weight of the composite dietary fiber, the content of total dietary fiber (TDF) in the composite dietary fiber is ≥70%, preferably 70-80%, and the proportion of soluble dietary fiber (SDF) is 20-50%.

7. A preparation method of a composite dietary fiber, characterized in that, The cereal dietary fiber described in any one of claims 1-3 is compounded with inulin to obtain the composite dietary fiber. Preferably, the compounding ratio of the cereal dietary fiber to inulin is 5-1:1, preferably 3-2:

1.

8. The application of the composite dietary fiber described in any one of claims 5-7 in products beneficial to human health. Preferably, the benefits to human health include one or more of anti-aging, antioxidant, regulating intestinal flora, reducing chronic inflammation, improving exercise ability and cognitive ability, and enhancing immunity.

9. The application according to claim 8, wherein It also meets one or more of the following limitations: (1) The application of the composite dietary fiber in products for increasing the content of short-chain fatty acids in the intestine. More preferably, it is to increase the content of total acid, acetic acid, propionic acid, and butyric acid; (2) The application of the composite dietary fiber in products for improving the abundances of Bifidobacterium and Faecalibacterium prausnitzii in the intestine, and in products for improving the diversity of intestinal flora; (3) The application of the composite dietary fiber in products for increasing the levels of SOD, T-AOC, and GSH in the blood, and in products for reducing the content of inflammatory markers produced in the intestine, such as calprotectin and LCN-2; (4) The use of the composite dietary fiber in a product for improving the immunity of aging mammals is preferably to increase the number of immune primitive cells (T Naive ) levels, reducing T cell CD4 + 、CD8 + T CM The proportion of subtypes can enhance the immunity of the elderly population.

10. A food product, characterized in that, It contains the cereal dietary fiber described in any one of claims 1-3 and / or the composite dietary fiber described in any one of claims 5-7. Preferably, the food is: flour products, meat products, rice products, baked foods, snack foods, nutritional foods, meal replacement products, fillings or solid beverages.

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