Method for preparing small-particle-size oat globulin and prepared small-particle-size oat globulin
By treating oat globulin with high molecular weight dextran, small-particle-size oat globulin is prepared, which solves the problem of easy aggregation of oat protein during extraction, processing or storage, and maintains its solubility and functional properties, making it suitable for food processing.
Patent Information
- Application Number
- CN202511795013.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-02
- Publication Date
- 2026-02-27
AI Technical Summary
Existing technologies are prone to aggregation during oat protein extraction, processing, or storage, leading to reduced solubility and loss of functional properties. Furthermore, existing methods for controlling particle size are energy-intensive and may cause protein denaturation or bitterness, which contradicts the trend towards clean labeling.
By mixing high molecular weight dextran with oat globulin, the probability of non-specific aggregation caused by collisions is reduced by occupying solvent space and restricting the degree of freedom of protein molecule diffusion. Furthermore, the protein conformation is stabilized through the entropy effect, thus preparing small-particle-size oat globulin.
It effectively prevents oat globulin from aggregating and oxidizing during extraction, processing, or storage, maintaining its solubility and functional properties to meet the needs of the food processing industry.
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Figure CN121569877A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application belongs to the technical field of food industry, and particularly relates to a method for preparing small-particle-size oat globulin and small-particle-size oat globulin prepared by the method. BACKGROUND
[0002] Oat protein is an important plant protein with high nutritional value and emulsifying properties, and is widely used in the food industry. However, oat protein is prone to aggregation during extraction, processing or storage, resulting in reduced solubility and loss of functional properties, especially under high temperature or high concentration conditions.
[0003] Particle size is a key factor determining the success or failure of oat protein application, which directly determines the solubility, smoothness, physical stability and functional properties of the protein solution. A small particle size of nanoscale is more ideal, which can avoid sedimentation, eliminate grittiness and prolong the shelf life. However, the existing particle size control technologies all have significant defects: high energy consumption and possible protein denaturation or re-aggregation in high-strength mechanical homogenization method, which is only a temporary solution; enzyme treatment is effective but produces unacceptable bitterness and changes the functional properties of the protein; pH adjustment method requires the use of large amounts of acid and alkali and raises safety concerns; and the addition of stabilizers violates the current market trend of clean label. Therefore, there is an urgent need to develop a new technology that is more efficient, economical and can balance flavor and clean label to solve this problem. SUMMARY
[0004] The present application provides a method for preparing small-particle-size oat globulin, comprising the following steps: Dissolve the oat globulin in a NaCl solution, then add dextran, mix well, heat in a water bath, then dialyze to remove salt ions, and centrifuge to remove dextran; freeze-dry the precipitate after centrifugation to obtain small-particle-size oat globulin.
[0005] In the above method, the mass ratio of oat globulin to dextran is 1:(2-10); preferably 1:6.
[0006] In the above method, the mass concentration of the NaCl solution is 2-10%; preferably 6%.
[0007] In the above method, the dextran is high-molecular-weight dextran; the molecular weight is ≥2000; in a specific embodiment, the molecular weight of the dextran can be preferably ≥200000.
[0008] In the above method, the water bath heating conditions are: heating at 30-40℃ for 2-10 h; preferably heating at 37℃ for 6 h.
[0009] In the above method, the dialysis conditions are: dialysis at 2~6℃ for 24~72 h; preferably: dialysis at 4℃ for 48 h.
[0010] In the above method, the centrifugation conditions are: centrifugation at 5000~10000 rpm for 5~15 min at 2~6℃; preferably: centrifugation at 8000 rpm for 10 min at 4℃.
[0011] This invention provides small-particle-size oat globulin prepared by the above method.
[0012] This invention provides the application of dextran in the preparation of small-particle-size oat globulin.
[0013] This invention provides the application of dextran in improving the stability of oat globulin; the application is to prevent oat globulin from aggregating and oxidizing during extraction, processing or storage, thereby improving its stability.
[0014] The beneficial effects of this invention are as follows: This invention utilizes high molecular weight dextran to treat oat globulin. By occupying solvent space and restricting the diffusion freedom of protein molecules, the dextran chains reduce the probability of non-specific aggregation caused by collisions. At the same time, through the entropy effect, it stabilizes the native conformation of the protein, avoiding irreversible aggregation of oat globulin due to the exposure of the hydrophobic core. This promotes the production of small-particle-size oat globulin, making it less prone to aggregation and oxidation during extraction, processing, or storage, thus better maintaining its solubility and functional properties. It has important application prospects in the food processing field. Attached Figure Description
[0015] Figure 1 This is a model diagram of NaCl-12S oat globulin-dextran; where the molecular ratio of 12S oat globulin to dextran is 1:0.
[0016] Figure 2 This is a model diagram of NaCl-12S oat globulin-dextran; where the molecular ratio of 12S oat globulin to dextran is 1:25.
[0017] Figure 3 This is a model diagram of NaCl-12S oat globulin-dextran; where the molecular ratio of 12S oat globulin to dextran is 1:50.
[0018] Figure 4 This is a model diagram of NaCl-12S oat globulin-dextran; in which the molecular ratio of 12S oat globulin to dextran is 1:75.
[0019] Figure 5This is a model diagram of NaCl-12S oat globulin-dextran; where the molecular ratio of 12S oat globulin to dextran is 1:100.
[0020] Figure 6 The mean square shift plot of 12S oat globulin is shown; among them, test examples 1 to 5 represent the molecular ratios of 12S oat globulin to dextran as 1:0, 1:25, 1:50, 1:75, and 1:100, respectively. Detailed Implementation
[0021] The proteins used in the following embodiments of the present invention are all oat globulins. Oat globulins are a component of oat protein, accounting for 70% to 80% of oat protein.
[0022] Glucan is a naturally occurring polysaccharide macromolecule widely found in plant cell walls (such as oat β-glucan). Glucan chains reduce the probability of non-specific aggregation caused by collisions by occupying solvent space and restricting the diffusion freedom of protein molecules. Simultaneously, they stabilize the native conformation of proteins through entropy effects, preventing irreversible aggregation caused by the exposure of hydrophobic cores. In this invention, glucan does not participate in the reaction; it merely influences the conformation of oat globulins through steric effects, promoting the formation of smaller oat globulin particles.
[0023] In this invention, the oat globulin used is prepared by the following method: Oat seeds were ground into powder using a grinder. The oat flour was defatted in hexane at a mass ratio of 1:10 for 6 hours (magnetically stirred for 3 hours, then allowed to stand for 3 hours). The precipitate was collected and dried overnight in a fume hood. The defatted oat flour was dispersed in distilled water at a mass ratio of 1:10, magnetically stirred until homogeneous, and the solution was adjusted to pH 10 to extract oat protein isolate. The solution was centrifuged at 5000 rpm for 15 minutes to obtain an oat protein solution. The oat protein solution was adjusted to pH 7 and centrifuged at 5000 rpm for 15 minutes to remove water-soluble albumin. The obtained protein precipitate was dispersed in 6% NaCl solution and stirred for 2 hours. The supernatant was collected after centrifugation at 5000 rpm for 15 minutes, the pH was adjusted to 4.5, and the solution was centrifuged at 5000 rpm for 15 minutes. The pH of the resulting oat globulin precipitate was readjusted to 7, and dialyzed using a 3.5 kDa membrane for 24 hours. The precipitate was freeze-dried to obtain oat globulin powder.
[0024] Other materials used in this invention, unless otherwise stated, are commercially available. Other terms used in this invention, unless otherwise specified, generally have the meanings commonly understood by those skilled in the art. The invention is further described in detail below with reference to specific embodiments and data. The following embodiments are merely illustrative and not intended to limit the scope of the invention in any way.
[0025] Example 1 The steps for preparing small-particle-size oat globulin are as follows: 1.00 g of oat globulin was dissolved in 100 mL of NaCl solution (6% by mass), then 2.00 g of dextran (molecular weight: 200,000) was added, mixed well, and heated in a 37 ℃ water bath for 6 h. The mixture was then dialyzed at 4 ℃ for 48 h to remove salt ions, and then centrifuged at 8000 rpm for 10 min at 4 ℃ to remove the dextran. The precipitate was freeze-dried for 48 h to obtain small-particle oat globulin.
[0026] Example 2 The steps for preparing small-particle-size oat globulin are as follows: 1.00 g of oat globulin was dissolved in 100 mL of NaCl solution (6% by mass), then 4.00 g of dextran (molecular weight: 200,000) was added, mixed well, and heated in a 37 ℃ water bath for 6 h. The mixture was then dialyzed at 4 ℃ for 48 h to remove salt ions, and then centrifuged at 8000 rpm for 10 min at 4 ℃ to remove dextran. The precipitate was freeze-dried for 48 h to obtain small-particle oat globulin.
[0027] Example 3 The steps for preparing small-particle-size oat globulin are as follows: 1.00 g of oat globulin was dissolved in 100 mL of NaCl solution (6% by mass), then 6.00 g of dextran (molecular weight: 200,000) was added, mixed well, and heated in a 37 ℃ water bath for 6 h. The mixture was then dialyzed at 4 ℃ for 48 h to remove salt ions, followed by centrifugation at 8000 rpm for 10 min at 4 ℃ to remove dextran. The precipitate was freeze-dried for 48 h to obtain small-particle oat globulin.
[0028] Example 4 The steps for preparing small-particle-size oat globulin are as follows: 1.00 g of oat globulin was dissolved in 100 mL of NaCl solution (6% by mass), then 8.00 g of dextran (molecular weight: 200,000) was added, mixed well, and heated in a 37 ℃ water bath for 6 h. The mixture was then dialyzed at 4 ℃ for 48 h to remove salt ions, and then centrifuged at 8000 rpm for 10 min at 4 ℃ to remove dextran. The precipitate was freeze-dried for 48 h to obtain small-particle oat globulin.
[0029] Example 5 The steps for preparing small-particle-size oat globulin are as follows: 1.00 g of oat globulin was dissolved in 100 mL of NaCl solution (6% by mass), then 10.00 g of dextran (molecular weight: 200,000) was added, mixed well, and heated in a 37 ℃ water bath for 6 h. The mixture was then dialyzed at 4 ℃ for 48 h to remove salt ions, and then centrifuged at 8000 rpm for 10 min at 4 ℃ to remove dextran. The precipitate was freeze-dried for 48 h to obtain small-particle oat globulin.
[0030] Comparative Example 1 The steps for preparing small-particle-size oat globulin are as follows: 1.00 g of oat globulin was dissolved in 100 mL of NaCl solution (6% by mass), heated in a 37 ℃ water bath for 6 h, dialyzed at 4 ℃ for 48 h to remove salt ions, and then centrifuged at 8000 rpm for 10 min at 4 ℃. The precipitate after centrifugation was freeze-dried for 48 h to obtain small-particle-size oat globulin.
[0031] Comparative Example 2 The steps for preparing small-particle-size oat globulin are as follows: 1.00 g of oat globulin was dissolved in 100 mL of NaCl solution (6% by mass), then 6.00 g of chitosan (molecular weight: 200,000) was added, mixed well, and heated in a 37 ℃ water bath for 6 h. The mixture was then dialyzed at 4 ℃ for 48 h to remove salt ions, and then centrifuged at 8000 rpm for 10 min at 4 ℃. The precipitate was freeze-dried for 48 h to obtain small-particle-size oat globulin.
[0032] Comparative Example 3 The steps for preparing small-particle-size oat globulin are as follows: 1.00 g of oat globulin was dissolved in 100 mL of NaCl solution (6% by mass), then 6.00 g of maltose was added, mixed well, and heated in a 37 ℃ water bath for 6 h. The mixture was then dialyzed at 4 ℃ for 48 h to remove salt ions, and then centrifuged at 8000 rpm for 10 min at 4 ℃. The precipitate was freeze-dried for 48 h to obtain small-particle oat globulin.
[0033] I. Particle Size Measurement The oat globulin powder obtained in the above-described embodiments was dissolved in 6% NaCl solution to achieve a final protein concentration of 1 mg / mL. Subsequently, the average particle size of the oat globulin was measured using a nanoparticle size analyzer at 25 °C.
[0034] The measurement results are shown in Table 1: Table 1. Particle size of oat globulin As shown in Table 1, treatment of oat globulin with high molecular weight dextran significantly reduces its particle size. This is because high molecular weight dextran alters the solution environment of oat globulin. The dextran chains reduce the probability of non-specific aggregation caused by collisions by occupying solvent space and restricting the diffusion freedom of protein molecules. Simultaneously, the entropy effect stabilizes the native conformation of the protein, preventing irreversible aggregation caused by the exposure of the hydrophobic core.
[0035] Chitosan carries a positive charge, while oat globulin carries a negative charge; the two are strongly attracted to each other, resulting in the formation of large aggregates. Furthermore, low molecular weight maltose may also cross-link with proteins, forming covalent aggregates, thus increasing the particle size of oat globulin.
[0036] Furthermore, as shown in Table 1, the particle size of oat globulin initially decreased and then increased with increasing glucan content. This indicates that the amount of glucan also significantly affects the particle size of oat globulin. When the mass ratio of oat globulin (raw material) to glucan is between 1:(4~6), the particle size of oat globulin can be significantly reduced.
[0037] II. Molecular Simulation Test The structure of 12S oat globulin was obtained from the protein library (Uniport); then, the molecular structures of water, NaCl, 12S oat globulin, and dextran were constructed in Materials Studio software; the above molecular structures were used to construct 12S oat globulin models with different crowding levels in the Amorphous Cell module; the molecular ratios of 12S oat globulin to dextran were 1:0, 1:25, 1:50, 1:75, and 1:100, respectively, corresponding to test examples 1 to 5.
[0038] In the molecular simulation, the molecular weight of the 12S oat globulin sequence used was 58.545 kDa, and the molecular weight of the dextran was 2434.13.
[0039] The geometric configuration of the constructed NaCl-12S oat globulin-dextran model was optimized and energy minimization was performed. The optimized structure was then subjected to annealing for structural relaxation. Finally, the relaxed structure was analyzed by molecular dynamics calculations in the Foricite module.
[0040] Analysis results as follows Figures 1-6 And as shown in Table 2: Table 2 Hydrogen bond parameters of 12S oat globulin As shown in Table 2, with the increase of the amount of dextran, the number of hydrogen bonds initially decreased and then increased, while the average bond length first lengthened and then shortened. At a given amount of dextran, the decrease in the number of hydrogen bonds, the lengthening of bond length, and the weakening of bond energy indicate that the intermolecular attraction between 12S oat globulin molecules is weakened, thereby inhibiting aggregation and enhancing diffusion.
[0041] from Figure 6 It can be seen that the mean square displacement of 12S oat globulin first increases and then decreases with the increase of the amount of dextran. At a certain amount of dextran, 12S oat globulin has a compact conformation and enhanced mobility.
[0042] The above results demonstrate from the perspective of molecular simulation tests that high molecular weight dextran can inhibit the aggregation of oat globulin and enhance its diffusion ability, thus playing an important role in maintaining the small particle size of oat globulin.
[0043] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any other way. Any person skilled in the art may make changes or modifications to the above-disclosed technical content to create equivalent embodiments. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the protection scope of the present invention.
Claims
1. A method for preparing small-particle-size oat globulin, characterized in that, The steps are as follows: Oat globulin was dissolved in NaCl solution, then dextran was added, mixed well, heated in a water bath, and then dialyzed to remove salt ions and centrifuged to remove dextran. The precipitate after centrifugation was freeze-dried to obtain small-particle-size oat globulin.
2. The method for preparing small-particle-size oat globulin according to claim 1, characterized in that, The mass ratio of oat globulin to dextran is 1:(2~10).
3. The method for preparing small-particle-size oat globulin according to claim 2, characterized in that, The mass ratio of oat globulin to dextran is 1:
6.
4. The method for preparing small-particle-size oat globulin according to claim 1, characterized in that, The NaCl solution has a mass concentration of 2-10%.
5. The method for preparing small-particle-size oat globulin according to claim 1, characterized in that, The molecular weight of the dextran is ≥2000.
6. The method for preparing small-particle-size oat globulin according to claim 5, characterized in that, The molecular weight of the dextran is ≥200,000.
7. The method for preparing small-particle-size oat globulin according to claim 1, characterized in that, The water bath heating conditions are: heating at 30~40 ℃ for 2~10 h.
8. The method for preparing small-particle-size oat globulin according to claim 1, characterized in that, The dialysis conditions are: dialysis at 2~6 ℃ for 24~72 h.
9. The method for preparing small-particle-size oat globulin according to claim 1, characterized in that, The centrifugation conditions are as follows: centrifugation at 5000-10000 rpm for 5-15 min at 2-6 ℃.
10. Small-particle-size oat globulin prepared by the method according to any one of claims 1 to 9.