CN114874301A - Preparation method and application of β-conglycinin composed of different subunits - Google Patents

Abstract

The invention discloses a preparation method of β-conglycinin composed of different subunits. The process includes: extracting defatted soybean flour at 45° C. and pH of 8.5, and using CaCl ₂ precipitation agent to prepare β-conglycinin. The application of β-conglycinin in the preparation of hypolipidemic drugs by precipitation of protein, and the preparation of different subunits to form β-conglycinin.

Description

Preparation method and application of β-conglycinin composed of different subunits

technical field

The invention relates to the technical field of preparation of β-conglycinin composed of different subunits, and more particularly to a preparation method and application of β-conglycinin composed of different subunits.

Background technique

The improvement of living standards makes consumers' demand for animal protein increase year by year, but due to the disadvantages of high fat, high calorie and high cholesterol, it will lead to an increase in hyperlipidemia and complications. At present, statins are mainly used as lipid-lowering drugs, but there will be a certain degree of side effects during the treatment. Therefore, in the age of green ideas and health, people are encouraged to look for new sources of plant protein. Soybeans are the most nutritious beans, with protein, fat and carbohydrate content of about 40%, 18% and 25%, respectively, and are known as "the king of beans". Soy protein is rich in amino acids (including 8 amino acids required by the human body), and its value is close to the essential amino acid requirements established by the World Health Organization and the United Nations Food and Agriculture Organization.

The importance of diet alone or in combination with medication for hyperlipidemia and hypercholesterolemia has been recognized, and dietary therapy has also been developed in this direction. Daily intake of 2.75% β-conglycinin reduces plasma TC and TG in high cholesterol diet-fed rats. Ferreira et al found that β-conglycinin had the same effect as fenofibrate and rosuvastatin in controlling plasma TG, HDL-C and TC, indicating that the binding of this protein to rosuvastatin altered the drug in vivo role in homeostasis. Liu et al compared the magnitude of the activity of β-conglycinin and SPI plasma TC reduction in hypercholesterolemic mice. β-Conglycinin was found to lower plasma cholesterol by inhibiting cholesterol absorption and increasing bile acid synthesis. In conclusion, the development of soy protein products has beneficial effects on a variety of diseases, especially for those patients for whom pharmacological approaches are ineffective or have adverse side effects.

Lovati et al. for the first time confirmed the role of β-conglycinin in health care in vivo. In this study, rats ingested β-conglycinin had a 35% lower plasma cholesterol (TC) concentration than the control group, a result that was as effective as rosuvastatin; it was further found that β-conglycinin Globulin significantly reduced triglyceride (TG) levels in rats; it was subsequently found that the α+α' subunit had a higher LDL receptor with higher up-regulation activity than the β subunit. In addition, Manzoni et al. assessed the ability of β-conglycinin-purified α' subunits to modulate LDL receptor activity by tracking the uptake and degradation of LDL in labeled human hepatoma HepG2 cells, and found that the α' subunit's effect on LDL receptors The upregulation of LDL receptors was significantly greater than that of control cells. Meanwhile, the subcellular localization of fluorescently labeled β-conglycinin in HepG2 cells further confirmed previous indirect evidence that the α′ subunit plays a key role in cellular cholesterol homeostasis, revealing that β-conglycinin Potential relationship to thioredoxin-1 (Trx1) and serum cyclophilin B (CypB). Fukui's study found that oral administration of α' subunits could significantly reduce plasma TC and TG levels in rats under the exclusion of isoflavones, confirming the role of β-conglycinin and α' subunits in cellular systems and The role of glycinin in dyslipidemia regulation. However, there are few reports on the control of protein subunits to study the hypolipidemic function of proteins.

Since the 1950s, people have been continuously researching the separation methods of β-conglycinin and glycinin, mainly including salting out method and alkali-soluble acid precipitation method. Alkali-soluble acid precipitation method is regarded as a classic separation method because of its better yield and purity. The β-conglycinin and glycinin fractions separated by Saio method are not completely separated, and the purity is low. However, the Thanh method will convert the β-conglycinin analog into free state, resulting in cross-contamination, resulting in a lower content of the resulting component. The Nagano method has the advantages of good separation effect and high purity, but the disadvantage is that the extraction rate is low.

Therefore, how to provide β-conglycinin composed of different subunits with high extraction rate is an urgent problem to be solved by those skilled in the art.

SUMMARY OF THE INVENTION

In view of this, the present invention provides a preparation method of β-conglycinin with different subunit compositions.

In order to achieve the above object, the present invention adopts the following technical solutions:

A preparation method of β-conglycinin composed of different subunits, comprising the steps of:

Defatted soybean meal was extracted at 45°C, pH 8.5, and β-conglycinin was precipitated with a _CaCl2 precipitant.

As a preferred technical solution of the above technical solution, the process of extracting defatted soybean meal at 45°C and pH of 8.5 includes:

1) Mix defatted soybean powder with water, adjust the pH value to 8.5, and stir at 45°C to obtain an extract;

2) Centrifuge the extract, add solid NaSHO ₃ to 0.98 g/L to the supernatant, adjust the pH to 6.4, and leave it overnight;

3) The supernatant was adjusted to pH 5.5, stirred for 0.5 h, centrifuged, the collected supernatant was diluted 2 times with pure water, adjusted to pH 5.0, centrifuged for 20 min, and the supernatant was taken.

As the preferred technical solution of the above technical solution, the process of precipitating β-conglycinin with CaCl ₂ precipitant is as follows: CaCl ₂ is dispersed in water at a ratio of 1:5 (m/v), dialyzed at 4°C for 48h, The dialyzed protein solution was freeze-dried to obtain β-conglycinin.

As a preferred technical solution of the above technical solution, the ratio of defatted soybean meal to water is 1:5 (m/v).

As a preferred technical solution of the above technical solution, the stirring is stirring for 2h.

As a preferred technical solution of the above technical solution, the centrifugation is 9000g, 4°C, and centrifugation for 30 minutes.

The application of the different subunit composition β-conglycinin prepared by the above preparation method in the preparation of hypolipidemic drugs.

It can be known from the above technical solutions that, compared with the prior art, the present invention provides a preparation method of β-conglycinin with different subunit compositions. By controlling the temperature and pH value during extraction, and adding a precipitating agent, Improved extraction efficiency.

Description of drawings

In order to explain the embodiments of the present invention or the technical solutions in the prior art more clearly, the following briefly introduces the accompanying drawings that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only It is an embodiment of the present invention. For those of ordinary skill in the art, other drawings can also be obtained according to the provided drawings without creative work.

Figure 1 shows the effects of different subunit constituent proteins on the oil red O staining of 3T3-L1 cells.

Figure 2 shows the effect of different subunit constituent protein treatments on lipid accumulation in 3T3-L1 preadipocytes, compared with the model group, ***p<0.001;

Figure 3 shows the effects of different subunit composition protein treatments on glucose uptake (A) and glycerol release (B) in 3T3-L1 preadipocytes; compared with the model group, **p<0.01, ***p< 0.001;

Figure 4 shows the effect of subunit composition on the water and oil absorption of SPI.

Detailed ways

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

Example 1 Preparation of β-conglycinin

Select special soybeans composed of different subunits, and the particles are plump, uniform in size, and consistent in color. The wild-type soybeans and soybeans composed of different subunits are peeled and then crushed and ground through an 80-mesh sieve; at room temperature, weigh a certain amount of soybeans The powder was mixed with n-hexane at a ratio of 1:5 (m/v) to prepare defatted soybean powder. The above defatted soybean powder was mixed with water at a ratio of 1:15 (m/v), and the pH was adjusted to 8.5 with 2M NaOH. After fully stirring at 45°C for 2h, centrifuge at 9000g, 4°C for 30min, take the supernatant, add solid NaSHO ₃ to 0.98g/L, adjust the pH to 6.4, overnight; adjust the pH of the supernatant to 5.5, stir for 0.5h, 9000g, 4°C, centrifugation for 30min, the collected supernatant was diluted 2 times with pure water, adjusted to pH 5.0, 6500g, 4°C, centrifugation for 20min, the precipitate was dispersed in water at a ratio of 1:5 (m/v), Dialyzed at 4°C for 48 h (water was changed at least 4 times during the period), and the fully dialyzed protein solution was freeze-dried and stored for later use.

Example 2 Influence of temperature, pH value and addition of precipitant on extraction rate

Select special soybeans composed of different subunits, and the particles are plump, uniform in size, and consistent in color. The wild-type soybeans and soybeans composed of different subunits are peeled and then crushed and ground through an 80-mesh sieve; at room temperature, weigh a certain amount of soybeans The powder is mixed with n-hexane in a ratio of 1:5 (m/v) to prepare defatted soybean powder, the above defatted soybean powder is mixed with water in a ratio of 1:15 (m/v), the pH is adjusted to 8.5 with 2M NaOH, and the temperature is adjusted to 8.5. Take 25°C, 35°C, 45°C, 55°C, and 65°C for single factor experiment, centrifuge at 9000g, 4°C for 30min, take the supernatant and add solid NaSHO ₃ to 0.98g/L, adjust pH to 6.4, overnight ; Adjust the pH of the supernatant to 5.5, stir for 0.5h, centrifuge at 9000g, 4°C for 30min, dilute the collected supernatant by 2 times with pure water, adjust the pH to 5.0, centrifuge at 6500g, 4°C for 20min, and remove the precipitation with pure water. The ratio of 1:5 (m/v) was dispersed in water, dialyzed at 4°C for 48h (the water was changed at least 4 times during the period), and the completely dialyzed protein solution was freeze-dried and stored for later use.

The effect of pH value on the extraction effect: Mix the above defatted soybean powder with water at a ratio of 1:15 (m/v), adjust the pH to 7.5, 8, 8.5, 9, 9.5 with 2M NaOH, and after fully stirring for 2 hours, extract The temperature was 45°C for single factor experiment, 9000g, 4°C, centrifugation for 30min, the supernatant was added to solid NaSHO ₃ to 0.98g/L, the pH was adjusted to 6.4, overnight; the supernatant was adjusted to pH 5.5, stirred for 0.5 h, 9000g, 4°C, centrifugation for 30min, the collected supernatant was diluted 2 times with pure water, adjusted to pH 5.0, 6500g, 4°C, centrifugation for 20min, and the precipitate was dispersed in the ratio of 1:5 (m/v). In water, dialyze at 4°C for 48h (the water was changed at least 4 times during the period), and freeze-dry the completely dialyzed protein solution and store it for later use.

The effect of the precipitant CaCl ₂ on the extraction effect: The above defatted soybean powder was mixed with water at a ratio of 1:15 (m/v), and the pH was adjusted to 8.5 with 2M NaOH. After fully stirring for 2 hours, the extraction temperature was 45 °C. Single factor experiment, 9000g, 4℃, centrifugation for 30min, take the supernatant and add solid NaSHO ₃ to 0.98g/L, then add 0, 5, 10, 20, 30mmol/L precipitant CaCl ₂ respectively, adjust pH to 6.4 , overnight; the pH of the supernatant was adjusted to 5.5, stirred for 0.5 h, centrifuged at 9000g, 4°C for 30min, the collected supernatant was diluted 2 times with pure water, adjusted to pH 5.0, centrifuged at 6500g, 4°C for 20min, the The precipitate was dispersed in water at a ratio of 1:5 (m/v), dialyzed at 4°C for 48 h (water was changed at least 4 times during the period), and the fully dialyzed protein solution was freeze-dried and stored for later use.

Effects of root extraction temperature, pH value and CaCl ₂ addition on protein extraction rate to determine optimal extraction conditions

The results are shown in Table 1 to Table 5;

Table 1

Extraction temperature/℃ Protein mass fraction % Extraction rate % 25 73.54 33.13 35 86.51 35.08 45 87.51 38.85 55 85.43 37.46 65 69.54 49.20

It can be seen from Table 1 that high temperature is conducive to protein dissolution, but too high will lead to its denaturation. As can be seen from Table 1, the protein content and extraction rate of β-conglycinin increased first and then decreased with the increase of temperature, and the best conditions were at 45 °C.

Table 2 The effect of pH value on β-conglycinin

It can be seen from Table 2 that the pH value of the solution prepared by the method of Thanh and Shibasaki is 8.0, but it is known that the isoelectric point of the main subunit of glycinin is 8.0-8.5. When the pH value is adjusted from 8.0 to 9.0, β-conglycinin The protein content and extraction of the protein first increased and then decreased. At pH 8.5, the protein content and extraction rate obtained by extraction were the highest,

Table 3 The effect of precipitation dose on β-conglycinin

Precipitation dose mmol/L Protein mass fraction % Extraction rate % 0 87.51 38.85 5 81.64 30.26 10 75.58 6.65 20 69.82 4.42 30 67.44 2.26

It can be seen from Table 3 that the increase of the precipitant CaCl ₂ leads to a decrease in the protein content of β-conglycinin, and it is found that the addition of 10 mmol/L CaCl ₂ has a great influence on the extraction rate of β-conglycinin.

Table 4 Box-Behnken Center Combination Test Results

Table 5 Analysis of variance of regression model equation

Tab.5Analysis of variance of the fitted regression model

source of variance sum of square degrees of freedom variance F value p salience regression model 15.20 9 1.69 84.92 <0.0001 ** A 0.02 1 0.02 0.86 0.3845 B 0.56 1 0.56 28.24 0.0011 ** C 3.71 1 3.71 186.65 <0.0001 ** AB 0.12 1 0.12 6.16 0.0421 * AC 0.17 1 0.17 8.66 0.0216 * BC 3.60 1 3.60 0.18 0.6833 A² 2.79 1 2.79 140.34 <0.0001 ** B² 6.28 1 6.28 315.95 <0.0001 ** C² 0.64 1 0.64 32.07 0.0008 ** residual 0.14 7 0.02 lack of fit 0.06 3 0.02 0.97 0.4886 pure error 0.08 4 0.02 total 15.34 16

Use Design-Expert 10.0.1 to analyze the test data and get the quadratic polynomial regression equation:

Y=39.806+0.04625*A+0.265*B-0.68125*C-0.175*AB+0.2075*AC+0.03*BC-0.81425*A2-1.22175*B2-0.38925*C2

where Y is the extraction rate, A is the extraction temperature, B is the pH value, and C is the _CaCl content.

As shown in Table 5, the F value in the analysis of variance is used to test the significance of the influence of each variable on the response value. The larger the F, the higher the significance of the corresponding variable. When the model significance test p<0.05, it indicates that the model is statistically significant. The order of the effect of process conditions on the extraction rate is: C>B>A, that is, CaCl ₂ content>pH value>extraction temperature. The coefficient of determination of the model, R ² =0.9909, indicates that the model has high significance, and the experimental model fits well with the real data.

In order to further accurately determine the optimal extraction method and maximize the extraction rate as the optimization objective, according to the running results of the Design-Expert 10.0.1 software, the optimal process is as follows: the extraction temperature is 46 °C, the pH value is 8.5, and the CaCl ₂ content is 0.9 mmol/L, the maximum extraction rate and mass fraction obtained were 40.088% and 89.85%, respectively.

Under these conditions, the experiment was repeated three times, and the extraction rate of No. 1 was finally measured to be 41.010%, and the mass fraction of β-conglycinin was 90.67%, which had little deviation from the predicted value (p>0.05), which indicated that the model was reliable. of. At this time, the extraction rate and quality fraction of No. 2 were 37.131% and 89.51%, the extraction rate and quality fraction of No. 3 were 38.499% and 90.03%, and the extraction rate and quality fraction of No. 4 were 39.054% and 88.27%, respectively. .

The subunit compositions of the four histones were measured as follows:

Table 6 Composition of four β-conglycinin subunits

protein number α' alpha beta 1 17.40 17.54 13.43 2 15.70 18.96 13.15 3 18.54 15.46 14.34 4 17.76 29.12 13.20

Example 2 Lipid-lowering property test

Culture of 3T3-L1 cells

3T3-L1 preadipocytes were cultured in DMEM containing 10% FBS and antibiotics (100 U/mL penicillin and 0.1 μg/mL streptomycin) at 37°C in a humidified atmosphere of 5% CO 2 20 . After complete confluency of preadipocytes and 2 days of exposure inhibition, change to differentiation medium I, including 0.5mM IBMX, 1 μM DEX, and 10 μg/mL INS and culture for 2 days (0-2 days), preadipocytes were mixed with 10DMEM in 10DMEM. % FBS plus 1 μM INS (2-4 days), cells were then cultured in DMEM medium supplemented with 10% FBS for an additional 4 days (4-8 days). Change the medium every 2d.

Determination of lipid accumulation

Oil red O staining (ORO) was used to determine excess lipid accumulation in mature adipocytes. Fully differentiated 3T3-L1 preadipocytes were washed twice with PBS and fixed with 4% paraformaldehyde for 30 min at 25°C. Fixed cells were successively washed three times with PBS buffer and then washed with 60% isopropanol for 5 min. Cells were stained with ORO solution for 20 min and then washed with distilled water. Finally, the cell morphology was observed and photographed using an inverted fluorescence microscope (DMIL LED Fluo, Germany).

Determination of triglyceride (TG) content

Triglyceride (TG) content in 3T3-L1 preadipocytes after 8 days of differentiation was measured by applying a triglyceride colorimetric assay kit (Asan Pharmaceutical Company, Ltd., Yeongcheon, Kyungbuk, Korea) according to the manufacturer's instructions. In summary, cells were washed twice with cold PBS, differentiated cells were lysed with TritonX-100 (50 mmol/L Tris, 0.15 mol/L NaCl, 10 mmol/L EDTA, 0.1% Tween-20), TG kit and BCA The kits were used to measure TG concentration and total protein concentration, respectively.

Determination of glucose uptake and glycerol release

Fully differentiated 3T3-L1 preadipocytes were cultured with DMEM and incubated with β-conglycinin at a concentration of 100 μg/mL for 1 day. Glucose uptake and glycerol release were determined with a glucose assay kit and a glycerol content assay kit according to the manufacturer's instructions. Before glycerol assay, lipase must be inactivated at 70 °C for 10 min and centrifuged at 5,000 rpm for 5 min at room temperature.

results and analysis

Cell differentiation examined by ORO staining is shown in Figure 1. As lipid accumulation occurred gradually, 3T3-L1 preadipocytes differentiated into mature adipocytes with several oil droplets after 8 days of induction. Compared with the normal group, mature adipocytes were larger, round, and filled with large lipid droplets (model group). After the addition of protein, the oil red O staining area was reduced, the color became lighter, and the mature adipocytes were less round than the model group. Preliminary results showed that No. 3 had more inhibitory effects than No. 1 than the other groups.

Cells treated with the protein showed lower intracellular lipid accumulation compared to the model group. This suggests that treatment inhibits differentiation. Furthermore, TG content in 3T3-L1 preadipocytes treated with different subunit constituent proteins was significantly (p<0.001) decreased by 33.25, 24.8, 44.36 and 33.57%, respectively. It was observed that protein No. 3 showed low TG accumulation, as shown in Figure 2;

The glucose concentration in the medium was also measured to evaluate the glucose uptake of 3T3-L1 preadipocytes, and the glucose consumption of the different subunit composition protein groups was significantly increased (p<0.01). Compared with the model group, the glucose intake increased to 0.86, 0.81, 0.98 and 0.82 times, respectively. In particular, No. 3 significantly increased glucose uptake in 3T3-L1 preadipocytes (p<0.001). The stability of blood glucose in vivo is mainly achieved by insulin regulation of glucose transport in adipose tissue and skeletal muscle tissue. Insulin sensitivity describes the degree of insulin resistance. The hallmarks of impaired insulin sensitivity are decreased insulin-stimulated glucose uptake and decreased insulin inhibition of adipose tissue lipolytic capacity. This suggests that protein 3 can promote glucose uptake and utilization by 3T3-L1 preadipocytes and has specific ameliorating effects on impaired insulin sensitivity and insulin resistance.

The main function of TG is to store and supply energy. Hydrolysis of triglyceride lipase releases glycerol and free fatty acids for use by body tissues. Thus, glycerol release represents lipolysis in adipocytes. We performed an in vitro assay of the amount of glycerol released into the culture medium from differentiated 3T3-L1 preadipocytes. The results showed that, compared with the normal group, in the extracellular medium of differentiated 3T3-L1 preadipocytes, the amount of glycerol released after treatment with different subunit constituent proteins was significantly increased. Compared with No. 1, the glycerol content of No. 3 was significantly lower than other groups (p<0.05), indicating that No. 3 protein could significantly inhibit the release of glycerol in adipocytes. Inhibition of adipocyte differentiation prevents adipose tissue expansion and can be used for obesity management; see Figure 3;

A linear regression equation was established according to the composition content of four protein subunits and triglyceride content (Y), Y=0.161α'-0.855α+0.193β+114.989, we found that α subunit was significantly negatively correlated with triglyceride content. Consistent with the above index results, a better lipid-lowering effect was shown when the α subunit content was the least.

Example 3 Water and oil absorption of different subunit constituent proteins

Determination of water and oil absorption

0.3g of protein sample was added to 3mL of water (oil) and placed in a 5mL centrifuge tube, shaken and mixed, and then allowed to stand for 30min, centrifuged at 10,000g for 30min, and the weight was recorded after adsorbing the unbound water (oil) on the surface. Water absorption capacity, WAC) was calculated by the mass of water adsorbed per gram of protein, and Oil absorption capacity (OAC) was calculated by the mass of oil adsorbed per gram of protein. The formula is as follows:

In the formula: m ₁ is the mass (g) of the protein sample;

m ₂ is the mass (g) of the protein sample added to the tube before centrifugation;

m ₃ is the mass (g) of the sample added to the tube after centrifugation;

m ₄ is the mass (g) of the protein sample;

m ₅ is the mass (g) of the protein sample added to the tube before centrifugation;

m ₆ is the mass (g) of the sample added to the tube after centrifugation.

The physicochemical properties of proteins are mainly the interactions between water and properties, surface properties and proteins. Water absorption is the interaction between protein and water, which is a measure of the adsorption capacity of protein products; oil absorption is the hydrophobic interaction between the non-polar regions of proteins and the non-polar aliphatic chains of lipids, indicating protein adsorption The size of the grease capacity. The comparison of water and oil absorption of SPI with different subunit compositions is shown in Figure 4. The water absorption of No. 3 (52.72%) and No. 4 (56.64%) is lower than that of No. 1 (78.35%), while the water absorption of No. 2 is lower than that of No. 2 (56.64%). No. 1 high, this result shows that the subunit composition has a certain influence on the water absorption capacity of the protein. The oil absorption of No. 3 (15%) was slightly higher than that of No. 1 (14.65%), while other histones had no significant difference (p>0.05). The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments, and the same and similar parts between the various embodiments can be referred to each other.

The above description of the disclosed embodiments enables any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims (7)

1. a kind of preparation method of different subunit composition β-conglycinin, is characterized in that, comprises the following steps: Defatted soybean meal was extracted at 45°C, pH 8.5, and β-conglycinin was precipitated with a _CaCl2 precipitant. 2. the preparation method of a kind of different subunit composition β-conglycinin according to claim 1, is characterized in that, at 45 DEG C, pH is that the process that defatted soybean meal is extracted under 8.5 comprises: 1) Mix defatted soybean powder with water, adjust the pH value to 8.5, and stir at 45°C to obtain an extract; 2) Centrifuge the extract, add solid NaSHO ₃ to 0.98 g/L to the supernatant, adjust the pH to 6.4, and leave it overnight; 3) The supernatant was adjusted to pH 5.5, stirred for 0.5 h, centrifuged, the collected supernatant was diluted 2 times with pure water, adjusted to pH 5.0, centrifuged for 20 min, and the supernatant was taken. 3. the preparation method of a kind of different subunit composition β-conglycinin according to claim ₂ , it is characterized in that, with CaCl precipitating agent is carried out to the process that β-conglycinin is precipitated as: by CaCl ₂ Disperse in water at a ratio of 1:5 (m/v), dialyze for 48h at 4°C, freeze-dry the completely dialyzed protein solution to obtain β-conglycinin. 4 . The method for preparing β-conglycinin composed of different subunits according to claim 2 , wherein the ratio of defatted soybean meal to water is 1:5 (m/v). 5 . 5. the preparation method of a kind of different subunit composition β-conglycinin according to claim 2, is characterized in that, described stirring is stirring 2h. 6 . The method for preparing β-conglycinin with different subunit compositions according to claim 2 , wherein the centrifugation is 9000 g at 4° C. for 30 min. 7 . 7. The application of the different subunit composition β-conglycinin prepared by the preparation method according to claim 1-6 in the preparation of a hypolipidemic drug.

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