Preparation method of ultrahigh pressure microjet homogenization-helicase modified bean dreg insoluble dietary fiber
By treating soybean residue using ultra-high pressure microfluidic homogenization combined with enzymatic methods, and employing stepwise enzymatic hydrolysis with heat-resistant α-amylase, neutral protease, amylase, and snail enzyme, the problem of low modification efficiency of insoluble dietary fiber in soybean residue was solved, achieving high-value utilization and green and safe resource utilization of soybean residue.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- NORTHEAST AGRICULTURAL UNIVERSITY
- Filing Date
- 2026-03-23
- Publication Date
- 2026-05-12
AI Technical Summary
In existing technologies, the modification methods for insoluble dietary fiber in soybean residue are inefficient, making it difficult to achieve high-value utilization, and often require the use of chemical additives, which affects its resource utilization effect.
Soybean residue was treated with a combination of ultra-high pressure microfluidic homogenization and enzymatic methods, including stepwise enzymatic hydrolysis by thermostable α-amylase, neutral protease, amyloglucosidase and snail enzyme, combined with ultra-high pressure microfluidic treatment, to prepare insoluble dietary fiber from soybean residue.
It significantly improves the yield and quality of insoluble dietary fiber from soybean residue, achieving green and efficient resource utilization, and is suitable for functional foods and special medical purpose formula foods.
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Figure CN122004485A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of deep processing technology of soybean residue resource utilization, and particularly relates to an insoluble dietary fiber from soybean residue and its preparation method. Background Technology
[0002] Dietary fiber (DF) refers to a class of carbohydrate polymers with a degree of polymerization (DP) ≥ 3 that cannot be digested and absorbed by the human small intestine, but have positive effects on health. It can be derived from natural plants or obtained through extraction or artificial synthesis. Its main components include cellulose, hemicellulose, pectin, and other polysaccharides, hence its reputation as the "seventh nutrient." Dietary fiber can be divided into two categories based on its solubility: soluble dietary fiber (SDF) and insoluble dietary fiber (IDF). Compared to soluble dietary fiber, insoluble dietary fiber mainly regulates intestinal function through physical mechanisms in the gastrointestinal system. It increases stool volume, promotes intestinal peristalsis, and effectively prevents and relieves constipation; it can also absorb and excrete harmful substances, helping to maintain colon health and reduce the risk of colorectal cancer. Furthermore, it enhances satiety, aids in weight management and blood sugar stability, and is a key dietary component for maintaining digestive health.
[0003] Soybean residue is a typical byproduct of soybean product processing, a solid residue left after making soy milk or tofu, mainly composed of soybean cell walls, incompletely extracted proteins, and carbohydrates. It is rich in dietary fiber, high-quality plant protein, various minerals, and essential bioactive components. Soybean residue is a good source of bioactive substances, playing a significant role in promoting intestinal peristalsis, assisting in regulating blood sugar and lipids, enhancing satiety to control weight, lowering cholesterol, improving immunity, and preventing chronic diseases. Therefore, targeted modification of soybean residue to significantly improve its insoluble dietary fiber content and functional properties is a key path to overcome its resource utilization bottleneck and achieve high-value, intensive processing, which is of great significance for promoting the industrial upgrading and sustainable utilization of soybean-based byproducts.
[0004] Ultra-high pressure microfluidic homogenization (UHPC) drives materials through microchannels under ultra-high pressure, generating physical effects such as shearing, impact, and cavitation. This disrupts the dense structure and intermolecular forces of the materials, triggering various physicochemical changes. UHPC is a green, efficient, and gentle technology that modifies materials while preserving active ingredients and without introducing chemical additives. In recent years, it has been successfully applied to the extraction and structural optimization of high-value-added components from various natural products. Meanwhile, snail enzymes, as a complex enzyme preparation, exhibit unique advantages in improving fiber purity and functional properties due to their ability to effectively hydrolyze cell wall polysaccharides and impurities. Currently, there are few reports on the synergistic effect of UHPC and enzymatic methods in the targeted modification of insoluble dietary fiber in soybean residue. Therefore, the innovation of this synergistic strategy is expected to provide a new path for the high-value development of soybean residue. Summary of the Invention
[0005] In view of this, the purpose of this invention is to provide a method for preparing insoluble dietary fiber from soybean residue, which adopts a preparation process of ultra-high pressure microfluidic homogenization combined with enzymatic method, which has the advantages of high efficiency modification, green safety and high yield, and is suitable for the preparation of raw materials in the fields of functional foods, special medical purpose formula foods and the like.
[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:
[0007] This invention provides a method for preparing insoluble dietary fiber from soybean residue, comprising the following steps: defatting soybean residue to obtain defatted soybean residue powder; thoroughly mixing the defatted soybean residue powder with deionized water and then subjecting it to ultra-high pressure microfluidic homogenization treatment to obtain a soybean residue powder-water suspension; subjecting the soybean residue powder-water suspension to enzymatic hydrolysis by high-temperature resistant α-amylase, neutral protease, and amyloglucosidase in sequence, followed by enzyme inactivation, and then enzymatic hydrolysis by snail enzyme, followed by enzyme inactivation and drying to obtain insoluble dietary fiber from soybean residue; the working pressure of the ultra-high pressure microfluidic homogenization treatment is 200~250MPa, and the treatment time is 20~25min.
[0008] Preferably, the amount of heat-resistant α-amylase added for hydrolysis is 1%~1.5% per 50g of soybean residue mixture, and the enzyme activity of the heat-resistant α-amylase is 2×104U / g; the temperature of the heat-resistant α-amylase hydrolysis is 75~95℃, the time is 1~1.5h, and the pH is 5.0~8.0.
[0009] Preferably, the amount of neutral protease added for hydrolysis is 2%~2.5% neutral protease per 50g of soybean residue mixture, and the enzyme activity of the neutral protease is 5×104U / g; the hydrolysis temperature of the neutral protease is 35~55℃, the time is 2~2.5h, and the pH is 6.5~8.0.
[0010] Preferably, the amount of amylase added for hydrolysis is 1%~1.5% amylase per 50g of soybean residue mixture, and the enzyme activity of the amylase is 2×104U / g; the hydrolysis temperature is 50~65℃, the time is 1~1.5h, and the pH is 4.0~5.5.
[0011] Preferably, the snail enzyme hydrolysis is performed by adding 1% to 3% snail enzyme per 50g of soybean residue mixture, and the snail enzyme activity is 3U / mg; the snail enzyme hydrolysis temperature is 40 to 60℃, the time is 1 to 4 hours, and the pH is 4.0 to 7.5.
[0012] Preferably, the raw material used for degreasing is petroleum ether, the material-to-liquid ratio for degreasing is 1g soybean residue: 10~15mL petroleum ether, and the degreasing time is 5~10h.
[0013] The present invention also provides insoluble dietary fiber from soybean residue prepared by the above preparation method.
[0014] The beneficial effects of this invention are:
[0015] This invention proposes a green modification process for insoluble dietary fiber from soybean residue based on ultra-high pressure microfluidic homogenization combined with enzymatic treatment. This method, without chemical additives, significantly improves the yield and quality of insoluble dietary fiber through the efficient synergy of physical shearing and enzymatic hydrolysis, achieving high-value utilization of soybean residue's insoluble dietary fiber resources. Compared to other dietary fiber modification and processing methods, the ultra-high pressure microfluidic homogenization process of this invention requires no chemical reagents and is pollution-free. Combined with enzymatic methods, it significantly increases the yield of insoluble dietary fiber from soybean residue, providing a green and feasible technical path for the efficient resource utilization of soybean residue.
[0016] The preparation method provided by this invention is simple to operate, green and safe, and achieves a yield of up to 87.20% of insoluble dietary fiber from soybean residue. The ultra-high pressure microfluidic homogenization combined with enzymatic method provided by this invention significantly enhances the physicochemical properties of insoluble dietary fiber from soybean residue. This not only effectively increases the economic added value of soybean residue, a byproduct of soybean processing, but also demonstrates great potential application value in functional foods, bio-based materials, and other fields. Attached Figure Description
[0017] Figure 1 The flowchart is shown in Example 1.
[0018] Figure 2 Results for the yield of insoluble dietary fiber from soybean residue;
[0019] Figure 3 Results for the insoluble dietary fiber content of soybean residue;
[0020] Figure 4The results show the water-holding capacity of insoluble dietary fiber in soybean residue.
[0021] Figure 5 The results show the oil-holding capacity of insoluble dietary fiber in soybean residue.
[0022] Figures 2-5 The lowercase letters in the equation indicate whether the difference between any two groups is significant. If the lowercase letters of any two groups are different, it indicates that there is a significant difference between the two groups. If the lowercase letters of any two groups are the same, it indicates that there is no significant difference between the two groups. Detailed Implementation
[0023] This invention provides a method for preparing insoluble dietary fiber from soybean residue, comprising the following steps: defatting soybean residue to obtain defatted soybean residue powder; thoroughly mixing the defatted soybean residue powder with deionized water and then subjecting it to ultra-high pressure microfluidic homogenization treatment to obtain a soybean residue powder-water suspension; subjecting the soybean residue powder-water suspension to enzymatic hydrolysis by high-temperature resistant α-amylase, neutral protease, and amyloglucosidase in sequence, followed by enzyme inactivation, and then enzymatic hydrolysis by snail enzyme, followed by enzyme inactivation and drying to obtain insoluble dietary fiber from soybean residue; the working pressure of the ultra-high pressure microfluidic homogenization treatment is 200~250MPa, and the treatment time is 20~25min.
[0024] This invention does not specifically limit the source of the soybean residue. In this invention, the solvent used for degreasing is preferably petroleum ether. Degreasing is performed according to national standard GB5009.6-2025, followed by filtration and air drying to obtain degreased soybean residue. After obtaining the degreased soybean residue, it is preferably first subjected to impurity removal and pulverization, and then subjected to ultra-high pressure micro-jet homogenization. This invention does not specifically limit the specific pulverization method, but preferably, the pulverized residue is passed through a 60-mesh sieve.
[0025] In this invention, after obtaining the soybean residue powder-water suspension, a stepwise directional enzymatic hydrolysis method is used to remove impurities such as starch and protein. First, a thermoresistant α-amylase hydrolysis is performed. The preferred amount of thermoresistant α-amylase added is 1%~1.5% per 50g of soybean residue, more preferably 1%~1.3% per 50g of soybean residue. The preferred enzyme activity of the thermoresistant α-amylase is 2×104U / g. The preferred temperature for the thermoresistant α-amylase hydrolysis is 75~95℃, more preferably 80~90℃. The preferred hydrolysis time is 1~1.5h, more preferably 1~1.3h. The preferred pH value for the thermoresistant α-amylase hydrolysis is 5.0~8.0, more preferably 5.5~7.0.
[0026] After the thermostable α-amylase hydrolysis is completed, the enzyme is preferably inactivated first, cooled to room temperature, and then the pH value is adjusted to 6.5-8.0, more preferably 7.0-8.0, before adding neutral protease for further hydrolysis. The preferred amount of neutral protease added is 2%-2.5% per 50g of soybean residue, more preferably 2%-2.3% per 50g of soybean residue. The preferred enzyme activity of the neutral protease is 5×10⁴ U / g. The preferred hydrolysis temperature is 35-55℃, more preferably 40-50℃, and the preferred hydrolysis time is 2-2.5h, more preferably 2-2.3h.
[0027] After neutral protease hydrolysis, the enzyme is preferably inactivated first, cooled to room temperature, and the pH adjusted to 4.0-5.5, more preferably 4.0-5.0, before adding amylase for enzymatic hydrolysis. The preferred amount of amylase added for hydrolysis is 1%-1.5% per 50g of soybean residue, more preferably 1%-1.3% per 50g of soybean residue. The preferred enzyme activity of the amylase is 2×10⁴ U / g. The preferred hydrolysis temperature is 50-65℃, more preferably 50-60℃, and the preferred hydrolysis time is 1-1.5h, more preferably 1-1.3h.
[0028] After the amylase hydrolysis is completed, the enzyme is inactivated, and the pH is adjusted to 4.0-7.5, more preferably 5.0-7.0. Then, the mixture is treated with snail enzyme. The preferred amount of snail enzyme added for the hydrolysis is 1%-3% per 50g of soybean residue mixture, more preferably 1.5%-2.5% per 50g of soybean residue mixture. The preferred enzyme activity of the snail enzyme is 3U / mg. The preferred hydrolysis temperature is 40-60℃, more preferably 45-55℃. The preferred hydrolysis time is 1-4h, more preferably 1-3h.
[0029] In this invention, after the snail enzyme hydrolysis is completed, it is preferable to perform enzyme inactivation treatment, followed by drying treatment to obtain insoluble dietary fiber from soybean residue. The drying temperature is preferably 60~85℃, more preferably 65~75℃, and the drying time is preferably 20~40h, more preferably 25~35h.
[0030] The present invention also provides insoluble dietary fiber from soybean residue prepared by the above preparation method.
[0031] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.
[0032] Unless otherwise specified, the following embodiments are all conventional methods.
[0033] The ultra-high pressure microjet used in the following examples is the HPW-10 type, developed by Shanghai Zhirui Precision Equipment Co., Ltd.; the high-temperature resistant α-amylase was purchased from Shanghai Maclean Biochemical Co., Ltd.; the neutral protease was purchased from Shandong Longkete Enzyme Preparation Co., Ltd.; the amylase was purchased from Shanghai Shifeng Co., Ltd.; and the snail enzyme was purchased from Solarbio Technology Co., Ltd.
[0034] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.
[0035] All the following experiments were performed in triplicate, and the data are expressed as mean ± standard deviation.
[0036] Example 1
[0037] A method for preparing insoluble dietary fiber from soybean residue:
[0038] (1) Degreasing: Add petroleum ether to soybean residue at a ratio of 1g soybean residue to 12 mL petroleum ether, stir and soak at room temperature for 6 hours, filter, and air dry to obtain degreased soybean residue;
[0039] (2) Remove impurities from defatted soybean residue, pulverize it, and pass it through a 60-mesh sieve;
[0040] (3) After thoroughly mixing the defatted soybean residue powder from (2) with deionized water at a ratio of 1:20, the mixture is subjected to ultra-high pressure micro-jet homogenization to obtain a soybean residue powder-water suspension. The ultra-high pressure micro-jet homogenization parameters are: working pressure of 200 MPa and processing time of 20 min.
[0041] (4) Stepwise directional enzymatic hydrolysis to remove starch, protein, etc.: Adjust the pH of the soybean residue powder-water suspension obtained in step (3) to 6.5, and then add thermostable α-amylase for enzymatic hydrolysis. The amount of thermostable α-amylase with an enzyme activity of 2×104 U / g is 1%, and the hydrolysis is carried out at 85℃ for 1 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated, cooled to room temperature, and the pH is adjusted to 7.5. Then, neutral protease is added for enzymatic hydrolysis. The amount of neutral protease with an enzyme activity of 5×104 U / g is 2%, and the hydrolysis is carried out at 45℃ for 2 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated, cooled to room temperature, and the pH is adjusted to 4.5. Then, amyloglucosidase is added for enzymatic hydrolysis. The amount of amyloglucosidase with an enzyme activity of 2×104 U / g is 1%, and the hydrolysis is carried out at 55℃ for 1 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated to obtain soybean residue product.
[0042] (5) Adjust the pH of the soybean residue product obtained in (4) to 6.0, then add snail enzyme. The amount of snail enzyme with an enzyme activity of 3 U / mg is 2%. Then enzymatically hydrolyze at 50℃ for 2 hours, and finally inactivate the enzyme in a boiling water bath for 10 minutes to obtain the product solution. After centrifugation, the precipitate is obtained.
[0043] (6) Sample drying: The product obtained in (5) was dried at a temperature of 65°C for 30 hours to obtain soybean residue insoluble dietary fiber.
[0044] The flowchart of the above preparation method is as follows: Figure 1 As shown.
[0045] Compare with Example 1
[0046] The difference from Example 1 is that steps (3) and (5) are not included, while the rest are the same as in Example 1.
[0047] Compare with Example 2
[0048] The difference from Example 1 is that step (5) is not included; the rest is the same as Example 1.
[0049] Compare with Example 3
[0050] The difference from Example 1 is that step (3) is not included; the rest is the same as Example 1.
[0051] Example 2
[0052] Determination of the yield of insoluble dietary fiber (IDF) in soybean residue
[0053] The IDF yield of Examples 1, 1, 2, and 3 was determined using the AOAC method. The insoluble dietary fiber content of soybean residue is expressed as a percentage of the remaining material after removing protein and ash.
[0054] Soybean residue IDF yield as follows Figure 2 As shown.
[0055] Depend on Figure 2 It can be seen that the yield of soybean residue IDF prepared in Example 1 was increased by 1.35 times, 1.18 times, and 1.12 times compared with that of Control Example 1, Control Example 2, and Control Example 3, respectively.
[0056] Example 3
[0057] Determination of IDF content in soybean residue
[0058] The IDF content of Example 1, Control Example 1, Control Example 2 and Control Example 3 was determined according to GB5009.88-2023.
[0059] Soybean pulp IDF content such as Figure 3 As shown.
[0060] Depend on Figure 3It can be seen that the content of IDF in soybean residue prepared in Example 1 was increased by 1.26 times, 1.18 times, and 1.09 times compared with that in Control Example 1, Control Example 2, and Control Example 3, respectively.
[0061] Example 4
[0062] Water-holding capacity (WHC) of soybean residue IDF
[0063] Accurately weigh 0.5 g of the insoluble dietary fiber prepared in Control Examples 1, 2, 3, and 1, denoted as M1, dissolve it in 15 mL of deionized water, stir uniformly at 25°C for 2 h, then let it stand for 10 h, centrifuge at 6000 rpm for 15 min, discard the supernatant, remove residual water from the centrifuge tube by adsorption with filter paper, and accurately weigh the precipitate, denoted as M2. Water-holding capacity is calculated using the following formula:
[0064] WHC(g / g) = (M2−M1) / M1
[0065] The water-holding capacity results of soybean residue IDF are as follows Figure 4 As shown.
[0066] Depend on Figure 4 It can be seen that the water-holding capacity of the soybean residue IDF prepared in Example 1 was increased by 1.58 times, 1.37 times, and 1.30 times compared with that of Control Example 1, Control Example 2, and Control Example 3, respectively.
[0067] Example 5
[0068] Determination of IDF oil holding capacity (OHC) of soybean residue
[0069] Accurately weigh 0.5 g of the insoluble dietary fiber prepared in Control Examples 1, 2, 3, and 1, denoted as M1, and dissolve it in 15 mL of soybean oil. Stir uniformly at 25°C for 2 hours, then let stand for 10 hours. Centrifuge at 6000 rpm for 15 minutes, discard the supernatant, and remove residual soybean oil from the centrifuge tube by adsorption with filter paper. Accurately weigh the precipitate mass, denoted as M2. The oil holding capacity is calculated using the following formula:
[0070] OHC(g / g) = (M2−M1) / M1
[0071] The results of the oil holding capacity of soybean residue IDF are as follows: Figure 5 As shown.
[0072] Depend on Figure 5 It can be seen that the oil holding capacity of the soybean residue IDF prepared in Example 1 was increased by 1.83 times, 1.15 times, and 1.53 times compared with the control examples 1, 2, and 3, respectively.
Claims
1. A method for preparing insoluble dietary fiber from soybean residue, characterized in that, The process includes the following steps: defatting soybean residue to obtain defatted soybean residue powder; thoroughly mixing the defatted soybean residue powder with deionized water and then subjecting it to ultra-high pressure microfluidic homogenization to obtain a soybean residue powder-water suspension; sequentially subjecting the soybean residue powder-water suspension to enzymatic hydrolysis by high-temperature resistant α-amylase, neutral protease, and amyloglucosidase, followed by enzyme inactivation, and then further enzymatic hydrolysis by snail enzyme, followed by enzyme inactivation and drying to obtain insoluble dietary fiber from soybean residue; the working pressure of the ultra-high pressure microfluidic homogenization is 200~250MPa, and the processing time is 20~25min.
2. The preparation method according to claim 1, characterized in that, The amount of heat-resistant α-amylase added for hydrolysis is 1%~1.5% per 50g of soybean residue mixture, and the enzyme activity of the heat-resistant α-amylase is 2×104U / g; the temperature of the heat-resistant α-amylase hydrolysis is 75~95℃, the time is 1~1.5h, and the pH is 5.0~8.
0.
3. The preparation method according to claim 1, characterized in that, The amount of neutral protease added for hydrolysis is 2%~2.5% per 50g of soybean residue mixture, and the enzyme activity of the neutral protease is 5×104U / g; the hydrolysis temperature of the neutral protease is 35~55℃, the time is 2~2.5h, and the pH is 6.5~8.
0.
4. The preparation method according to claim 1, characterized in that, The amount of amylase added for hydrolysis is 1%~1.5% amylase per 50g of soybean residue mixture, and the enzyme activity of the amylase is 2×104U / g; the hydrolysis temperature is 50~65℃, the time is 1~1.5h, and the pH is 4.0~5.
5.
5. The preparation method according to claim 1, characterized in that, The snail enzyme hydrolysis is performed by adding 1% to 3% snail enzyme per 50g of soybean residue mixture, and the snail enzyme activity is 3U / mg; the snail enzyme hydrolysis temperature is 40 to 60℃, the time is 1 to 4 hours, and the pH is 4.0 to 7.
5.
6. The preparation method according to claim 1, characterized in that, The raw material used for degreasing is petroleum ether, the material-to-liquid ratio for degreasing is 1g soybean residue: 10~15mL petroleum ether, and the degreasing time is 5~10h.
7. The soybean residue insoluble dietary fiber prepared by the preparation method according to any one of claims 1-6.