Pickering nano-emulsion stabilized by soybean protein fibrils as well as preparation method and application of Pickering nano-emulsion

Abstract

The embodiment of the invention discloses a Pickering nano-emulsion stabilized by soybean protein fibrils as well as a preparation method and application thereof, relates to the technical field of medicines, and aims to provide a soybean protein fibril nano-emulsion specially used for improving the lipid digestibility of old people. The preparation method comprises the following steps: dispersing soybean protein isolate in water to obtain soybean protein dispersion liquid; adding protease into the soybean protein dispersion liquid to carry out a restrictive enzymatic hydrolysis reaction; after the reaction is completed, the hydrolysis degree is controlled to be 1%-10%, heating enzyme deactivation treatment, cooling treatment, centrifugal treatment and drying treatment are sequentially conducted, and the polypeptide nanoparticles are obtained; adding the polypeptide nano-particles into water to obtain a polypeptide nano-particle dispersion liquid, adjusting the pH value of the polypeptide nano-particle dispersion liquid, and sequentially performing hydration treatment, high-temperature heating treatment, cooling treatment, centrifugal treatment and drying treatment to obtain soybean protein fibril freeze-dried powder; adding the soybean protein fibril freeze-dried powder into water to serve as a water phase for later use; and uniformly mixing the water phase and the oil phase in proportion, and sequentially carrying out pre-emulsification treatment and ultrasonic treatment to obtain the Pickering nano-emulsion stabilized by the soybean protein fibrils.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine, in particular to a Pickering nanoemulsion stabilized by soybean protein fibrils and a preparation method and application thereof. BACKGROUND

[0002] Precise regulation of lipid digestion rate is of great significance for improving postprandial metabolic response and enhancing the delivery efficiency of lipid-soluble bioactive substances. The structure of the Pickering emulsion interface protein is a decisive factor in regulating lipid digestion kinetics, which regulates the digestion process through the dual mechanisms of physical barrier and dynamic response. The thickness of the adsorbed layer and the aggregation state of the protein formed at the oil-water interface directly dominate the interface mechanical strength: a dense and ordered protein network can inhibit lipase from approaching the lipid droplet core through steric hindrance effect, significantly delaying the initial rate of lipolysis; while a loose or low-stability protein layer is easily disturbed by the gastrointestinal environment, resulting in the failure of the interface protection function and accelerating the release of lipids. This regulation strategy based on the structure of the interface protein, namely interface engineering, provides a new way to enhance the absorption capacity of nutrients for people of different ages.

[0003] With the global population structure accelerating towards aging, achieving healthy aging has become a major social challenge. The demand for nutrients and bioactive substances of the elderly increases significantly, but the decline in the function of the digestive system (such as reduced gastric acid secretion and reduced pancreatic enzyme activity) leads to a sharp decline in nutrient absorption efficiency, and this contradiction is particularly pronounced in lipid nutrients. SUMMARY

[0004] The main purpose of the present application is to provide a Pickering nanoemulsion stabilized by soybean protein fibrils and a preparation method and application thereof, aiming to provide a soybean protein fibril nanoemulsion specially used to improve the lipid digestion rate of the elderly.

[0005] To achieve the above-mentioned purpose, the technical solutions adopted by the embodiments of the present application are as follows: In a first aspect, the present application provides a preparation method of a Pickering nanoemulsion stabilized by soybean protein fibrils, comprising the following steps: dispersing soybean protein isolate in water to obtain a soybean protein dispersion; adding protease to the soybean protein dispersion to perform a limited enzymatic hydrolysis reaction; after the reaction is completed, controlling the degree of hydrolysis to be 1% to 10%, and sequentially performing heat deactivation treatment, cooling treatment, centrifugal treatment and drying treatment to obtain polypeptide nanoparticles; adding the polypeptide nanoparticles to water to obtain a polypeptide nanoparticle dispersion, adjusting the pH, and sequentially performing hydration treatment, high-temperature heating treatment, cooling treatment, centrifugal treatment and drying treatment to obtain a soybean protein fibril freeze-dried powder; The soybean protein fibril freeze-dried powder is added into water as a water phase for standby; After the water phase and the oil phase are mixed uniformly in proportion, pre-emulsification treatment and ultrasonic treatment are sequentially performed to obtain a Pickering nanoemulsion stabilized by soybean protein fibrils.

[0006] As some optional embodiments of the present application, the mass-volume ratio of the soybean protein isolate to the water is 1 g-10 g:100 mL; The protease is a flavor protease, and the mass of the flavor protease is 0.2wt%-1wt% of the soybean protein isolate.

[0007] As some optional embodiments of the present application, the dispersion time for dispersing the soybean protein isolate in water to obtain a soybean protein dispersion liquid is 1 h-2 h, and stirring is performed at a rate of 200 rpm-400 rpm during dispersion; The parameters of the limited enzymolysis reaction are 50°C, 10 min-60 min; The parameters of the heat-killing enzyme treatment are 70°C-90°C, 5 min-60 min; The parameters of the cooling treatment are ice water bath for 10 min-20 min; The parameters of the drying treatment are vacuum freeze-drying for 24 h; The parameters of the centrifugal treatment are 5000 rpm-10000 rpm, 1-3 times, 10 min-30 min / time.

[0008] As some optional embodiments of the present application, the particle size of the polypeptide nanoparticles is 50 nm-200 nm.

[0009] As some optional embodiments of the present application, the mass-volume ratio of the polypeptide nanoparticles to the water is 1 g-10 g:100 mL; During the hydration treatment, the stirring speed is maintained at 130 rpm-180 rpm, and the parameters of the hydration treatment are 4°C, 2 h-24 h, and pH=2-4; The parameters of the high-temperature heating treatment are 80°C-100°C, 45 min-60 min, 90 min-120 min, or 180 min-240 min; The parameters of the centrifugal treatment are 5000 rpm-10000 rpm, 10 min-60 min, and 15°C-25°C; The parameters of the cooling treatment are ice water bath for 10 min-20 min; The parameters of the drying treatment are vacuum freeze-drying for 24 h; The protein fibril length in the soybean protein fibril freeze-dried powder is 0nm-100nm, 300nm-400nm or 700nm-900nm.

[0010] As some optional embodiments of the present application, the mass-volume ratio of the soybean protein fibril freeze-dried powder to the water is 1g-10g:100mL, and the stirring rate during mixing is 100rpm-150rpm.

[0011] As some optional embodiments of the present application, the oil phase comprises one of medium-chain triglycerides, corn oil and soybean oil. The mixing ratio of the oil phase to the water phase is 1:9. The pre-emulsification treatment refers to high-speed shearing treatment, and the parameters of the high-speed shearing are 9000rpm-15000rpm and 1min-5min. The parameters of the ultrasonic treatment are 200W-800W and 5min-20min.

[0012] In a second aspect, the embodiments of the present application further provide a Pickering nanoemulsion stabilized by soybean protein fibrils, which is prepared by the method as described above.

[0013] As some optional embodiments of the present application, the particle size of the Pickering nanoemulsion is 100nm-250nm.

[0014] In a third aspect, the embodiments of the present application further provide an application of the Pickering nanoemulsion stabilized by soybean protein fibrils as described above, which is used for preparing a delivery carrier of drugs, nutrients and fat-soluble active substances in the field of food or medicine.

[0015] Compared with the prior art, the present application has the following beneficial effects: The Pickering nanoemulsion stabilized by soybean protein fibrils provided by the present application uses soybean protein fibrils with different lengths as Pickering stabilizers, and all of them show the effect of significantly improving the lipid digestion rate under the digestion model of the elderly. If stabilized by longer soybean protein fibrils, it has the effect of significantly delaying lipids and shows the characteristics of sustained release, and is suitable for food that needs to provide energy continuously and avoid rapid increase of blood sugar and blood lipids, such as special diet food for diabetic patients or sports nutrition. If stabilized by shorter soybean protein fibrils, it can significantly improve the free fatty acid release rate (see the data in the embodiments, close to 80%), and is suitable for solving the problem of insufficient absorption of lipid nutrients caused by reduced digestive function in the elderly. It can be seen that the Pickering nanoemulsion stabilized by soybean protein fibrils provided by the present application provides a way to regulate the digestion characteristics of the emulsion by adjusting the fibril length, and can achieve the directional design of the nutrient release mode of the emulsion. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 Transmission electron microscopy images of soy protein isolate and soy protein fibrils prepared for Example 1-3 and Comparative Example 1 of the present application; Figure 2 Transmission electron microscopy images of emulsion interface morphology prepared for Example 1-3 and Comparative Example 1 of the present application.

[0017] Figure 3 Free fatty acid release profiles of emulsions prepared for Example 1-3 and Comparative Example 1 of the present application under an elderly digestion model. DETAILED DESCRIPTION

[0018] It should be understood that the specific embodiments described herein are merely exemplary and not intended to limit the application.

[0019] The present application research found that the length of the fiber is a key factor to regulate its interface behavior and the final digestion kinetics: longer fibers tend to form more dense, higher mechanical strength network structure at the interface, which may delay the lipolysis process; while shorter fibers may provide different interface coverage density, affecting the enzyme adsorption efficiency at the initial stage of digestion. Therefore, the present application controls the average length of protein fibrils, thus regulates the interface characteristics and digestion responsiveness of Pickering emulsion, in order to provide a certain basis for designing suitable emulsion delivery system for the elderly population.

[0020] Specifically, the present application provides a preparation method of Pickering nanoemulsion stabilized by soy protein fibrils, comprising the following steps: Step 1: dispersing soybean protein isolate in water to obtain a soybean protein dispersion; adding protease to the soybean protein dispersion to perform a limited enzyme hydrolysis reaction; after the reaction is completed, controlling the degree of hydrolysis to be 1%~10% (preferably 2%~10%), and sequentially performing heat enzyme inactivation treatment, cooling treatment, centrifugal treatment and drying treatment to obtain polypeptide nanoparticles.

[0021] The mass-volume ratio of the soybean protein isolate to the water in step 1 is 1g~10g:100mL; preferably 2g~6g:100mL.

[0022] The protease is a flavor protease, and the mass of the flavor protease is 0.2wt%~1wt% of the soybean protein isolate, preferably 0.3wt%~0.8wt%.

[0023] The dispersion time when the soybean protein isolate is dispersed in water to obtain a soybean protein dispersion is 1h~2h, and stirring is performed at a rate of 200rpm~400rpm during dispersion; The parameters of the restriction enzyme reaction are: 50℃, 10min~60min; The parameters of the heat enzyme treatment are: 70℃~90℃, 5min~60min; The parameters of the cooling treatment are: ice water bath 10min-20min; The parameters of the drying treatment are: vacuum freeze-drying 24h; The parameters of the centrifugal treatment are: 5000rpm~10000rpm, 1~3 times, 10min~30min / time.

[0024] The particle size of the polypeptide nanoparticles is 50nm~200nm, preferably 80nm~100nm.

[0025] Step 2: add the polypeptide nanoparticles to water to obtain a polypeptide nanoparticle dispersion, adjust the pH, and then sequentially perform hydration treatment, high-temperature heating treatment, cooling treatment, centrifugal treatment, and drying treatment to obtain a soybean protein fibril freeze-dried powder.

[0026] In step 2, the mass-volume ratio of the polypeptide nanoparticles to the water is 1g~10g:100mL, preferably 1g~3g:100mL.

[0027] During the hydration treatment, the stirring speed is maintained at 130rpm~180rpm (preferably 140rpm~170rpm), and the parameters of the hydration treatment are: 4℃, 2h-24h, pH=2~4 (preferably 4℃, 4h~12h, pH=2).

[0028] The parameters of the high-temperature heating treatment are: 80℃~100℃, 45min~60min, 90min~120min, or 180min~240min; preferably 90℃, 45min~60min, 90min~120min, or 180min~240min.

[0029] The parameters of the centrifugal treatment are: 5000rpm~10000rpm, 10min~60min, 15℃~25℃; preferably 9000rpm, 15min, 20℃.

[0030] The parameters of the cooling treatment are: ice water bath 10min~20min; The parameters of the drying treatment are: vacuum freeze-drying 24h; The length of the protein fibrils in the soybean protein fibril freeze-dried powder is 0nm~100nm, 300nm~400nm, or 700nm~900nm.

[0031] It needs to be further explained that in step 2, by controlling the high-temperature heat treatment of the nanoparticle dispersion liquid for three different times, and then cooling it, the fibrous process is interrupted, and protein fibrils of different lengths are obtained.

[0032] Step 3: Add the soy protein fibril freeze-dried powder to water as a water phase for standby.

[0033] In step 3, the mass-volume ratio of the soy protein fibril freeze-dried powder to the water is 1g-10g:100mL, preferably 1g-5g:100mL; the stirring rate during mixing is 100rpm-150rpm, preferably 110rpm-130rpm.

[0034] Step 4: After the water phase and the oil phase are mixed uniformly in proportion, pre-emulsification treatment and ultrasonic treatment are carried out in sequence to obtain a Pickering nanoemulsion stabilized by soy protein fibrils.

[0035] In step 4, the oil phase includes one of medium-chain triglycerides, corn oil, and soybean oil; preferably medium-chain triglycerides.

[0036] The mixing ratio of the oil phase to the water phase is 1:9. The pre-emulsification treatment refers to high-speed shearing treatment, and the parameters of the high-speed shearing are 9000rpm-15000rpm, 1min-5min; preferably 10000rpm, 3min.

[0037] The parameters of the ultrasonic treatment are 200W-800W, 5min-20min, preferably 300-600W, 10min.

[0038] The particle size of the Pickering nanoemulsion is 100nm-250nm.

[0039] The application of the above-mentioned Pickering nanoemulsion stabilized by soy protein fibrils is used to prepare a delivery carrier for drugs, nutrients, and fat-soluble active substances in the food field or the medical field; that is, the Pickering nanoemulsion stabilized by soy protein fibrils can be applied to a delivery carrier for drugs, nutrients, and fat-soluble active substances in the food and medical fields, and is particularly suitable for the field of nutrition supplementation for the elderly, providing a basis for solving the malnutrition problem of the elderly in terms of lipids.

[0040] In order to facilitate those skilled in the art to understand the technical solutions of the present application, the present application will be further described in detail in conjunction with the specific embodiments: It should be noted that the following processes are not specifically detailed, which can be implemented or understood by referring to the prior art by those skilled in the art. The reagents or instruments not specified by the manufacturer are considered to be conventional products that can be purchased on the market.

[0041] Example 1: Preparation of soy protein fibrils: Soy protein isolate was dissolved in deionized water (40 mg / ml), and the pH was adjusted to 7 using 1M NaOH. 0.5% flavor protease was added, and the enzyme was hydrolyzed in a constant temperature shaker at 50°C for 0.5h. The enzyme was inactivated and centrifuged, and the supernatant was freeze-dried to obtain peptide nanoparticles. The freeze-dried powder was dissolved in deionized water (20 mg / ml), and the pH was adjusted to 2 using 6M HCl. The nanoparticles were hydrated overnight. The nanoparticle solution was heated in a constant temperature water bath at 90°C for 45 min, cooled in an ice bath, and centrifuged at 9000 rpm for 15 min at 20°C. The supernatant was freeze-dried to obtain a soy protein fibril powder sample.

[0042] Preparation of Pickering nanoemulsion: 1g of soy protein fibril powder sample was dissolved in 45mL of deionized water, and the mixture was stirred at 500rpm by a magnetic stirrer at room temperature for 1h, and hydrated overnight at 4°C to obtain an aqueous phase system of the emulsion; medium-chain triglycerides were used as the oil phase (5g). The aqueous phase and the oil phase were mixed, and then the oil-water two-phase was pre-emulsified using a high-speed shearing machine, with a shearing speed of 12000 rpm and a shearing time of 3 min. The crude emulsion was further treated by ultrasonic, and ice bath was used to avoid excessive heating during the treatment. The ultrasonic conditions were total time of 10 min, pulse time of 1.0 s, intermittent time of 1.0 s, and power of 400 W. A Pickering nanoemulsion stabilized by soy protein isolate was obtained.

[0043] Example 2: Preparation of soy protein fibrils: Soy protein isolate was dissolved in deionized water (40 mg / ml), and the pH was adjusted to 7 using 1M NaOH. 0.5% flavor protease was added, and the enzyme was hydrolyzed in a constant temperature shaker at 50°C for 0.5h. The enzyme was inactivated and centrifuged, and the supernatant was freeze-dried to obtain peptide nanoparticles. The freeze-dried powder was dissolved in deionized water (20 mg / ml), and the pH was adjusted to 2 using 6M HCl. The nanoparticles were hydrated overnight. The nanoparticle solution was heated in a constant temperature water bath at 90°C for 120 min, cooled in an ice bath, and centrifuged at 9000 rpm for 15 min at 20°C. The supernatant was freeze-dried to obtain a soy protein fibril powder sample.

[0044] Preparation of Pickering nanoemulsion: 1 g of soy protein fibril powder sample was dissolved in 45 mL of deionized water, stirred at 500 rpm by a magnetic stirrer at room temperature for 1 h, and fully hydrated at 4 °C overnight to obtain the aqueous phase system of the emulsion; medium-chain triglycerides were used as the oil phase (5 g). The aqueous phase was mixed with the oil phase, and then the oil-water two-phase was pre-emulsified using a high-speed shearing machine, the shearing condition was 12000 rpm for 3 min. Further ultrasonic treatment was carried out on the coarse emulsion, and ice bath treatment was carried out during the treatment to avoid excessive heating, the ultrasonic condition was 10 min in total, 1.0 s in pulse time, 1.0 s in intermittent time, and 400 W in power, to obtain the Pickering nanoemulsion stabilized by soy protein isolate.

[0045] Example 3: Preparation of soy protein fibril: Soy protein isolate was dissolved in deionized water (40 mg / ml), the pH was adjusted to 7 using 1M NaOH, 0.5% flavor protease was added, and the enzyme was hydrolyzed at 50°C in a constant temperature shaker for 0.5 h, the enzyme was inactivated and centrifuged, the supernatant was freeze-dried to obtain peptide nanoparticles. The obtained freeze-dried powder was dissolved in deionized water (20 mg / ml), the pH was adjusted to 2 using 6M HCl, and the hydration was carried out overnight. The nanoparticle solution was heated in a constant temperature water bath at 90°C for 240 min, cooled in an ice bath, centrifuged at 9000 rpm for 15 min at 20°C, and the obtained supernatant was freeze-dried, which was the soy protein fibril powder sample.

[0046] Preparation of Pickering nanoemulsion: 1 g of soy protein fibril powder sample was dissolved in 45 mL of deionized water, stirred at 500 rpm by a magnetic stirrer at room temperature for 1 h, and fully hydrated at 4 °C overnight to obtain the aqueous phase system of the emulsion; medium-chain triglycerides were used as the oil phase (5 g). The aqueous phase was mixed with the oil phase, and then the oil-water two-phase was pre-emulsified using a high-speed shearing machine, the shearing condition was 12000 rpm for 3 min. Further ultrasonic treatment was carried out on the coarse emulsion, and ice bath treatment was carried out during the treatment to avoid excessive heating, the ultrasonic condition was 10 min in total, 1.0 s in pulse time, 1.0 s in intermittent time, and 400 W in power, to obtain the Pickering nanoemulsion stabilized by soy protein isolate.

[0047] Comparative Example 1 1 g of soybean protein isolate was dissolved in 45 mL of deionized water, stirred at 500 rpm by a magnetic stirrer at room temperature for 1 h, and fully hydrated at 4 °C overnight to obtain an aqueous phase system of emulsion; medium-chain triglycerides were used as the oil phase (5 g). The aqueous phase and the oil phase were mixed, and then the oil-water two-phase was pre-emulsified using a high-speed shearing machine, the shearing condition was 12000 rpm for 3 min. Further ultrasonic treatment was carried out on the coarse emulsion, and ice bath treatment was carried out during the treatment to avoid excessive heating, the ultrasonic condition was 10 min in total, 1.0 s in pulse time, 1.0 s in intermittent time, and 400 W in power, to obtain a Pickering nanoemulsion stabilized by soybean protein fibrils.

[0048] In the above examples and comparative examples, the emulsion particle size was measured as follows: The particle size of the Pickering nanoemulsion stabilized by soybean protein fibrils was measured by Mastersizer 2000, and the average particle size of the emulsion (nm) was obtained.

[0049] In the following examples, the determination method of free fatty acid was as follows: The method of "Static in vitro Digestion Model for the General Elderly Population: International Consensus on INFOGEST" was used to simulate the gastrointestinal digestion process. After adding lipase during the intestinal digestion process, timing was immediately carried out, 0.25 M NaOH was added dropwise during the digestion process to keep the pH value at 7.0, and the consumption volume and time of NaOH were recorded, and the digestion was continued for 2 h. The free fatty acid release amount (FFA) ) can be obtained by the following formula: ; In the formula, represents the amount of NaOH used (L), represents the concentration of NaOH (M), represents the average relative molecular mass of the oil (g·mol -1 ), represents the total mass of the oil in the reaction system (g).

[0050] In the following examples, the measurement method of fiber length was as follows: Image J software was used to measure the length manually according to the scale length.

[0051] In the following examples, the determination method of initial digestion rate was as follows: The free fatty acid release curve was fitted in origin (R R2 > 0.999), and the formula was as follows: ; In the formula, xThe value represents the digestion time; all other parameters are fitting constants. Taking the first derivative of the equation, the slope obtained when x is 0 min represents the initial lipid digestion rate (in % / min).

[0052] The methods for observing the microstructure of the emulsion interface in the following embodiments are as follows: Prepare a diluent for the emulsion sample and a phosphotungstic acid solution (1%, w / v) using deionized water. Place one drop (approximately 10 μL) of the diluted emulsion sample onto a copper grid and allow it to adsorb at room temperature for 2 minutes. Carefully remove the sample with filter paper, then gently heat and dry it with an alcohol lamp for 1 minute. Place one drop (approximately 10 μL) of the phosphotungstic acid solution onto the copper grid, stain for 2 minutes, and then blot dry with filter paper. After drying overnight, the emulsion interface morphology can be observed.

[0053] The results obtained using the above methods are shown in Table 1 and... Figures 1-3 As shown.

[0054] Table 1: ; Depend on Figure 1 As shown in Examples 1-3, the length of soybean protein fibrils increases with heating time and changes from short-curved to long-straight, resulting in changes in protein structure. The length obtained is positively correlated with heating time.

[0055] Depend on Figure 2 As shown, the emulsion interface protein structures of Examples 1-3 are all stabilized by different numbers of soybean protein fibers, while Comparative Example 1 is stabilized by spherical particles.

[0056] Depend on Figure 3 As shown, the initial rate of Comparative Example 1 was significantly higher than that of Examples 2 and 3, indicating that long fibers have the ability to delay lipid digestion; the release rate of free fatty acids in Comparative Example 1 was significantly lower than that of Examples 1, 2 and 3, indicating that Pickering emulsion stabilized by soybean protein fibrils has the ability to significantly improve lipid digestion in an elderly digestive model.

[0057] As shown in Table 1, a stable emulsion can be prepared from soybean protein fiber. The emulsion particle size is small, around 200 nm. This indicates that the only factor affecting lipid digestibility is the interfacial protein structure of the emulsion, i.e., fiber length. Compared with Comparative Example 1, the initial digestibility of the emulsion stabilized by soybean protein fiber continuously decreases, while the final value of free fatty acid release increases significantly.

[0058] Based on this, it can be seen that the technical solution described in this application has the following advantages: (1) The Pickering nanoemulsion stabilized by soybean protein fibrils provided in the present application uses soybean protein fibrils with different lengths as Pickering stabilizers for the first time, and all have the effect of significantly improving the lipid digestion rate under the digestion model of the elderly.

[0059] (2) The Pickering nanoemulsion stabilized by soybean protein fibrils provided in the present application has the effect of significantly delaying lipid when stabilized by longer soybean protein fibrils, has a sustained-release property, and is suitable for food that needs to provide energy continuously and avoid rapid rise of blood sugar and blood lipid, such as special diet food for diabetic patients or sports nutrition.

[0060] (3) The Pickering nanoemulsion stabilized by soybean protein fibrils provided in the present application can significantly improve the free fatty acid release rate (see the data in the examples, close to 80%) when stabilized by shorter soybean protein fibrils, and is suitable for solving the problem of insufficient absorption of lipid nutrition caused by reduced digestive function in the elderly.

[0061] (4) The Pickering nanoemulsion stabilized by soybean protein fibrils provided in the present application provides a method for regulating the digestion characteristics of the emulsion by adjusting the length of the fibrils, which can realize the directional design of the nutrition release mode of the emulsion.

[0062] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A process for the preparation of a Pickering nanoemulsion stabilized by soy protein fibrils, characterized in that, The method comprises the following steps: dispersing soybean protein isolate in water to obtain a soybean protein dispersion; adding protease to the soybean protein dispersion to perform a limited enzymatic hydrolysis reaction; after the reaction is completed, controlling the degree of hydrolysis to be 1%-10%, and sequentially performing heat-enzyme-killing treatment, cooling treatment, centrifugal treatment, and drying treatment to obtain polypeptide nanoparticles; adding the polypeptide nanoparticles to water to obtain a polypeptide nanoparticle dispersion, adjusting the pH, and sequentially performing hydration treatment, high-temperature heating treatment, cooling treatment, centrifugal treatment, and drying treatment to obtain a soybean protein fibril freeze-dried powder; adding the soybean protein fibril freeze-dried powder to water as an aqueous phase for standby; mixing the aqueous phase and an oil phase uniformly according to a proportion, and sequentially performing pre-emulsification treatment and ultrasonic treatment to obtain a Pickering nanoemulsion stabilized by soybean protein fibrils.

2. The method for preparing a Pickering nanoemulsion stabilized by soy protein fibrils according to claim 1, characterized in that, The mass-volume ratio of the soybean protein isolate to the water is 1g-10g:100mL. The protease is a flavor protease, and the mass of the flavor protease is 0.2wt%-1wt% of the soybean protein isolate.

3. The method for preparing a Pickering nanoemulsion stabilized by soybean protein fibrils according to claim 1, characterized in that, The dispersion time when the soybean protein isolate is dispersed in water to obtain the soybean protein dispersion is 1h-2h, and stirring is performed at a rate of 200rpm-400rpm during the dispersion; The parameters of the limited enzymatic hydrolysis reaction are 50°C, and 10min-60min. The parameters of the heat-enzyme-killing treatment are 70°C-90°C, and 5min-60min. The parameters of the cooling treatment are ice water bath for 10min-20min. The parameters of the drying treatment are vacuum freeze-drying for 24h. The parameters of the centrifugal treatment are 5000rpm-10000rpm, 1-3 times, and 10min-30min / time.

4. The method for preparing a Pickering nanoemulsion stabilized by soybean protein fibrils according to claim 1, characterized in that, The particle size of the polypeptide nanoparticles is 50nm-200nm.

5. The method for preparing a Pickering nanoemulsion stabilized by soybean protein fibrils according to claim 1, characterized in that, The mass-volume ratio of the polypeptide nanoparticles to the water is 1g-10g:100mL. During the hydration treatment, the stirring speed is maintained at 130rpm-180rpm, and the parameters of the hydration treatment are 4°C, 2h-24h, and pH=2-4. The parameters of the high-temperature heating treatment are 80°C-100°C, 45min-60min, 90min-120min, or 180min-240min. The parameters of the centrifugal treatment are 5000rpm-10000rpm, 10min-60min, and 15°C-25°C. The parameters of the cooling treatment are ice water bath for 10min-20min. The parameters of the drying treatment are vacuum freeze-drying for 24h. The length of the protein fibrils in the soybean protein fibril freeze-dried powder is 0nm-100nm, 300nm-400nm, or 700nm-900nm.

6. The method for preparing a Pickering nanoemulsion stabilized by soybean protein fibrils according to claim 1, characterized in that, The mass-volume ratio of the soybean protein fibril freeze-dried powder to the water is 1g-10g:100mL, and the stirring rate during the mixing is 100rpm-150rpm.

7. The method for preparing a Pickering nanoemulsion stabilized by soybean protein fibrils according to claim 1, characterized in that, The oil phase comprises one of medium-chain triglycerides, corn oil, and soybean oil. The mixing ratio of the oil phase to the water phase is 1:9; The pre-emulsification treatment refers to high-speed shearing treatment, and the parameters of the high-speed shearing are 9000 rpm-15000 rpm and 1 min-5 min; The parameters of the ultrasonic treatment are 200 W-800 W and 5 min-20 min.

8. A Pickering nanoemulsion stabilized by soy protein fibrils, characterized in that, Obtained by the method according to any one of claims 1-7.

9. The Pickering nanoemulsion stabilized by soy protein fibrils according to claim 8, characterized in that, The particle size of the Pickering nanoemulsion is 100 nm-250 nm.

10. Use of a Pickering nanoemulsion stabilized by soy protein fibrils as claimed in claim 8, characterized in that, A delivery carrier for preparing a drug, a nutrient substance, a fat-soluble active substance in the field of food or the field of medicine.

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