Construction method and application of amylopectin ternary nano assembly with high assembly index

Abstract

The invention relates to the field of bioengineering, in particular to a construction method and application of an amylopectin ternary nano assembly with a high assembly index. The invention provides a construction method of a ternary nano assembly. The construction method comprises the following steps: S1, extracting amylopectin of bulk crops by adopting an ethanol sedimentation method; s2, extracting endogenous lipids of fruits of completely mature bulk crops by adopting a supercritical carbon dioxide extraction method; s3, extracting endogenous proteins of fruits of immature bulk crops by adopting an isoelectric precipitation method; s4, the amylopectin, the endogenous lipid and the endogenous protein are subjected to a self-assembly reaction, and the ternary nano assembly is obtained. The particle size of the ternary complex can reach nanometer, and the ternary complex is high in CI, high in SDS content, low in multi-stage digestion rate, stable in blood sugar release process and good in chewiness.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of bioengineering, and particularly relates to a method for constructing high-assembly-index amylopectin ternary nanoassemblies and application thereof. BACKGROUND

[0002] Common staple crops include corn, waxy corn, wheat, potato, barley, rice, sorghum, and the yield accounts for more than 90% of the global food, which is the most important source of nutrition for the global population. China is the world's largest staple crop food producer, so further development and utilization of staple crops are of great significance. The staple crops grown in the black soil in northern China generally have high yield (≥1 ton / acre), low cost (≤5 yuan / kg), good flavor and texture, and good color, so they have been widely loved by consumers as high-quality resources. In addition, the fat, protein and starch contents of unripe staple crop fruits are greater than 0.1%, 6% and 65% respectively. However, the fat content of fully mature staple crop fruits is usually greater than 5%. However, due to the high mineral content, the cracking rate is high, the shelf life is short, and a large number of cracked and unsold staple crop fruits are discarded every year, resulting in waste of starch, protein and lipid resources. Based on the above analysis, the construction of ternary assemblies of starch, endogenous lipid and endogenous protein, which are the most important nutrients that can be absorbed and utilized by humans in staple crop fruits, can provide new strategic nutritional resources for humans.

[0003] Starch is a semi-crystalline superpolymer composed of amylose and amylopectin. Amylopectin is a macromolecular structure with high branching, and the proportion of α-1,6 glycosidic bonds is about 5%. The proportion of amylopectin in staple crops is usually 70-90%, and the proportion of amylopectin in natural waxy corn starch can reach more than 99%, which is the most important fine structure in starch structure. The side chains of amylopectin can form a double helix structure through intermolecular interaction, which further affects the formation of slow digestible starch (SDS) and resistant starch (RS) structures. However, RS is extremely difficult to release glucose, which cannot meet the nutritional needs of the human body, so this patent focuses on the formation rate of SDS structure that can slow down the multi-level digestion rate and slowly release nutrients, which can improve and prevent sugar metabolism disorders through dietary approach. In addition, industrially extracted starch usually has low purity, and the surface will adsorb a large amount of endogenous lipid and protein, so after heat processing of starch-based food raw materials, starch and endogenous lipid and protein can spontaneously form ternary complexes through non-covalent bonds during the retrogradation process. This phenomenon can greatly change the digestibility, but there are few reports on the slow digestion SDS structure content of ternary complexes.

[0004] Compared with the strong polar and nonpolar interaction between amylose helix structure and lipid, protein ligand, due to the short chain length and the high steric hindrance caused by the branched structure of amylopectin, the self-assembly ability of amylopectin-lipid, protein complex is significantly reduced. Therefore, the self-complexing index (CI) of amylopectin-fatty acid or amylopectin-protein complex is significantly lower than that of amylose complex, but both of them show similar digestibility. And the ultrasonic technology can significantly reduce the complex particle size of starch and its related derivatives and significantly improve the purity. However, so far, there are still few patents about whether the endogenous ternary nano-assembly of amylopectin has a lower nano-particle size and better slow digestibility after ultrasonic.

[0005] The commonly used methods for constructing starch ternary complex with anti-digestibility are mainly about amylose, such as rapid viscosity analyzer, double screw extrusion method, high pressure microfluidization method, etc. However, the CI of the ternary complex prepared by the above methods is low, the time is long, the generated complex has low CI, poor anti-enzymatic property, and more type I complex, the constructed complex usually does not have nano-particle size and slow digestibility, and the mechanical equipment requirement is high. Sometimes it needs to be combined with chemical means to assist in reducing the digestibility. The preparation methods of starch-lipid nano-assembly reported in many previous patents and articles mainly involve single long-time water bath magnetic stirring, enzyme degradation, and chemical reagent assisted assembly, but the time is long, a large amount of chemical reagents are added, and the focus is on anti-digestibility while ignoring the nutritional absorption demand of the population, and the chewability is poor, which cannot guarantee the CI of the complex. At present, there is no patent about the development of a new method for quickly constructing large-scale amylopectin-endogenous lipid-endogenous protein nano-assembly with slow digestibility, good chewability and high CI. SUMMARY

[0006] Therefore, the present application provides a method for constructing high-assembly-index amylopectin ternary nano-assembly and its application. The particle size of the ternary complex of the present application can reach nano, the CI is high, and the multi-stage digestion rate is low and the blood glucose release process is smooth.

[0007] In order to achieve the above-mentioned application purposes, the present application provides the following technical solutions:

[0008] The present application provides a method for constructing ternary nano-assembly, comprising the following steps:

[0009] S1: using ethanol sedimentation method to extract amylopectin of large-scale crops;

[0010] S2: using supercritical carbon dioxide extraction method to extract endogenous lipid of fully mature fruits of large-scale crops;

[0011] S3: extracting endogenous proteins of immature fruits of a bulk crop by isoelectric precipitation;

[0012] S4: performing a self-assembly reaction on the amylopectin, the endogenous lipid, and the endogenous protein to obtain the ternary nano-assembly.

[0013] In some embodiments of the present application, in the above construction method, the bulk crop includes one or more of waxy corn, corn, wheat, potato, barley, rice, and sorghum.

[0014] In some embodiments of the present application, in the above construction method, the ethanol precipitation method includes the following steps: after the bulk crop is completely gelatinized, centrifugation is performed, and the precipitate is collected, and then the ethanol precipitation method is used to obtain the amylopectin.

[0015] In some embodiments of the present application, in the above construction method, the complete gelatinization includes the following steps: the bulk crop and water are mixed at a ratio of 1-6:100 (w / w), heated in a water bath at 100 ℃ for 20 min, and then cooled to 25 ℃ after complete gelatinization.

[0016] In some embodiments of the present application, in the above construction method, the centrifugation is performed at a speed of 5000xg for 10 min.

[0017] In some embodiments of the present application, in the above construction method, the ethanol precipitation method includes the following steps: the precipitate collected in the above step is mixed with ethanol at a ratio of 90% (w / w), ultrasonically dispersed at room temperature for 10 min at 100-200 W, centrifuged at 5000xg for 10 min, and washed with ethanol for 3 times to remove ethanol, and then the amylopectin is obtained.

[0018] In some embodiments of the present application, in the above construction method, the supercritical carbon dioxide extraction method includes the following steps: after the fruits of the completely mature bulk crop are extracted, separated, and purified, the endogenous lipid is obtained.

[0019] In some embodiments of the present application, in the above construction method, the extraction is performed at a pressure of 10-40 Mpa and a temperature of 50-80 ℃, the separation is performed at a pressure of 2-20 Mpa and a temperature of 50-80 ℃, and the purification is performed by a rotary evaporation method at a temperature of 50-80 ℃.

[0020] In some embodiments of the present application, in the above construction method, the isoelectric precipitation method includes the following steps: the fruits of the immature bulk crop are mixed with distilled water, centrifuged, and the supernatant is collected and isoelectrically precipitated, and then the endogenous protein is obtained after dialysis.

[0021] In some embodiments of the present application, in the above method, the mass ratio of the immature fruits of the bulk crop to the distilled water is 1:20.

[0022] In some embodiments of the present application, in the above method, the centrifugal speed is 6000xg and the time is 20 min.

[0023] In some embodiments of the present application, in the above method, after the supernatant is collected, the pH is adjusted to 4.0-6.0 with 2M hydrochloric acid for isoelectric precipitation.

[0024] In some embodiments of the present application, in the above method, the mass ratio of the amylopectin, the endogenous lipid and the endogenous protein is 1:(0.05-0.15):(0.05-0.20).

[0025] In some embodiments of the present application, in the above method, the mass ratio of the amylopectin, the endogenous lipid and the endogenous protein is 1:(0.065-0.12):(0.07-0.17).

[0026] In some embodiments of the present application, in the above method, the mass ratio of the amylopectin, the endogenous lipid and the endogenous protein is 1:(0.075-0.10):(0.09-0.13).

[0027] In some embodiments of the present application, in the above method, the temperature of the self-assembly reaction is 80-100℃, the power of the ultrasonic is 300-800 W, and the time is 0.2-3h.

[0028] In some embodiments of the present application, in the above method, the temperature of the self-assembly reaction is 85-95℃, the power of the ultrasonic is 400-700 W, and the time is 1.0-2.2h.

[0029] In some embodiments of the present application, in the above method, the temperature of the self-assembly reaction is 88-92℃, the power of the ultrasonic is 500-600 W, and the time is 1.4-1.8h.

[0030] In some embodiments of the present application, in the above method, after the self-assembly reaction, a drying step is further included.

[0031] In some embodiments of the present application, in the above method, the drying is vacuum freeze drying; the specific parameters are that the material layer temperature is 30-60℃, the vacuum degree is 30 Pa, the in-machine cold trap temperature is -80℃, the temperature rising speed is 2-10℃ / min, the freeze drying is 24h, and the freeze drying is performed under the conditions to freeze dry the water mass percentage content to ≤10%.

[0032] The application also provides the ternary nano-assembly obtained by the construction method.

[0033] The application also provides the application of the ternary nano-assembly in any of the following aspects.

[0034] ), the preparation of products for improving and / or preventing glucose metabolism disorders; and / or

[0035] ), the preparation of products for preventing and / or relieving type II diabetes; and / or

[0036] ), the preparation of products for preventing and / or relieving hyperlipidemia; and / or

[0037] ), the preparation of products for preventing and / or relieving hypertension; and / or

[0038] ), the preparation of products for preventing and / or relieving obesity; and / or

[0039] I) the preparation of products for preventing and / or relieving swallowing disorders.

[0040] The beneficial effects of the application include:

[0041] (1) In the extraction of amylopectin, the extraction of endogenous proteins, the extraction of endogenous lipids, and the construction of ternary nano-assembly of amylopectin, the power, processing time, temperature, and rotation speed of the instrument are all part of the overall technical solution. They support each other in function and have an interactive relationship. The above-mentioned coordination can solve the technical problems of the application and achieve the technical effects of the application. The key to achieving the key of the ternary assembly of amylopectin of bulk crops reaching the nanoscale, significantly improving slow digestion, CI, anti-thermal decomposition, anti-enzymatic decomposition, and texture characteristics, slowly releasing glucose, improving texture characteristics (chewing), and good particle stability.

[0042] (2) The application is based on the construction of slow-digesting ternary nano-assembly of amylopectin of bulk crops by high-power ultrasonic assistance. The process is simple and convenient, time-consuming is short, and no toxic and harmful reagents are used in the construction process. The preparation process is greatly optimized. Compared with other methods, the patent greatly reduces the particle size while saving process cost and increasing self-assembly speed, thereby greatly increasing the self-assembly index. The digestion curve is smooth, and the nutrition can be provided continuously and slowly. The taste is greatly improved, and the environment is not polluted. The cost is low, and it is suitable for industrial automatic large-scale production. BRIEF DESCRIPTION OF DRAWINGS

[0043] ​​​​​​In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the embodiments or prior art description will be briefly introduced as follows.

[0044] Figure 1 A two-dimensional infrared contrast schematic diagram of waxy corn amylopectin ternary nanoassemblies and waxy corn amylopectin paste in Example 1 is shown in the figure.

[0045] Figure 2 A LOS digestion kinetics contrast schematic diagram of waxy corn amylopectin ternary nanoassemblies and waxy corn amylopectin paste in Example 1 is shown in the figure.

[0046] Figure 3 A texture characteristic curve contrast schematic diagram of wheat amylopectin ternary nanoassemblies and wheat amylopectin paste in Example 2 is shown in the figure.

[0047] Figure 4 A particle size contrast schematic diagram of wheat amylopectin ternary nanoassemblies in Example 2 and potato amylopectin ternary nanoassemblies in Example 3 is shown in the figure.

[0048] Figure 5 A micro-morphology contrast schematic diagram of barley amylopectin ternary nanoassemblies in Example 4 and rice amylopectin ternary nanoassemblies in Example 5 is shown in the figure.

[0049] Figure 6 An XRD contrast schematic diagram of barley amylopectin ternary nanoassemblies in Example 4 and sorghum amylopectin ternary nanoassemblies in Example 6 is shown in the figure.

[0050] Figure 7 A DSC contrast schematic diagram of waxy corn amylopectin ternary nanoassemblies in Example 1 and corn starch ternary micro-nanoassemblies in Comparative Example 1 is shown in the figure.

[0051] Figure 8 A mean square nano-surface roughness contrast schematic diagram of potato amylopectin ternary nanoassemblies in Example 3 and potato starch ternary micro-nanoassemblies in Comparative Example 2 is shown in the figure.

[0052] Figure 9 A freeze-thaw stability contrast schematic diagram of rice amylopectin ternary nanoassemblies in Example 5 and rice starch ternary micro-nanoassemblies in Comparative Example 3 is shown in the figure.

[0053] Figure 10 An X-ray scattering characteristic distance contrast schematic diagram of crystalline lamella and amorphous region of rice amylopectin ternary nanoassemblies constructed by ultrasonic-assisted technology in Example 4 and nano-barley starch ternary complexes prepared by ultrasonic-ball milling technology in Comparative Example 4 is shown in the figure. DETAILED DESCRIPTION

[0054] The application discloses a construction method of high-assembly-index branched amylopectin ternary nanoassemblies and application thereof.

[0055] It should be understood that the expression "one or more of" includes individually each of the objects recited after the expression and various combinations of two or more of the recited objects, unless otherwise understood from the context and usage. The expression "and / or", in connection with three or more recited objects, should be understood to have the same meaning, unless otherwise understood from the context.

[0056] The use of the expressions "comprising", "having" or "including", including their grammatical variations, should be generally understood as open-ended and non-limiting, for example, not excluding additional unrecited elements or steps, unless otherwise specifically stated or understood from the context.

[0057] It should be understood that the order of steps or order of performing certain actions is immaterial so long as the application remains operable. Moreover, two or more steps or actions can be conducted simultaneously.

[0058] The use of any and all examples, or exemplary language herein, for example, "for instance" or "such as", is intended merely to better illustrate the application and does not indicate that further unclaimed claims are limited to those items listed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the application.

[0059] Further, the numerical ranges and parameters setting forth the broadest scope of the application are approximations, and the numerical values set forth in the detailed description are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Therefore, unless otherwise specified, all ranges disclosed herein are to be understood to be approximations, and the numerical values underlying any such ranges are intended to be considered accurate within the context of the application. Herein, "about" or "approximately" means within 10% or 5% or 1% or 0.5% of a stated value or range.

[0060] The application provides a construction method of slow-digesting bulk branched amylopectin-endogenous lipid-endogenous protein nanoassemblies, comprising the following steps:

[0061] A) using an ethanol sedimentation method to extract branched amylopectin from bulk crops in large quantities;

[0062] B) using a carbon dioxide supercritical extraction method to extract endogenous lipids from fully matured fruits of bulk crops, and using an isoelectric deposition method to extract endogenous proteins from immature fruits of bulk crops;

[0063] C) Self-assembly reaction of bulk amylopectin with endogenous protein, endogenous lipid, and the obtained assembly is subjected to high-power ultrasonic to obtain endogenous ternary nano-assembly of bulk amylopectin;

[0064] D) The endogenous ternary nano-assembly of bulk amylopectin is dried, and the obtained product is obtained after screening and packaging.

[0065] Preferably, in the construction method of the slow-digesting bulk amylopectin-endogenous lipid-endogenous protein nano-assembly, the bulk amylopectin is extracted from waxy corn, corn, wheat, potato, barley, rice, and sorghum.

[0066] Preferably, the method for extracting the bulk amylopectin is as follows: after the bulk amylopectin sample is completely gelatinized by adding water, the precipitate is extracted by the ethanol precipitation method according to the principle that the branched-chain molecular weight is greater than that of amylose.

[0067] Preferably, in the method for extracting the bulk amylopectin, the starch sample is mixed with distilled water at a ratio of 1-6:100 (w / w), heated at 100 ℃ for 20 min in a water bath (preferably in a large water bath), completely gelatinized, cooled to 25 ℃, centrifuged at 5000×g for 10 min, and the precipitate is taken out, and the step is repeated twice; the branched-chain starch precipitate is mixed with ethanol at a ratio of 90% (w / w), ultrasonically dispersed at 100-200 W at room temperature for 10 min (preferably in a large ultrasonic cleaning machine), centrifuged at 5000×g for 10 min, and the precipitate is washed with ethanol for 3 times to obtain the branched-chain starch.

[0068] Preferably, the method for extracting the endogenous lipid in the fruit of the bulk crop is as follows: the fully mature bulk crop fruit is fed into a carbon dioxide supercritical extraction instrument, and the crude oil is extracted by the method of extraction followed by separation, and then the endogenous lipid is purified and collected.

[0069] Preferably, in the method for extracting the endogenous lipid in the bulk crop, the extraction is carried out at a pressure of 10-40 Mpa and a temperature of 50-80 ℃, preferably in an extraction tank; the separation is carried out at a pressure of 2-20 Mpa and a temperature of 50-80 ℃, preferably in a separation kettle; and the purification is carried out by rotary evaporation at a temperature of 50-80 ℃, preferably in a rotary evaporator.

[0070] Preferably, the method for extracting the endogenous protein in the fruit of the bulk crop is as follows: the immature bulk crop fruit is mixed with distilled water, centrifuged, and the supernatant is isoelectrically precipitated, and then the endogenous protein is obtained after dialysis.

[0071] Preferably, in the method for extracting endogenous proteins in bulk crop fruit powder, the bulk crop fruit powder is mixed with distilled water at a ratio of 1:20 (w / w), centrifuged at 6000 x g for 20 min, the supernatant is adjusted to pH 4.0-6.0 with 2M hydrochloric acid (HCl) for isoelectric precipitation, and after isoelectric precipitation, centrifuged at 8000 x g for 30 min; the obtained precipitate is redissolved in distilled water, and the endogenous globulin and albumin are dialyzed with a 7-11 kDa dialysis membrane.

[0072] Preferably, the method for constructing slow-digesting bulk crop amylopectin-endogenous protein-endogenous lipid nanoassemblies is: adding endogenous lipids and proteins in proportion after mixing bulk crop starch with distilled water, and preferably performing the self-assembly reaction quickly in a high-precision magnetic stirrer; and the obtained bulk crop amylopectin ternary assembly is treated by high-power ultrasonic (preferably in an ultrasonic cell disruptor) to obtain amylopectin ternary nanoassemblies.

[0073] Preferably, in the method for constructing slow-digesting bulk crop amylopectin ternary nanoassemblies, the bulk crop amylopectin is mixed with distilled water at a ratio of 1-6:30 (w / w) in a magnetic stirrer, and lipids and proteins are added at a mass ratio of amylopectin:endogenous lipids:endogenous proteins of 1:0.05-0.15:0.05-0.20, the treatment temperature is 80-100 ℃, the instrument speed is 100-400 rpm, the treatment time is 10-30 min (magnetic stirrer parameters (for constructing ternary assemblies)), the ultrasonic power of the nano-sized ternary assemblies is 300-800 W, and the treatment time is 0.2-3 h (ultrasonic parameters (for reducing particle size and increasing CI)); more preferably, the amylopectin is mixed with distilled water at a ratio of 2-5:30 (w / w), the lipids and proteins are added at a ratio of 1:0.065-0.12:0.07-0.17 with reference to the mass of amylopectin, the treatment temperature is 85-95 ℃, the instrument speed is 150-350 rpm, the treatment time is 15-25 min, the ultrasonic power is 400-700 W, and the treatment time is 1.0-2.2 h; most preferably, the amylopectin is mixed with distilled water at a ratio of 3-4:30 (w / w), the lipids and proteins are added at a ratio of 1:0.075-0.10:0.09-0.13, the treatment temperature is 88-92 ℃, the instrument speed is 200-300 rpm, the treatment time is 18-22 min, the ultrasonic power is 500-600 W, and the treatment time is 1.4-1.8 h.

[0074] Preferably, in the method for constructing the slowly digestible bulk crop amylopectin ternary nano-assemblies, the drying is vacuum freeze drying; and the specific parameters are: material layer temperature of 30-60 DEG C, vacuum degree of 30 Pa, machine internal cold trap temperature of-80 DEG C, temperature rising speed of 2-10 DEG C / min, freeze drying for 24 h, and freeze drying to moisture mass percentage of ≤10% under the above conditions.

[0075] In the processes of amylopectin extraction, endogenous protein extraction, endogenous lipid extraction and amylopectin ternary nano-assemblies construction, the power, processing time, temperature, rotation speed and the like of the instrument all belong to the overall technical solution, and are mutually supported and interacted in function, and the above-mentioned cooperation can solve the technical problems of the present application and achieve the technical effects of the present application, that is, the particle size of the bulk crop amylopectin ternary assemblies reaches the nanometer level, the slow digestibility, CI, anti-thermal decomposition, anti-enzymatic hydrolysis and texture characteristics are significantly improved, the glucose can be slowly released, the texture characteristics (chewing property) are improved, and the particle stability is good.

[0076] The present application constructs slowly digestible bulk crop amylopectin ternary nano-assemblies based on high-power ultrasonic assistance, and the process method is simple and convenient, time-consuming is short, no toxic and harmful reagents are used in the construction process, the preparation process is greatly optimized, compared with other methods, the present application greatly reduces the particle size while saving process cost and increasing self-assembly speed, thereby greatly increasing the self-assembly index, increasing the SDS structure, and providing nutrition slowly and continuously, greatly improving the taste, and not polluting the environment, low cost, and suitable for industrial automatic large-scale production.

[0077] A method for constructing slowly digestible bulk crop amylopectin-endogenous lipid-endogenous protein nano-assemblies, and the nano-assemblies are prepared according to the method, wherein: the purity of the amylopectin is 91.56-98.93%, the purity of the endogenous lipid is 89.95-94.33%, the purity of the endogenous protein is 88.01-93.94%, the two-dimensional infrared full spectrum of the ternary nano-assemblies all presents obvious ternary compound peaks, the SDS content is 37.71-43.28, the logarithmic slope mode (LOS) digestion kinetics shows two-stage digestibility, the first-stage digestion kinetics rate (k1) is 2.43-2.92*10 -2 min - , the second-stage digestion kinetics rate (k2) is 1.44-2.17*10 - ² min - , the particle morphology presents irregular polygon, the particle surface is relatively compact, the number of enzymatic cavities is small, the particle size is 99.35-155.62 nm, the CI is 66.80-82.58%, and the molecular weight is 2.55-5.25*10 7g / mol, the characteristic distance of the crystalline lamella and amorphous region is 6.66-10.37 Å, all are V-type crystal structure, the crystallinity is 18.87-23.68%, the mean square nanometer surface roughness is 8.01-12.17 nm, the gelatinization temperature is 92.12-98.65 ℃, the hardness is 33.66-55.15 g, the cohesion is 0.55-0.81, the elasticity is 0.52-0.88, and the three freeze-thaw water release rate is in the range of 10.66-22.37% to 20.48-36.67%.

[0078] The slowly digestible major amylopectin-endogenous lipid-endogenous protein nanoassembly can be used as a precise nutrition element to regulate the dietary structure, realize the control of the blood glucose level of the body, and is expected to become one of important means for preventing the occurrence of glucose metabolism disorder.

[0079] At present, although the starches of major crops such as corn, wheat, barley, rice, sorghum, waxy corn, potato and the like have a certain anti-digestibility, they cannot be eaten directly, and must be processed to be completely gelatinized before being edible. However, the spatial configuration of the starch after heat processing is irreversibly changed, the particle size is greatly increased, and the surface layer of the gelatinized starch granule has a cavity, so that the digestive enzymes can quickly and rapidly enzymatically hydrolyze from the inside of the granule, so that the digestibility is greatly increased, thereby leading to the risk of too fast digestion rate and glucose metabolism disorder. In addition, the high polymerization force of the original starch molecular crosslinking network makes it difficult to remove the endogenous proteins and fats attached to the surface of the starch granule in the mass production of starch. Therefore, a small amount of amylose-lipid-protein assembly is preferentially formed in the industrialized heat processing of starch, which improves the flavor, texture, taste, sensory characteristics and anti-digestibility of the starch paste. The starch ternary assembly granule is composed of a poor stability type I complex and a good stability type IIb complex, and therefore has a “multi-system supermolecular structure”. The above characteristics make the ternary assembly have a large particle size, about 800-10000 nm, an irregular morphology and a sponge-like appearance. In order to solve this problem, the present application provides a construction method of slowly digestible major amylopectin-endogenous lipid-endogenous protein nanoassembly.

[0080] The present application greatly reduces the construction steps, and the amylopectin, lipid and protein are self-assembled after high-temperature gelatinization, supplemented with ultrasonic treatment, and the branched amylopectin is self-assembled efficiently in a medium of distilled water without adding too many reagents, so that the branched amylopectin has high purity, stable particle size of nanometer (below 200 nm), no sudden nutrient release in the enzyme hydrolysis stage, and the prepared product reaches the quality of batch application while saving raw materials. At the same time, the emulsifying activity and antioxidant property of endogenous protein and lipid can increase the preservative property of the starch-lipid-protein, and therefore, the present application is even superior to the existing level in some aspects.

[0081] The present application discloses the following technical effects:

[0082] The present application provides a construction method of slowly-digestible bulk branched amylopectin-endogenous lipid-endogenous protein nanometer assembly, and the method belongs to a physical heat treatment process, and has the advantages of simple preparation mode, suitability for mass production, green and environmentally-friendly method, high CI of the nanometer assembly particles, uniformity, significantly improved anti-hydrolysis property, and good stability in cold storage. In addition, in vivo experiments after rat gavage show that eating the endogenous ternary compound can relieve the rising rate of blood glucose index, increase the acid production efficiency of probiotic fermentation, thereby improving the intestinal pH environment to synergistically proliferate probiotics, and reduce blood lipids to reduce the body fat rate, so that the present application can prevent or relieve type II diabetes, high blood lipids, high blood pressure and obesity.

[0083] In the examples 1 to 6, comparative examples 1 to 4 and effect examples of the present application, the raw materials and reagents used can be purchased from the market.

[0084] The present application will be further described below in combination with examples:

[0085] Example 1: Construction of slowly-digestible waxy corn amylopectin endogenous ternary nanometer assembly by high-power ultrasonic assisted technology

[0086] Extraction of waxy corn starch amylopectin: 100 g of corn starch was added to 3 kg of distilled water, and gelatinized in a large water bath at 100 DEG C for 20 min, cooled to 25 DEG C, and the gelatinized liquid was taken and centrifuged at 5000xg for 10 min to take the precipitate, and the step was repeated twice, and 80 g of the precipitate was added to 720 g of anhydrous ethanol, and dispersed by ultrasonic treatment in a large ultrasonic cleaning machine at 150 W for 10 min, and centrifuged at 5000xg for 10 min, and the precipitate was washed for 3 times, and 52.95 g of branched amylopectin was collected.

[0087] Extraction of endogenous lipid from mature waxy corn: 2 kg of fully mature corn kernels were dried at 55 ℃ for 24 h, ground and sieved through a 50 mesh sieve. 1 kg of the corn powder was placed in the extraction tank of a supercritical CO2 extraction instrument, and the pressure was set to 25 MPa (extraction tank) and 12 MPa (separation kettle), and the temperature was set to 65 ℃. The crude lipid was extracted. The crude lipid was purified by rotary evaporation at 65 ℃, and 79.68 g of endogenous lipid was obtained.

[0088] Extraction of endogenous protein from immature waxy corn: 2 kg of corn kernels were dried, ground and sieved. 500 g of immature corn powder was mixed with 10 kg of distilled water in a blender (400 rpm, 20 min), and the suspension was centrifuged at 6000 x g for 20 min. The supernatant was collected. The supernatant was adjusted to pH 5.0 with 2 M HCl to precipitate the protein, and centrifuged (6000 x g, 20 min). The precipitate was resuspended and dialyzed through a 9 kDa dialysis membrane to obtain 18.16 g of endogenous globulin and albumin.

[0089] Construction of waxy corn amylopectin-endogenous lipid-endogenous protein nanoassembly: 50 g of corn amylopectin was placed in 700 g of distilled water, and 3.75 g of endogenous lipid and 4.75 g of endogenous protein were added to the pool, and treated at 90 ℃ for 20 min; the rotation speed was maintained at 250 rpm, and after cooling to room temperature, the sample was dehydrated using a vacuum filtration machine, and the wet sample was collected. The wet sample was added with excess water and ultrasonicated in a cell disruptor at 520 W for 1.5 h.

[0090] Collection of ternary nanoassembly: the free lipid and protein were removed by washing with 50% ethanol for 3 times. The flocculent was dried by freeze vacuum drying machine (45 ℃, heating rate 6 ℃ / min, cold trap-80 ℃, vacuum degree 30 Pa, 24 h) to a moisture content of ≤10%. The freeze-dried sample was ground and sieved through an 800 mesh sieve, and packaged to obtain the ternary nanoassembly, and the packaged ternary complex was 56.87 g.

[0091] The waxy corn amylopectin ternary nanoassembly of the present application was detected for in vitro digestion characteristics, complex characteristics, multi-level spatial structure, and physicochemical properties. The purity of the waxy corn amylopectin was 98.93%, the purity of the endogenous lipid was 94.33%, and the purity of the endogenous protein was 93.94%. The two-dimensional infrared full spectrum of the ternary nanoassembly showed obvious ternary complex peaks, the SDS content was 43.28, and the LOS digestion kinetics showed two-stage digestibility, k1 was 2.43 x 10 - ² min - ¹, and k2 was 1.44 x 10 - ² min -¹, although the particle morphology is irregular polygon, it does not lose the morphology order, the particle surface is smooth and tight, the average particle size is 99.87 nm, CI is 82.58%, the molecular weight is 5.25×10 7 g / mol, the characteristic distance between the crystalline lamella and the amorphous region is 6.66Å, it is V-type crystalline structure, the crystallinity is 23.68%, the mean square nanometer surface roughness is 8.01 nm, the gelation temperature is 98.65℃, the hardness is 33.66g, the cohesion is 0.55, the elasticity is 0.52, and the water release rate after three freeze-thaw cycles is in the range of 10.66~22.37%.

[0092] Example 2 Construction of endogenous ternary nano-assembly of slowly digestible wheat amylopectin by high-power ultrasonic assisted technology

[0093] Extraction of wheat starch amylopectin: 3 kg of distilled water was added to 200 g of wheat starch, and the mixture was pasted in a large water bath at 100℃ for 20 min. The pasting liquid was cooled to 25℃, and then centrifuged at 5000×g for 10 min. The sediment was collected and repeated twice. 100 g of the sediment was added to 900 g of anhydrous ethanol, and then dispersed by ultrasonic in a large ultrasonic cleaning machine at 160 W for 10 min. The mixture was centrifuged at 5000×g for 10 min, and the sediment was washed for three times. Finally, 133.65 g of amylopectin was collected.

[0094] Extraction of endogenous lipid from mature wheat: 4 kg of fully matured wheat grains were dried at 55℃ for 24 h, and then ground and sieved through a 50 mesh sieve. 2 kg of the wheat powder was placed in the extraction tank of a supercritical CO2 extraction instrument, and the pressure was set to 30 MPa (extraction tank) and 15 MPa (separation kettle), and the temperature was set to 70℃. The crude lipid was obtained by extraction. The crude lipid was purified by rotary evaporation at 70℃, and 166.31 g of endogenous lipid was obtained.

[0095] Extraction of endogenous protein from immature wheat: 4 kg of immature wheat was dried, ground and sieved. 2 kg of the immature wheat powder was mixed with 40 kg of distilled water in a blender (400 rpm, 20 min), and then the suspension was centrifuged at 6000×g for 20 min. The supernatant was collected. The supernatant was adjusted to pH 5.5 with 2 M HCl to precipitate the protein, and then centrifuged (6000×g, 20 min). The precipitate was resuspended and dialyzed through a 9.5 kDa dialysis membrane, and 40.17 g of endogenous globulin and albumin was obtained.

[0096] Construction of wheat amylopectin-endogenous lipid-endogenous protein nanosassembly: wheat amylopectin 100 g was placed in 1 kg distilled water, and 9 g of endogenous lipid and 11 g of endogenous protein were continuously added to the pool, and treated at 91 ℃ for 20 min; the rotation speed was maintained at 280 rpm, and after cooling to room temperature, vacuum filtration was used for dehydration, and the wet sample was collected. The wet sample was added with excess water and ultrasonicated in a cell disrupter at 580 W for 1.7 h.

[0097] Collection of ternary nanosassembly: free lipid, protein was removed by washing 3 times with 50% ethanol. The flocculent was dried by freeze vacuum dryer (50 ℃, heating rate 7 ℃ / min, cold trap-80 ℃, vacuum degree 30 Pa, 24 h) to a moisture content of ≤10%. The freeze-dried sample was ground and passed through an 800-mesh sieve, and the ternary nanosassembly was obtained after packaging, and the ternary complex 115.17 g was obtained after packaging.

[0098] The wheat amylopectin ternary nanosassembly of the application was detected for in vitro digestion characteristics, complex characteristics, multi-level spatial structure, and physicochemical properties. The purity of amylopectin was 96.05%, the purity of endogenous lipid was 93.11%, and the purity of endogenous protein was 92.92%. The two-dimensional infrared full spectrum of the ternary nanosassembly showed obvious ternary complex peaks, the SDS content was 42.56, the LOS digestion kinetics showed two-stage digestion, k1 was 2.50 × 10 - ² min - ¹, k2 was 1.65 × 10 - ² min - ¹, the particle morphology showed irregular polygon, the morphology uniformity was high, the particle surface was smooth and tight, the average particle size was 99.35 nm, the CI was 80.81%, the molecular weight was 5.20 × 10 7 g / mol, the characteristic distance between the crystalline lamella and the amorphous region was 6.88 Å, it was a V-type crystalline structure, the crystallinity was 22.15%, the mean square nanosurface roughness was 8.81 nm, the gelatinization temperature was 97.95 ℃, the hardness was 35.17 g, the cohesion was 0.59, the elasticity was 0.62, and the three freeze-thaw water separation rate was in the range of 12.37~25.57%.

[0099] Example 3 Construction of slowly digestible potato amylopectin endogenous ternary nanosassembly by high-power ultrasonic assisted technology

[0100] Extraction of amylopectin from potato starch: 150 g of potato starch was added to 1.5 kg of distilled water, and gelatinized in a large water bath at 100 °C for 20 min. The gelatinized solution was cooled to 25 °C, and then centrifuged at 5000 x g for 10 min. This step was repeated twice, and the precipitate was collected. The precipitate was added to 792 g of anhydrous ethanol, and dispersed in a large ultrasonic cleaner at 100 W for 10 min. The mixture was centrifuged at 5000 x g for 10 min, and the precipitate was washed three times. The amylopectin was collected as 70.44 g.

[0101] Extraction of endogenous lipids from mature potato: 3 kg of fully mature potato was dried at 55 °C for 24 h, and then ground and sieved through a 50 mesh screen. 1.5 kg of the potato powder was placed in the extraction tank of a supercritical CO2 extraction instrument, and the pressure was set to 10 MPa (extraction tank) and 2 MPa (separation kettle). The temperature was set to 50 °C, and the crude lipids were extracted. The crude lipids were purified by rotary evaporation at 50 °C, and 124.08 g of endogenous lipids were obtained.

[0102] Extraction of endogenous proteins from immature potato: 3 kg of potato was dried, ground, and sieved. 1.5 kg of the immature potato powder was mixed with 30 kg of distilled water in a blender (400 rpm, 20 min), and the suspension was centrifuged at 6000 x g for 20 min. The supernatant was collected. The supernatant was adjusted to pH 4.0 with 2 M HC1 to precipitate the proteins, and then centrifuged (6000 x g, 20 min). The precipitate was resuspended, and then dialyzed through a 7 kDa dialysis membrane to obtain 29.97 g of endogenous globulin and albumin.

[0103] Construction of potato amylopectin-endogenous lipid-endogenous protein nanoassemblies: 70 g of amylopectin was placed in 2.1 kg of distilled water, and 3.5 g of endogenous lipids and 3.5 g of endogenous proteins were added to the pool. The mixture was treated at 80 °C for 10 min, and the rotation speed was maintained at 100 rpm. After cooling to room temperature, the mixture was dehydrated using a vacuum filtration machine, and the wet sample was collected. The wet sample was added to excess water, and then sonicated in a cell disrupter at 300 W for 0.2 h.

[0104] Collection of ternary nanoassemblies: The ternary nanoassemblies were washed three times with 50% ethanol to remove free lipids and proteins. The flocculent was dehydrated by freeze-drying in a freeze-drying machine (30 °C, heating rate of 2 °C / min, cold trap at -80 °C, vacuum degree of 30 Pa, and time of 24 h) to a water content of ≤10%. The freeze-dried sample was ground and sieved through an 800 mesh screen, and then packaged to obtain the ternary nanoassemblies. The packaged ternary nanoassemblies were obtained as 72.66 g.

[0105] The potato amylopectin ternary nano-assemblies are detected for in-vitro digestion characteristics, complex characteristics, multi-level spatial structure, physical and chemical characteristics, the purity of amylopectin is 91.56%, the purity of endogenous lipid is 89.95%, the purity of endogenous protein is 88.99%, the two-dimensional infrared full spectrum of the ternary nano-assemblies presents obvious ternary complex peaks, the SDS content is 37.71, the LOS digestion kinetics shows two-stage digestion, k1 is 2.92*10 - ² min - ¹, k2 is 2.17*10 - ² min - ¹, the particle morphology presents irregular polygon, the morphology uniformity is poor, the particle surface is accompanied by part of the milky protrusion, the fractal dimension is higher, but is more compact, the average particle size is 155.62 nm, CI is 66.80%, the molecular weight is 2.55*10 7 g / mol, the characteristic distance between the crystalline lamella and the amorphous region is 10.37 Å, it is a V-type crystalline structure, the crystallinity is 18.87%, the mean square nano-surface roughness is 12.17 nm, the gelatinization temperature is 92.12 ℃, the hardness is 55.15 g, the cohesion is 0.81, the elasticity is 0.88, and the three-time freeze-thaw water extraction rate is in the interval of 20.48-36.67%.

[0106] Example 4: High-power ultrasonic assisted technology for constructing slowly digestible barley amylopectin endogenous ternary nano-assemblies

[0107] Extracting barley starch amylopectin: 5 kg of distilled water is added to 300 g of barley starch, and the starch is gelatinized in a large water bath at 100 ℃ for 20 min. The gelatinized liquid is cooled to 25 ℃, and then centrifuged at 5000* g for 10 min to obtain the precipitate. The above step is repeated twice, and then 201.13 g of the precipitate is added to 2538 g of anhydrous ethanol. The mixture is dispersed by ultrasonic in a large ultrasonic cleaning machine at 200 W for 10 min, and then centrifuged at 5000* g for 10 min. The above step is repeated three times, and then 190.65 g of amylopectin is collected.

[0108] Extracting mature barley endogenous lipid: 5 kg of completely mature barley is dried at 55 ℃ for 24 h, and then ground and sieved through a 50 mesh sieve. 2.5 kg of the barley powder is placed in the extraction tank of a supercritical CO2 extraction instrument, and the pressure is set to 15 MPa (extraction tank) and 7 MPa (separation kettle), and the temperature is set to 60 ℃. The crude lipid is obtained by extraction. The crude lipid is purified by rotary evaporation at 60 ℃, and 196.01 g of endogenous lipid is obtained.

[0109] Extraction of endogenous proteins from immature barley: 5 kg of barley was dried, ground and sieved. 2.5 kg of immature barley powder was mixed with 50 kg of distilled water in a blender (400 rpm, 20 min), and the supernatant was obtained by centrifugation of the suspension at 6000 x g for 20 min. The supernatant was precipitated by adjusting the pH to 6 with 2 M HCl, and the precipitate was centrifuged (6000 x g, 20 min), resuspended, and dialyzed through a 11 kDa dialysis membrane to obtain 40.14 g of endogenous globulin and albumin.

[0110] Construction of barley amylopectin-endogenous lipid-endogenous protein nanoassemblies: 150 g of amylopectin was placed in 750 g of distilled water, and 22.5 g of endogenous lipid and 30 g of endogenous protein were added to the pool, which was treated at 100 °C for 30 min; the rotation speed was maintained at 400 rpm, and after cooling to room temperature, the wet sample was dehydrated using a vacuum filtration machine. The wet sample was added to an excess of water and ultrasonicated in a cell disrupter at 800 W for 3 h.

[0111] Collection of ternary nanoassemblies: free lipids and proteins were removed by washing with 50% ethanol for 3 times. The flocculent was dehydrated by freeze-drying in a freeze-drying machine (60 °C, heating rate 10 °C / min, cold trap -80 °C, vacuum degree 30 Pa, 24 h) to a moisture content of ≤ 10%. The freeze-dried sample was ground and sieved through an 800 mesh sieve, and the ternary nanoassemblies were obtained after packaging, and the ternary complex was obtained after packaging, and the weight of the ternary complex was 195.00 g.

[0112] The barley amylopectin ternary nanoassemblies of the present application were detected for in vitro digestion characteristics, complex characteristics, multi-level spatial structure, physicochemical properties, and the purity of the amylopectin was 92.98%, the purity of the endogenous lipid was 91.07%, and the purity of the endogenous protein was 88.01%. The two-dimensional infrared full spectrum of the ternary nanoassemblies showed obvious ternary complex peaks, the SDS content was 39.05, the LOS digestion kinetics showed two-stage digestion, k1 was 2.69 x 10 - ² min - ¹, k2 was 2.10 x 10 - ² min - ¹, showed irregularly aggregated particle morphology, the particle surface was accompanied by protrusions, a small amount of enzymatic cavities were visible, the roughness was high, but the overall was tight, the average particle size was 146.65 nm, the CI was 73.91%, and the molecular weight was 3.13 x 10 7g / mol, the characteristic distance of crystalline lamella and amorphous region is 9.11 Å, V-type crystal structure, crystallinity is 20.12%, the mean square nanometer surface roughness is 10.88 nm, gelatinization temperature is 92.92℃, hardness is 52.20 g, cohesion is 0.72, elasticity is 0.75, and the water release rate after three freeze-thaw cycles is in the range of 19.39~33.20%.

[0113] Example 5 High-power ultrasonic-assisted technology for constructing slow digestion rice amylopectin endogenous ternary nano-assembly

[0114] Extraction of rice starch amylopectin: 20 kg of distilled water was added to 400 g of rice starch, and the mixture was pasted in a large water bath at 100℃ for 20 min. After cooling to 25℃, the pasting liquid was taken and centrifuged at 5000xg for 10 min. This step was repeated twice, and 320 g of the precipitate was collected and added to 2880 g of anhydrous ethanol. The mixture was ultrasonically dispersed in a large ultrasonic cleaner at 130 W for 10 min, and then centrifuged at 5000xg for 10 min. The precipitate was washed three times, and 285.21 g of amylopectin was collected.

[0115] Extraction of mature rice endogenous lipid: 8 kg of completely mature rice was dried at 55℃ for 24 h, ground and sieved through a 50 mesh sieve. 4 kg of rice powder was placed in the extraction tank of a supercritical CO2 extraction instrument, and the pressure was set to 15 MPa (extraction tank) and 8 MPa (separation kettle), and the temperature was set to 60℃. The crude lipid was obtained by extraction. The crude lipid was purified by rotary evaporation at 60℃, and 300.11 g of endogenous lipid was obtained.

[0116] Extraction of immature rice endogenous protein: 8 kg of rice was dried, ground and sieved. 4 kg of immature rice powder was mixed with 80 kg of distilled water in a blender (400 rpm, 20 min), and the suspension was centrifuged at 6000xg for 20 min. The supernatant was collected. The supernatant was adjusted to pH 4.5 with 2 M HCl to precipitate the protein, and centrifuged (6000xg, 20 min). The precipitate was resuspended and dialyzed through an 8 kDa dialysis membrane to obtain 82.18 g of endogenous globulin and albumin.

[0117] Construction of rice amylopectin-endogenous lipid-endogenous protein nano-assembly: 250 g of amylopectin was placed in 3750 g of distilled water, and 16.25 g of endogenous lipid and 17.50 g of endogenous protein were added to the pool, and the mixture was treated at 85℃ for 13 min. The rotation speed was maintained at 150 rpm, and after cooling to room temperature, the mixture was dehydrated using a vacuum filter, and the wet sample was collected. The wet sample was added with excess water and ultrasonicated in a cell disruptor at 400 W for 1 h.

[0118] Collect ternary nano-assemblies: use 50% ethanol to wash 3 times to remove free lipids, proteins. The flocculation is dried by freeze vacuum dryer (40 ℃, the temperature rising speed is 4 ℃ / min, the cold trap is -80 ℃, the vacuum degree is 30 Pa, 24 h) to the moisture mass percentage content ≤10%. The freeze-dried sample is ground and passed through an 800 mesh sieve, and the ternary nano-assemblies are obtained after packaging, and the ternary complex 271.64 g is obtained after packaging.

[0119] The amylopectin ternary nano-assemblies of the rice are detected for in vitro digestion characteristics, complex characteristics, multi-level spatial structure, and physicochemical properties. The purity of the amylopectin is 94.55%, the purity of the endogenous lipids is 92.61%, and the purity of the endogenous proteins is 90.18%. The two-dimensional infrared full spectrum of the ternary nano-assemblies presents obvious ternary complex peaks. The SDS content is 41.33. The LOS digestion kinetics shows two-stage digestibility. k1 is 2.60×10 - ² min - ¹, k2 is 1.90×10 - ² min - ¹. The particle morphology is not obviously broken, is in irregular morphology, has poor morphology uniformity, has low particle surface roughness, has fewer enzymatic cavities, has an average particle size of 105.91 nm, has a CI of 75.11%, has a molecular weight of 3.33×10 7 g / mol, has a characteristic distance of 8.75 Å between the crystalline lamella and the amorphous region, has a V-type crystalline structure, has a crystallinity of 20.97%, has a mean square nanosurface roughness of 9.10 nm, has a gelatinization temperature of 94.13℃, has a hardness of 50.67 g, has a cohesion of 0.68, has an elasticity of 0.70, and has a three-time freeze-thaw water separation rate in the interval 17.61~32.66%.

[0120] Example 6: Construction of slow-digesting sorghum amylopectin endogenous ternary nano-assemblies by high-power ultrasonic assisted technology

[0121] Extracting sorghum starch amylopectin: 9 kg distilled water is added to 450 g rice starch, and the starch is gelatinized in a large water bath at 100 ℃ for 20 min. The gelatinized liquid is cooled to 25 ℃, and the precipitate is obtained by centrifugation at 5000×g for 10 min. The step is repeated twice, and 360 g of the precipitate is added to 3240 g of anhydrous ethanol. The mixture is dispersed by ultrasonic in a large ultrasonic cleaning machine at 180 W for 10 min, and then centrifuged at 5000×g for 10 min. The precipitate is washed 3 times, and 311.80 g of amylopectin is collected.

[0122] Extraction of endogenous lipids from mature sorghum: 12 kg of fully mature sorghum was dried at 55 ℃ for 24 h, ground, and passed through a 50-mesh sieve. 6 kg of sorghum powder was placed in the extraction vessel of a supercritical CO2 extractor, with the pressure set at 35 MPa (extraction vessel) and 18 MPa (separation vessel), and the temperature at 70 ℃ for extraction to obtain crude lipids. The crude lipids were purified by rotary evaporation at 70 ℃, yielding 400.23 g of endogenous lipids.

[0123] Extraction of endogenous proteins from immature sorghum: 12 kg of sorghum was dried, ground, and sieved. 6 kg of immature sorghum powder was mixed with 120 kg of distilled water in a mixer (400 rpm, 20 min). The suspension was centrifuged at 6000×g for 20 min, and the supernatant was collected. The supernatant was adjusted to pH 5.7 with 2 M HCl to precipitate the proteins. The mixture was centrifuged (6000×g, 20 min), the precipitate was reconstituted, and dialyzed through a 10 kDa dialysis membrane to obtain 133.46 g of endogenous globulins and albumin.

[0124] Construction of sorghum amylopectin-endogenous lipid-endogenous protein nanoassemblies: 300 g of amylopectin was placed in 1800 g of distilled water, and 36 g of endogenous lipids and 51 g of endogenous protein were added to the same container. The mixture was treated at 95 ℃ for 25 min, with the rotation speed maintained at 350 rpm. After cooling to room temperature, the samples were dehydrated using a vacuum filter, and the wet samples were collected. Excess water was added to the wet samples, and the samples were sonicated at 700 W for 2.2 h in a cell disruptor.

[0125] Ternary nanoassemblies were collected by washing three times with 50% ethanol to remove free lipids and proteins. The flocculent material was then dried and dehydrated in a freeze-dryer (57 ℃, heating rate 8 ℃ / min, cold trap -80 ℃, vacuum degree 30 Pa, 24 h) until the moisture content was ≤ 10%. The freeze-dried sample was ground, passed through an 800-mesh sieve, and packaged to obtain ternary nanoassemblies. 370.75 g of the ternary composite was obtained after packaging.

[0126] The sorghum amylopectin ternary nanoassemblies of this invention were tested for in vitro digestibility, complex characteristics, multi-level spatial structure, and physicochemical properties. The amylopectin purity was 95.05%, the endogenous lipid purity was 91.97%, and the endogenous protein purity was 91.08%. The two-dimensional infrared full spectrum of the ternary nanoassemblies showed a clear ternary complex peak, the SDS content was 41.85%, and the LOS digestion kinetics showed two-stage digestibility with a k1 of 2.54 × 10⁻⁶. - ² min - ¹, k² is 1.78 × 10 - ² min -¹, the particle morphology is relatively complete, the particles are in a random state, the particle morphologies differ greatly, no obvious enzymolysis channels are observed on the particle surface, the fractal dimension is low, the average particle size is 101.99 nm, the CI is 76.42%, the molecular weight is 4.01 x 10 7 g / mol, the characteristic distance between the crystalline lamella and the amorphous region is 7.65 Å, it is a V-type crystalline structure, the crystallinity is 21.72%, the mean square nanosurface roughness is 8.95 nm, the gelation temperature is 95.89 °C, the hardness is 46.23 g, the cohesiveness is 0.64, the elasticity is 0.65, and the water release rate after three freeze-thaw cycles is in the range of 15.82~30.44%.

[0127] Comparative Example 1 Construction of Endogenous Ternary Micro-nano Assembly of Corn Starch by High-power Ultrasonic Assisted Technology

[0128] Extraction of endogenous lipid from mature corn: 2 kg of completely mature corn kernels were dried at 55 °C for 24 h, ground and sieved through a 50 mesh sieve. 1 kg of corn powder was placed in a supercritical CO2 extraction instrument extraction tank, the pressure was set to 25 MPa (extraction tank) and 12 MPa (separation kettle), and the temperature was 65 °C. Crude lipid was obtained by extraction. The crude lipid was purified by rotary evaporation at 65 °C to obtain 72.55 g of endogenous lipid.

[0129] Extraction of endogenous protein from immature corn: 2 kg of corn kernels were dried, ground and sieved. 500 g of immature corn powder was mixed with 10 kg of distilled water in a blender (400 rpm, 20 min), and the supernatant was obtained by centrifugation of the suspension at 6000 x g for 20 min. The supernatant was precipitated by adjusting the pH to 5.0 with 2 M HCl, centrifuged (6000 x g, 20 min), and the precipitate was redissolved and dialyzed through a 9 kDa dialysis membrane to obtain 17.15 g of endogenous globulin and albumin.

[0130] Construction of corn starch-endogenous lipid-endogenous protein nano assembly: 50 g of corn starch was placed in 700 g of distilled water, and 3.75 g of endogenous lipid and 4.75 g of endogenous protein were added to the tank, which was treated at 90 °C for 20 min; the rotation speed was maintained at 250 rpm, and after cooling to room temperature, the wet sample was collected by vacuum filtration. The wet sample was added with excess water and ultrasonicated in a cell disruptor at 520 W for 1.5 h.

[0131] Collection of ternary micro-nano assembly: 3 times of 50% ethanol washing to remove free lipids, proteins. The flocculation was dried by freeze vacuum dryer (45 ℃, heating rate 6 ℃ / min, cold trap-80 ℃, vacuum degree 30 Pa, 24 h) to moisture content ≤10%. The freeze-dried sample was ground and passed through an 800-mesh sieve, and the ternary nano assembly was obtained after packaging, and the ternary complex 51.85 g was obtained after packaging.

[0132] The corn starch ternary micro-nano assembly was detected for in vitro digestion characteristics, complex characteristics, multi-level spatial structure, and physicochemical properties. The endogenous lipid purity was 88.71%, the endogenous protein purity was 86.52%, the two-dimensional infrared full spectrum of the ternary nano assembly showed obvious ternary complex peaks, the SDS content was 32.28, the LOS digestion kinetics showed two-stage digestion, k1 was 3.17×10 - ² min - ¹, k2 was 2.01×10 - ² min - ¹, the particle morphology showed irregular polygon and loss of sequence, presented broken, high roughness, small enzyme-cavity on the surface, the average particle size was 299.15 nm, CI was 61.44%, the molecular weight was 1.86×10 7 g / mol, the characteristic distance between the crystalline lamella and the amorphous region was 12.51 Å, it was a V-type crystalline structure, the crystallinity was 17.12%, the mean square nanosurface roughness was 12.04 nm, the gelatinization temperature was 92.08 ℃, the hardness was 95.74 g, the cohesion was 0.85, the elasticity was 0.90, and the three freeze-thaw water extraction rate was in the range of 27.59~40.88%.

[0133] Comparative Example 2 High-power ultrasonic assisted technology to construct endogenous ternary micro-nano assembly of potato starch

[0134] Extraction of endogenous lipids from mature potatoes: 3 kg of fully mature potatoes were dried at 55 ℃ for 24 h, ground and passed through a 50-mesh sieve. 1.5 kg of potato powder was placed in a supercritical CO2 extraction instrument extraction tank, the pressure was set to 10 MPa (extraction tank) and 2 MPa (separation kettle), the temperature was 50 ℃, and the crude lipid was extracted. The crude lipid was purified by rotary evaporation at 50 ℃, and 119.76 g of endogenous lipid was obtained.

[0135] Extraction of immature potato endogenous protein: 3 kg of potato was dried, ground and sieved. 1.5 kg of immature potato powder was mixed with 30 kg of distilled water in a blender (400 rpm, 20 min), and the suspension was centrifuged at 6000 x g for 20 min. The supernatant was collected. The supernatant was precipitated with 2 M HCl to pH 4.0, centrifuged (6000 x g, 20 min), and the precipitate was resuspended and dialyzed through a 7 kDa dialysis membrane to obtain 32.72 g of endogenous globulin and albumin.

[0136] Construction of potato starch-endogenous lipid-endogenous protein micro-nano assembly: 70 g of amylopectin was placed in 2.1 kg of distilled water, and 3.5 g of endogenous lipid and 3.5 g of endogenous protein were added to the pool, and treated at 80 ℃ for 10 min; the rotation speed was maintained at 100 rpm, and after cooling to room temperature, vacuum filtration was used for dehydration, and the wet sample was collected. The wet sample was added with excess water and ultrasonicated in a cell crusher at 300 W for 0.2 h.

[0137] Collection of ternary micro-nano assembly: free lipid and protein were removed by washing with 50% ethanol for 3 times. The flocculent was dried by freeze vacuum drying machine (30 ℃, heating rate 2 ℃ / min, cold trap-80 ℃, vacuum degree 30 Pa, 24 h) to a moisture content of ≤10%. The freeze-dried sample was ground and sieved through an 800 mesh sieve, and the ternary nano assembly was obtained after packaging, and the ternary complex was obtained after packaging, with a weight of 72.66 g.

[0138] The mature potato amylopectin ternary nano assembly was detected for in vitro digestion characteristics, complex characteristics, multi-level spatial structure, and physicochemical properties. The purity of endogenous lipid was 89.95%, the purity of endogenous protein was 88.99%, the two-dimensional infrared full spectrum of the ternary nano micro assembly showed obvious ternary complex peaks, the SDS content was 28.72, the LOS digestion kinetics showed two-stage digestion, k1 was 4.19 x 10 - ² min - ¹, k2 was 2.88 x 10 - ² min - ¹, there were a certain number of holes on the surface of the particles, a large number of lacteal protrusions, a sponge-like structure, a high fractal dimension of the surface layer, a high overall roughness, an average particle size of 332.61 nm, a CI of 59.75%, a molecular weight of 1.50 x 10 7 g / mol, a characteristic distance between crystalline lamellae and amorphous regions of 12.68 Å, a V-type crystalline structure, a crystallinity of 16.09%, a mean square nano surface roughness of 14.07 nm, a gelatinization temperature of 90.79 ℃, a hardness of 110.41 g, a cohesion of 1.03, an elasticity of 1.10, and a three-time freeze-thaw water release rate in the range of 33.65-45.52%.

[0139] Comparative Example 3 High-power ultrasonic-assisted technology for constructing endogenous ternary micro-nano assembly of rice starch

[0140] Extraction of mature rice endogenous lipid: 8 kg of completely mature rice was dried at 55 °C for 24 h, ground and passed through a 50 mesh sieve. 4 kg of rice powder was placed in a supercritical CO2 extraction instrument extraction tank, the pressure was set to 15 MPa (extraction tank) and 8 MPa (separation kettle), and the temperature was 60 °C. The crude lipid was obtained by extraction. The crude lipid was purified by rotary evaporation at 60 °C to obtain 289.77 g of endogenous lipid.

[0141] Extraction of immature rice endogenous protein: 8 kg of rice was dried, ground and sieved. 4 kg of immature rice powder was mixed with 80 kg of distilled water in a blender (400 rpm, 20 min), and the suspension was centrifuged at 6000 x g for 20 min. The supernatant was collected. The supernatant was precipitated with 2 M HC1 to pH 4.5, centrifuged (6000 x g, 20 min), and the precipitate was redissolved and dialyzed through an 8 kDa dialysis membrane to obtain 90.09 g of endogenous globulin and albumin.

[0142] Construction of rice starch-endogenous lipid-endogenous protein micro-nano assembly: 250 g of amylopectin was placed in 3750 g of distilled water, and 16.25 g of endogenous lipid and 17.50 g of endogenous protein were added to the pool, and treated at 85 °C for 13 min; the rotation speed was maintained at 150 rpm, and after cooling to room temperature, vacuum filtration was used for dehydration, and the wet sample was collected. The wet sample was added with excess water and ultrasonicated in a cell disrupter at 400 W for 1 h.

[0143] Collection of ternary micro-nano assembly: free lipid and protein were removed by washing with 50% ethanol for 3 times. The flocculent was dried by freeze vacuum drying machine (40 °C, heating rate 4 °C / min, cold trap -80 °C, vacuum degree 30 Pa, 24 h) to a moisture content of ≤10%. The freeze-dried sample was ground and passed through an 800 mesh sieve, and the ternary nano assembly was obtained after packaging. The packaged ternary complex weighed 255.65 g.

[0144] The extracted rice starch ternary micro-nano assembly was detected for in vitro digestion characteristics, complex characteristics, multi-level spatial structure, and physicochemical properties. The purity of endogenous lipid was 91.96%, the purity of endogenous protein was 91.33%, the two-dimensional infrared full spectrum of the ternary nano micro-assembly showed obvious ternary complex peaks, the SDS content was 31.66, and the LOS digestion kinetics showed two-stage digestion, k1 was 3.83 x 10 - ² min - ¹, and k2 was 2.51 x 10- 2 min - 1, the particles showed obvious broken state, the surface had a large number of small and large holes and grooves, a large number of milky protruding structures, a loose sponge-like structure, the average particle size was 286.56 nm, the CI was 62.29%, the molecular weight was 1.68 x 10 7 g / mol, the characteristic distance between the crystalline lamella and the amorphous region was 11.71 Å, it was a V-type crystalline structure, the crystallinity was 16.55%, the mean square nanosurface roughness was 12.20 nm, the gelation temperature was 91.20 °C, the hardness was 103.77 g, the cohesion was 0.94, the elasticity was 0.99, and the water release rate after three freeze-thaw cycles was in the range of 30.91~42.04%.

[0145] Comparative Example 4 Preparation of nano-barley amylopectin ternary complex by ultrasonic-ball milling technology

[0146] Extraction of endogenous lipids from barley: 5 kg of barley was peeled, dried at 60 °C for 24 h, ground, and sieved through a 50 mesh sieve. 2.5 kg of the powder was placed in a large extraction tank in a carbon dioxide supercritical extraction instrument. The instrument was started and the pressure parameters of the extraction tank and the separation kettle were set to 27 Mpa and 7 Mpa, respectively. The temperature of the extraction tank and the separation kettle was 45 °C. The collected crude oil was purified using a rotary evaporator (60 °C) to obtain 155.10 g of endogenous lipids with impurities removed.

[0147] Extraction of endogenous proteins from barley: 5 kg of barley was peeled, dried at 60 °C for 24 h, ground, and sieved through a 50 mesh sieve. 2.5 kg of the powder was mixed with 25 kg of distilled water in a blender (500 rpm, 30 min). The mixed suspension was centrifuged at 5000 x g for 15 min, and the supernatant after centrifugation was collected. The supernatant was adjusted to pH 4.4 with 2M HC1 at 4 °C for isoelectric precipitation. After complete precipitation, the precipitated sample was centrifuged (6000 x g, 20 min), and the supernatant was poured out. The precipitate was redissolved with distilled water. The endogenous globulin and albumin solution (36.67 g) was dialyzed using an 8 kDa dialysis membrane.

[0148] Construction of barley starch-endogenous lipid-endogenous protein nanoassemblies: 150 g of barley starch was placed in an ultrasonic cell disruptor sample cell, 4.2 kg of distilled water was added to the cell, and the starch was gelatinized at a power of 470 W and a temperature of 90 ℃ for 25 min; 45 g of endogenous lipid and 75 g of endogenous protein were then added to the cell, and the power was maintained at 530 W; the temperature was 93 ℃ for 25 min, and the sample was dehydrated using a vacuum filter press, and a wet sample was collected. The free lipid and protein were removed by washing the sample three times with 30% ethanol, and the sample was centrifuged at 5000 x g for 15 min. The flocculent precipitate was dehydrated by freeze-drying in a vacuum drying machine (70 ℃, cold trap -70 ℃, vacuum degree 33 Pa, 24 h) to a moisture content of 6%. The freeze-dried sample was placed in a light ball mill and ball-milled at a speed of 250 r / min for 170 h. The resulting complex was sieved through a 700-mesh sieve, and the ternary complex was packaged to obtain 170.91 g of the ternary complex.

[0149] The barley starch ternary nanoassemblies were detected for in vitro digestion characteristics, complex characteristics, multi-level spatial structure, and physicochemical properties. The purity of the endogenous lipid was 88.74%, the purity of the endogenous protein was 87.60%, the two-dimensional infrared full spectrum of the ternary nanoassemblies showed obvious ternary complex peaks, the SDS content was 29.77, the LOS digestion kinetics showed two-stage digestibility, k1 was 4.55 x 10 - ² min - ¹, k2 was 3.76 x 10 - ² min - ¹, showed a random dispersion of porous particles, the particle surface was accompanied by a large number of protrusions, a large number of enzymatic cavities were visible, the roughness was high, the average particle size was 195.75 nm, the CI was 52.80%, the molecular weight was 1.62 x 10 7 g / mol, the characteristic distance between the crystalline lamellae and the amorphous region was 18.97 Å, it was a V-type crystalline structure, the crystallinity was 15.71%, the mean square nanosurface roughness was 22.31 nm, the gelatinization temperature was 90.14 ℃, the hardness was 50.15 g, the cohesion was 0.44, the elasticity was 0.31, and the three freeze-thaw water separation rates were in the range of 27.76-50.88%.

[0150] Effect example

[0151] Figure 1 The two-dimensional infrared comparison diagram of the waxy maize amylopectin ternary nanoassemblies of Example 1 and the waxy maize amylopectin paste is shown in FIG. 1. Figure 1 As can be seen from FIG. 1, the waxy maize amylopectin endogenous ternary nanoassemblies show obvious ligand residue peaks and more strong infrared group information, indicating that the complex is not only successfully constructed, but also has a more compact and ordered semi-crystalline structure.

[0152] Figure 2 The Los digestion kinetics comparison diagram of the waxy corn amylopectin ternary nanoassembly and the waxy corn amylopectin paste of Example 1 is shown in Figure Figure 1 It can be seen that the endogenous ternary nanoassembly of amylopectin has lower in vitro digestibility, more stable digestion stage, and lower multi-stage digestion kinetics rate than the starch paste, indicating that it can stably reduce blood sugar and provide nutrition slowly and continuously.

[0153] Figure 3 The texture profile curve comparison diagram of the wheat amylopectin ternary nanoassembly and the wheat amylopectin paste of Example 2 is shown in Figure Figure 3 It can be seen that the wheat amylopectin ternary nanoassembly has lower texture kinetics curve, lower hardness, cohesiveness, elasticity, etc. than the wheat amylopectin paste, and has better chewiness and taste.

[0154] Figure 4 The particle size comparison diagram of the wheat amylopectin ternary nanoassembly of Example 2 and the potato amylopectin ternary nanoassembly of Example 3 is shown in Figure Figure 4 It can be seen that both of them show nano-particle size, and the wheat amylopectin ternary nanoassembly is smaller than the potato amylopectin ternary nanoassembly, indicating that its structure is more compact and the particle is more compact.

[0155] Figure 5 The micro-morphology comparison diagram of the barley amylopectin ternary nanoassembly of Example 4 and the rice amylopectin ternary nanoassembly of Example 5 is shown in Figure Figure 5 It can be seen that the rice amylopectin ternary nanoassembly is smaller than the barley amylopectin ternary nanoassembly, has fewer holes, more uniform morphology, compact and smooth particles, and lower roughness.

[0156] Figure 6 The XRD comparison diagram of the barley amylopectin ternary nanoassembly of Example 4 and the sorghum amylopectin ternary nanoassembly of Example 6 is shown in Figure Figure 6 It can be seen that both of them show V-type crystal structure, and the crystallinity of the sorghum amylopectin ternary nanoassembly is higher than that of the barley amylopectin ternary nanoassembly, and the content of the paracrystalline is less, indicating that the number of internal enzymatic sites is less.

[0157] Figure 7 The DSC comparison diagram of the waxy corn amylopectin ternary nanoassembly of Example 1 and the corn starch ternary micro-nano assembly of Comparative Example 1 is shown in Figure Figure 7 It can be seen that the waxy corn amylopectin ternary nanoassembly has stronger heat resistance, heat stability, and higher particle stability than the corn starch ternary micro-nano assembly.

[0158] Figure 8For example 3 potato amylopectin ternary nano-assembly and the mean square nano-surface roughness of the comparative example 2 potato starch ternary micro-nano-assembly are compared, as shown in the schematic diagram of Figure 8 It can be seen that the mean square nano-surface roughness of the potato amylopectin ternary nano-assembly is lower than that of the potato starch ternary micro-nano-assembly, indicating that the size of the "water stop" is lower and the size of the enzymatic channel is smaller.

[0159] Figure 9 For example 5 rice amylopectin ternary nano-assembly and the freeze-thaw stability of the comparative example 3 rice starch ternary micro-nano-assembly are compared, as shown in the schematic diagram of Figure 9 It can be seen that the freeze-thaw water separation of the rice amylopectin ternary nano-assembly is weaker than that of the rice starch ternary micro-nano-assembly, indicating that the shelf life is longer and the texture change during storage is smaller.

[0160] Figure 10 For example 4 rice amylopectin ternary nano-assembly constructed by ultrasonic auxiliary technology and the characteristic distance of crystalline lamella and amorphous region obtained by X-ray scattering of the comparative example 4 nano-barley starch ternary compound prepared by ultrasonic-ball milling technology are compared, as shown in the schematic diagram of Figure 10 It can be seen that the diffraction distance of the rice amylopectin ternary assembly is obviously shortened compared with the barley starch ternary compound, and the characteristic distance of the crystalline lamella and the amorphous region is greatly reduced, thereby improving the slow digestion.

[0161] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A method for constructing ternary nanoassemblies, characterized in that, Includes the following steps: S1: Extraction of amylopectin from major crops using ethanol precipitation method; S2: Endogenous lipids were extracted from the fruits of fully mature bulk crops using supercritical carbon dioxide extraction. S3: Extracting endogenous proteins from immature fruits of major crops using isoelectric point precipitation method; S4: The amylopectin, the endogenous lipids, and the endogenous proteins are subjected to a self-assembly reaction to obtain the ternary nanoassembly; S1, S2 and S3 are not ranked in any particular order.

2. The construction method as described in claim 1, characterized in that, The major crops include one or more of the following: waxy corn, corn, wheat, potato, barley, rice, and sorghum.

3. The construction method as described in claim 1 or 2, characterized in that, The ethanol precipitation method includes the following steps: after the bulk crop is completely gelatinized, it is centrifuged, the precipitate is collected, and the amylopectin is obtained by ethanol precipitation.

4. The construction method according to any one of claims 1 to 3, characterized in that, The supercritical carbon dioxide extraction method includes the following steps: extracting, separating, and purifying the fruits of the fully mature bulk crops to obtain the endogenous lipids.

5. The construction method according to any one of claims 1 to 4, characterized in that, The isoelectric point precipitation method includes the following steps: mixing the immature fruits of the bulk crop with distilled water, centrifuging, collecting the supernatant isoelectric precipitation, and dialysis to obtain the endogenous protein.

6. The construction method according to any one of claims 1 to 5, characterized in that, The mass ratio of the amylopectin, the endogenous lipids, and the endogenous proteins is 1:(0.05~0.15):(0.05~0.20).

7. The construction method according to any one of claims 1 to 6, characterized in that, The self-assembly reaction is carried out at a temperature of 80~100℃, with an ultrasonic power of 300~800 W and a duration of 0.2~3h.

8. The construction method according to any one of claims 1 to 7, characterized in that, The self-assembly reaction is followed by a drying step.

9. The ternary nanoassembly obtained by the construction method according to any one of claims 1 to 8.

10. The application of the ternary nanoassembly as described in claim 9 in any of the following: ( ), and prepare products that improve and / or prevent disorders of glucose metabolism; and / or ( In the preparation of products for the prevention and / or relief of type 2 diabetes; and / or ( In the preparation of products for the prevention and / or relief of hyperlipidemia; and / or ( In the preparation of products for the prevention and / or relief of hypertension; and / or ( In the preparation of products for the prevention and / or relief of obesity; and / or ( I) In the preparation of products for the prevention and / or relief of swallowing disorders.

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