A high-satiety chicken-based small molecule peptide and its application in chicken meal replacement powder

Abstract

This invention provides a high-satiety chicken small molecule peptide and its application in chicken meal replacement powder. First, a gastric self-expanding microsphere is prepared, with yeast β-glucan and hydroxypropyl methylcellulose as the wall material and modified chicken small molecule peptide as the core material. The microsphere absorbs water and expands in the acidic environment of the stomach, generating a feeling of fullness. In the intestinal environment, it slowly releases the modified chicken small molecule peptide, promoting absorption and regulating intestinal health. Furthermore, solid-state fermentation of mixed grain powder using Aspergillus oryzae is used to prepare the chicken meal replacement powder. The addition of the gastric self-expanding microsphere and fermented mixed grain powder further enhances satiety, meeting the market demand for healthy meal replacement foods.

Description

Technical Field

[0001] This invention belongs to the field of food processing technology, specifically relating to a high-satiety chicken small molecule peptide and its application in chicken meal replacement powder. Background Technology

[0002] With the fast pace of modern life, meal replacement foods have become an important choice for health management due to their convenience. While whole-grain meal replacement products are rich in dietary fiber and minerals, they have significant drawbacks in practical application: First, the nutrient ratio is unbalanced, lacking high-quality protein and functional active ingredients; second, the product form is monotonous, with a rough texture, poor solubility and mixing properties, and a tendency to clump and separate, severely impacting the user experience; third, conventional meal replacement powders rely on the physical expansion effect of crude fiber to achieve a feeling of fullness, but this feeling of fullness is short-lived and cannot meet the needs of long-term dietary control. Furthermore, existing technologies attempt to improve the nutritional function of meal replacement powders by enzymatically hydrolyzing animal proteins (such as chicken breast) to prepare small-molecule peptides; however, these small-molecule peptides are easily digested by the stomach, resulting in insufficient satiety and failing to achieve targeted and sustained release of nutrients. At the same time, the inherent anti-nutritional factors (such as phytic acid and trypsin inhibitors) and the large-molecule crude fiber structure in whole-grain bases significantly reduce the bioavailability of protein and trace elements, further weakening the nutritional efficacy of meal replacement products.

[0003] Existing technologies mostly focus on physical modification (such as emulsification and ultrasonic treatment) to improve solubility, or on enhancing protein absorption through a single enzymatic hydrolysis process. However, such solutions cannot systematically address the core challenges of meal replacement powders, such as maintaining satiety, slow release of nutrients, and degradation of anti-nutritional factors. Summary of the Invention

[0004] Technical Problem to be Solved: To address the aforementioned technical problems, the purpose of this invention is to provide a high-satiety chicken small molecule peptide and its application in chicken meal replacement powder. Firstly, a gastric self-expanding microsphere is prepared and coated with the chicken small molecule peptide. The wall material of the gastric self-expanding microsphere is yeast β-glucan and hydroxypropyl methylcellulose, and the core material is modified chicken small molecule peptide. The microsphere absorbs water and expands in the acidic environment of the stomach, generating a feeling of fullness. In the intestinal environment, it slowly releases the chicken small molecule peptide, promoting absorption and regulating intestinal health. Secondly, aspergillus oryzae is used to perform solid-state fermentation of mixed grain powder before it is used to prepare chicken meal replacement powder. The addition of the gastric self-expanding microsphere and fermented mixed grain powder helps to enhance satiety and meet the market demand for healthy meal replacement foods.

[0005] Technical solution: A high satiety chicken small molecule peptide, wherein the high satiety chicken small molecule peptide is a gastric self-expanding microsphere containing modified chicken small molecule peptide.

[0006] The wall material of the self-expanding microspheres in the stomach is yeast β-glucan and hydroxypropyl methylcellulose, and the core material is modified chicken small molecule peptides. The preparation steps of the self-expanding microspheres in the stomach encapsulating the modified chicken small molecule peptides are as follows:

[0007] S1. Dissolve yeast β-glucan in water, stir well, and prepare a yeast β-glucan solution with a concentration of 1-2%;

[0008] S2. Dissolve hydroxypropyl methylcellulose in water and stir evenly at 70-80℃ to prepare a hydroxypropyl methylcellulose solution with a concentration of 1.5-2.5%;

[0009] S3. Mix the yeast β-glucan solution and the hydroxypropyl methylcellulose solution, homogenize, and prepare the wall material solution;

[0010] S4. Dissolve the modified chicken small molecule peptides in water to prepare a modified chicken small molecule peptide solution with a concentration of 5-6%;

[0011] S5. Add the modified chicken small molecule peptide solution to the wall material solution and stir to form an emulsion;

[0012] S6. Add the emulsion dropwise to corn oil containing 0.5% CaCl2 and stir to form a W / O emulsion;

[0013] S7. Add 1% citric acid to the W / O emulsion and cure at room temperature for 30 minutes to obtain microspheres. After washing with deionized water and freeze-drying, the self-expanding microspheres in the stomach are obtained.

[0014] Furthermore, in step S3, the volume ratio of yeast β-glucan solution to hydroxypropyl methylcellulose solution is 1:(2-2.5). Furthermore, the preparation steps of the modified chicken small molecule peptide in step S4 are as follows:

[0015] Step 1: Remove connective tissue from chicken breast, mince it into a paste, add water, homogenize, heat to boiling for 10-20 minutes, cool to 45-55℃, add immobilized protease, enzymatically hydrolyze at 45-55℃ for 4-5 hours, inactivate the enzyme, cool to room temperature, centrifuge at 7000-8000 r / min for 10-15 minutes, collect the supernatant, freeze dry to obtain chicken small molecule peptide freeze-dried powder;

[0016] Step 2: Dissolve the lyophilized chicken small molecule peptide powder and palmitic acid in anhydrous ethanol, add EDC and NHS, adjust the pH to 7.5-8.5, stir at 40-50℃ for 6-8 hours, precipitate with 95% ethanol, centrifuge, and lyophilize to obtain the modified chicken small molecule peptide.

[0017] Furthermore, in step 1, the mass-to-volume ratio of minced meat to water is 1:(2-2.5); and the amount of immobilized protease added is 0.8-1 mg / mL.

[0018] Furthermore, the preparation steps of the immobilized protease are as follows:

[0019] Step 1: Weigh 1g of sodium alginate and dissolve it in 50mL of water. Stir and dissolve in a 37℃ water bath for 1 hour to prepare a 2% sodium alginate solution.

[0020] Step 2: Dilute the mixed protease solution (alkaline protease: flavor protease = 1:1) 5-10 times with borate-sodium hydroxide buffer solution at pH 2.3. Take 5 mL of the diluted enzyme solution and add it to the sodium alginate solution. Stir well and let stand to obtain the sodium alginate solution containing the enzyme solution.

[0021] Step 3: Dissolve 0.75g of chitosan in 50mL of 5% acetic acid solution, add 5mL of 2% CaCl2 solution, and mix thoroughly in a 37℃ water bath to obtain a chitosan solution;

[0022] Step 4: Add the sodium alginate solution containing the enzyme solution to 100 mL of chitosan solution at a rate of 3 drops / s, let it solidify for 2 min to form gel microspheres, then add 1 mL of 0.5% glutaraldehyde solution, stir for 1.5 h, let it stand overnight at 4 °C, wash, and freeze dry to obtain the immobilized protease.

[0023] Furthermore, in step 2, the mass-to-volume ratio of the chicken small molecule peptide lyophilized powder to anhydrous ethanol is 1 g:(10-20) mL; and the molar ratio of the chicken small molecule peptide lyophilized powder to palmitic acid is 1:(1-2).

[0024] Furthermore, in step 2, the mass ratio of EDC to palmitic acid is 1:(1-1.2); and the molar ratio of EDC to NHS is (1-1.5):1.

[0025] Furthermore, in step S5, the volume ratio of the modified chicken small molecule peptide solution to the wall material solution is 1:(1-2).

[0026] The application of the aforementioned high-satiety chicken small molecule peptides in chicken meal replacement powder.

[0027] The specific preparation steps for the above-mentioned chicken meal replacement powder are as follows:

[0028] (1) Emulsification: Dissolve 60-80 parts of high-satiety chicken small molecule peptides, 3-4 parts of mixed fat-soluble vitamins, 2-3 parts of pectin, 4-5 parts of β-cyclodextrin, and 8-10 parts of tagatose in 100-120 parts of water, shear and homogenize to make an emulsion, and store it at 4℃ after stabilization; (2) Preparation of fermented mixed grain powder: Grind soybean powder, yam and potato into powder in a mass ratio of 1:1:1, add water to adjust the moisture content to 45-50%, sterilize, inoculate with Aspergillus oryzae spore suspension at an inoculation amount of 4-6%, ferment for 48-72 hours, dry and sieve to obtain fermented mixed grain powder;

[0029] (3) Ultrasound: Mix the emulsion and fermented grain powder in a ratio of (2-3):1, add sweetener, and sonicate to obtain a mixture;

[0030] (4) Freeze-dry and grind into powder to obtain chicken meal replacement powder.

[0031] Beneficial effects:

[0032] 1. This invention prepares a high-satiety chicken-based small molecule peptide (i.e., a self-expanding microsphere in the stomach), with yeast β-glucan and hydroxypropyl methylcellulose as the wall material. Yeast β-glucan has good biocompatibility, stability, and a certain degree of viscosity, and can form a stable three-dimensional gel network with hydroxypropyl methylcellulose, providing support and protection for the microspheres and preventing premature destruction of the core material by gastric acid. Hydroxypropyl methylcellulose has good film-forming properties, water absorption, and swelling properties, allowing the microspheres to rapidly absorb water and swell in a gastric acid environment, generating an immediate feeling of satiety through mechanical compression of the stomach wall and prolonging the duration of the feeling of satiety. In addition, hydroxypropyl methylcellulose is a cellulose derivative, and the hydroxypropyl and methyl groups introduced into its molecular chain give it pH sensitivity and swelling properties, allowing it to expand in a gastric acid environment (pH 5.5). In the 1.0-3.0 range, a tight gel network is formed through hydrogen bonds, which delays the dissolution of the core material. In the intestine (pH≥5.0), the alkaline environment breaks the hydrogen bonds and triggers rapid disintegration. The synergistic design of the two not only solves the problem of short-lived satiety, but also achieves targeted delivery of nutrients. It is an innovative strategy for regulating satiety in meal replacement products.

[0033] 2. The core material of the self-expanding microspheres in the stomach prepared in this invention is modified chicken small molecule peptides. Firstly, chicken is used as the animal protein source. Immobilized enzymes promote the hydrolysis of animal protein into small molecule peptides. The resulting small molecule peptides (molecular weight < 3kDa) not only enhance flavor and provide antioxidant effects but also promote the digestion and absorption of animal protein. The chicken small molecule peptides obtained after enzymatic hydrolysis are then esterified. The modified chicken small molecule peptides are effectively protected in the acidic environment of the stomach due to the hydrogen bonding cross-linking of the wall material, delaying their release. Upon entering the intestine (pH ≥ 5.0), the wall material gel network disintegrates, and the core material is rapidly released and targeted for absorption, preventing premature digestion in the stomach and thus effectively prolonging the feeling of fullness. Furthermore, the modified chicken small molecule peptides have excellent acid resistance and lipid solubility, can penetrate the intestinal mucosal barrier, and have improved bioavailability compared to unmodified peptides, promoting digestion and absorption.

[0034] 3. The yeast β-glucan and hydroxypropyl methylcellulose in the gastric self-expanding microsphere wall material prepared in this invention are both hydrophilic polymer materials, which are beneficial to promote dissolution. However, during the reconstitution stage (neutral water environment), they only undergo preliminary water absorption and expansion and do not immediately form a dense gel network. Therefore, they do not hinder the solubility during reconstitution. The dense gel is mainly formed in the stomach (acidic environment), which avoids prematurely affecting the reconstitution performance.

[0035] 4. Conventional meal replacement powders often contain coarse fiber, large particles, and anti-nutritional factors, making them difficult to digest and absorb. Therefore, this invention adds fermented whole grain powder to chicken meal replacement powder, offering the following advantages: 1) Solid-state fermentation of the whole grain powder using Aspergillus oryzae decomposes anti-nutritional factors and produces active substances such as γ-aminobutyric acid (GABA) and antioxidant peptides; 2) Aspergillus oryzae secretes phytase, which decomposes the anti-nutritional factor phytic acid into inositol and phosphate, significantly reducing its content; the protease secreted by Aspergillus oryzae degrades trypsin inhibitors and soybean lectins, promoting the digestion and absorption of the whole grain powder; 3) After fermentation, microorganisms secrete proteases... Digestive enzymes such as amylase and lipase break down large molecules (such as protein, starch, and cellulose) in grains into smaller molecules (such as amino acids, small peptides, oligosaccharides, and short-chain fatty acids) that are more easily absorbed. The digestibility and absorption efficiency of its nutrients is significantly better than that of unfermented grain powder. It can also significantly reduce the particle size and roughness of grain powder and reduce clumping caused by large particle aggregation. 4) Aspergillus oryzae fermented grain powder may contain both prebiotics (oligosaccharides) and functional enzymes (such as acidic proteases). The two work together to promote the proliferation of bifidobacteria and improve intestinal health. At the same time, oligosaccharides also have better hydrophilicity and can disperse quickly when in contact with water, reducing clumping caused by surface tension differences. Attached image description:

[0036] Figure 1The solubility diagrams are for the chicken meal replacement powders prepared in Example 5 and Comparative Examples 5-7. Detailed Implementation

[0037] The present invention will be further described below with reference to the accompanying drawings and embodiments. The following embodiments are illustrative of the present invention, but the present invention is not limited to the following embodiments:

[0038] The preparation steps of modified chicken small molecule peptides are as follows:

[0039] Step 1: Weigh 1g of sodium alginate and dissolve it in 50mL of water. Stir and dissolve in a 37℃ water bath for 1 hour to prepare a 2% sodium alginate solution.

[0040] Step 2: Dilute the mixed protease solution (alkaline protease: flavor protease = 1:1) 5 times with borate-sodium hydroxide buffer solution at pH 2.3. Take 5 mL of the diluted enzyme solution and add it to the sodium alginate solution. Stir well and let stand to obtain the sodium alginate solution containing the enzyme solution.

[0041] Step 3: Dissolve 0.75g of chitosan in 50mL of 5% acetic acid solution, add 5mL of 2% CaCl2 solution, and mix thoroughly in a 37℃ water bath to obtain a chitosan solution;

[0042] Step 4: Add the sodium alginate solution containing the enzyme solution to 100 mL of chitosan solution at a rate of 3 drops / s, let it solidify for 2 min to form gel microspheres, then add 1 mL of 0.5% glutaraldehyde solution, stir for 1.5 h, let it stand overnight at 4 °C, wash, and freeze dry to obtain the immobilized protease.

[0043] Step 5: Remove connective tissue from chicken breast, mince it into a paste, add water (the mass-to-volume ratio of meat paste to water is 1:2), homogenize, heat to boiling for 15 minutes, cool to 45℃, add 1 mg / mL immobilized protease, enzymatically hydrolyze at 50℃ for 4 hours, inactivate the enzyme, cool to room temperature, centrifuge at 7000 r / min for 15 minutes, collect the supernatant, and freeze-dry to obtain chicken small molecule peptide freeze-dried powder; Step 6: Dissolve chicken small molecule peptide freeze-dried powder and palmitic acid in anhydrous ethanol (the mass-to-volume ratio of chicken small molecule peptide freeze-dried powder to anhydrous ethanol is 1 g:15 mL, and the molar ratio of chicken small molecule peptide freeze-dried powder to palmitic acid is 1:1). Add EDC and NHS (the mass ratio of EDC to palmitic acid is 1:1, and the molar ratio of EDC to NHS is 1:1). Adjust the pH to 8.0, stir at 40℃ for 8 hours, precipitate with 95% ethanol, centrifuge, and freeze-dry to obtain modified chicken small molecule peptides.

[0044] The prepared modified chicken small molecule peptides were used in the preparation of gastric self-expanding microspheres in subsequent examples.

[0045] Example 1

[0046] A method for preparing gastric self-expanding microspheres includes the following steps:

[0047] S1. Dissolve yeast β-glucan in water, stir well, and prepare a 2% yeast β-glucan solution;

[0048] S2. Dissolve hydroxypropyl methylcellulose in water and stir evenly at 80°C to prepare a 1.5% hydroxypropyl methylcellulose solution;

[0049] S3. Mix yeast β-glucan solution and hydroxypropyl methylcellulose solution at a volume ratio of 1:2, homogenize, and obtain wall material solution;

[0050] S4. Dissolve the modified chicken small molecule peptide in water to prepare a 5% modified chicken small molecule peptide solution;

[0051] S5. Add the modified chicken small molecule peptide solution to the wall material solution at a volume ratio of 1:2, stir, and form an emulsion;

[0052] S6. Add the emulsion dropwise to corn oil containing 0.5% CaCl2 and stir to form a W / O emulsion;

[0053] S7. Add 1% citric acid to the W / O emulsion and cure at room temperature for 30 minutes to obtain microspheres. After washing with deionized water and freeze-drying, the self-expanding microspheres in the stomach are obtained.

[0054] Example 2

[0055] A method for preparing gastric self-expanding microspheres includes the following steps:

[0056] S1. Dissolve yeast β-glucan in water, stir well, and prepare a 2% yeast β-glucan solution;

[0057] S2. Dissolve hydroxypropyl methylcellulose in water and stir evenly at 80°C to prepare a 1.5% hydroxypropyl methylcellulose solution;

[0058] S3. Mix yeast β-glucan solution and hydroxypropyl methylcellulose solution at a volume ratio of 1:2.5, homogenize, and obtain wall material solution;

[0059] S4. Dissolve the modified chicken small molecule peptide in water to prepare a 5% modified chicken small molecule peptide solution;

[0060] S5. Add the modified chicken small molecule peptide solution to the wall material solution at a volume ratio of 1:2, stir, and form an emulsion;

[0061] S6. Add the emulsion dropwise to corn oil containing 0.5% CaCl2 and stir to form a W / O emulsion;

[0062] S7. Add 1% citric acid to the W / O emulsion and cure at room temperature for 30 minutes to obtain microspheres. After washing with deionized water and freeze-drying, the self-expanding microspheres in the stomach are obtained.

[0063] Example 3

[0064] A method for preparing gastric self-expanding microspheres includes the following steps:

[0065] S1. Dissolve yeast β-glucan in water, stir well, and prepare a 2% yeast β-glucan solution;

[0066] S2. Dissolve hydroxypropyl methylcellulose in water and stir evenly at 80°C to prepare a 1.5% hydroxypropyl methylcellulose solution;

[0067] S3. Mix yeast β-glucan solution and hydroxypropyl methylcellulose solution at a volume ratio of 1:2, homogenize, and obtain wall material solution;

[0068] S4. Dissolve the modified chicken small molecule peptide in water to prepare a 5% modified chicken small molecule peptide solution;

[0069] S5. Add the modified chicken small molecule peptide solution to the wall material solution at a volume ratio of 1:1, stir, and form an emulsion;

[0070] S6. Add the emulsion dropwise to corn oil containing 0.5% CaCl2 and stir to form a W / O emulsion;

[0071] S7. Add 1% citric acid to the W / O emulsion and cure at room temperature for 30 minutes to obtain microspheres. After washing with deionized water and freeze-drying, the self-expanding microspheres in the stomach are obtained.

[0072] Example 4

[0073] A method for preparing gastric self-expanding microspheres includes the following steps:

[0074] S1. Dissolve yeast β-glucan in water, stir well, and prepare a 2% yeast β-glucan solution;

[0075] S2. Dissolve hydroxypropyl methylcellulose in water and stir evenly at 80°C to prepare a 1.5% hydroxypropyl methylcellulose solution;

[0076] S3. Mix yeast β-glucan solution and hydroxypropyl methylcellulose solution at a volume ratio of 1:2, homogenize, and obtain wall material solution;

[0077] S4. Dissolve the modified chicken small molecule peptide in water to prepare a 5% modified chicken small molecule peptide solution;

[0078] S5. Add the modified chicken small molecule peptide solution to the wall material solution at a volume ratio of 1:1.5, stir, and form an emulsion;

[0079] S6. Add the emulsion dropwise to corn oil containing 0.5% CaCl2 and stir to form a W / O emulsion;

[0080] S7. Add 1% citric acid to the W / O emulsion and cure at room temperature for 30 minutes to obtain microspheres. After washing with deionized water and freeze-drying, the self-expanding microspheres in the stomach are obtained.

[0081] Comparative Example 1

[0082] The difference between this comparative example and Example 2 is that the amount of hydroxypropyl methylcellulose added is too low, as detailed below:

[0083] A method for preparing gastric self-expanding microspheres includes the following steps:

[0084] S1. Dissolve yeast β-glucan in water, stir well, and prepare a 2% yeast β-glucan solution;

[0085] S2. Dissolve hydroxypropyl methylcellulose in water and stir evenly at 80°C to prepare a 1.5% hydroxypropyl methylcellulose solution;

[0086] S3. Mix yeast β-glucan solution and hydroxypropyl methylcellulose solution at a volume ratio of 1:1, homogenize, and obtain wall material solution;

[0087] S4. Dissolve the modified chicken small molecule peptide in water to prepare a 5% modified chicken small molecule peptide solution;

[0088] S5. Add the modified chicken small molecule peptide solution to the wall material solution at a volume ratio of 1:2, stir, and form an emulsion;

[0089] S6. Add the emulsion dropwise to corn oil containing 0.5% CaCl2 and stir to form a W / O emulsion;

[0090] S7. Add 1% citric acid to the W / O emulsion and cure at room temperature for 30 minutes to obtain microspheres. After washing with deionized water and freeze-drying, the self-expanding microspheres in the stomach are obtained.

[0091] Comparative Example 2

[0092] The difference between this comparative example and Example 2 is that it uses unmodified chicken small molecule peptides, as detailed below:

[0093] A method for preparing gastric self-expanding microspheres includes the following steps:

[0094] S1. Dissolve yeast β-glucan in water, stir well, and prepare a 2% yeast β-glucan solution;

[0095] S2. Dissolve hydroxypropyl methylcellulose in water and stir evenly at 80°C to prepare a 1.5% hydroxypropyl methylcellulose solution;

[0096] S3. Mix yeast β-glucan solution and hydroxypropyl methylcellulose solution at a volume ratio of 1:2.5, homogenize, and obtain wall material solution;

[0097] S4. Dissolve the chicken small molecule peptides in water to prepare a 5% chicken small molecule peptide solution;

[0098] S5. Add the chicken small molecule peptide solution to the wall material solution at a volume ratio of 1:2, stir, and form an emulsion;

[0099] S6. Add the emulsion dropwise to corn oil containing 0.5% CaCl2 and stir to form a W / O emulsion;

[0100] S7. Add 1% citric acid to the W / O emulsion and cure at room temperature for 30 minutes to obtain microspheres. After washing with deionized water and freeze-drying, the self-expanding microspheres in the stomach are obtained.

[0101] Comparative Example 3

[0102] The difference between this comparative example and Example 2 is that yeast β-glucan was not added, as detailed below:

[0103] A method for preparing gastric self-expanding microspheres includes the following steps:

[0104] S1. Dissolve hydroxypropyl methylcellulose in water and stir evenly at 80°C to prepare a 1.5% hydroxypropyl methylcellulose solution, which is the wall material solution;

[0105] S2. Dissolve the modified chicken small molecule peptide in water to prepare a 5% modified chicken small molecule peptide solution;

[0106] S3. Add the modified chicken small molecule peptide solution to the wall material solution at a volume ratio of 1:2, stir, and form an emulsion;

[0107] S4. Add the emulsion dropwise to corn oil containing 0.5% CaCl2 and stir to form a W / O emulsion;

[0108] S5. Add 1% citric acid to the W / O emulsion and cure at room temperature for 30 min to obtain microspheres. After washing with deionized water and freeze-drying, the self-expanding microspheres in the stomach are obtained.

[0109] Comparative Example 4

[0110] The difference between this comparative example and Example 2 is that hydroxypropyl methylcellulose was not added, as detailed below:

[0111] A method for preparing gastric self-expanding microspheres includes the following steps:

[0112] S1. Dissolve yeast β-glucan in water, stir well, and prepare a 2% yeast β-glucan solution, which is the wall material solution;

[0113] S2. Dissolve the modified chicken small molecule peptide in water to prepare a 5% modified chicken small molecule peptide solution;

[0114] S3. Add the modified chicken small molecule peptide solution to the wall material solution at a volume ratio of 1:2, stir, and form an emulsion;

[0115] S4. Add the emulsion dropwise to corn oil containing 0.5% CaCl2 and stir to form a W / O emulsion;

[0116] S5. Add 1% citric acid to the W / O emulsion and cure at room temperature for 30 min to obtain microspheres. After washing with deionized water and freeze-drying, the self-expanding microspheres in the stomach are obtained.

[0117] Performance testing:

[0118] (1) Expansion rate

[0119] Using the volumetric method, the particle size of the prepared gastric self-expanding microspheres was first determined by a laser particle size analyzer, and the microsphere volume V0 was calculated. The microspheres were then placed in simulated gastric fluid and oscillated at a constant temperature of 37°C. The microspheres were removed periodically, and the microsphere volume V was calculated. t The expansion rate of the microspheres is calculated using the following formula:

[0120] Expansion rate (%) = (V t -V0) / V0×100%

[0121] Table 1. Expansion rates of the self-expanding gastric microspheres prepared in Examples 1-4 and Comparative Examples 1-4

[0122] 30min 1h 2h Example 1 315％ 388％ 420％ Example 2 350％ 395％ 442％ Example 3 284％ 326％ 390％ Example 4 302％ 344％ 406％ Comparative Example 1 212％ 285％ 358％ Comparative Example 2 342％ 379％ 421％ Comparative Example 3 301％ 348％ 390％ Comparative Example 4 157％ 182％ 210％

[0123] As shown in Table 1, hydroxypropyl methylcellulose in the wall material of the self-expanding microspheres in the stomach plays the main role in expansion. In Example 2, the expansion rate of the microspheres after soaking in simulated gastric juice for 2 hours reached as high as 442%, which produced an immediate feeling of fullness by mechanically compressing the stomach wall and prolonging the satiety time. In Comparative Example 1, the amount of hydroxypropyl methylcellulose added was too low, resulting in a significant decrease in the expansion rate. The expansion rate of the microspheres prepared by adding unmodified chicken small molecule peptides in Comparative Example 2 was not much different from that in the examples. The expansion rate of the microspheres in Comparative Example 3 without yeast β-glucan was slightly reduced. The expansion rate of the microspheres in Comparative Example 4 without hydroxypropyl methylcellulose was the lowest.

[0124] (2) In vitro simulation experiment

[0125] A certain amount of self-expanding microspheres from the stomach were taken, and the wall material structure was destroyed using a strong alkaline solution to release all the core material. The total concentration of modified chicken small molecule peptides in the microspheres was determined by ultraviolet spectrophotometry. An equal amount of microspheres were placed in simulated gastric fluid and incubated at 37°C with shaking for 4 hours, with samples taken every 2 hours. The microspheres treated with gastric fluid were transferred to simulated intestinal fluid and incubated at 37°C with shaking for 4 hours. The concentration of modified chicken small molecule peptides in the solution was measured and the release rate was calculated.

[0126] Table 2. Release rates of modified chicken small molecule peptides in the gastric self-expanding microspheres prepared in Examples 1-4 and Comparative Examples 1-4.

[0127]

[0128]

[0129] As shown in Table 2, yeast β-glucan can form a more stable three-dimensional gel network with hydroxypropyl methylcellulose, providing support and protection for the microspheres and preventing gastric acid from prematurely damaging the core material. However, the mechanical strength of microspheres made by embedding modified chicken small molecule peptides with yeast β-glucan alone or hydroxypropyl methylcellulose alone is lower than that of the composite wall material. This causes the single wall material to disintegrate too quickly in the gastric acid environment, resulting in the premature release of the core material. The release rate of modified chicken small molecule peptides increases in the stomach but decreases in the intestine.

[0130] Therefore, the gastric self-expanding microspheres prepared in Example 2 were selected for the subsequent preparation of chicken meal replacement powder.

[0131] Example 5

[0132] The specific preparation steps for chicken meal replacement powder are as follows:

[0133] S1. Emulsification: 80g of the self-expanding microspheres in the stomach prepared in Example 2, 3g of mixed fat-soluble vitamins, 3g of pectin, 4g of β-cyclodextrin, and 8g of tagatose were dissolved in 100g of water, and the mixture was sheared and homogenized to form an emulsion. After stabilization, the emulsion was stored at 4°C.

[0134] S2. Preparation of fermented mixed grain powder: Grind soybean flour, yam, and potato into powder at a mass ratio of 1:1:1, add water to adjust the moisture content to 45%, sterilize, inoculate with Aspergillus oryzae spore suspension at an inoculation rate of 6%, ferment for 60 hours, dry, and sieve to obtain fermented mixed grain powder; S3. Ultrasound: Mix the emulsion and fermented mixed grain powder at a ratio of 3:1, add sweetener, and sonicate to obtain a mixture;

[0135] S4. Freeze-dry and grind into powder to obtain chicken meal replacement powder.

[0136] Comparative Example 5

[0137] The difference between this comparative example and Example 5 is that modified chicken small molecule peptides were directly added, as detailed below:

[0138] The specific preparation steps for chicken meal replacement powder are as follows:

[0139] S1. Emulsification: Dissolve 60g of modified chicken small molecule peptides, 3g of mixed fat-soluble vitamins, 3g of pectin, 4g of β-cyclodextrin and 8g of tagatose in 100g of water, shear and homogenize to make an emulsion, and store at 4℃ after stabilization.

[0140] S2. Preparation of fermented mixed grain powder: Grind soybean flour, yam, and potato into powder at a mass ratio of 1:1:1, add water to adjust the moisture content to 45%, sterilize, inoculate with Aspergillus oryzae spore suspension at an inoculation rate of 6%, ferment for 60 hours, dry, and sieve to obtain fermented mixed grain powder; S3. Ultrasound: Mix the emulsion and fermented mixed grain powder at a ratio of 3:1, add sweetener, and sonicate to obtain a mixture;

[0141] S4. Freeze-dry and grind into powder to obtain chicken meal replacement powder.

[0142] Comparative Example 6

[0143] The difference between this comparative example and Example 5 is that unmodified chicken small molecule peptides were directly added, as detailed below:

[0144] The specific preparation steps for chicken meal replacement powder are as follows:

[0145] S1. Emulsification: Dissolve 60g of chicken small molecule peptides, 3g of mixed fat-soluble vitamins, 3g of pectin, 4g of β-cyclodextrin, and 8g of tagatose in 100g of water, shear and homogenize to form an emulsion, and store at 4℃ after stabilization.

[0146] S2. Preparation of fermented mixed grain powder: Grind soybean flour, yam, and potato into powder at a mass ratio of 1:1:1, add water to adjust the moisture content to 45%, sterilize, inoculate with Aspergillus oryzae spore suspension at an inoculation rate of 6%, ferment for 60 hours, dry, and sieve to obtain fermented mixed grain powder; S3. Ultrasound: Mix the emulsion and fermented mixed grain powder at a ratio of 3:1, add sweetener, and sonicate to obtain a mixture;

[0147] S4. Freeze-dry and grind into powder to obtain chicken meal replacement powder.

[0148] Comparative Example 7

[0149] The difference between this comparative example and Example 5 is that whole grain powder is directly added, as detailed below:

[0150] The specific preparation steps for chicken meal replacement powder are as follows:

[0151] S1. Emulsification: 80g of the self-expanding microspheres in the stomach prepared in Example 2, 3g of mixed fat-soluble vitamins, 3g of pectin, 4g of β-cyclodextrin, and 8g of tagatose were dissolved in 100g of water, and the mixture was sheared and homogenized to form an emulsion. After stabilization, the emulsion was stored at 4°C.

[0152] S2. Preparation of mixed grain flour: Grind soybean flour, yam, and potato into flour in a mass ratio of 1:1:1;

[0153] S3. Ultrasound: Mix the emulsion and grain powder in a 3:1 ratio, add sweetener, and sonicate to obtain a mixture;

[0154] S4. Freeze-dry and grind into powder to obtain chicken meal replacement powder.

[0155] Performance index measurement:

[0156] (1) Solubility

[0157] The solubility of different chicken meal replacement powders was compared by mixing them with warm water at 50-60℃.

[0158] Depend on Figure 1 It can be seen that the addition of gastric self-expanding microspheres and fermented grain powder in the chicken meal replacement powder of Example 5 resulted in good solubility, while the solubility of Comparative Examples 5 and 6 decreased because the chicken small molecule peptides were not coated inside the microspheres. In Comparative Example 7, unfermented grain powder was added directly, and the presence of crude fiber and anti-nutritional factors greatly hindered the dissolution process.

[0159] (2) Antioxidant activity

[0160] ①DPPH free radical scavenging rate

[0161] Prepare a 0.2 mmol / L DPPH solution. Prepare a 5 mg / mL chicken meal replacement powder solution using 75% ethanol as the solvent. Mix the DPPH solution and chicken meal replacement powder solution for 2 min, centrifuge, and collect the supernatant. Add the supernatant to a 96-well microplate, mix for 60 s, and incubate at room temperature in the dark for 30 min. Measure the absorbance at 517 nm and calculate the DPPH free radical scavenging rate using the following formula:

[0162] DPPH free radical scavenging rate (%) = 1 - (A1 - A2) / A3 × 100

[0163] In the formula, A1 is the absorbance of the sample; A2 is the absorbance of the blank sample; and A3 is the absorbance of the control sample.

[0164] ②ABTS+ free radical scavenging rate

[0165] The ABTS+ radical solution was diluted to an absorbance of 0.70 ± 0.02 at 734 nm. A chicken meal replacement powder solution with a concentration of 5 mg / mL was prepared using 75% ethanol as the solvent. The ABTS+ radical solution and the chicken meal replacement powder solution were mixed for 2 min, centrifuged, and the supernatant was collected. The ABTS+ radical scavenging rate was calculated using the following formula:

[0166] ABTS+ free radical scavenging rate (%) = (A0-A1) / A0×100

[0167] In the formula, A0 is the absorbance of the blank sample; A1 is the absorbance of the sample.

[0168] Table 3. Antioxidant activity of chicken meal replacement powders prepared in Example 5 and Comparative Examples 5-7

[0169] DPPH free radical scavenging rate (%) ABTS+ free radical scavenging rate (%) Example 5 79.71 67.82 Comparative Example 5 75.02 63.53 Comparative Example 6 73.68 61.21 Comparative Example 7 69.87 54.07

[0170] As shown in Table 3, enzymatic hydrolysis of chicken meat to prepare small molecule peptides and fermentation of mixed grain powder can also decompose the macromolecules in mixed grains into more easily absorbed antioxidant peptides, which have good antioxidant effects. However, the free radical scavenging rates of DPPH and ABTS+ in the comparative example are lower than those in the example.

[0171] (3) Anti-nutritional factor content

[0172] Trypsin inhibitors: enzyme activity inhibition method;

[0173] Phytic acid: Spectrophotometry;

[0174] Soybean lectin: ELISA quantification.

[0175] Table 4. Content of anti-nutritional factors in the chicken meal replacement powders prepared in Example 5 and Comparative Examples 5-7

[0176] Trypsin inhibitor (TIU / g) Phytic acid (g / 100g) Soybean lectin (HU / mg) Example 5 4 0.23 Not detected Comparative Example 5 5 0.26 Not detected Comparative Example 6 6 0.21 Not detected Comparative Example 7 30 1.52 2.0

[0177] As shown in Table 4, fermenting the mixed grain flour helps reduce the content of anti-nutritional factors and promotes digestion and absorption. However, in Comparative Example 7, which directly added unfermented mixed grain flour, the contents of trypsin inhibitor, phytic acid, and soybean lectin were all higher than those in the Example.

[0178] (4) Sensory test of satiety

[0179] Instructions for use: Eat a light diet the night before the experiment, fast for 12 hours and participate in the experiment on an empty stomach. On the morning of the experiment, consume the chicken meal replacement powder prepared in Example 5 and Comparative Examples 5-7 at 8:00 am (50g of meal replacement powder is mixed with warm water to make 200mL and consumed within 5 minutes).

[0180] Sixty healthy adults, half male and half female, aged 45-55, were selected and randomly divided into four groups. Each group consumed one of the four types of chicken meal replacement powders on an empty stomach according to the consumption instructions (each type of meal replacement powder was consumed one day apart). The order of consumption was randomly assigned. The time of the first feeling of hunger was recorded and the average value was calculated.

[0181] Table 5. Time to first hunger pangs for chicken meal replacement powders prepared in Example 5 and Comparative Examples 5-7

[0182] Example 5 Comparative Example 5 Comparative Example 6 Comparative Example 7 Time to first feeling of hunger (h) <![CDATA[4.8±0.6 a ]]> <![CDATA[3.2±0.5 b ]]> <![CDATA[2.9±0.7 b ]]> <![CDATA[2.5±0.4 b ]]>

[0183] As shown in Table 5, the initial hunger time in Example 5 was significantly longer than that in other groups. This indicates that in the embodiments of the present invention, modified chicken small molecule peptides were embedded in self-expanding microspheres in the stomach, and the esterified modified peptides were released in the intestines in a targeted manner, continuously providing amino acid signals to inhibit hunger hormones. In addition, the soluble fiber and prebiotics in the fermented grain powder could also prolong the transmission of intestinal satiety signals. The synergistic effect of the self-expanding microspheres in the stomach and the fermentation process resulted in the prepared chicken meal replacement powder having a high satiety.

[0184] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention, or modify them into equivalent embodiments, without departing from the spirit and technical essence of the present invention. Therefore, any simple modifications, equivalent substitutions, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention, without departing from the content of the technical solutions of the present invention, shall still fall within the scope of protection of the present invention.

Claims

1. A high satiety chicken small molecule peptide, characterized in that: The highly satiating chicken small molecule peptides are in-stomach self-expanding microspheres containing modified chicken small molecule peptides. The wall material of the self-expanding microspheres in the stomach is yeast β-glucan and hydroxypropyl methylcellulose, and the core material is modified chicken small molecule peptides; The preparation steps of the gastric self-expanding microspheres containing the modified chicken small molecule peptide are as follows: S1. Dissolve yeast β-glucan in water, stir well, and prepare a yeast β-glucan solution with a concentration of 1-2%; S2. Dissolve hydroxypropyl methylcellulose in water and stir evenly at 70-80℃ to prepare a hydroxypropyl methylcellulose solution with a concentration of 1.5-2.5%; S3. Mix the yeast β-glucan solution and the hydroxypropyl methylcellulose solution, homogenize, and prepare the wall material solution; S4. Dissolve the modified chicken small molecule peptides in water to prepare a modified chicken small molecule peptide solution with a concentration of 5-6%; S5. Add the modified chicken small molecule peptide solution to the wall material solution and stir to form an emulsion; S6. Add the emulsion dropwise to corn oil containing 0.5% CaCl2 and stir to form a W / O emulsion; S7. Add 1% citric acid to the W / O emulsion and cure at room temperature for 30 min to obtain microspheres. After washing with deionized water and freeze-drying, the self-expanding microspheres in the stomach are obtained. In step S3, the volume ratio of yeast β-glucan solution to hydroxypropyl methylcellulose solution is 1:(2-2.5). The preparation steps of the modified chicken small molecule peptide in step S4 are as follows: Step 1: Remove connective tissue from chicken breast, mince it into a paste, add water, homogenize, heat to boiling for 10-20 minutes, cool to 45-55℃, add immobilized protease, enzymatically hydrolyze at 45-55℃ for 4-5 hours, inactivate the enzyme, cool to room temperature, centrifuge at 7000-8000 r / min for 10-15 minutes, collect the supernatant, freeze dry to obtain chicken small molecule peptide freeze-dried powder; Step 2: Dissolve the lyophilized chicken small molecule peptide powder and palmitic acid in anhydrous ethanol, add EDC and NHS, adjust the pH to 7.5-8.5, stir at 40-50℃ for 6-8 hours, precipitate with 95% ethanol, centrifuge, and lyophilize to obtain modified chicken small molecule peptide.

2. The chicken small molecule peptide with high satiety according to claim 1, characterized in that: In step 1, the mass-to-volume ratio of minced meat to water is 1:(2-2.5); the amount of immobilized protease added is 0.8-1 mg / mL.

3. The chicken-based small molecule peptide with high satiety according to claim 1, characterized in that, In step 2, the mass-to-volume ratio of chicken small molecule peptide freeze-dried powder to anhydrous ethanol is 1 g: (10-20) mL; the molar ratio of chicken small molecule peptide freeze-dried powder to palmitic acid is 1: (1-2).

4. The chicken-based small molecule peptide with high satiety according to claim 1, characterized in that, In step 2, the mass ratio of EDC to palmitic acid is 1:(1-1.2); the molar ratio of EDC to NHS is (1-1.5):

1.

5. The chicken-based small molecule peptide with high satiety according to claim 1, characterized in that, In step S5, the volume ratio of the modified chicken small molecule peptide solution to the wall material solution is 1:(1-2).

6. The application of the high-satiety chicken small molecule peptide according to any one of claims 1-5 in chicken meal replacement powder.

7. The application according to claim 6, characterized in that, The specific preparation steps of the chicken meal replacement powder are as follows: (1) Emulsification: Dissolve the high-satiety chicken small molecule peptides, mixed fat-soluble vitamins, pectin and β-cyclodextrin in water, cut and homogenize to make an emulsion, and let it stand for storage; (2) Preparation of fermented mixed grain powder: Soybean flour, yam and potato are ground into powder, water is added to adjust the moisture content, sterilized, inoculated with Aspergillus oryzae spore suspension, fermented, dried and sieved to obtain fermented mixed grain powder; (3) Ultrasound: Mix the emulsion with fermented grain powder, add sweetener, and sonicate to obtain a mixture; (4) Freeze-dry and grind into powder to obtain chicken meal replacement powder.

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