Blood sugar regulation dietary carrier based on multi-target hypoglycemic peptide and preparation method thereof

Abstract

The invention discloses a blood sugar regulation dietary carrier based on multi-target hypoglycemic peptides and a preparation method thereof, and belongs to the technical field of food science and nutrition engineering. The invention aims to solve the problems of low utilization rate of existing pine protein, poor activity of peptide products, single target spot and mismatching of delivery systems. The method comprises the following steps: 1, ultrasonic composite enzymolysis extraction; 2, gradient grading purification; 3, vacuum concentration and freeze drying; 4, liquefying the auxiliary materials; and 5, carrying out gradient variable-temperature baking. The sequence of the hypoglycemic peptide prepared by the method is NTDVQKLEHIFGAH, the thermal stability reaches 130 DEG C, and the hypoglycemic peptide has the multi-target synergistic effects of enhancing intestinal barriers, improving glycolipid metabolism, inhibiting inflammation, enhancing insulin sensitivity and the like. The thermally stable carrier matrix constructed by the method disclosed by the invention can realize efficient delivery and function synergy of active peptides, and a brand new solution is provided for dietary intervention of diabetes mellitus.

Description

Technical Field

[0001] The present invention relates to the preparation of a blood sugar regulating dietary carrier based on multi-target glucose-lowering peptides and a preparation method thereof, belonging to the field of A23L 33 / 18 food science and nutritional engineering technology, and is particularly suitable for blood sugar regulation in patients with type II diabetes. Background Art

[0002] While current dietary regulation approaches based on protein modification have shown potential health benefits, and dietary interventions based on food-derived active peptides have attracted considerable attention due to their high safety and good compliance, current technologies still face the following key bottlenecks: ① Limited raw material utilization: Mainstream glucose-lowering peptides are mostly derived from conventional raw materials such as milk protein and soy protein. Pinus lanceolata protein contains 18 amino acids, of which the proportion of functional amino acids such as glutamic acid (12.7%) and arginine (9.3%) is significantly higher than that of soy protein. However, its dense globular structure leads to low efficiency of conventional enzymatic hydrolysis. ② Functional singleness defect: Existing glucose-lowering peptides mostly focus on a single mechanism of action, such as α-glucosidase inhibition or DPP-IV inhibition, making it difficult to achieve multi-target synergistic regulation. Clinical studies have shown that the onset of type 2 diabetes usually involves multiple pathways such as insulin resistance, inflammation, and intestinal flora disorders, and single-mechanism products cannot meet the needs of comprehensive regulation; ③ Insufficient adaptability of the delivery system: Traditional peptide carriers are mainly liquid beverages or capsules, which have problems such as peptide degradation (activity loss >40% after 6 months of storage at room temperature) and poor palatability. Although baking solid carriers can improve stability, high-temperature processing can easily destroy the structure-activity relationship of the peptide (such as disulfide bond cleavage rate >60%), resulting in loss of functional activity. Summary of the Invention

[0003] The present invention aims to solve the problems of existing hypoglycemic foods, such as limited raw material utilization, single functional ingredients, and ineffective utilization of active ingredients. It provides a blood sugar regulating dietary carrier based on multi-target hypoglycemic peptides and a preparation method thereof, which is carried out according to the following steps:

[0004] 1. Petroleum ether was added to Pinus lanceolata kernel oil cake at a ratio of 1:10 w / v to remove oil, and distilled water was added to the defatted powder at a material-liquid ratio of 1:50-1:100. The powder was treated with an ultrasonic power of 200-300 W for 30-40 min, and then enzymatic hydrolysis was performed to break the cell wall. The composite enzymatic hydrolysis conditions were as follows: at 45-55° C. and pH 7.0-9.5, 7000 U / g of a composite protease consisting of papain, alkaline protease, and pepsin at a mass ratio of (2-4):(4-8):1 was added according to the mass of the substrate, and enzymatic hydrolysis was performed for 2-2.5 h to obtain an enzymatic hydrolyzate.

[0005] Second, the enzymatic hydrolysate was inactivated in a 90°C water bath for 15 minutes, concentrated to 1 / 5 of the original volume using a rotary evaporator (vacuum degree -0.095 MPa, 50°C), and then centrifuged at 8000×g for 20 minutes. The supernatant was collected to obtain the hypoglycemic peptide crude extract;

[0006] 3. Gradient purification of the hypoglycemic peptide crude liquid was performed. The primary purification was performed using a DEAE-52 ion exchange chromatography column (2.6×40 cm) with a linear gradient elution of 0.1-0.5 M NaCl solution at a flow rate of 3 mL / min, and 2-4 times the column volume of the eluate was collected; the secondary purification was performed using a Sephadex G50 gel chromatography column (1.6×60 cm) with isocratic elution of ultrapure water at a flow rate of 3 mL / min, and 1-3 times the column volume of the elution peak was collected to obtain the purified solution;

[0007] Fourth, the purified solution was concentrated (vacuum degree -0.095MPa, 50°C) and vacuum freeze-dried (cold trap temperature -55°C, vacuum degree 10Pa) for 48 hours to obtain a solid powder of the hypoglycemic peptide;

[0008] 5. 10-15% avocado oil, 5-10% whole egg liquid, 10% sugar-free dark chocolate particles (D90 ≤ 2mm), and 3-5% skim milk powder are liquefied at 100-150° C. for 5-10 minutes until completely liquid as a liquid phase, and then mixed with a solid phase matrix including 30-40% pine nut protein powder, 10-15% almond powder, and 3-5% erythritol, and the functional added glucose-lowering peptide is 10-20% of the total mass of the carrier;

[0009] 6. The dietary carrier is baked at a gradient temperature: in the shaping stage, the hot air flow rate is 3.5-4.0 m / s, and the upper and lower fires are baked at 180°C for 1-2 minutes to form a dense shaping layer; in the reaction stage, the hot air flow rate is adjusted to 1.0-1.5 m / s, and the upper and lower fires are baked at 120°C for 15-20 minutes to complete the Maillard reaction; in the coating stage, the hot air flow rate is restored to 2.0-2.5 m / s, and the upper and lower fires are baked at 180°C for 1-2 minutes to form an aroma coating layer, and the environment is cooled to obtain a multi-target hypoglycemic peptide blood sugar regulation dietary carrier.

[0010] The beneficial effects of the present invention are:

[0011] 1. The breakthrough use of Pinus sylvestris nut protein, a unique plant protein resource in my country, as raw material is rich in multiple functional amino acids compared to conventional raw materials. Using ultrasound-assisted composite enzymatic hydrolysis and directional purification technology, highly active hypoglycemic peptides were extracted from Pinus sylvestris nut protein for the first time. This fills a technological gap in the preparation of highly active hypoglycemic peptides from nuts, avoids the allergen risk of traditional animal-derived hypoglycemic peptides (such as whey protein peptides), and solves the problems of low hydrolysis degree and poor activity of peptides in existing technologies.

[0012] 2. The use of ultrasound (20-40kHz, 200-300W) assisted multi-enzyme synergy (papain, alkaline protease, pepsin) directed enzymatic hydrolysis technology effectively solved the problem of low enzymatic hydrolysis efficiency caused by the dense globular structure of Pinus lanceolata protein, significantly improved the hydrolysis degree of hypoglycemic peptides (20.69% to 27.91%), and successfully released highly active hypoglycemic peptide fragments, solving the problems of low efficiency and poor activity of single technologies (such as pure enzymatic hydrolysis or ultrasound); combined with an ion exchange-gel chromatography two-stage purification system, a characteristic peptide segment (NTDVQKLEHIFGAH) with multi-target regulatory functions was accurately obtained.

[0013] 3. An innovative gradient temperature baking process was designed to construct a heat-stable carrier matrix. While ensuring the taste and quality of the dietary carrier, it achieved efficient delivery and functional synergy of highly active hypoglycemic peptides, effectively overcoming the problem of traditional baking processes damaging the activity of peptides.

[0014] 4. The glucose-lowering peptides in the obtained dietary carriers act on α-glucosidase (IC 50 =5.21 mg / mL) and α-amylase (IC 50 =1.71mg / mL) had a significant inhibitory effect, and the dietary carrier had a variety of multi-target regulatory functions, including upregulating the abundance of intestinal short-chain fatty acids to enhance intestinal barrier function (abundance increased by ≥20%), regulating lipid metabolism, regulating sugar metabolism levels, inhibiting the release of inflammatory factors, and improving insulin sensitivity, etc., which can achieve comprehensive regulation of blood sugar disorders.

[0015] 5. Realize high-value utilization of resources and industrial extension, turn Pinus elata nut oil cake (traditional waste) into treasure, reduce production costs and promote the recycling of forest resources.

[0016] 6. It is green, safe and has commercial potential. It is prepared entirely by physical biomimetic methods without the need for organic solvents and complies with FDA / GB 2760 food safety standards.

[0017] Figures in the specification

[0018] Figure 1 This is a diagram of ion exchange chromatography purification of the technology of the present invention in Example 1;

[0019] Figure 2 This is a diagram of gel chromatography column purification of the present invention in Example 2;

[0020] Figure 3 The dietary carrier prepared in Example 4 restores lipid metabolism disorders in type II diabetic mice;

[0021] Figure 4 The restorative effect of the dietary carrier prepared in Example 5 on glucose metabolism disorder in type II diabetic mice;

[0022] Figure 5 The dietary carrier prepared in Example 6 has a restorative effect on the liver and kidney tissue morphology of type II diabetic mice;

[0023] Figure 6 Effects of the dietary carrier prepared in Example 8 on the species composition of the intestinal flora of type II diabetic mice (phylum, class, order, family);

[0024] Figure 7 Effects of the dietary carrier prepared in Example 10 on the characteristic species of intestinal flora in type II diabetic mice;

[0025] Figure 8 This is a TG-DSC thermogravimetric analysis graph of the glucose-lowering peptide added to the dietary carrier prepared in Example 11;

[0026] Figure 9 It is the technical roadmap of the method of the present invention. DETAILED DESCRIPTION

[0027] Specific embodiment 1: This embodiment is a blood sugar regulating dietary carrier based on multi-target hypoglycemic peptides and its preparation method, which is carried out according to the following steps:

[0028] 1. Petroleum ether was added to Pinus lanceolata kernel oil cake at a ratio of 1:10 w / v to remove oil, and distilled water was added to the defatted powder at a material-liquid ratio of 1:50-1:100. The powder was treated with an ultrasonic power of 200-300 W for 30-40 min, and then enzymatic hydrolysis was performed to break the cell wall. The composite enzymatic hydrolysis conditions were as follows: at 45-55° C. and pH 7.0-9.5, 7000 U / g of a composite protease consisting of papain, alkaline protease, and pepsin at a mass ratio of (2-4):(4-8):1 was added according to the mass of the substrate, and enzymatic hydrolysis was performed for 2-2.5 h to obtain an enzymatic hydrolyzate.

[0029] Second, the enzymatic hydrolysate was inactivated in a 90°C water bath for 15 minutes, concentrated to 1 / 5 of the original volume using a rotary evaporator (vacuum degree -0.095 MPa, 50°C), and then centrifuged at 8000×g for 20 minutes. The supernatant was collected to obtain the hypoglycemic peptide crude extract;

[0030] 3. Gradient purification of the hypoglycemic peptide crude liquid was performed. The primary purification was performed using a DEAE-52 ion exchange chromatography column (2.6×40 cm) with a linear gradient elution of 0.1-0.5 M NaCl solution at a flow rate of 3 mL / min, and 2-4 times the column volume of the eluate was collected; the secondary purification was performed using a Sephadex G50 gel chromatography column (1.6×60 cm) with isocratic elution of ultrapure water at a flow rate of 3 mL / min, and 1-3 times the column volume of the elution peak was collected to obtain the purified solution;

[0031] Fourth, the purified solution was concentrated (vacuum degree -0.095MPa, 50°C) and vacuum freeze-dried (cold trap temperature -55°C, vacuum degree 10Pa) for 48 hours to obtain a solid powder of the hypoglycemic peptide;

[0032] 5. 10-15% avocado oil, 5-10% whole egg liquid, 10% sugar-free dark chocolate particles (D90 ≤ 2mm), and 3-5% skim milk powder are liquefied at 100-150° C. for 5-10 minutes until completely liquid as a liquid phase, and then mixed with a solid phase matrix including 30-40% pine nut protein powder, 10-15% almond powder, and 3-5% erythritol, and the functional added glucose-lowering peptide is 10-20% of the total mass of the carrier;

[0033] 6. The dietary carrier is baked at a gradient temperature: in the shaping stage, the hot air flow rate is 3.5-4.0 m / s, and the upper and lower fires are baked at 180°C for 1-2 minutes to form a dense shaping layer; in the reaction stage, the hot air flow rate is adjusted to 1.0-1.5 m / s, and the upper and lower fires are baked at 120°C for 15-20 minutes to complete the Maillard reaction; in the coating stage, the hot air flow rate is restored to 2.0-2.5 m / s, and the upper and lower fires are baked at 180°C for 1-2 minutes to form an aroma coating layer, and the environment is cooled to obtain a multi-target hypoglycemic peptide blood sugar regulation dietary carrier.

[0034] The ion exchange chromatography purification elution curve under this embodiment is shown in the attached figure. Figure 1 shown.

[0035] Specific embodiment 2: This embodiment differs from specific embodiment 1 in that the amount of eluent used in step 3 using the Sephadex G50 gel chromatography column is 4 times the column volume, and the elution flow rate is 2.5 mL / min. Other aspects are the same as specific embodiment 1. The gel chromatography exchange column purification elution curve under this embodiment is shown in the attached figure. Figure 2 As shown, the glucose-lowering peptide prepared at a concentration of 10 mg / mL had an inhibition rate of 38.49% on α-glucosidase and 16.79% on α-amylase.

[0036] Specific embodiment 3: This embodiment differs from specific embodiment 1 in that the ultrasonic power was set to 150W and the enzymatic hydrolysis time was extended to 4 hours. The degree of hydrolysis of the resulting active hypoglycemic peptide was 21.69% (determined by the pH-star method). Mass spectrometry analysis showed the molecular weight of the target peptide to be 1567.82 Da.

[0037] Specific embodiment 4: This embodiment differs from the specific embodiment 1 in that: 30% pine nut protein powder (red pine protein, pine pine protein), 10% glucose-lowering peptide powder of the embodiment 1, 12% avocado oil, and 10% dark chocolate particles (D90=1.8mm) are mixed. The dietary carrier prepared in this way regulates lipid metabolism in type II diabetic mice as shown in the attached figure. Figure 3 As shown, (A): total cholesterol level; (B): high-density lipoprotein cholesterol level; (C): low-density lipoprotein cholesterol level; (D): triglyceride level; (E): adiponectin level; (F): leptin level. The prepared dietary vehicle significantly reduced serum triglyceride (48.22%), total cholesterol (40.24%), low-density lipoprotein cholesterol (44.19%), and leptin (72.63%) levels in type 2 diabetic mice, while increasing high-density lipoprotein cholesterol (141.63%) and adiponectin (78.10%) levels.

[0038] Specific embodiment 5: This embodiment differs from specific embodiment 1 in that: 20% pine nut protein powder (red pine protein, pine pine protein), 20% almond powder, 15% glucose-lowering peptide powder of embodiment 1, 8% avocado oil, and 5% dark chocolate particles (D90=1.8mm) are mixed. The dietary carrier prepared in this way regulates glucose metabolism in type II diabetic mice as shown in the attached figure. Figure 4 Figures show (A): Hexokinase activity; (B): Pyruvate kinase activity; (C): Phosphofructokinase activity; (D): Glucose-6-phosphate dehydrogenase activity; (E): Glycated hemoglobin level; (F): Insulin level. The dietary vehicle activated the activities of the key glycolytic enzymes hexokinase (3.12-fold), pyruvate kinase (0.28-fold), and phosphofructokinase (0.26-fold) in type 2 diabetic mice, and reduced the activity of glucose-6-phosphate dehydrogenase (20.30%), thereby inhibiting the compensatory activation of the pentose phosphate pathway (PPP). Furthermore, the dietary vehicle reduced Glycated hemoglobin levels (28.53%) and serum insulin levels (69.96%) in type 2 diabetic mice.

[0039] Specific embodiment 6: This embodiment differs from specific embodiment 1 in that a composite protease consisting of papain, alkaline protease, and pepsin in a mass ratio of (1-3): (3-6): (2-4) is used at 7000 U / g and enzymatic hydrolysis is carried out for 2 hours. The effect of the dietary carrier prepared in this way on the organ tissue morphology of type II diabetic mice is shown in the attached figure. Figure 5Figures 24 and 25 show liver morphology analysis (A) in the control group; (B) in the type 2 diabetic mice group; and (C) in the dietary vehicle-treated group. Kidney morphology analysis: (D) in the control group; (E) in the type 2 diabetic mice group; and (F) in the dietary vehicle-treated group. Red circles represent glomeruli. The dietary vehicle effectively alleviated reduced hepatic glycogen deposition, hepatocyte hyalinization, central venous blood cell aggregation, significant dilation of the hepatic sinusoids, blurred hepatic plate boundaries, and the appearance of watery vacuoles within hepatocytes in type 2 diabetic mice. It also restored the number of Kupffer cells in the livers of type 2 diabetic mice and improved liver function. It also effectively reduced inflammatory cells in the central veins of the liver and improved dilation of the hepatic sinusoids. Furthermore, the dietary vehicle effectively alleviated glomerular hypertrophy, enlarged cysts, thickening of the glomerular capillary basement membrane (GBM), increased intercapillary matrix, narrowing of the vascular lumen, desquamation and edema of renal tubular epithelial cells, renal interstitial fibrosis, and inflammatory cell infiltration in type 2 diabetic mice.

[0040] Specific embodiment seven: This embodiment differs from specific embodiment one in that baking was performed at a constant temperature of 180°C for 15 minutes. The dietary carrier exhibits a non-uniform carbonized layer on the surface, with deep carbonization visible at the edges. Gas chromatography-mass spectrometry detected unusually high concentrations of pyrolysis products, including furans (2.8 mg / kg) and acrylamide (1.2 mg / kg). Sensory evaluation confirmed a typical burnt bitterness and a smoky flavor derived from lignin degradation. Changes in physical and chemical properties indicate a late-stage Maillard reaction and excessive pyrolysis, resulting in a complete loss of edible value.

[0041] Specific embodiment eight: This embodiment differs from specific embodiment one in that the raw material ratio of the dietary carrier is: pine nut protein powder (20%), hypoglycemic peptide (30%), avocado oil (20%), sugar-free dark chocolate (10%), and whole egg liquid (10%). After the base materials are mixed, they are cooked at 150°C for 10 minutes to form a soft material carrier. The effect of this dietary carrier on the species abundance of the intestinal flora of type II diabetic mice is shown in the attached figure. Figure 6As shown, (A): relative abundance of phylum; (B): relative abundance of class; (C): relative abundance of order; (D): relative abundance of family. At the phylum level, it significantly reduced the abundance ratio of Firmicutes / Bacteroidetes in the intestines of type 2 diabetic mice in a dose-dependent manner. This was mainly due to the upregulation of the abundance of Firmicutes, which can produce short-chain fatty acids (SCFAs, acetate and butyrate) to strengthen the intestinal barrier. At the class level, it upregulated the relative abundance of Clostridia (phylum Bacteroidetes), which can produce SCFAs, inhibit inflammatory responses, and regulate metabolism. Dietary intervention reduced the relative abundance of Bacilli in the intestines of type 2 diabetic mice by more than 50%, indicating that it can alleviate metabolic acidosis caused by D-lactic acid accumulation. At the phenotypic level, the dietary vector reduced the relative abundance of Erysipelotrichia (-96.60%) and increased the relative abundance of Bacteroidales (+76.43%) in the intestines of type 2 diabetic mice, suggesting that it can reduce insulin resistance through immune regulation. At the phylogenetic level, the dietary vector significantly increased the relative abundance of Muribaculaceae and Lachnospiraceae in the intestines of type 2 diabetic mice, suggesting that it can activate the insulin receptor / phosphatidylinositol-3-kinase / protein kinase B (IRS / PI3K / Akt) signaling pathway or inhibit inflammatory responses by regulating the Th17 / Treg balance.

[0042] Specific embodiment nine: This embodiment differs from specific embodiment one in that the composite enzyme used in step two is composed only of papain and alkaline protease, and the hydrolysis degree of the prepared hypoglycemic peptide is 18.94%. The peptide sequence confirmed by LC-MS / MS is SDDVLEAAFNTDVQKLEHIFGAH, and the half-maximal inhibitory concentration for α-amylase is 25.91±2.43 mg / mL; the half-maximal inhibitory concentration for α-glucosidase is 24.89±3.61 mg / mL.

[0043] Specific embodiment 10: This embodiment differs from specific embodiment 1 in that the baking process is as follows: setting stage: 185℃ / 4.0m / s / 1min; reaction stage: 115℃ / 1.0m / s / 20min; coating stage: 175℃ / 2.5m / s / 1.5min. The effect of this dietary carrier on the characteristic species of intestinal flora in type II diabetic mice is shown in the figure below. Figure 7As shown in the figure, (A) is the principal component analysis between groups; (B) is the principal component analysis of species; and (C) is the genus-level species composition heat map used for species clustering. The dietary carrier can increase the relative abundance of Muribaculaceae, Limosilactobacillus, and Ligilactobacillus in the advocacy, and can treat type 2 diabetes by reducing oxidative stress and inflammation. The dietary carrier can significantly reduce the number of pathogenic bacteria in the intestine (Turicibacter: -54.80%; Staphylococcus: -30.91%) and effectively alleviate adipose tissue inflammation, insulin resistance, and bile acid metabolism disorders caused by pathogens.

[0044] Specific embodiment 11: The peptide powder obtained in Example 1 was subjected to differential scanning calorimetry (DSC) to detect that its melting peak exceeded 130°C. The specific results are shown in the attached Figure 8 , confirming its high temperature resistance. After application to baking carriers, the activity retention rate was verified by in vitro enzyme activity >91.2%.

[0045] Specific embodiment 12: According to the purification process of embodiment 1, a continuous flow centrifuge (8000×g) and a spray drying tower (inlet air 180°C / outlet air 80°C) are connected to achieve 200L of enzymatic hydrolysate per hour, and the peptide powder production capacity reaches 1.5kg / h.

Claims

1. A blood sugar regulating dietary carrier based on multi-target glucose-lowering peptides and a preparation method thereof, characterized in that: The following steps are involved: (1) Pretreatment and enzymatic hydrolysis of raw materials: Add petroleum ether to the pine kernel oil cake at a ratio of 1:10 (w / v) for defatting, take the defatted powder and add distilled water at a material-liquid ratio of 1:50-1:100 (g / mL), and treat it at an ultrasonic power of 200-300 W for 30-40 min; then, add 7000 U / g of a composite protease composed of papain, alkaline protease, and pepsin at a mass ratio of (2-4):(4-8):1 according to the mass of the substrate at 45-55 °C and pH 7.0-9.5, and enzymatic hydrolysis is carried out for 2.0-2.5 h to obtain an enzymatic solution; (2) Inactivation and crude extraction: The enzymatic hydrolysate obtained in step (1) was inactivated in a 90°C water bath for 15 min, concentrated to 1 / 5 of the original volume using a rotary evaporator at a vacuum degree of -0.095 MPa and 50°C, and then centrifuged at 8000×g for 20 min. The supernatant was collected to obtain a crude extract of hypoglycemic peptide; (3) Gradient purification: The crude liquid obtained in step (2) was subjected to DEAE-52 ion exchange chromatography and Sephadex G50 gel chromatography in sequence, wherein the primary purification was performed using a 0.1-0.5 M NaCl solution at a flow rate of 3 mL / min for gradient elution over 2-4 column volumes, and the secondary purification was performed using ultrapure water at a flow rate of 3 mL / min for elution over 1-3 column volumes to obtain a purified solution; (4) Drying treatment: The purified solution was concentrated at a vacuum degree of -0.095 MPa and 50°C, and then vacuum freeze-dried at a cold trap temperature of -55°C and a vacuum degree of 10 Pa for 48 hours to obtain a solid powder of the pine gum hypoglycemic peptide; (5) Matrix preparation: 10-15% avocado oil, 5-10% whole egg liquid, 10% sugar-free dark chocolate particles (D90 ≤ 2 mm), and 3-5% skim milk powder are liquefied at 100-150°C for 5-10 minutes to form a liquid phase, which is then mixed with 30-40% pine nut protein powder, 10-15% almond powder, and 3-5% erythritol as a solid matrix, and 10-20% of the total mass of the carrier is added with hypoglycemic peptide powder; (6) Gradient temperature baking: A three-stage hot air baking process is adopted, including the shaping stage: hot air flow rate of 3.5-4.0 m / s, baking at 180°C for 1-2 min; reaction stage: adjusting the hot air flow rate to 1.0-1.5 m / s, baking at 120°C for 15-20 min; coating stage: restoring the hot air flow rate to 2.0-2.5 m / s, baking at 180°C for 1-2 min, and cooling the environment to 25±2°C to obtain a multi-target hypoglycemic peptide blood glucose regulation dietary carrier.

2. The blood sugar regulating dietary carrier based on multi-target glucose-lowering peptides and the preparation method thereof according to claim 1, characterized in that: In the step (1), the ultrasonic treatment frequency is 40 kHz, and the defatted powder is passed through an 80-mesh sieve (particle size ≤ 180 μm).

3. The blood sugar regulating dietary carrier based on multi-target glucose-lowering peptides and the preparation method thereof according to claim 1, characterized in that: In the step (3), the size of the DEAE-52 ion exchange chromatography column is 2.6×40 cm, and the size of the Sephadex G50 gel chromatography column is 1.6×60 cm.

4. The blood sugar regulating dietary carrier based on multi-target glucose-lowering peptides and the preparation method thereof according to claim 1, characterized in that: The amino acid sequence of the solid powder of the Pinus lanceolata glucose-lowering peptide in step (4) is shown in SEQ ID NO: 1, which is NTDVQKLEHIFGAH, confirmed by LC-MS / MS, and has a thermal stability of up to 130° C., as verified by DSC.

5. The blood sugar regulating dietary carrier based on multi-target glucose-lowering peptides and the preparation method thereof according to claim 1, characterized in that: The glucose-lowering peptide in the dietary carrier is effective for α-glucosidase (IC 50 =5.21 mg / mL) and α-amylase (IC 50 =1.71 mg / mL) had a significant inhibitory effect.

6. The blood sugar regulating dietary carrier based on multi-target glucose-lowering peptides and the preparation method thereof according to claim 1, characterized in that: The dietary carrier obtained in step (6) has at least two of the following multi-target regulatory functions: (1) Enhance intestinal barrier function by upregulating the abundance of intestinal short-chain fatty acids, with the abundance increased by ≥20%; (2) Regulate lipid metabolism, reduce serum triglyceride ≥30%, total cholesterol ≥35%, low-density lipoprotein cholesterol ≥40%, and leptin ≥60%, while increasing high-density lipoprotein cholesterol ≥80% and adiponectin ≥60%; (3) Regulates glucose metabolism: increases hexokinase activity by 3 times, pyruvate kinase activity by 0.2 times, phosphofructokinase activity by 0.25 times, and decreases glucose-6-phosphate dehydrogenase activity by ≥15%; (4) Inhibition of TNF-α and IL-6 inflammatory cytokine release by ≥35%, upregulation of cystathionine metabolic abundance by ≥20%, and upregulation of Limosilactobacillus and Ligilactobacillus bacterial abundance by 15%; (5) Improve insulin sensitivity, reduce serum insulin levels by ≥40%, and reduce fasting blood glucose levels by 35%.

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