A hypoglycemic zero residue composite powder, a preparation method and application thereof

Abstract

This invention discloses a hypoglycemic compound powder containing burdock, its preparation method, and its applications. The preparation method includes: enzymatic hydrolysis of raw materials, two-stage mixed-culture fermentation, and microencapsulation. This invention employs a CO2-based cell-wall breaking and enzymatic hydrolysis synergistic technology to significantly improve the extraction efficiency of active ingredients such as diosgenin and polysaccharides from burdock; through two-stage mixed-culture directional fermentation, lipid degradation is achieved, generating functional factors; and microencapsulation using burdock fiber, fucoidan, and peanut peptides as composite packaging materials improves product stability and bioavailability. The product of this invention exhibits a multi-mechanism synergistic hypoglycemic effect, significant hypoglycemic efficacy, good stability, and promising application prospects.

Description

Technical Field

[0001] This invention relates to the field of functional food processing technology, specifically to a hypoglycemic compound powder, its preparation method, and its application. Background Technology

[0002] Bulbs, or tubers, are the bulbils that grow in the leaf axils of the yam plant (Dioscorea opposita Thunb.), a member of the Dioscoreaceae family. Commonly known as yam beans or yam eggs, they are an important component of the above-ground parts of the yam plant, yielding 200-300 kg per mu (approximately 0.16 acres), making them an extremely abundant resource. However, due to their small size and low commercial value, in traditional yam cultivation and processing, except for a small amount kept for seed, the vast majority are discarded as waste or used as fertilizer, resulting in serious resource waste and environmental pollution. In recent years, with the deepening research on plants that are both food and medicine, the nutritional and medicinal value of bulbils has gradually attracted attention. Studies have shown that bulbils are rich in polysaccharides, flavonoids, saponins, allantoin, taurine, various essential amino acids, and trace elements (iron, zinc, copper, manganese, selenium, etc.). The content of polysaccharides, flavonoids, polyphenols, and allantoin is higher than that of yam tubers. Modern pharmacological studies have confirmed that bulbils possess various biological activities such as hypoglycemic, antioxidant, and anti-fatigue effects. Therefore, bulbils are a functional food ingredient with great development potential. Currently, there are some reports on the development and utilization technologies of bulbils, such as the process of preparing yam bean porridge, yam bean beverages, and yam bean preserves using yam beans as raw materials. However, this technology only uses traditional processing methods such as steaming, grinding, and sugar boiling, without stepwise extraction and targeted enrichment of the active ingredients in bulbils. The resulting products have low levels of functional components and unclear functional characteristics.

[0003] Currently, technologies for developing and utilizing residual ions share the following common problems: First, the extraction and utilization rates of active ingredients are low; second, most methods employ simple crushing and mixing or one-time extraction, resulting in low levels of functional components in the products; third, while existing technologies may combine residual ions with other raw materials, these are often simple additives without synergistic design; and fourth, product forms are mainly concentrated in traditional food sectors, lacking the development of modern functional food dosage forms. Therefore, developing a functional food product and its preparation method that can achieve high-value utilization of residual ion functional components and possess clear synergistic effects has significant practical and economic value. Summary of the Invention

[0004] Technical problems to be solved: This invention aims to solve the technical problems of low extraction efficiency, low bioavailability and poor product stability of the active ingredients in the prior art. This invention improves the content of functional substances through compound fermentation and improves product stability and hypoglycemic effect through encapsulation process, so as to achieve synergistic hypoglycemic effect of multiple mechanisms, and the product has excellent flavor and stability.

[0005] Technical solution: A method for preparing a hypoglycemic compound powder, comprising the following steps: S1. Raw material pretreatment: The residual particles are washed, dried and then pulverized at low temperature. The residual particles are sieved to obtain residual particle powder. The residual particle powder is put into the reaction vessel, CO2 is introduced to maintain the pressure, and then the pressure is released to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add the complex enzyme buffer to the reaction vessel, adjust the pH value, stir evenly to start enzymatic hydrolysis, slowly pass CO2 during the process, raise the temperature to inactivate the enzyme, centrifuge and take the supernatant enzymatic hydrolysate. S3. Fermentation: Cool the enzymatic hydrolysate, inoculate it with a suspension of Yersinia lipophila, and carry out aerobic fermentation under stirring to obtain the first stage fermentation broth. Inoculate the mixed bacterial suspension into the first stage fermentation broth and carry out anaerobic fermentation to obtain the final fermentation broth. Separate the final fermentation broth through a membrane and then concentrate it through a nanofiltration membrane to obtain the concentrated fermentation broth. S4. Formulation: The mixture of brown algae oligosaccharide and peanut peptide with water is ultrasonically added to the concentrated fermentation broth and stirred to react. Then, burdock fiber powder is added, stirred and homogenized, and spray-dried to obtain the oligomeric complex powder.

[0006] Furthermore, in step S1, the pressure is increased to 10-20 MPa and maintained for 5-15 minutes.

[0007] Furthermore, in step S2, the mass ratio of the complex enzyme buffer to the residual powder is (7-8):(2-3); the concentration of the complex enzyme buffer is 2.5-5 wt.%, and the buffer is phosphate buffer; the pH is adjusted to 6.5-7.5; the enzymatic hydrolysis temperature is 35-45℃, and the time is 1-2 h; CO2 is introduced to maintain the pressure at 2-5 MPa; the temperature is raised to 80-90℃, and the enzyme inactivation time is 10-20 min.

[0008] Furthermore, the complex enzyme buffer comprises, by weight fraction: 5-15 parts neutral protease, 3-8 parts lipase, 2-6 parts pullulanase, 5-10 parts amylase, 8-15 parts cellulase, and 3-7 parts debranching enzyme. Furthermore, the concentration of the *Yarrowia lipophila* suspension in step S3 is 1 × 10⁻⁶. 7 -1×10 8 The inoculum concentration is CFU / mL, and the inoculum size is 5-8 wt.% of the enzymatic hydrolysate. The stirring speed is 150-200 rpm. The temperature for aerobic fermentation is 30-35℃, and the time is 18-24 h. The inoculum size for mixed bacterial suspension is 10-15 wt.%. The temperature for anaerobic fermentation is 35-38℃, and the time is 24-36 h.

[0009] Furthermore, the mixed bacterial suspension includes Propionibacterium acnes, Lactobacillus reuteri, and Bifidobacterium, with a colony count ratio of (1-2):(2-3):(1-2); the concentration of the mixed bacterial suspension is 1×10⁻⁶. 6 -1×10 8CFU / mL; membrane separation uses a 0.22μm microporous membrane, and nanofiltration membrane has a molecular weight cutoff of 600-1000 Da.

[0010] Furthermore, in step S4, the mass ratio of the mixed liquid to the concentrated fermentation liquid and burdock fiber powder is (50-70):(40-60):(2-3); the ultrasonic power is 150-250W, and the time is 10-15min; the stirring speed is 200-300rpm, and the time is 30-60min; the homogenization speed is 1000-2000rpm, and the time is 10-15min.

[0011] Furthermore, in step S4, the content of brown algae oligosaccharides in the mixture is 2-4 wt.%, and the content of peanut peptides is 1-3 wt.%.

[0012] A hypoglycemic compound powder prepared by any of the above preparation methods.

[0013] Furthermore, the application of the aforementioned hypoglycemic granule compound powder in the preparation of functional foods and hypoglycemic drugs. Beneficial effects

[0014] This invention utilizes a CO2-enzymatic hydrolysis synergistic technology. CO2 is introduced into the intercellular spaces of the raw material cells, creating a microscopic explosion effect within the cells. This physically disrupts the cellulose-starch complex structure, increasing the release of functional components and fully exposing the binding sites required for subsequent enzymatic hydrolysis. The continuous introduction of CO2 not only improves the efficiency of enzymatic hydrolysis but also inhibits oxidation reactions, preventing the degradation and loss of heat-sensitive active ingredients such as diosgenin and allantoin. Compared to traditional enzymatic hydrolysis or water extraction methods, this invention improves extraction efficiency and lays a material foundation for the high-value utilization of zinc resources.

[0015] This invention introduces four functional strains: Yersinia lipolyticis, Propionibacterium, Lactobacillus reuteri, and Bifidobacterium. This staged, multi-strain fermentation not only achieves efficient conversion of lipocytes and enrichment of functional factors through biotransformation, but also promotes glucagon-like peptide-1 secretion and regulates pancreatic β-cell function through metabolites. At the same time, the synergistic effect of multiple mechanisms, such as Bifidobacterium improving insulin resistance, significantly enhances the product's hypoglycemic efficacy and intestinal health regulation function, forming a synergistic hypoglycemic system.

[0016] This invention utilizes enzymatic hydrolysis to provide a more easily metabolizable substrate for subsequent fermentation. By reducing starch content through enzymatic hydrolysis, some starch is converted into resistant starch. The slow fermentation and metabolism of resistant starch provides a continuous and stable energy source for the mixed-culture fermentation in the later stage, avoiding the drastic early pH drop caused by the rapid acid production of ordinary starch fermentation. In addition, in the first stage, Yersinia lipolyticis efficiently degrades fatty acids after lipase hydrolysis, eliminating lipid interference and generating short-chain fatty acid precursors. The enzymatic hydrolysis in the early stage significantly improves fermentation efficiency and the yield of functional factors.

[0017] This invention uses burdock dietary fiber, fucoidan oligosaccharides, and peanut peptides as composite packaging materials, achieving multiple functional integrations of encapsulation protection, synergistic hypoglycemic effects, and probiotic enhancement. Burdock dietary fiber not only serves as a structural support material to enhance the mechanical strength of the microcapsules but also slows down the rate of glucose formation due to its α-glucosidase inhibitory activity. Fucoidan oligosaccharides, as prebiotics, promote the colonization of beneficial intestinal bacteria and regulate intestinal barrier function. Peanut peptides provide enteric protection, preventing the active ingredients from being destroyed by gastric acid while enhancing the body's immunity. This encapsulation system effectively solves the technical challenges of easy oxidation and destruction of active ingredients, ensuring the full absorption and utilization of hypoglycemic factors at the site of action, while significantly improving the product's flavor characteristics and storage stability.

[0018] This invention constructs a synergistic hypoglycemic system with multi-target synergistic effects. The active ingredients such as diosgenin and polysaccharides in the tubercle can directly inhibit α-glucosidase activity, delay the absorption of carbohydrates in the intestine, and reduce postprandial blood glucose peaks. The short-chain fatty acids such as propionic acid produced by fermentation metabolism can inhibit hepatic gluconeogenesis, promote the secretion of glucagon-like peptide-1 (GLP-1) in the intestine, enhance insulin sensitivity, and improve glucose metabolism. Probiotics can regulate the intestinal microecological balance and promote the absorption of active ingredients in the intestine. The components form a synergistic effect, which is more significant and stable in hypoglycemic effect than a single hypoglycemic component, and has no side effects of chemical drugs, making it suitable for long-term consumption. Detailed Implementation

[0019] The present invention will be further described below with reference to embodiments. These embodiments are illustrative of the present invention, but the present invention is not limited to these embodiments: Example 1

[0020] A method for preparing a hypoglycemic compound powder includes the following steps: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 300g of residual particle powder into the reaction vessel, introduce CO2 and pressurize it to 15MPa for 10min, and then release the pressure to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add 700g of complex enzyme buffer to the reaction vessel. The complex enzyme buffer concentration is 3wt.%, the solvent is phosphate buffer, and the complex enzyme composition is 10 parts of neutral protease, 5 parts of lipase, 4 parts of pullulanase, 7 parts of amylase, 12 parts of cellulase, and 5 parts of debranching enzyme. Adjust the pH to 7.0, stir evenly, control the temperature at 40℃ and hydrolyze for 1.5h. During this period, slowly introduce CO2, maintain the pressure at 3MPa, raise the temperature to 85℃ to inactivate the enzyme for 15min, centrifuge and collect the supernatant enzymatic hydrolysate. S3. Fermentation: Cool 500g of enzymatic hydrolysate and inoculate with 25g of *Yersinia lipophila* suspension. Perform aerobic fermentation at 35℃ for 18 hours with stirring at 180 rpm to obtain the first-stage fermentation broth. Inoculate the first-stage fermentation broth with 50g of a mixed bacterial suspension and perform anaerobic fermentation at 37℃ for 24 hours to obtain the final fermentation broth. The mixed bacterial suspension includes *Propionibacterium acnes*, *Lactobacillus reuteri*, and *Bifidobacterium*, with a colony count ratio of 1:2:1. The concentration of the mixed bacterial suspension is 1×10⁻⁶. 7 The final fermentation broth was separated by passing CFU / mL through a 0.22μm microporous membrane and then concentrated through a 1000Da nanofiltration membrane to obtain concentrated fermentation broth. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated fermentation broth and stir at 200rpm for 40min. Then add 4g of burdock fiber powder, stir, homogenize at 1500rpm for 10min, and spray dry to obtain oligomeric complex powder. Example 2

[0021] A method for preparing a hypoglycemic compound powder includes the following steps: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 200g of residual particle powder into the reaction vessel, introduce CO2 and pressurize it to 15MPa for 10min, and then release the pressure to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add 800g of complex enzyme buffer to the reaction vessel. The complex enzyme buffer concentration is 3wt.%, the solvent is phosphate buffer, and the complex enzyme composition is 10 parts of neutral protease, 5 parts of lipase, 4 parts of pullulanase, 7 parts of amylase, 12 parts of cellulase, and 5 parts of debranching enzyme. Adjust the pH to 7.0, stir evenly, control the temperature at 40℃ and hydrolyze for 1.5h. During this period, slowly introduce CO2, maintain the pressure at 3MPa, raise the temperature to 85℃ to inactivate the enzyme for 15min, centrifuge and collect the supernatant enzymatic hydrolysate. S3. Fermentation: Cool 500g of enzymatic hydrolysate and inoculate with 25g of *Yersinia lipophila* suspension. Perform aerobic fermentation at 35℃ for 18 hours with stirring at 180 rpm to obtain the first-stage fermentation broth. Inoculate the first-stage fermentation broth with 50g of a mixed bacterial suspension and perform anaerobic fermentation at 37℃ for 24 hours to obtain the final fermentation broth. The mixed bacterial suspension includes *Propionibacterium acnes*, *Lactobacillus reuteri*, and *Bifidobacterium*, with a colony count ratio of 1:2:1. The concentration of the mixed bacterial suspension is 1×10⁻⁶. 7 The final fermentation broth was separated by passing CFU / mL through a 0.22μm microporous membrane and then concentrated through a 1000Da nanofiltration membrane to obtain concentrated fermentation broth. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated fermentation broth and stir at 200rpm for 40min. Then add 4g of burdock fiber powder, stir, homogenize at 1500rpm for 10min, and spray dry to obtain oligomeric complex powder. Example 3

[0022] A method for preparing a hypoglycemic compound powder includes the following steps: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 300g of residual particle powder into the reaction vessel, introduce CO2 and pressurize it to 15MPa for 10min, and then release the pressure to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add 700g of complex enzyme buffer to the reaction vessel. The complex enzyme buffer concentration is 3wt.%, the solvent is phosphate buffer, and the complex enzyme composition is 10 parts of neutral protease, 5 parts of lipase, 4 parts of pullulanase, 7 parts of amylase, 12 parts of cellulase, and 5 parts of debranching enzyme. Adjust the pH to 7.0, stir evenly, control the temperature at 40℃ and hydrolyze for 1.5h. During this period, slowly introduce CO2, maintain the pressure at 5MPa, raise the temperature to 85℃ to inactivate the enzyme for 15min, centrifuge and collect the supernatant enzymatic hydrolysate. S3. Fermentation: Cool 500g of enzymatic hydrolysate and inoculate with 25g of *Yersinia lipophila* suspension. Perform aerobic fermentation at 35℃ for 18 hours with stirring at 180 rpm to obtain the first-stage fermentation broth. Inoculate the first-stage fermentation broth with 50g of a mixed bacterial suspension and perform anaerobic fermentation at 37℃ for 24 hours to obtain the final fermentation broth. The mixed bacterial suspension includes *Propionibacterium acnes*, *Lactobacillus reuteri*, and *Bifidobacterium*, with a colony count ratio of 1:2:1. The concentration of the mixed bacterial suspension is 1×10⁻⁶. 7 The final fermentation broth was separated by passing CFU / mL through a 0.22μm microporous membrane and then concentrated through a 1000Da nanofiltration membrane to obtain concentrated fermentation broth. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated fermentation broth and stir at 200rpm for 40min. Then add 4g of burdock fiber powder, stir, homogenize at 1500rpm for 10min, and spray dry to obtain oligomeric complex powder. Example 4

[0023] A method for preparing a hypoglycemic compound powder includes the following steps: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 300g of residual particle powder into the reaction vessel, introduce CO2 and pressurize it to 15MPa for 10min, and then release the pressure to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add 700g of complex enzyme buffer to the reaction vessel. The complex enzyme buffer concentration is 3wt.%, the solvent is phosphate buffer, and the complex enzyme composition is 10 parts of neutral protease, 5 parts of lipase, 4 parts of pullulanase, 7 parts of amylase, 12 parts of cellulase, and 5 parts of debranching enzyme. Adjust the pH to 7.0, stir evenly, control the temperature at 40℃ and hydrolyze for 1.5h. During this period, slowly introduce CO2, maintain the pressure at 3MPa, raise the temperature to 85℃ to inactivate the enzyme for 15min, centrifuge and collect the supernatant enzymatic hydrolysate. S3. Fermentation: Cool 500g of enzymatic hydrolysate and inoculate with 40g of *Yeastra lipolytica* suspension. Perform aerobic fermentation at 35℃ for 18h under stirring at 180rpm to obtain the first-stage fermentation broth. Inoculate the first-stage fermentation broth with 50g of a mixed bacterial suspension and perform anaerobic fermentation at 37℃ for 24h to obtain the final fermentation broth. The mixed bacterial suspension includes *Propionibacterium acnes*, *Lactobacillus reuteri*, and *Bifidobacterium*, with a colony count ratio of 1:2:1. The concentration of the mixed bacterial suspension is 1×10⁻⁶. 7 The final fermentation broth was separated by passing CFU / mL through a 0.22μm microporous membrane and then concentrated through a 1000Da nanofiltration membrane to obtain concentrated fermentation broth. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated fermentation broth and stir at 200rpm for 40min. Then add 4g of burdock fiber powder, stir, homogenize at 1500rpm for 10min, and spray dry to obtain oligomeric complex powder. Example 5

[0024] A method for preparing a hypoglycemic compound powder includes the following steps: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 300g of residual particle powder into the reaction vessel, introduce CO2 and pressurize it to 15MPa for 10min, and then release the pressure to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add 700g of complex enzyme buffer to the reaction vessel. The complex enzyme buffer concentration is 3wt.%, the solvent is phosphate buffer, and the complex enzyme composition is 10 parts of neutral protease, 5 parts of lipase, 4 parts of pullulanase, 7 parts of amylase, 12 parts of cellulase, and 5 parts of debranching enzyme. Adjust the pH to 7.0, stir evenly, control the temperature at 40℃ and hydrolyze for 1.5h. During this period, slowly introduce CO2, maintain the pressure at 3MPa, raise the temperature to 85℃ to inactivate the enzyme for 15min, centrifuge and collect the supernatant enzymatic hydrolysate. S3. Fermentation: Cool 500g of enzymatic hydrolysate and inoculate with 25g of *Yeastra lipolytica* suspension. Perform aerobic fermentation at 35℃ for 18 hours with stirring at 180 rpm to obtain the first-stage fermentation broth. Inoculate the first-stage fermentation broth with 50g of a mixed bacterial suspension and perform anaerobic fermentation at 37℃ for 36 hours to obtain the final fermentation broth. The mixed bacterial suspension includes *Propionibacterium acnes*, *Lactobacillus reuteri*, and *Bifidobacterium*, with a colony count ratio of 1:2:1. The concentration of the mixed bacterial suspension is 1×10⁻⁶. 7 The final fermentation broth was separated by passing CFU / mL through a 0.22μm microporous membrane and then concentrated through a 1000Da nanofiltration membrane to obtain concentrated fermentation broth. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated fermentation broth and stir at 200rpm for 40min. Then add 4g of burdock fiber powder, stir, homogenize at 1500rpm for 10min, and spray dry to obtain oligomeric complex powder. Example 6

[0025] A method for preparing a hypoglycemic compound powder includes the following steps: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 300g of residual particle powder into the reaction vessel, introduce CO2 and pressurize it to 15MPa for 10min, and then release the pressure to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add 700g of complex enzyme buffer to the reaction vessel. The complex enzyme buffer concentration is 3wt.%, the solvent is phosphate buffer, and the complex enzyme composition is 10 parts of neutral protease, 5 parts of lipase, 4 parts of pullulanase, 7 parts of amylase, 12 parts of cellulase, and 5 parts of debranching enzyme. Adjust the pH to 7.0, stir evenly, control the temperature at 40℃ and hydrolyze for 1.5h. During this period, slowly introduce CO2, maintain the pressure at 3MPa, raise the temperature to 85℃ to inactivate the enzyme for 15min, centrifuge and collect the supernatant enzymatic hydrolysate. S3. Fermentation: Cool 500g of enzymatic hydrolysate and inoculate with 25g of *Yersinia lipophila* suspension. Perform aerobic fermentation at 35℃ for 18 hours with stirring at 180 rpm to obtain the first-stage fermentation broth. Inoculate the first-stage fermentation broth with 50g of a mixed bacterial suspension and perform anaerobic fermentation at 37℃ for 24 hours to obtain the final fermentation broth. The mixed bacterial suspension includes *Propionibacterium acnes*, *Lactobacillus reuteri*, and *Bifidobacterium*, with a colony count ratio of 1:2:1. The concentration of the mixed bacterial suspension is 1×10⁻⁶. 7 The final fermentation broth was separated by passing CFU / mL through a 0.22μm microporous membrane and then concentrated through a 1000Da nanofiltration membrane to obtain concentrated fermentation broth. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated fermentation broth and stir at 200rpm for 40min. Then add 6g of burdock fiber powder, stir, homogenize at 1500rpm for 10min, and spray dry to obtain oligomeric complex powder. Comparative Example 1

[0026] The difference between this comparative example and Example 1 is that fermentation is not performed, as detailed below: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 300g of residual particle powder into the reaction vessel, introduce CO2 and pressurize it to 15MPa for 10min, and then release the pressure to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add 700g of complex enzyme buffer to the reaction vessel. The complex enzyme buffer concentration is 3wt.%, the solvent is phosphate buffer, and the complex enzyme composition is 10 parts of neutral protease, 5 parts of lipase, 4 parts of pullulanase, 7 parts of amylase, 12 parts of cellulase, and 5 parts of debranching enzyme. Adjust the pH to 7.0, stir evenly, control the temperature at 40℃ and hydrolyze for 1.5h. During this period, slowly introduce CO2, maintain the pressure at 3MPa, raise the temperature to 85℃ to inactivate the enzyme for 15min, centrifuge and collect the supernatant enzyme hydrolysate, separate it through a 0.22μm microporous membrane, and then concentrate it through a 1000Da nanofiltration membrane to obtain concentrated enzyme hydrolysate. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated enzymatic hydrolysate and stir at 200rpm for 40min. Then add 4g of burdock fiber powder, stir, homogenize at 1500rpm for 10min, and spray dry to obtain oligomeric complex powder. Comparative Example 2

[0027] The difference between this comparative example and Example 1 is that it does not use *Yarrowia lipolyticis* for fermentation, as detailed below: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 300g of residual particle powder into the reaction vessel, introduce CO2 and pressurize it to 15MPa for 10min, and then release the pressure to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add 700g of complex enzyme buffer to the reaction vessel. The complex enzyme buffer concentration is 3wt.%, the solvent is phosphate buffer, and the complex enzyme composition is 10 parts of neutral protease, 5 parts of lipase, 4 parts of pullulanase, 7 parts of amylase, 12 parts of cellulase, and 5 parts of debranching enzyme. Adjust the pH to 7.0, stir evenly, control the temperature at 40℃ and hydrolyze for 1.5h. During this period, slowly introduce CO2, maintain the pressure at 3MPa, raise the temperature to 85℃ to inactivate the enzyme for 15min, centrifuge and collect the supernatant enzymatic hydrolysate. S3. Fermentation: Cool 500g of enzymatic hydrolysate, inoculate with 50g of mixed bacterial suspension, and anaerobic ferment at 37℃ for 24h to obtain the final fermentation broth. The mixed bacterial suspension includes Propionibacterium acnes, Lactobacillus reuteri, and Bifidobacterium, with a colony count ratio of 1:2:1, and the concentration of the mixed bacterial suspension is 1×10⁻⁶. 7 The final fermentation broth was separated by passing CFU / mL through a 0.22μm microporous membrane and then concentrated through a 1000Da nanofiltration membrane to obtain concentrated fermentation broth. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated fermentation broth and stir at 200rpm for 40min. Then add 4g of burdock fiber powder, stir, homogenize at 1500rpm for 10min, and spray dry to obtain oligomeric complex powder. Comparative Example 3

[0028] The difference between this comparative example and Example 1 is that no mixed bacteria fermentation is added, as detailed below: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 300g of residual particle powder into the reaction vessel, introduce CO2 and pressurize it to 15MPa for 10min, and then release the pressure to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add 700g of complex enzyme buffer to the reaction vessel. The complex enzyme buffer concentration is 3wt.%, the solvent is phosphate buffer, and the complex enzyme composition is 10 parts of neutral protease, 5 parts of lipase, 4 parts of pullulanase, 7 parts of amylase, 12 parts of cellulase, and 5 parts of debranching enzyme. Adjust the pH to 7.0, stir evenly, control the temperature at 40℃ and hydrolyze for 1.5h. During this period, slowly introduce CO2, maintain the pressure at 3MPa, raise the temperature to 85℃ to inactivate the enzyme for 15min, centrifuge and collect the supernatant enzymatic hydrolysate. S3. Fermentation: Cool 500g of enzymatic hydrolysate, inoculate with 25g of Yersinia lipophila suspension, and carry out aerobic fermentation at 35℃ for 18h under stirring at 180rpm to obtain fermentation broth. Separate the fermentation broth through a 0.22μm microporous membrane, and then concentrate it through a 1000Da nanofiltration membrane to obtain concentrated fermentation broth. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated fermentation broth and stir at 200rpm for 40min. Then add 4g of burdock fiber powder, stir, homogenize at 1500rpm for 10min, and spray dry to obtain oligomeric complex powder. Comparative Example 4

[0029] The difference between this comparative example and Example 1 is that it does not involve enzymatic hydrolysis, as detailed below: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 300g of residual particle powder into the reaction vessel, introduce CO2 and pressurize it to 15MPa for 10min, and then release the pressure to obtain the treated residual particle powder. S2. Add 700g of phosphate buffer to the reaction vessel, stir in a water bath for 1.5h, and centrifuge to collect the supernatant; S3. Fermentation: Cool 500g of supernatant, inoculate with 25g of *Yersinia lipolyticis* suspension, and perform aerobic fermentation at 35℃ for 18h under stirring at 180rpm to obtain the first-stage fermentation broth. Inoculate the first-stage fermentation broth with 50g of mixed bacterial suspension, and perform anaerobic fermentation at 37℃ for 24h to obtain the final fermentation broth. The mixed bacterial suspension includes *Propionibacterium acnes*, *Lactobacillus reuteri*, and *Bifidobacterium*, with a colony count ratio of 1:2:1, and the concentration of the mixed bacterial suspension is 1×10⁻⁶. 7 The final fermentation broth was separated by passing CFU / mL through a 0.22μm microporous membrane and then concentrated through a 1000Da nanofiltration membrane to obtain concentrated fermentation broth. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated fermentation broth and stir at 200rpm for 40min. Then add 4g of burdock fiber powder, stir, homogenize at 1500rpm for 10min, and spray dry to obtain oligomeric complex powder. Comparative Example 5

[0030] The difference between this comparative example and Example 1 is that CO2 is not introduced during the enzymatic hydrolysis process, as detailed below: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 300g of residual particle powder into the reaction vessel, introduce CO2 and pressurize it to 15MPa for 10min, and then release the pressure to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add 700g of complex enzyme buffer to the reaction vessel. The complex enzyme buffer concentration is 3wt.%, the solvent is phosphate buffer, and the complex enzyme composition is 10 parts of neutral protease, 5 parts of lipase, 4 parts of pullulanase, 7 parts of amylase, 12 parts of cellulase, and 5 parts of debranching enzyme. Adjust the pH to 7.0, stir evenly, control the temperature at 40℃ for 1.5h for enzymatic hydrolysis, raise the temperature to 85℃ for 15min to inactivate the enzyme, centrifuge and collect the supernatant enzymatic hydrolysate. S3. Fermentation: Cool 500g of enzymatic hydrolysate and inoculate with 25g of *Yersinia lipophila* suspension. Perform aerobic fermentation at 35℃ for 18 hours with stirring at 180 rpm to obtain the first-stage fermentation broth. Inoculate the first-stage fermentation broth with 50g of a mixed bacterial suspension and perform anaerobic fermentation at 37℃ for 24 hours to obtain the final fermentation broth. The mixed bacterial suspension includes *Propionibacterium acnes*, *Lactobacillus reuteri*, and *Bifidobacterium*, with a colony count ratio of 1:2:1. The concentration of the mixed bacterial suspension is 1×10⁻⁶. 7 The final fermentation broth was separated by passing CFU / mL through a 0.22μm microporous membrane and then concentrated through a 1000Da nanofiltration membrane to obtain concentrated fermentation broth. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated fermentation broth and stir at 200rpm for 40min. Then add 4g of burdock fiber powder, stir, homogenize at 1500rpm for 10min, and spray dry to obtain oligomeric complex powder. Comparative Example 6

[0031] The difference between this comparative example and Example 1 is that the raw material pretreatment does not involve CO2 pressurization, as detailed below: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 300g of residual particle powder into the reaction vessel. S2. Enzymatic hydrolysis: Add 700g of complex enzyme buffer to the reaction vessel. The complex enzyme buffer concentration is 3wt.%, the solvent is phosphate buffer, and the complex enzyme composition is 10 parts of neutral protease, 5 parts of lipase, 4 parts of pullulanase, 7 parts of amylase, 12 parts of cellulase, and 5 parts of debranching enzyme. Adjust the pH to 7.0, stir evenly, control the temperature at 40℃ and hydrolyze for 1.5h. During this period, slowly introduce CO2, maintain the pressure at 3MPa, raise the temperature to 85℃ to inactivate the enzyme for 15min, centrifuge and collect the supernatant enzymatic hydrolysate. S3. Fermentation: Cool 500g of enzymatic hydrolysate and inoculate with 25g of *Yersinia lipophila* suspension. Perform aerobic fermentation at 35℃ for 18 hours with stirring at 180 rpm to obtain the first-stage fermentation broth. Inoculate the first-stage fermentation broth with 50g of a mixed bacterial suspension and perform anaerobic fermentation at 37℃ for 24 hours to obtain the final fermentation broth. The mixed bacterial suspension includes *Propionibacterium acnes*, *Lactobacillus reuteri*, and *Bifidobacterium*, with a colony count ratio of 1:2:1. The concentration of the mixed bacterial suspension is 1×10⁻⁶. 7 The final fermentation broth was separated by passing CFU / mL through a 0.22μm microporous membrane and then concentrated through a 1000Da nanofiltration membrane to obtain concentrated fermentation broth. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated fermentation broth and stir at 200rpm for 40min. Then add 4g of burdock fiber powder, stir, homogenize at 1500rpm for 10min, and spray dry to obtain oligomeric complex powder. Comparative Example 7

[0032] The difference between this comparative example and Example 1 is that burdock fiber powder is not added, as detailed below: S1. Raw material pretreatment: Wash the residual particles, dry them, pulverize them at low temperature, and sieve them to obtain residual particle powder. Put 300g of residual particle powder into the reaction vessel, introduce CO2 and pressurize it to 15MPa for 10min, and then release the pressure to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add 700g of complex enzyme buffer to the reaction vessel. The complex enzyme buffer concentration is 3wt.%, the solvent is phosphate buffer, and the complex enzyme composition is 10 parts of neutral protease, 5 parts of lipase, 4 parts of pullulanase, 7 parts of amylase, 12 parts of cellulase, and 5 parts of debranching enzyme. Adjust the pH to 7.0, stir evenly, control the temperature at 40℃ and hydrolyze for 1.5h. During this period, slowly introduce CO2, maintain the pressure at 3MPa, raise the temperature to 85℃ to inactivate the enzyme for 15min, centrifuge and collect the supernatant enzymatic hydrolysate. S3. Fermentation: Cool 500g of enzymatic hydrolysate and inoculate with 25g of *Yersinia lipophila* suspension. Perform aerobic fermentation at 35℃ for 18 hours with stirring at 180 rpm to obtain the first-stage fermentation broth. Inoculate the first-stage fermentation broth with 50g of a mixed bacterial suspension and perform anaerobic fermentation at 37℃ for 24 hours to obtain the final fermentation broth. The mixed bacterial suspension includes *Propionibacterium acnes*, *Lactobacillus reuteri*, and *Bifidobacterium*, with a colony count ratio of 1:2:1. The concentration of the mixed bacterial suspension is 1×10⁻⁶. 7 The final fermentation broth was separated by passing CFU / mL through a 0.22μm microporous membrane and then concentrated through a 1000Da nanofiltration membrane to obtain concentrated fermentation broth. S4. Formulation: Add 4g of brown algae oligosaccharide and 3g of peanut peptide to 97g of water and sonicate at 200W for 10min to obtain a mixed solution. Add the mixture to 80g of concentrated fermentation broth, stir at 200rpm for 40min, homogenize at 1500rpm for 10min, and spray dry to obtain the oligomeric complex powder.

[0033] Performance testing Assay for α-glucosidase inhibitory activity The inhibitory activity of each sample against α-glucosidase was determined using the PNPG method. The specific steps are as follows: Take 50 μL of sample solutions of different concentrations, add 50 μL of α-glucosidase solution, pre-incubate at 37℃ for 10 min, add 100 μL of PNPG solution, react at 37℃ for 20 min, terminate the reaction by adding 10 μL of 1 mol / L Na2CO3 solution, measure the absorbance at a wavelength of 405 nm, and calculate the inhibition rate.

[0034]

[0035] The results are shown in Table 1. The inhibition rate of α-glucosidase in the examples was significantly higher than that in the comparative examples, and the IC50 value was also lower. The IC50 value of Comparative Example 1 (unfermented) was 1.85 mg / mL, which was significantly higher than that of Example 1 (0.52 mg / mL), with an inhibition rate of 65.8%, which was 43.8% lower than that of Example 1. This indicates that the two-stage mixed fermentation significantly improved the inhibitory activity of α-glucosidase, mainly because fermentation produced more short-chain fatty acids and functional metabolites. The IC50 value of Comparative Example 4 (unenzymatic hydrolysis) was 1.45 mg / mL, which was higher than that of Example 1, indicating that the CO2 cell wall disruption-enzymatic hydrolysis synergistic technology played a key role in releasing active ingredients and improving fermentation efficiency. The IC50 value of Comparative Example 5 (enzymatic hydrolysis without CO2) was higher than that of Example 1, and the inhibition rate was also significantly lower than that of Example 1, confirming the protective effect of CO2 in the enzymatic hydrolysis process and avoiding the degradation of heat-sensitive active ingredients. The data of Comparative Example 6 (raw material without CO2 pressurization) shows that CO2 explosion pretreatment plays an important role in destroying cell wall structure and improving the release of active ingredients.

[0036] Table 1. Inhibitory activity of each sample against α-glucosidase

[0037] In vitro simulated digestion stability test The study simulates the gastrointestinal digestive environment to examine the stability and activity retention rate of the samples during digestion.

[0038] Simulated gastric juice (SGF): 0.2% NaCl, adjusted to pH 1.2, with pepsin (3.2 g / L). Simulated intestinal fluid (SIF): 0.05 mol / L KH2PO4, adjusted to pH 7.5, with the addition of pancreatic enzyme (10 g / L). Experimental procedure: Add the sample solution to simulated gastric juice, incubate at 37°C for 2 hours, adjust the pH to 7.5, add simulated intestinal juice, incubate at 37°C for 4 hours, measure the change in α-glucosidase inhibitory activity before and after digestion, and calculate the activity retention rate.

[0039] The results are shown in Table 2. In the examples, the retention rates after gastric digestion were all above 90%, and the retention rates after intestinal digestion were all above 85%. In Comparative Example 7, without the addition of burdock fiber, the retention rate after intestinal digestion decreased from 88.6% in Example 1 to 61.2%, indicating that burdock fiber plays a crucial role in protecting the active ingredients in intestinal fluid.

[0040] Table 2. Retention rate of bioactive components after in vitro simulated digestion

[0041] Blood sugar lowering effect evaluation SPF-grade male SD rats, weighing 180-220g, were purchased and acclimatized for one week. A diabetic model was established by feeding them a high-fat, high-sugar diet for four weeks, followed by intraperitoneal injection of streptozotocin (STZ) 35mg / kg. Successful model establishment was defined as a fasting blood glucose level >16.7mmol / L. Rats were divided into groups: a normal control group was administered distilled water by gavage; a model control group was administered distilled water by gavage; and a positive control group was administered metformin 200mg / kg by gavage. The example and comparative studies used 500mg / kg. Ten rats were assigned to each group, and the administration was continuous for four weeks. Fasting blood glucose (FBG) was measured weekly.

[0042] The results are shown in Table 3. Example 1 showed a 49.1% reduction in blood glucose, compared to only 13.7% in Comparative Example 1, fully demonstrating the core role of the two-stage mixed-culture fermentation in lowering blood glucose. Comparative Example 2 showed a 29.5% reduction, significantly lower than that of Example 1, indicating that the first-stage fat degradation laid the foundation for subsequent blood glucose lowering, demonstrating the synergistic effect of the mixed bacteria. Compared to Example 1, Comparative Example 3 showed a 13% reduction, confirming the combined effect of Propionibacterium acnes, Lactobacillus reuteri, and Bifidobacterium. Comparative Example 4 showed a 19.1% reduction in blood glucose, indicating that the CO2 cell wall disruption-enzymatic hydrolysis synergistic technology is crucial for the release of active ingredients.

[0043] Table 3. Changes in fasting blood glucose levels in rats of each group (mmol / L)

[0044] 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 method for preparing a hypoglycemic compound powder, characterized in that, Includes the following steps: S1. Raw material pretreatment: The residual particles are washed, dried and then pulverized at low temperature. The residual particles are sieved to obtain residual particle powder. The residual particle powder is put into the reaction vessel, CO2 is introduced and pressurized to maintain the pressure, and then the pressure is released to obtain the treated residual particle powder. S2. Enzymatic hydrolysis: Add the complex enzyme buffer to the reaction vessel, adjust the pH value, stir evenly to start enzymatic hydrolysis, slowly pass CO2 during the process, heat up to inactivate the enzyme, centrifuge and take the supernatant enzymatic hydrolysate. S3. Fermentation: Cool the enzymatic hydrolysate, inoculate it with a suspension of Yersinia lipophila, and carry out aerobic fermentation under stirring to obtain the first stage fermentation broth. Inoculate the mixed bacterial suspension into the first stage fermentation broth and carry out anaerobic fermentation to obtain the final fermentation broth. Separate the final fermentation broth through a membrane and then concentrate it through a nanofiltration membrane to obtain the concentrated fermentation broth. S4. Formulation: The mixture of brown algae oligosaccharide and peanut peptide with water is ultrasonically added to the concentrated fermentation broth and stirred to react. Then, burdock fiber powder is added, stirred and homogenized, and spray-dried to obtain the oligomeric complex powder.

2. The method for preparing a hypoglycemic compound powder according to claim 1, characterized in that: In step S1, the pressure is increased to 10-20 MPa and maintained for 5-15 minutes.

3. The method for preparing a hypoglycemic compound powder according to claim 1, characterized in that: In step S2, the mass ratio of the complex enzyme buffer to the residual powder is (7-8):(2-3); the concentration of the complex enzyme buffer is 2.5-5 wt.%, and the buffer is phosphate buffer; the pH value is adjusted to 6.5-7.5; the enzymatic hydrolysis temperature is 35-45℃, and the time is 1-2 h; CO2 is introduced to maintain the pressure at 2-5 MPa; the temperature is raised to 80-90℃, and the enzyme inactivation time is 10-20 min.

4. The method for preparing a hypoglycemic compound powder according to claim 1, characterized in that: The complex enzyme buffer contains, by weight fraction, 5-15 parts neutral protease, 3-8 parts lipase, 2-6 parts pullulanase, 5-10 parts amylase, 8-15 parts cellulase, and 3-7 parts debranching enzyme.

5. The method for preparing a hypoglycemic compound powder according to claim 1, characterized in that: In step S3, the concentration of the *Yarrowia lipophila* suspension is 1 × 10⁻⁶. 7 -1×10 8 The inoculum concentration is CFU / mL, and the inoculum size is 5-8 wt.% of the enzymatic hydrolysate. The stirring speed is 150-200 rpm. The temperature for aerobic fermentation is 30-35℃, and the time is 18-24 h. The inoculum size for mixed bacterial suspension is 10-15 wt.%. The temperature for anaerobic fermentation is 35-38℃, and the time is 24-36 h.

6. The method for preparing a hypoglycemic compound powder according to claim 1, characterized in that: The mixed bacterial suspension includes Propionibacterium acnes, Lactobacillus reuteri, and Bifidobacterium, with a colony count ratio of (1-2):(2-3):(1-2); the concentration of the mixed bacterial suspension is 1×10⁻⁶. 6 -1×10 8 CFU / mL; membrane separation uses a 0.22μm microporous membrane, and nanofiltration membrane has a molecular weight cutoff of 600-1000 Da.

7. The method for preparing a hypoglycemic compound powder according to claim 1, characterized in that: In step S4, the mass ratio of the mixed liquid to the concentrated fermentation liquid and burdock fiber powder is (50-70):(40-60):(2-3); the ultrasonic power is 150-250W and the time is 10-15min; the stirring speed is 200-300rpm and the time is 30-60min; the homogenization speed is 1000-2000rpm and the time is 10-15min.

8. The method for preparing a hypoglycemic compound powder according to claim 1, characterized in that: In step S4, the mixture contains 2-4 wt.% brown algae oligosaccharides and 1-3 wt.% peanut peptides.

9. A hypoglycemic compound powder prepared by any one of the preparation methods according to claims 1-8.

10. The application of the hypoglycemic compound powder according to claim 9 in the preparation of functional foods and hypoglycemic drugs.

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