Diabetes intervention beverage based on trifocal-microbial axis theory and preparation method

Abstract

The invention relates to the technical field of functional food, in particular to a diabetes intervention beverage based on a triple-jiao-microbial axis theory and a preparation method of the diabetes intervention beverage based on the triple-jiao-microbial axis theory. Saccharomyces boulardii, lactobacillus plantarum DR7 and ackermansiella muciniphila AM06 are subjected to solid-liquid two-phase stepped fermentation to prepare the lactobacillus plantarum and ackermansiella muciniphila. The traditional Chinese medicine tri-jiao syndrome differentiation and the modern intestinal flora theory are fused, the raw materials are converted into a product rich in lovastatin, 1-deoxynojirimycin and other active ingredients through multi-strain synergistic fermentation, the product can act on multiple glycometabolism targets and intestinal microecology at the same time, the multiple effects of reducing blood sugar, improving insulin resistance, adjusting flora and the like are achieved, and the product is suitable for being used as a health-care food. And a new dietary solution is provided for intervention of the type 2 diabetes mellitus.

Description

Technical Field

[0001] This invention relates to the field of functional food technology, and in particular to a diabetes intervention beverage based on the tri-joule-microbe axis theory and its preparation method. Background Technology

[0002] Type 2 diabetes is a globally prevalent chronic metabolic disease with a continuously rising incidence rate, posing a serious public health problem. While current first-line treatments effectively control blood sugar, long-term use is generally limited by factors such as gastrointestinal intolerance, risk of hypoglycemia, weight gain, and the inability to prevent the progressive decline of pancreatic β-cell function.

[0003] In recent years, with the development of microbiome research, studies have confirmed that the occurrence and development of type 2 diabetes mellitus (T2DM) are closely related to the dysregulation of the "gut microbiota-metabolites-host" signaling axis. Patients generally exhibit a series of microecological imbalances, such as decreased abundance of beneficial bacteria producing short-chain fatty acids, abnormally active branched-chain amino acid synthesis and metabolism, and impaired intestinal barrier function leading to increased lipopolysaccharide translocation and chronic low-grade inflammation, which in turn exacerbate systemic insulin resistance.

[0004] In Traditional Chinese Medicine (TCM) theory, the "Triple Burner" governs the flow of Qi and body fluids. Its functional description—"the upper burner is like mist, the middle burner like a fermenting vat, and the lower burner like a drainage ditch"—highly aligns with the modern medical "intestine-liver-pancreas-system" axis. This suggests that holistic, targeted regulation of the "Triple Burner-microbe axis" could potentially improve glucose and lipid metabolism and homeostasis across multiple pathways, thereby compensating for the limitations of single-target chemical drugs.

[0005] However, most of the blood sugar-lowering functional foods or health products currently on the market focus only on a single active ingredient and generally suffer from the following defects: First, they lack compound formulations based on systems theory, making it difficult to achieve synergistic effects on multiple targets; second, they lack stable and controllable biotransformation processes, resulting in large quality differences between batches of products; and third, they lack a quality marker system that is closely related to the core efficacy and can be quantitatively monitored.

[0006] The present invention aims to solve the technical problems existing in the prior art. To this end, a diabetes intervention beverage based on the tri-john-microbe axis theory and its preparation method are proposed. Summary of the Invention

[0007] The purpose of this invention is to provide a diabetes intervention beverage based on the tri-joule-microbe axis theory and its preparation method, so as to solve the technical problems existing in the prior art.

[0008] By adopting the above technical solution, the present invention has the following beneficial effects:

[0009] This invention provides a fermented beverage for diabetes intervention. The beverage uses budding purple yam as the core solid matrix, combined with Poria cocos, kudzu root, hawthorn, chickpea germ, buckwheat sprout, mulberry leaf, oat germ, and bitter melon as raw materials. After being inoculated with Ganoderma lucidum water for germination, it is prepared by sequentially inoculating with Saccharomyces boulardii, Lactobacillus plantarum DR7, and Akkermansia muciniphila AM06 for solid-state and liquid-state two-phase step fermentation. The beverage contains ≥0.30% (w / w) lovastatin, ≥0.05% (w / w) 1-deoxynojirimycin, and anthocyanin retention ≥85%. Preferably, the budding purple yam is obtained by inoculating with Ganoderma lucidum water at 15℃ for 24 hours, with a sprout length of 1-3 cm and a sprout weight ≥12% of the total weight of the raw materials.

[0010] As a further aspect of the present invention, the total duration of the stepped fermentation does not exceed 48 hours. Specifically, the solid-state fermentation stage involves inoculating 3% *Saccharomyces boulardii* and fermenting for 6 hours, followed by inoculating 2% *Lactobacillus plantarum* DR7 and continuing fermentation for 12 hours. The liquid-state fermentation stage involves inoculating 1.5% *Ackermania mutans* AM06 and fermenting under anaerobic conditions at 35°C for 18 hours.

[0011] As a further aspect of the present invention: the viable count of Akkermansia muciniphila in the beverage is 1.0×10^6-1.0×10^8 cells / mL, and the lead (Pb) content is ≤0.18mg / kg.

[0012] This invention provides a method for preparing the above-mentioned fermented beverage for diabetes intervention, comprising the following steps:

[0013] a. Wash and cut the purple yam into pieces, and use Ganoderma lucidum water to promote germination at 15℃ for 24 hours to obtain the germinating purple yam;

[0014] b. Mix the budding purple yam with Poria cocos, kudzu root, hawthorn, chickpea germ, buckwheat sprout, mulberry leaf, oat germ and bitter melon, and carry out solid-state two-stage fermentation;

[0015] c. Add Ganoderma lucidum water to the solid fermentation material, adjust the pH of the system to 5.0-5.4, and inoculate with Akkermansia melanophilus AM06 for liquid anaerobic fermentation;

[0016] d. The liquid fermentation product is subjected to low-temperature cold soaking, filtration, instantaneous sterilization, and aseptic filling to obtain the final product. As a further aspect of this invention: the Ganoderma lucidum water is sterilized through a 0.22μm filter membrane, wherein the polysaccharide content is ≥2.5% w / v, and the pH value is 6.0-6.5.

[0017] As a further aspect of the present invention: the germination-promoting conditions in step a are to make the purple yam buds grow to 1-3cm and the bud weight ≥ 12% of the total weight of the raw materials.

[0018] As a further aspect of the present invention: the solid-state two-stage fermentation in step b specifically involves: first, inoculating with Saccharomyces boulardii and fermenting for 6 hours under aeration conditions of 28°C, 85% relative humidity; then inoculating with Lactobacillus plantarum DR7 and continuing fermentation for 12 hours under the same temperature and humidity conditions.

[0019] As a further aspect of the present invention: the low-temperature cold soaking in step d is soaking at 4°C for 6 hours; the instantaneous sterilization is maintaining at 65°C for 30 seconds.

[0020] This invention provides the use of the above-mentioned diabetes intervention fermented beverage in the preparation of functional foods or special medical purpose formulations for reducing fasting blood glucose, improving insulin resistance, or increasing the relative abundance of Akkermansia muciniphila in the intestine.

[0021] Compared with the prior art, the beneficial effects of the present invention are:

[0022] 1. This product innovatively combines the TCM theory of "Triple Energizer" with the modern "Gut Microbiota-Gut-Pancreatic Axis" theory. The formulation is based on a "Meridian-Microbiota-Target" network, achieving synergistic compatibility of nine food and medicinal substances. Through a single formulation, the product simultaneously targets multiple key targets and processes, including α-glucosidase inhibition, GLP-1 receptor activation, AMPK signaling pathway regulation, DPP-4 inhibition, and gut microbiota remodeling. This overcomes the limitations of single-component or single-target products, achieving synergistic intervention for type 2 diabetes mellitus involving glucose and lipid metabolism disorders, insulin resistance, and gut microbiota dysbiosis.

[0023] 2. A unique biotransformation process is employed, involving a "solid-liquid two-phase step fermentation" and a "yeast-Lactobacillus-Ackermania three-stage inoculation." This process, through the sequential action of microorganisms, not only efficiently converts macromolecules in the raw materials into small-molecule active ingredients but also significantly improves the yield and bioavailability of the target active ingredients. For example, compared to conventional fermentation, this process can increase the yield of lovastatin several times over while effectively retaining heat-sensitive components such as 1-deoxynojirimycin and anthocyanins, achieving a balance between "enhanced efficacy" and "stability."

[0024] 3. A complete and scalable standardized process flow has been established, from raw material germination and multi-strain fermentation to post-processing, with a total time controllable within 48 hours and low energy consumption. More importantly, a "PAG three-dimensional index" centered on "potency-availability-protection" has been proposed as a quality control concept, and a comprehensive quality marker system covering key active ingredients, microbiological indicators, and safety indicators has been established. Combined with blockchain coding technology, real-time monitoring and traceability of key parameters throughout the production process have been achieved, ensuring the stability and consistency of product batches.

[0025] 4. All raw materials are selected from food-grade pharmaceutical substances, and after microbial fermentation and transformation, the risk of potential adverse reactions is further reduced. Animal toxicology experiments show that its maximum tolerated dose is high, and no liver or kidney toxicity was observed. Preliminary clinical trials show that while the product significantly improves glycated hemoglobin and postprandial blood glucose, the incidence of gastrointestinal side effects is extremely low, and patient compliance is high, providing a safe basis for long-term dietary intervention.

[0026] 5. The final product can be made into a liquid beverage for direct consumption, or it can be encapsulated and spray-dried into an instant powder. The powder has low moisture content, good stability, and rapid dispersibility after reconstitution, making it easy to store and transport. It can also be used as a raw material for functional foods or special medical foods in different forms such as solid beverages and capsules, meeting the needs of different consumption scenarios and applications. Detailed Implementation

[0027] Embodiments of the present invention are described in detail below, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The following embodiments are exemplary and intended to explain the present invention, and should not be construed as limiting the present invention.

[0028] The following disclosure provides numerous different embodiments or examples for implementing various structures of the invention. To simplify the disclosure, specific examples of components and arrangements are described below. These are merely examples and are not intended to limit the scope of the invention. Furthermore, reference numerals and / or letters may be repeated in different examples. Such repetition is for simplification and clarity and does not in itself indicate a relationship between the various embodiments and / or arrangements discussed.

[0029] This invention proposes a "microbial community-mediated regulation of the three jiaos" strategy. Using resistant starch and mucopolysaccharides from germinating purple yam as the core, it acts on the upper jiao, selectively proliferating Akkermansia muciniphila and promoting glucagon-like peptide-1 (GLP-1) secretion. Mulberry leaves rich in 1-deoxynojirimycin (1-DNJ) and bitter melon rich in polypeptide P act on the middle jiao, inhibiting α-glucosidase activity. Poria cocos, kudzu root, and hawthorn act on the lower jiao, improving the insulin signaling pathway. Through a two-phase solid-state and liquid-state fermentation process, the macromolecules of the raw materials are converted into easily absorbed small-molecule active substances, and beneficial components such as lovastatin and γ-aminobutyric acid (GABA) are enriched.

[0030] Example 1: Laboratory-scale preparation

[0031] According to the formula shown in Table 1, a total of 6 kg of raw materials were accurately weighed. Purple yam was cut into small pieces of approximately 10 mm³, and *Ganoderma lucidum* water was added at a material-to-liquid ratio of 1:1.2 (w / v). Germination was promoted at 15℃ in the dark for 24 hours, and the sprout length was measured to be approximately 2 cm, with the sprout weight reaching 13% of the raw material weight. The germinating purple yam was mixed with other raw materials, and first inoculated with 3% *Saccharomyces boulardii* CNCMI-745, fermenting for 6 hours at 28℃, 85% relative humidity, and aeration. Then, 2% *Lactobacillus plantarum* DR7 was inoculated, and fermentation continued for 12 hours. Eight times the volume of *Ganoderma lucidum* water was added to the solid fermentation material to adjust the pH to 5.2, and 1.5% *Ackermania amphiphila* AM06 was inoculated. Fermentation was carried out at 35℃ under anaerobic conditions for 18 hours. The fermentation broth was cold-soaked at 4℃ for 6 hours, filtered, and then sterilized at 65℃ for 30 seconds. The resulting beverage was aseptically bottled. Test results: Lovastatin content 0.35% (w / w), 1-deoxynojirimycin content 0.07% (w / w), anthocyanin retention rate 88.2%, Akkermansia myxophilus viable count 2.1×10^7 cells / mL, lead content 0.05 mg / kg.

[0032] Example 2: Pilot-scale preparation

[0033] Production was scaled up using a 500L solid-state fermenter and a 1m³ anaerobic liquid-state fermenter. The raw material ratios and process flow were the same as in Example 1, with the stirring speed adjusted to 12 rpm to prevent cell damage. The final product yield was 92%, and the relative standard deviation of the key active ingredient content from the laboratory scale was less than 5%, demonstrating the feasibility of process scale-up.

[0034] Example 3: Preparation of Powder Formulations

[0035] The fermentation broth obtained in Example 2 was encapsulated with 30% (w / w) maltodextrin and dried using spray drying technology with an inlet air temperature of 180°C and an outlet air temperature of 85°C to obtain an instant powder with a moisture content of 3.8%. This powder contained 0.06% (w / w) 1-deoxynojirimycin and 0.32% (w / w) lovastatin, with a dispersibility time of 15 seconds.

[0036] Table 1: Beverage formulation composition (based on the preparation of 1000 mL of final product)

[0037]

[0038] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0039] Although embodiments of the invention have been shown and described, those skilled in the art will understand that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents.

Claims

1. A fermented beverage for diabetes intervention, characterized in that, Using purple yam in its germination stage as a solid substrate, and combining it with Poria cocos, kudzu root, hawthorn, chickpea germ, buckwheat sprout, mulberry leaf, oat germ, and bitter melon as raw materials, the beverage is prepared by sequentially inoculating *Ganoderma lucidum* (a type of mushroom) with water to promote germination, followed by solid-state and liquid-state two-phase step fermentation with *Saccharomyces boulardii*, *Lactobacillus plantarum* DR7, and *Ackermania muscaria* AM06. The beverage contains ≥0.30% (w / w) lovastatin, ≥0.05% (w / w) 1-deoxynojirimycin, and anthocyanin retention rate ≥85%.

2. The fermented beverage for diabetes intervention according to claim 1, characterized in that, The budding purple yam was obtained by promoting germination of Ganoderma lucidum in water at 15℃ for 24 hours. Its bud length is 1-3cm and the bud weight is ≥12% of the total weight of the raw material.

3. The fermented beverage for diabetes intervention according to claim 1 or 2, characterized in that, The total fermentation time of the step fermentation shall not exceed 48 hours; the solid fermentation stage is to first inoculate with 3% Saccharomyces boulardii and ferment for 6 hours, and then inoculate with 2% Lactobacillus plantarum DR7 and continue fermentation for 12 hours; the liquid fermentation stage is to inoculate with 1.5% Akkermansia muciniphila AM06 and ferment under anaerobic conditions at 35°C for 18 hours.

4. The diabetes intervention fermented beverage according to any one of claims 1 to 3, characterized in that, The beverage contained 1.0×10^6-1.0×10^8 cells / mL of viable Akkermansia muciniphila and ≤0.18mg / kg of lead.

5. A method for preparing a fermented beverage for diabetes intervention as described in any one of claims 1 to 4, characterized in that, Includes the following steps: a. Wash and cut the purple yam into pieces, and use Ganoderma lucidum water to promote germination at 15℃ to obtain purple yam in the germination stage; b. Mix the budding purple yam with Poria cocos, kudzu root, hawthorn, chickpea germ, buckwheat sprout, mulberry leaf, oat germ and bitter melon, and carry out solid-state two-stage fermentation; c. Add Ganoderma lucidum water to the solid fermentation material, adjust the pH of the system to 5.0-5.4, and inoculate with Akkermansia melanophilus AM06 for liquid anaerobic fermentation; d. The liquid fermentation product is post-processed to obtain the beverage.

6. The method according to claim 5, characterized in that, The Ganoderma lucidum water is sterilized by a 0.22μm filter membrane, and has a polysaccharide content of ≥2.5%w / v and a pH value of 6.0-6.

5.

7. The method according to claim 5, characterized in that, The germination time mentioned in step a is 24 hours. After germination, the purple yam sprouts are 1-3 cm long and the sprout weight is ≥ 12% of the total weight of the raw materials.

8. The method according to any one of claims 5 to 7, characterized in that, The solid-state two-stage fermentation described in step b is as follows: first, fermentation is carried out for 6 hours under conditions of 28°C, 85% relative humidity, and ventilation by inoculating with Saccharomyces boulardii; then, fermentation is continued for 12 hours by inoculating with Lactobacillus plantarum DR7 under the same temperature and humidity.

9. The method according to any one of claims 5 to 8, characterized in that, The post-processing described in step d includes cold soaking at 4°C for 6 hours, filtration, instantaneous sterilization at 65°C for 30 seconds, and aseptic filling.

10. The use of the diabetes intervention fermented beverage as described in any one of claims 1 to 4 in the preparation of functional foods or special medical purpose formulations for reducing fasting blood glucose, improving insulin resistance, or increasing the relative abundance of Akkermansia muciniphila in the intestine.

Images