A microbial preparation and application thereof in preparation of a spleen-invigorating and dampness-dispelling fermentation product

Abstract

The present application belongs to the technical field of microbial fermentation, and particularly relates to a microbial preparation and application thereof in preparation of a spleen-strengthening and dampness-dispelling fermentation product. The microbial preparation comprises Lactobacillus rhamnosus LRB-2 and Lactobacillus plantarum YSH08. Lactobacillus rhamnosus LRB-2 and Lactobacillus plantarum YSH08 exhibit a significant interspecies synergistic effect in co-culture in a spleen-strengthening and dampness-dispelling composition matrix. The fermentation product obtained by enzymolysis and inoculation of the above microbial preparation has a high content of total polyphenols, total flavonoids and polysaccharides, can significantly improve the clearance rate of DPPH free radicals, hydroxyl free radicals, superoxide anion free radicals and ABTS cationic free radicals, improve the antioxidant capacity, can achieve the effects of lowering blood lipids and lowering blood sugar by inhibiting the activity of pancreatic lipase and the activity of alpha-glucosidase, and can improve the state of white and greasy tongue fur, tongue body tooth marks and cracks in the spleen deficiency and dampness accumulation population.

Description

Technical Field

[0001] This invention belongs to the field of microbial fermentation technology, specifically relating to a microbial preparation and its application in the preparation of spleen-strengthening and dampness-removing fermented products. Background Technology

[0002] Medicinal and edible compound formulas, following the principles of traditional Chinese medicine compatibility, are rich in natural active components such as polyphenols, flavonoids, and polysaccharides, making them excellent fermentation substrates. However, the synergistic effect of compound fermentation is specific to the microbial strain and substrate, lacking universality across different formulas: a combination of microbial strains that significantly enhances the effect in one formula often yields poor results in others; each formula requires its own specific microbial strain combination to achieve the dissociation, transformation, and enrichment of active ingredients. The multiple medicinal herbs in a compound formula lead to a complex substrate composition, with significant differences in the composition, proportion, and physicochemical properties of active ingredients among different formulas. Tannins and strong antioxidants in some formulas can significantly inhibit microbial growth and metabolism; simultaneously, active substances within the substrate are prone to oxidation and polymerization, hindering the release of active ingredients. Furthermore, the active ingredients in compound formulas have a bidirectional regulatory effect on microbial growth; differences in the types and proportions of components directly affect cell proliferation and metabolic efficiency, further widening the differences in fermentation effects among different formulas.

[0003] At the application level, there is no universal adaptability for the synergistic growth of strains. There is no universal microbial community combination suitable for all compound substrates, and not all strains can form a synergistic symbiotic system in the same fermentation substrate. Different microbial strains exhibit significant differences in metabolic pathways, nutrient preferences, and environmental tolerance characteristics. When any strain is mixed and co-cultured, antagonistic problems such as competition for nutrient substrates, mutual inhibition of metabolites, and mismatched growth cycles easily occur, severely disrupting the overall homeostasis of the microbial community. In particular, the complex composition of medicinal and edible compound substrates, containing active ingredients such as tannins and polyphenols, has differentiated growth-promoting or inhibitory effects on different strains, further exacerbating the difficulty of microbial community adaptation.

[0004] Therefore, the core of efficient fermentation of food-medicine homology compounds lies in the precise matching of specific compound substrates with exclusive synergistic strains. Currently, the industry lacks mature systems for evaluating strain interactions and screening substrate compatibility, making it impossible to selectively identify stable symbiotic microbial communities that are compatible with specific compounds, thus hindering the precise matching of strains and substrates. This core technological bottleneck results in low fermentation conversion efficiency and limited efficacy enhancement, severely restricting the industrialization of the food-medicine homology fermentation industry. Summary of the Invention

[0005] Therefore, the present invention aims to provide a microbial preparation in which, compared with single bacteria, the two bacteria Lactobacillus rhamnosus LRB-2 and Lactobacillus plantarum YSH08 exhibit a significant interspecies synergistic proliferation effect. In addition, the microbial preparation can significantly increase the release of effective total polyphenols, total flavonoids and polysaccharides. At the same time, it can significantly improve the DPPH free radical scavenging rate, hydroxyl free radical scavenging rate, superoxide anion free radical scavenging rate and pancreatic lipase inhibition rate.

[0006] The second objective of this invention is to provide a fermented product that is both food and medicine, which can effectively enrich active ingredients and has higher content of total polyphenols, total flavonoids and polysaccharides.

[0007] The third objective of this invention is to provide a food-medicine homologous fermented product that can significantly improve the scavenging rates of DPPH free radicals, hydroxyl free radicals, superoxide anion free radicals, and ABTS cation free radicals, and has excellent antioxidant activity.

[0008] The fourth objective of this invention is to provide a fermented product that is both food and medicine, which can effectively inhibit the activity of pancreatic lipase and α-glucosidase, and has good potential for regulating blood lipids and blood sugar.

[0009] To solve the aforementioned technical problems / achieve the aforementioned objectives, or at least partially solve the aforementioned technical problems / achieve the aforementioned objectives, in a first aspect, the present invention provides a microbial preparation, the active ingredient of which includes *Lactobacillus rhamnosus* (…). Lacticaseibacillus rhamnosus LRB-2 and Lactobacillus plantarum ( Lactiplantibacillus plantarum YSH08;

[0010] The Lactobacillus rhamnosus LRB-2 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 39596, deposit date July 6, 2026, and deposit address No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

[0011] The preservation number of the Lactobacillus plantarum YSH08 is CGMCC No. 36182.

[0012] Preferably, the volume ratio of Lactobacillus rhamnosus LRB-2 to Lactobacillus plantarum YSH08 is (0.05-0.3):(0.05-0.3).

[0013] Specifically, in some embodiments of the present invention, the volume ratio of Lactobacillus rhamnosus LRB-2 to Lactobacillus plantarum YSH08 is 1:1.

[0014] Preferably, the viable count of the *Lactobacillus rhamnosus* LRB-2 is 1.0 × 10⁻⁶.8 CFU / mL or higher; the viable count of the *Lactobacillus plantarum* YSH08 is 1.0 × 10⁻⁶. 8 CFU / mL or higher.

[0015] Preferably, the microbial preparation further includes adjuvants for maintaining the survival of microorganisms.

[0016] In a second aspect, the present invention provides the application of the above-mentioned microbial preparation in the fermentation of food-medicine homologous compositions or in the preparation of food-medicine homologous fermented products.

[0017] In a third aspect, the present invention provides a medicinal and edible fermented product, which is prepared by sequentially enzymatically hydrolyzing and inoculating the above-mentioned microbial preparations from a medicinal and edible composition.

[0018] Preferably, the medicinal and edible composition comprises, by weight, 4-10 parts of foxnut, 3-8 parts of coix seed, 2-6 parts of red adzuki bean and 2-6 parts of dried tangerine peel.

[0019] More preferably, the medicinal and edible composition comprises, by weight, 4-8 parts of foxnut, 3-6 parts of coix seed, 3-6 parts of red adzuki bean and 2-5 parts of dried tangerine peel.

[0020] Specifically, in some embodiments of the present invention, the food-medicine composition comprises, by weight, 6 parts of Euryale ferox, 5 parts of Coix lacryma-jobi, 4 parts of red adzuki bean and 4 parts of dried tangerine peel.

[0021] Preferably, the microbial preparation includes Lactobacillus rhamnosus (Lactobacillus casei). Lacticaseibacillus rhamnosus LRB-2 and Lactobacillus plantarum ( Lactiplantibacillus plantarum YSH08;

[0022] The Lactobacillus rhamnosus LRB-2 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 39596, deposit date July 6, 2026, and deposit address No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

[0023] The preservation number of the Lactobacillus plantarum YSH08 is CGMCC No. 36182.

[0024] Preferably, the inoculation amount of the microbial preparation is 0.05-0.3%; specifically, in some embodiments of the present invention, the inoculation amount of the microbial preparation is 0.2%, and the inoculation amount is a volume ratio, that is, the volume ratio of the microbial preparation to the fermentation system.

[0025] Preferably, the volume ratio of Lactobacillus rhamnosus LRB-2 to Lactobacillus plantarum YSH08 is (0.05-0.3):(0.05-0.3).

[0026] More preferably, the volume ratio of Lactobacillus rhamnosus LRB-2 to Lactobacillus plantarum YSH08 is 1:1.

[0027] Preferably, the viable count of the *Lactobacillus rhamnosus* LRB-2 is 1.0 × 10⁻⁶. 8 CFU / mL or higher; the viable count of the *Lactobacillus plantarum* YSH08 is 1.0 × 10⁻⁶. 8 CFU / mL or higher.

[0028] Preferably, before inoculating with the microbial agent, the single strain needs to be activated. Specifically, the strain is inoculated into MRS medium and incubated at 35°C for 24 hours, and then passaged three times to fully activate the strain to a viable count of 1.0 × 10⁻⁶. 8 CFU / mL or higher.

[0029] A further preferred embodiment of the MRS culture medium is as follows: the culture medium formulation consists of 10.0 g / L peptone, 5.0 g / L beef extract, 4.0 g / L yeast extract, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 2 g / L triammonium hydrogen citrate, 5.0 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.05 g / L manganese sulfate, and 1 g / L Tween-80. The above MRS culture medium is then added to 1 L of pure water and autoclaved at 121°C for 15 min.

[0030] Preferably, the enzyme preparations used for enzymatic hydrolysis include cellulase, hemicellulase, pectinase, and acidic protease.

[0031] Preferably, the amount of cellulase added is 0.2-0.5%, the amount of hemicellulase added is 0.2-0.5%, the amount of pectinase added is 0.2-0.4%, and the amount of acidic protease added is 0.1-0.3%.

[0032] More preferably, the amount of cellulase added is 0.3%, the amount of hemicellulase added is 0.3%, the amount of pectinase added is 0.3%, and the amount of acidic protease added is 0.15%; the amount added is a weight ratio, which refers to the percentage of the weight of the enzyme preparation to the weight of the food-medicine homology composition.

[0033] More preferably, the cellulase activity is 10000U / mL-20000U / mL, the hemicellulase activity is 50000U / g-60000U / g, the pectinase activity is 60000U / mL-70000U / mL, and the acidic protease activity is 100000U / mL-200000 U / mL.

[0034] Specifically, in some embodiments of the present invention, the cellulase activity is 11,000 U / mL, the hemicellulase activity is 50,000 U / g, the pectinase activity is 60,000 U / mL, and the acidic protease activity is 100,000 U / mL.

[0035] In a fourth aspect, the present invention provides a method for preparing a medicinal and edible homologous fermented product, comprising the following steps: (1) mixing the medicinal and edible homologous composition with water evenly, adding an enzyme preparation for enzymatic hydrolysis, and sterilizing under high pressure steam after the enzymatic hydrolysis is completed to obtain an enzymatic hydrolysate; (2) inoculating the enzymatic hydrolysate with a microbial preparation for fermentation to obtain a fermented product.

[0036] Preferably, the solid-liquid ratio of the food-medicine homology composition to water is 1:(10-25); more preferably, the solid-liquid ratio of the food-medicine homology composition to water is 1:15.

[0037] Preferably, the enzymatic hydrolysis in step (1) is carried out in a water bath shaker, with the water bath temperature being 45-55℃, the rotation speed being 100-180 rpm, and the hydrolysis time being 2-5 h; specifically, in some embodiments of the present invention, the water bath temperature is 50℃, the rotation speed is 150 rpm, and the hydrolysis time is 3 h.

[0038] Preferably, the high-pressure steam sterilization temperature in step (1) is 110-120℃ and the time is 10-20min; specifically, in some embodiments of the present invention, the high-pressure steam sterilization temperature is 115℃ and the time is 15min.

[0039] Preferably, the fermentation temperature in step (2) is 30-40℃ and the time is 20-30h; specifically, in some embodiments of the present invention, the fermentation temperature is 37℃ and the time is 24h.

[0040] In a fifth aspect, the present invention provides an application of the above-mentioned food-medicine homologous fermented product or the food-medicine homologous fermented product obtained by the above preparation method in the preparation of functional products.

[0041] Preferably, the functional products include products for lowering blood sugar, products for lowering blood lipids, products for strengthening the spleen and removing dampness, and antioxidant products.

[0042] Compared with the prior art, the technical solution of the present invention has the following advantages:

[0043] 1. This invention provides a microbial preparation comprising Lactobacillus rhamnosus ( Lacticaseibacillus rhamnosus LRB-2 and Lactobacillus plantarum ( Lactiplantibacillus plantarumYSH08, after verification, showed that the colony counts of two strains, Lactobacillus rhamnosus LRB-2 and Lactobacillus plantarum YSH08, were significantly higher than the predicted values ​​when fermented for 8 hours in MRS medium and a spleen-strengthening and dampness-removing composition substrate, indicating an interspecies synergistic effect.

[0044] 2. This invention provides a microbial preparation that, after fermenting a spleen-strengthening and dampness-removing composition matrix, can significantly increase the content of core functional components such as total polyphenols, total flavonoids, and polysaccharides in the fermented product; at the same time, it can effectively enhance the fermented product's inhibition of pancreatic lipase activity and the scavenging rates of DPPH free radicals, hydroxyl free radicals, and superoxide anion free radicals.

[0045] 3. This invention provides a medicinal and edible fermented product, which is prepared by using a medicinal and edible spleen-strengthening and dampness-removing composition as raw material, and then inoculating it with Lactobacillus rhamnosus LRB-2 and Lactobacillus plantarum YSH08 for co-fermentation after enzymatic hydrolysis. The fermented product has significantly increased content of total polyphenols, total flavonoids and polysaccharides, and has excellent enrichment effect of active ingredients.

[0046] 4. This invention provides a medicinal and edible fermented product with excellent antioxidant properties. It can significantly improve the scavenging rates of DPPH free radicals, hydroxyl free radicals, superoxide anion free radicals, and ABTS cation free radicals, effectively scavenging free radicals in the body and can be widely used in various antioxidant products.

[0047] 5. This invention provides a medicinal and edible fermented product that can effectively inhibit the biological activity of α-glucosidase and pancreatic lipase, and has good hypoglycemic and lipid-lowering effects. It can be used in functional products that lower blood sugar and blood lipids. Moreover, human trials have verified that it has the effect of strengthening the spleen and removing dampness.

[0048] 6. The microbial preparation fermentation spleen-strengthening and dampness-removing composition matrix provided by the present invention can be precisely adapted. Under the premise of ensuring the content and functional activity of active ingredients in fermentation products, it can effectively reduce the dosage of fermentation strains and microbial preparations (the addition amount is only 0.1%), while significantly shortening the overall fermentation cycle (fermentation time is 24 hours), reducing fermentation energy consumption and production costs.

[0049] Instructions for the preservation of microbial strains:

[0050] Biological material: Lactobacillus rhamnosus LRB-2,

[0051] Accession number: CGMCC No. 39596;

[0052] Classification and nomenclature: Lactobacillus rhamnosus ( Lacticaseibacillus rhamnosus );

[0053] The deposit date is July 6, 2026;

[0054] Preservation institution: China General Microbiological Culture Collection Center, China Committee on the Preservation and Management of Microbial Culture Collections;

[0055] Abbreviation of depositary institution: CGMCC;

[0056] Location of the depository: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. Attached Figure Description

[0057] Figure 1 The graph shows the colony counts of different strains in experimental groups 1-4 after 8 hours in MRS medium; where A represents experimental group 1; B represents experimental group 2; C represents experimental group 3; and D represents experimental group 4.

[0058] Figure 2 The colony counts of different strains in experimental group 5 in MRS medium at 8 h (3 independent replicate experiments).

[0059] Figure 3 Colony counts of different bacterial strains fermented spleen-strengthening and dampness-removing enzymatic hydrolysate for 8 hours;

[0060] Figure 4 Figure showing the total polyphenol, flavonoid, and polysaccharide content in fermented products of medicinal and edible origin obtained from different strains;

[0061] Figure 5 The results of DPPH radical scavenging rate, hydroxyl radical scavenging rate, superoxide anion radical scavenging rate, and ABTS cationic radical scavenging rate of fermented products obtained from different strains are shown in the figure.

[0062] Figure 6 Figure showing the inhibition rate of α-glucosidase by fermented medicinal and edible products obtained from different strains;

[0063] Figure 7 Figure showing the inhibition rate of pancreatic lipase by fermented products of different strains of medicinal and edible materials;

[0064] Figure 8 Comparison of tongue appearance before and after children took the spleen-strengthening and dampness-removing fermented product in a human trial.

[0065] Figure 9 Comparison of tongue appearance before and after taking the spleen-strengthening and dampness-removing fermented product in adult subjects. Detailed Implementation

[0066] The following non-limiting embodiments are intended to provide a more comprehensive understanding of the invention by those skilled in the art, but do not limit the invention in any way. The following description is merely an exemplary illustration of the scope of protection claimed by the invention. Those skilled in the art can make various changes and modifications to the invention based on the disclosed content, and these should also fall within the scope of protection claimed by the invention. When numerical ranges are given in the embodiments, it should be understood that, unless otherwise stated in the invention, the two endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by those skilled in the art to which this invention pertains.

[0067] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all chemical reagents used in the embodiments of the present invention are obtained through conventional commercial channels. Products from different manufacturers do not have a significant impact on the effectiveness.

[0068] Euryale ferox, Coix lacryma-jobi, and red adzuki beans were purchased from Anhui Jiahe Traditional Chinese Medicine Technology Co., Ltd.; dried tangerine peel was purchased from Shanghai Ruiling Biotechnology Co., Ltd. Cellulase, pectinase, hemicellulase, and acidic protease were purchased from Xiasheng (Beijing) Biotechnology Development Co., Ltd., with cellulase activity of 11,000 U / mL, hemicellulase activity of 50,000 U / g, pectinase activity of 60,000 U / mL, and acidic protease activity of 100,000 U / mL.

[0069] Lactobacillus plantarum XD117, Lactiplantibacillus plantarum It is deposited at the China Center for Type Culture Collection, accession number CCTCC NO: M 20241730, and has been disclosed in patent CN119662456A.

[0070] Lactobacillus plantarum XD087, Lactiplantibacillus plantarum It is deposited at the China Center for Type Culture Collection, accession number CCTCC NO: M 20241729, and has been disclosed in patent CN119662456A.

[0071] Lactobacillus plantarum YSH08, Lactiplantibacillus plantarum It is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 36182 and has been published in Chinese patent CN121736957A.

[0072] Lactobacillus reuteri XS82, Latin name Limosilactobacillus reuteriIt is deposited at the China General Microbiological Culture Collection Center (CGMCC) on October 15, 2025, with accession number CGMCC No. 36181. The depository is located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. It has been published in patent CN121736956A.

[0073] Lactobacillus rhamnosus, derived from the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC 1.12734, has the Latin name... Lacticaseibacillus rhamnosus The original number was LRa05, and it was a commercially available strain.

[0074] Preparation Example 1: Activation and Cultivation of Microbial Strains

[0075] Activation of lactic acid bacteria: *Lactobacillus rhamnosus* LRB-2, *Lactobacillus plantarum* YSH08, *Lactobacillus plantarum* XD087, *Lactobacillus plantarum* XD117, *Lactobacillus rhamnosus* LRa05, and *Lactobacillus reuteri* XS82 were inoculated into MRS medium and cultured at 35℃ for 24 h. The culture was subcultured three times, and the bacterial suspension concentration was adjusted to 1.0 × 10⁻⁶. 8 CFU / mL.

[0076] The composition and preparation method of MRS medium are as follows: peptone 10.0 g / L, beef extract 5.0 g / L, yeast extract 4.0 g / L, glucose 20.0 g / L, dipotassium hydrogen phosphate 2.0 g / L, triammonium hydrogen citrate 2 g / L, sodium acetate 5.0 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.05 g / L, and Tween-80 1 g / L. Take the above MRS medium, add 1 L of pure water, and autoclave at 121℃ for 15 min for later use.

[0077] Example 1: Validation of interspecies synergistic effects - strain screening

[0078] Experimental Groups:

[0079] Experimental Group 1: Single Bacterial Group A: Inoculated with 0.1% (v / v) Lactobacillus reuteri XS82; Single Bacterial Group B: Inoculated with 0.1% (v / v) Lactobacillus plantarum YSH08; Co-culture Group: Inoculated with the above two bacteria at a ratio of 0.1%:0.1% (v / v), with a total inoculation amount of 0.2%.

[0080] Experimental Group 2: Single Bacterial Group A: Inoculated with 0.1% (v / v) Lactobacillus rhamnosus LRB-2; Single Bacterial Group B: Inoculated with 0.1% (v / v) Lactobacillus plantarum XD087; Co-culture Group: Inoculated with the above two bacteria at a ratio of 0.1%:0.1% (v / v), with a total inoculation amount of 0.2%.

[0081] Experimental Group 3: Single Bacterial Group A: Inoculated with 0.1% (v / v) Lactobacillus rhamnosus LRB-2; Single Bacterial Group B: Inoculated with 0.1% (v / v) Lactobacillus plantarum XD117; Co-culture Group: Inoculated with the above two bacteria at a ratio of 0.1%:0.1% (v / v), with a total inoculation amount of 0.2%.

[0082] Experimental Group 4: Single Bacterial Group A: Inoculated with 0.1% (v / v) Lactobacillus rhamnosus LRa05; Single Bacterial Group B: Inoculated with 0.1% (v / v) Lactobacillus plantarum YSH08; Co-culture Group: Inoculated with the above two bacteria at a ratio of 0.1%:0.1% (v / v), with a total inoculation amount of 0.2%.

[0083] Experimental Group 5: Single Bacterial Group A: Inoculated with 0.1% (v / v) Lactobacillus rhamnosus LRB-2; Single Bacterial Group B: Inoculated with 0.1% (v / v) Lactobacillus plantarum YSH08; Co-culture Group: Inoculated with the above two bacteria at a ratio of 0.1%:0.1% (v / v), with a total inoculation amount of 0.2%.

[0084] Each group was inoculated into MRS medium and incubated at 37°C. Samples were taken after 8 hours, and the viable count was determined by plate counting method.

[0085] The colony counts of the 5 strains are shown in the figure. Figure 1-2 The results showed that, under the MRS medium system, the co-culture of *Lactobacillus rhamnosus* LRB-2 and *Lactobacillus plantarum* YSH08 in experimental group 5 exhibited a synergistic growth effect between species, with the actual colony count exceeding the theoretical prediction obtained by adding the single-strain colony counts after 8 hours. In contrast, the actual co-culture colony counts of the co-culture groups in experimental groups 1-4 were all lower than the predicted values, showing no synergistic effect and even exhibiting inter-strain inhibition. This indicates that the inter-species synergistic effect of lactic acid bacteria co-culture is significantly strain-specific, and synergistic growth is not achieved at the genus level. Only the specific pairing of *Lactobacillus rhamnosus* LRB-2 and *Lactobacillus plantarum* YSH08 under MRS culture conditions resulted in a mutually beneficial growth-promoting phenomenon.

[0086] Example 2: Verification of the interspecies synergistic effect of the fermented spleen-strengthening and dampness-removing composition.

[0087] Experimental Groups:

[0088] Single-strain group A: Inoculated with 0.1% (v / v) Lactobacillus rhamnosus LRB-2.

[0089] Single-strain group B: Inoculated with 0.1% (v / v) Lactobacillus plantarum YSH08.

[0090] Two-bacterial co-culture group: The two bacteria were inoculated at a ratio of 0.1%:0.1% (v / v), with a total inoculation amount of 0.2%.

[0091] Each group was inoculated into a spleen-strengthening and dampness-removing food-derived enzymatic hydrolysate (preparation method as in Example 3), and incubated at 37°C. Samples were taken after 8 hours, and the viable count was determined by plate counting method.

[0092] See the graphs for the viable cell count results of each group. Figure 3 ,Depend on Figure 3 The results showed that after fermentation for 8 hours in the enzymatic hydrolysate of the spleen-strengthening and dampness-removing composition, the colony count of the specific combination of *Lactobacillus rhamnosus* LRB-2 and *Lactobacillus plantarum* YSH08 was 1132±79×10⁻⁶. 6 The CFU / mL level was significantly higher than the predicted value (611 ± 64 × 10⁻⁶). 6 The CFU / mL indicates that Lactobacillus rhamnosus LRB-2 and Lactobacillus plantarum YSH08 exhibit an intermediate synergistic growth effect under the conditions of the spleen-strengthening and dampness-removing enzymatic hydrolysate, and the two strains can promote each other's growth.

[0093] Example 3: Preparation of Spleen-Strengthening and Dampness-Removing Fermented Product

[0094] Preparation method of spleen-strengthening and dampness-removing fermented products

[0095] Step 1: Preparation of Spleen-Strengthening and Dampness-Removing Enzymatic Hydrolysate: The spleen-strengthening and dampness-removing composition (6 parts of Euryale ferox, 5 parts of Coix lacryma-jobi, 4 parts of red adzuki bean, and 4 parts of dried tangerine peel) was pulverized and mixed with water at a material-to-liquid ratio of 1:15 (m / v). 0.3% cellulase, 0.3% hemicellulase, 0.3% pectinase, and 0.15% acidic protease were added (all percentages by weight of the spleen-strengthening and dampness-removing composition; for example, if the composition weighs 100g, the cellulase addition is 0.3g). The mixture was then enzymatically hydrolyzed in a 50℃ water bath for 3 hours. After hydrolysis, the mixture was sterilized by high-pressure steam at 115℃ for 15 minutes to obtain the enzymatic hydrolysate.

[0096] Step 2: Activation and inoculation of the strain: Same as in preparation example 1.

[0097] Step 3: Fermentation: Set up 4 groups of experiments:

[0098] Blank control group (unfermented group): sterile food and medicine homologous enzymatic hydrolysate, without inoculation.

[0099] Single-strain group A: Inoculated with 0.2% (v / v) Lactobacillus rhamnosus LRB-2.

[0100] Single-strain group B: Inoculated with 0.2% (v / v) Lactobacillus plantarum YSH08.

[0101] Two-bacterial co-culture group: The two bacteria were inoculated at a ratio of 0.1%:0.1% (v / v), with a total inoculation amount of 0.2%.

[0102] Each group was incubated at 37℃ for 24 hours. After fermentation, the fermentation was terminated by high-pressure steam sterilization at 115℃ for 15 minutes, yielding a spleen-strengthening and dampness-removing fermented product.

[0103] Experimental Example 1: Determination of the main active ingredients in a spleen-strengthening and dampness-removing fermented product

[0104] Samples to be tested: Centrifuge the spleen-strengthening and dampness-removing fermented material at 8000 rpm for 10 min, collect the supernatant, and obtain the corresponding fermentation supernatant as the samples to be tested.

[0105] 1.1 Determination of polysaccharide content

[0106] Polysaccharide content was determined using the phenol-sulfuric acid method. Glucose standard curve: Prepare a 1 mg / mL glucose solution, diluted to concentrations of 0, 0.1, 0.2, 0.3, 0.4, and 0.5 mg / mL. Add 200 µL of glucose solution to six 2 mL Eppendorf tubes. Add 200 µL of 5% phenol to each Eppendorf tube. Transfer 1 mL of concentrated sulfuric acid to each Eppendorf tube and shake rapidly to ensure thorough mixing. Incubate the Eppendorf tubes in a 100℃ water bath for 20 min, then cool to room temperature under running water for 5 min. Measure the OD value of the reaction system at 490 nm. Calculate the polysaccharide content in the solution based on the standard curve. For the blank control group, use distilled water instead of the standard sugar solution.

[0107] 1.2 Determination of total flavonoid content

[0108] Prepare a 1 mg / mL rutin solution. Dissolve the rutin standard in 50% ethanol and dilute to concentrations of 0, 0.2, 0.4, 0.6, 0.8, and 1 mg / mL. Take six 2 mL Eppendorf tubes and add 100 μL of the rutin solution, 1.2 mL of water, and 100 μL of 5% sodium nitrite. React at room temperature for 5 min, then add 100 μL of aluminum nitrate and react at room temperature for another 5 min. Finally, add 500 μL of 4% sodium hydroxide and react at room temperature for 15 min. Spot 200 μL of each solution using a pipette. Measure the OD value at 510 nm. Calculate the total flavonoid content in the solution based on the standard curve. Use distilled water instead of the standard rutin solution for the blank control group.

[0109] 1.3 Determination of total polyphenol content

[0110] Prepare a 1 mg / mL gallic acid solution and dilute it to concentrations of 0, 0.1, 0.2, 0.3, 0.4, and 0.5 mg / mL. Take six 5 mL Eppendorf tubes, add 50 μL of gallic acid solution and 450 μL of water, then add 750 μL of Folin-Ciocalteu and react at room temperature for 5 min. Next, add 500 μL of 20% sodium carbonate and 3.25 mL of water and react at room temperature for 1 hour. Spot 200 μL of each solution using a pipette. Measure the OD value at 760 nm. Calculate the total polyphenol content in the solution based on the standard curve. For the blank control group, use distilled water instead of the standard gallic acid solution.

[0111] 1.4 Effects of Fermentation on Active Ingredients in the Spleen-Strengthening and Dampness-Removing Composition

[0112] Polyphenols, total flavonoids, and polysaccharides are key functional active substances in food and medicine homology, and fermentation processes can significantly alter the dissolution levels of their active ingredients. The effects of bacterial fermentation on the active ingredients in the spleen-strengthening and dampness-removing composition are as follows: Figure 4 As shown, by Figure 4 The results showed that the polyphenol content in the co-culture group reached 5.2142±0.0596 mg / g, significantly higher than the blank control group (1.8257±0.0418 mg / g) by 185.60%; the total flavonoid content in the co-culture group reached 3.5507±0.0902 mg / g, significantly higher than the blank control group (0.7653±0.0139 mg / g) by 363.96%; and the polysaccharide content in the co-culture group reached 261.1274±14.2350 mg / g, significantly higher than the blank control group (101.8160±6.0850 mg / g) by 156.47%. Furthermore, the total polyphenol, total flavonoid, and polysaccharide contents in the co-culture group were significantly higher than those in the single-strain group. Therefore, the co-fermentation of the two strains can effectively promote the release of polyphenols, total flavonoids, and polysaccharides from the spleen-strengthening and dampness-removing substrate, confirming a significant synergistic effect between the two strains.

[0113] Experimental Example 2: Determination of Antioxidant Levels

[0114] Samples to be tested: Centrifuge the spleen-strengthening and dampness-removing fermented material at 8000 rpm for 10 min, collect the supernatant, and obtain the corresponding fermentation supernatant as the samples to be tested.

[0115] 2.1 DPPH free radical scavenging rate:

[0116] Take 1 mL of sample extract into a 2 mL centrifuge tube, add 1 mL of 0.2 mmol / L DPPH ethanol solution (prepared with anhydrous ethanol), vortex to mix, and incubate at room temperature in the dark for 30 min. Use anhydrous ethanol instead of DPPH solution as the control group, and use anhydrous ethanol instead of the sample as the blank group. Take 200 μL of the reaction solution into a 96-well microplate and measure the absorbance at 517 nm. The calculation formula is as follows:

[0117] R1 = [1 - (AB) / C] × 100%

[0118] In the formula: R1: DPPH free radical scavenging rate, %; A: OD value of experimental group; B: OD value of control group; C: OD value of blank group.

[0119] 2.2 Hydroxyl radical scavenging rate:

[0120] Take 1 mL of sample extract into a 10 mL test tube, add 1 mL of 6 mmol / L FeSO4 solution and 1 mL of 6 mmol / L H2O2 solution sequentially, vortex to mix, and let stand for 10 min (for the blank group, use an equal volume of ultrapure water instead of the sample). Then add 1 mL of 6 mmol / L salicylic acid-ethanol solution, vortex to mix, and let stand for 30 min. Take 200 μL of the reaction solution into an ELISA plate and measure the absorbance at 510 nm. Calculate using the following formula:

[0121] R2 = (1 - A / B) × 100%

[0122] In the formula: R2: hydroxyl radical scavenging rate, %; A: OD510nm of the experimental group; B: OD510nm of the control group.

[0123] 2.3 Superoxide anion radical scavenging rate:

[0124] Mix 1 mL of sample extract with 4.5 mL of 0.05 mol / L Tris-HCl buffer (pH 8.2) and preheat in a 25°C water bath for 20 min. Add 0.3 mL of 0.05 mol / L pyrogallol solution and react precisely at 25°C for 4 min. Immediately afterward, add 1 mL of 8 mmol / L HCl to terminate the reaction. Measure the absorbance at 325 nm. Use ultrapure water instead of pyrogallol as the control group (B) and use ultrapure water instead of the sample as the blank group (C). The calculation formula is as follows:

[0125] R3 = [1 - (AB) / C] × 100%

[0126] In the formula, R3: superoxide anion radical scavenging rate; A: absorbance of the experimental group; B: absorbance of the control group (pure water instead of pyrogallol); C: absorbance of the blank group (pure water instead of the sample).

[0127] 2.4 ABTS radical scavenging rate:

[0128] Equal volumes of 7.4 mmol / L ABTS solution and 2.6 mmol / L K₂S₂O₈ solution were mixed and incubated in brown centrifuge tubes at room temperature in the dark for 12 h to allow the ABTS solution to settle. + • Ensure complete formation. Before use, dilute the above working solution with 95% ethanol to an OD734nm of 0.70 ± 0.02. Take 0.2 mL of sample extract and add 0.8 mL of diluted ABTS. + • After vortexing and mixing the working solution, react at room temperature for 6 min (the blank group is replaced with 95% ethanol instead of the sample). Take 200 μL of the reaction solution into an ELISA plate and measure the absorbance at 734 nm. The calculation formula is as follows:

[0129] R4 = (1 - A / B) × 100%

[0130] In the formula, R4: ABTS cation radical scavenging rate, %; A: OD734nm of the experimental group; B: OD734nm of the control group.

[0131] 2.5 Effects of Fermentation on the Antioxidant Active Components of the Spleen-Strengthening and Dampness-Removing Composition

[0132] DPPH radicals, hydroxyl radicals, superoxide anion radicals, and ABTS cationic radicals are core indicators for evaluating in vitro antioxidant activity. Each corresponds to a different type of free radical, and combined detection can comprehensively evaluate the antioxidant activity of a test substance. DPPH is a stable free radical scavenger and is commonly used to evaluate the free radical scavenging ability of substances in vitro. Hydroxyl radicals are currently known as the strongest oxidants; they react with almost every type of organic matter in cells, such as sugars, amino acids, and nucleic acids, making them extremely destructive and a major contributor to tissue peroxidation and cell membrane damage. Superoxide anions are one of the most common reactive oxygen species (ROS) in living organisms; excessive accumulation leads to oxidative stress, causing cell damage, aging, and various diseases. ABTS cationic radicals are often used to evaluate the total antioxidant capacity of a sample in scavenging free radicals and are commonly used to determine the total antioxidant capacity of plant extracts and foods.

[0133] The effect of fermentation on the antioxidant active ingredients of the spleen-strengthening and dampness-removing composition, such as Figure 5 As shown, by Figure 5The results showed that after co-culturing the spleen-strengthening and dampness-removing composition, the DPPH free radical scavenging rate reached 50.74%±0.83%, which was 92.85% higher than that of the blank control group (26.31%±2.73%); the hydroxyl free radical scavenging rate reached 64.33%±0.16%, which was 13.74% higher than that of the blank control group (56.56%±0.25%); the superoxide anion free radical scavenging rate reached 70.1%±0.44%, which was 42.85% higher than that of the blank control group (49.07%±2.2%); and the ABTS cationic free radical scavenging rate reached 39.12%±1.18%, which was 476.14% higher than that of the blank control group (6.79%±1.14%). Among them, the DPPH free radical, hydroxyl free radical, and superoxide anion free radical scavenging rates were all significantly higher than those of the single-strain fermentation group, indicating that co-fermentation of the two strains can effectively improve the ability of fermentation products to scavenge free radicals in vivo and significantly enhance the antioxidant capacity of fermentation products.

[0134] Experiment Example 3: Detection of lipid-lowering and blood glucose-lowering indicators

[0135] Samples to be tested: Centrifuge the spleen-strengthening and dampness-removing fermented material at 8000 rpm for 10 min, collect the supernatant, and obtain the corresponding fermentation supernatant as the samples to be tested.

[0136] 3.1 α-Glucosidase Inhibition Rate Experiment

[0137] Take 0.2 mL of sample and mix with 1 mL of α-glucosidase solution (enzyme activity 0.1 U / mL, prepared with 0.1 mol / L PBS buffer, pH 6.8), and pre-incubate at 37℃ for 10 min. Add 0.5 mL of 5 mmol / L PNPG substrate solution, and continue the reaction at 37℃ for 10 min. Terminate the reaction by adding 1 mL of 0.1 mol / L Na2CO3 solution, and measure the absorbance at 405 nm. The experimental groups are set up as follows: Experimental group (A) is sample + enzyme solution + substrate; experimental background group (B) uses ultrapure water instead of enzyme solution; blank group (C) uses ultrapure water instead of sample; blank background group (D) uses ultrapure water to replace both sample and enzyme solution. The calculation formula is as follows:

[0138] Inhibition rate (%) = [1 - (AB) / (CD)] × 100%

[0139] In the formula: A: absorbance of the experimental group; B: absorbance of the experimental background group (pure water replaces enzyme solution); C: absorbance of the blank group (pure water replaces sample); D: absorbance of the blank background group (pure water replaces sample and enzyme solution).

[0140] 3.2. Pancreatic lipase inhibition rate experiment

[0141] Take 0.5 mL of sample, 0.5 mL of pancreatic lipase solution (5 mg / mL, prepared with 25 mmol / L PBS buffer, pH 7.4), and 0.5 mL of PBS buffer, and pre-incubate at 37°C for 10 min. Add 0.5 mL of 11.2 mmol / L p-NPB substrate solution (dissolved with isopropanol), gently vortex to mix, and react at 37°C for 20 min. Measure the absorbance at 405 nm. The experimental groups and calculation formulas are the same as for α-glucosidase inhibition rate.

[0142] 3.3 Effect of fermentation on the α-glucosidase inhibition rate of the spleen-strengthening and dampness-removing composition

[0143] Alpha-glucosidase is a key carbohydrate hydrolase, crucial for regulating blood sugar levels. It hydrolyzes carbohydrates into glucose, which is then absorbed by the body. Studies have shown that inhibiting alpha-glucosidase activity can slow down carbohydrate digestion and absorption, thereby regulating blood sugar levels and lowering blood sugar.

[0144] The effect of fermentation on the α-glucosidase inhibition rate of the spleen-strengthening and dampness-removing composition, such as Figure 6 .like Figure 6 The results showed that, compared with the blank control group, the α-glucosidase inhibition rate of the co-culture group after fermentation of the spleen-strengthening and dampness-removing composition reached 44.07%±1.81%, which was 125.77% higher than that of the unfermented group, and there was no significant difference compared with the single-strain group. The results indicate that the interspecies synergistic effect of the two strains did not further amplify the in vitro inhibitory effect.

[0145] 3.4 Effect of fermentation on the pancreatic lipase inhibition rate of the spleen-strengthening and dampness-removing composition

[0146] Pancreatic lipase is a key enzyme in the intestine that catalyzes the hydrolysis of dietary triglycerides. When its activity is inhibited, the process of fat breakdown and digestion is hindered, which can reduce the body's absorption and utilization of dietary fat. Therefore, the pancreatic lipase inhibition rate is an important in vitro evaluation indicator for assessing the potential lipid-lowering activity of a sample.

[0147] The effect of fermentation on the pancreatic lipase inhibition rate of the spleen-strengthening and dampness-removing composition, such as Figure 7 .like Figure 7 The results showed that after co-culturing the spleen-strengthening and dampness-removing composition, the pancreatic lipase inhibition rate reached 71.49%±1.16%, which was 10.45 times higher than that of the unfermented group (6.84%±0.85%), and significantly higher than that of the single-strain group. This indicates that microbial fermentation is an effective means to improve the pancreatic lipase inhibition rate of fermented products, and the co-fermentation of Lactobacillus rhamnosus LRB-2 and Lactobacillus plantarum YSH08 further leveraged their synergistic advantages to achieve a better inhibitory effect.

[0148] In summary, the fermentation treatment of the spleen-tonifying and dampness-removing composition by *Lactobacillus rhamnosus* LRB-2 and *Lactobacillus plantarum* YSH08 can enhance the release of active substances such as polyphenols, flavonoids, and polysaccharides in the fermented product. It also significantly improves the DPPH radical scavenging rate, hydroxyl radical scavenging rate, superoxide anion radical scavenging rate, and ABTS cationic radical scavenging rate of the fermented product, thereby enhancing its antioxidant level. Furthermore, the above indicators of the co-fermentation system of the two strains are significantly higher than those of single-strain fermentation. These results indicate that the two-strain combination exhibits interspecies synergistic interaction in the spleen-tonifying and dampness-removing beverage matrix. The mutualistic interaction between the strains not only promotes bacterial proliferation but also further drives the generation and release of functional active substances.

[0149] Simultaneously, the synergistic effect of the two strains on pancreatic lipase inhibition activity was enhanced, confirming that co-fermentation can reshape the metabolic characteristics of the substrate and synergistically amplify the lipid-regulating and hypoglycemic activities of the spleen-tonifying and dampness-removing beverage in vitro, demonstrating a significant functional advantage compared to single-strain fermentation. These results also suggest that a reasonable combination of strains can fully tap the functional potential of traditional Chinese medicine substrates, providing experimental basis for the selection of strain combinations for spleen-tonifying and dampness-removing fermented beverages.

[0150] Experiment Example 4: Human Trial Wear

[0151] When the spleen is deficient, its function of transporting and transforming fluids declines, leading to impaired metabolism and distribution of body fluids. This results in the stagnation of dampness and cold turbidity within the body. The dampness rises and steams the tongue, causing it to swell and become enlarged due to the moisture. The tongue is also pressed against the teeth, forming teeth marks. Simultaneously, the accumulation of dampness and turbidity on the tongue surface manifests as a thick, greasy white coating. The teeth marks primarily reflect the underlying deficiency of spleen qi, while the thick, greasy white coating is an external sign of the excessive accumulation of cold and dampness within the body. Therefore, in this human trial, the tongue coating was used as a characteristic indicator to verify the spleen-strengthening and dampness-removing effects.

[0152] 4.1 Basic Information of Subjects: Subjects may experience symptoms such as a sticky feeling in the mouth, headache, body heaviness, abnormal bowel movements, and loss of appetite. The core criterion for inclusion is a yellow and greasy tongue coating, with or without teeth marks or cracks on the tongue. Subjects with severe heart, liver, or kidney disease, immunodeficiency, allergies, or who are pregnant or lactating are excluded. Adult subjects must have a yellow and greasy tongue coating, which may be accompanied by teeth marks or cracks; children must have a thick and greasy yellow tongue coating to be included in this trial.

[0153] 4.2 Grouping: Subjects were divided into an adult group and a children group, with 8 subjects in each group. The spleen-strengthening and dampness-removing fermented product prepared from the co-culture group of the two bacteria in Example 3 was separated into solid and liquid components. The supernatant obtained was used for human trials. The administration method was as follows: twice a day, divided into morning and afternoon doses, 50 mL each time, for 10 consecutive days.

[0154] 4.3 Observation Indicators: During the trial period, subjects avoided spicy, greasy, sweet, and raw / cold foods and maintained a stable daily routine. Before and after the trial, the subjects' tongues were observed under standardized photographic conditions. The thickness and color of the tongue coating, teeth marks, and cracks were observed and photographed. Simultaneously, any adverse reactions such as abdominal distension, diarrhea, or nausea were observed and recorded throughout the trial.

[0155] 4.4 Trial Results

[0156] The tongue appearance comparison images of the children's group and the adult group after taking Jianpi Qushi Yin (a traditional Chinese medicine formula) for 10 consecutive days are as follows. Figure 8-9 As shown. Figure 8 It was found that all children had thick, greasy tongue coating before the trial, mainly thick and greasy white coating, with some tongues appearing slippery and swollen, the coating covering the entire tongue surface, and significant coating accumulation at the root of the tongue. Among the children, the thick, greasy coating significantly subsided and the tongue surface became clean after taking the Jianpi Qushi Yin (a traditional Chinese medicine formula) for 10 days on days 1-3, 5, and 7, showing the most obvious effect. After taking the Jianpi Qushi Yin for 10 days on days 4, 6, and 8, the thick, greasy coating thinned, with only a small amount remaining, indicating a good improvement effect.

[0157] like Figure 9 As shown, before the trial, the main tongue appearance of the adult group was characterized by teeth marks, a thick white or yellow greasy coating, and cracks on the tongue surface. After 10 days of trial, the teeth marks on the tongue of the adult subjects were reduced, the thick greasy coating thinned and disappeared, the cracks on the tongue surface became shallower, and the tongue appearance signs improved to varying degrees.

[0158] Therefore, the composition of the present invention has the effect of strengthening the spleen and removing dampness. During the trial period, the subjects did not have obvious adverse reactions, and the product has good safety.

[0159] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A microbial preparation, characterized in that, Its active ingredients include Lactobacillus rhamnosus ( Lacticaseibacillus rhamnosus LRB-2 and Lactobacillus plantarum ( Lactiplantibacillus plantarum YSH08; The Lactobacillus rhamnosus LRB-2 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 39596, deposit date July 6, 2026, and deposit address No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing. The Lactobacillus plantarum YSH08 has the accession number CGMCC No.36182.

2. The application of the microbial preparation according to claim 1 in the fermentation of food-medicine homology compositions or in the preparation of food-medicine homology fermented products.

3. A fermented product that is both food and medicine, characterized in that, The fermented product is prepared by sequentially enzymatically hydrolyzing and inoculating the microbial preparation of claim 1 with the medicinal and edible homology composition; the medicinal and edible homology composition includes, by weight, 4-10 parts of Euryale ferox, 3-8 parts of Coix lacryma-jobi, 2-6 parts of red adzuki bean and 2-6 parts of dried tangerine peel.

4. The fermented product of medicinal and edible origin according to claim 3, characterized in that, The enzyme preparations used in the enzymatic hydrolysis include cellulase, hemicellulase, pectinase, and acidic protease. The amount of cellulase added is 0.2-0.5%, the amount of hemicellulase added is 0.2-0.5%, the amount of pectinase added is 0.2-0.4%, and the amount of acidic protease added is 0.1-0.3%.

5. The fermented product of medicinal and edible origin according to claim 3, characterized in that, The inoculum amount of the microbial preparation is 0.05-0.3%; the volume ratio of the inoculum amount of Lactobacillus rhamnosus LRB-2 and Lactobacillus plantarum YSH08 is (0.05-0.3):(0.05-0.3).

6. The fermented product of medicinal and edible origin according to claim 3, characterized in that, The viable counts of *Lactobacillus rhamnosus* LRB-2 or *Lactobacillus plantarum* YSH08 were both above 1.0 × 10⁻⁶. 8 CFU / mL or higher.

7. The method for preparing the fermented medicinal and edible products according to any one of claims 3-6, characterized in that, The steps include: (1) mixing the food-medicine homology composition with water and adding an enzyme preparation for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, the mixture is sterilized by high-pressure steam to obtain the enzymatic hydrolysate; (2) inoculating the enzymatic hydrolysate with the microbial preparation of claim 1 for fermentation to obtain the fermented product.

8. The method for preparing the fermented product of medicinal and edible origin according to claim 7, characterized in that, The enzymatic hydrolysis in step (1) is carried out in a water bath shaker, with the water bath temperature being 45-55℃, the rotation speed being 100-180rpm, and the hydrolysis time being 2-5h; the high-pressure steam sterilization temperature is 110-120℃, and the time is 10-20min; the fermentation temperature in step (2) is 30-40℃, and the time is 20-30h.

9. The application of the medicinal and edible fermented product according to any one of claims 3-6 or the medicinal and edible fermented product obtained by the preparation method according to any one of claims 7-8 in the preparation of functional products; The functional products include products for lowering blood sugar, products for lowering blood lipids, products for strengthening the spleen and removing dampness, and antioxidant products.

10. A functional product, characterized in that, This includes the medicinal and edible fermented product as described in any one of claims 3-6 or the medicinal and edible fermented product obtained by the preparation method described in any one of claims 7-8.

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