Application of bacillus coagulans probiotics in prevention and treatment of hyperuricemia

Abstract

The application discloses application of a Bacillus coagulans probiotic in prevention and treatment of hyperuricemia and belongs to the technical field of microorganisms. The application provides a new use of the Bacillus coagulans probiotic, the probiotic has a good inhibitory effect on in-vitro xanthine oxidase, has a good uric acid reducing effect on hyperuricemia HK-2 cells, regulates uric acid transport proteins and reduces inflammation, can inhibit uric acid generation of hyperuricemia, increase excretion and reduce inflammation, and thus has a good application prospect and value in prevention and treatment of hyperuricemia.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology, specifically relating to the application of Bacillus coagulans postbiotics in the prevention and treatment of hyperuricemia. Background Technology

[0002] Hyperuricemia (HUA) is a serious disease that severely endangers human health. It is primarily caused by increased uric acid production or impaired uric acid excretion, leading to excessive uric acid in the blood. It is the fourth most common basic metabolic disease after hypertension, hyperlipidemia, and hyperglycemia. Hyperuricemia is diagnosed when two fasting blood uric acid levels are higher than 420 µmol / L for men and higher than 360 µmol / L for women. Normally, the uric acid level in the human body is in dynamic equilibrium, due to a balance between uric acid production, absorption, excretion, and breakdown. Under certain circumstances, this balance can be disrupted, resulting in abnormal uric acid levels and potentially causing hyperuricemia.

[0003] Hyperuricemia has a series of complications, the most common being joint damage, commonly known as gout, caused by the deposition of urate crystals in the joints. Secondly, kidney damage is also a common complication, resulting from the continuous deposition of urate crystals in the renal tubules and subsequent inflammation of the kidney tissue. In addition, hyperuricemia is closely associated with diabetes, hypertension, obesity, cardiovascular and cerebrovascular diseases, and chronic kidney disease.

[0004] Currently, there are three main types of drugs for treating hyperuricemia (HUA): those that reduce uric acid production (XOD inhibitors), those that increase uric acid excretion (URAT1 inhibitors), and those that promote the oxidative breakdown of uric acid (uricase preparations). Common XOD inhibitors include allopurinol and febuxostat. Allopurinol can treat both primary and secondary gout, and it is also effective in treating HUA after liver transplantation and calcium oxalate stones. Febuxostat is used to treat secondary HUA. Both have the same mechanism: inhibiting xanthine oxidase and preventing uric acid synthesis, thereby reducing uric acid production and lowering blood uric acid levels. URAT1 inhibitors are transport proteins that increase uric acid excretion, and include benzbromarone and probenecid. Benzbromarone effectively inhibits the expression levels of URAT1 and GLUT9 proteins, significantly increasing the renal uric acid excretion rate and significantly decreasing the serum uric acid concentration, thus gaining widespread use among HUA patients. Probenecid inhibits the kidneys' clearance of organic anions and impairs the reabsorption of urate in the renal tubules. It not only promotes uric acid excretion but also reduces uric acid excretion in the renal tubules. Uricase works by promoting the oxidative decomposition of uric acid into allantoin and hydrogen peroxide to treat gout atrophic uric acid (HUA). Studies have shown that exogenous uricase can accelerate the dissolution of tophi and cure gout patients who are resistant to or contraindicated with uric acid-lowering drugs. Currently, raburicase is commonly used; it converts uric acid into soluble allantoin for excretion. As a recombinant uricase, it has a faster effect on relieving HUA compared to allopurinol. However, these drugs all have certain toxic side effects, such as hypersensitivity reactions in the cardiovascular and renal systems, impaired kidney function, and heart failure. Summary of the Invention

[0005] This invention provides the application of Bacillus coagulans postbiotic in the preparation of formulations with xanthine oxidase inhibitory activity.

[0006] This invention provides the application of Bacillus coagulans postbiotic in the preparation of formulations with uric acid-lowering effects.

[0007] This invention provides the application of Bacillus coagulans postbiotic in the preparation of formulations that reduce inflammation caused by high uric acid.

[0008] This invention provides the application of Bacillus coagulans postbiotic in the preparation of formulations for the prevention and treatment of hyperuricemia.

[0009] In this invention, the Bacillus coagulans postbiotic is prepared by the following method: Inoculate Bacillus coagulans culture into MRS medium and incubate at 30–40°C for 12–48 h until the bacterial count in the culture reaches at least 1.6 × 10⁻⁶. 9CFU / mL; then sterilize, centrifuge, collect the supernatant, ultrafilter, and obtain Bacillus coagulans postgenetic agent.

[0010] In the above technical solution, the Bacillus coagulans is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC No. 39337.

[0011] In the above technical solution, the bacterial content in the Bacillus coagulans bacterial solution is at least 1.0 × 10⁻⁶. 9 CFU / mL; the inoculum size is 1-5%, V / V.

[0012] In the above technical solution, a 5KD molecular weight membrane core is used for ultrafiltration to obtain a cutoff component of <5KD.

[0013] The beneficial effects of this invention are as follows: This invention provides a novel use for Bacillus coagulans postbiotics, which exhibit good inhibitory effects on xanthine oxidase in vitro and good uric acid-lowering effects on hyperuricemic HK-2 cells. It also regulates uric acid transport proteins and reduces inflammation; furthermore, it can inhibit uric acid production in hyperuricemia, increase excretion, and reduce its inflammatory response, thus showing good application prospects and value in the prevention and treatment of hyperuricemia. Attached Figure Description

[0014] Figure 1 The colony morphology of strain HYPostbio-Y1 is shown.

[0015] Figure 2 The image shows the microscopic morphology of strain HYPostbio-Y1.

[0016] Figure 3 The effect of post-genes on HK-2 cell viability.

[0017] Figure 4 This is due to the inhibitory effect of post-biotics on uric acid levels.

[0018] Figure 5 The effect of metagenerogen on the secretion of inflammatory factors by hyperuricemic HK-2 cells; where A is IL-1β, B is IL-6, and C is TNF-α.

[0019] Figure 6Western blot images of URAT1 protein expression under different treatment conditions (A); the effect of metagener on URAT1 protein expression in cells (B); Western blot images of GLUT9 protein expression under different treatment conditions (C); the effect of metagener on GLUT9 protein expression in cells (D); Western blot images of OAT1 protein expression under different treatment conditions (E); the effect of metagener on OAT1 protein expression in cells (F).

[0020] Figure 7 The effect of postbiotics on body weight in mice with hyperuricemia.

[0021] Figure 8 Effects of metabiotics on serum uric acid levels in hyperuricemic mice (A); Effects of metabiotics on fecal uric acid levels in hyperuricemic mice (B).

[0022] Figure 9 Effects of metabiotics on serum xanthine oxidase activity in hyperuricemic mice (A); Effects of metabiotics on hepatic adenosine deaminase activity in hyperuricemic mice (B).

[0023] Figure 10 Effects of metabiotics on serum creatinine levels in hyperuricemic mice (A); Effects of metabiotics on serum urea nitrogen levels in hyperuricemic mice (B).

[0024] Figure 11 Effects of postbiotics on serum IL-1β in hyperuricemic mice (A); Effects of postbiotics on serum TNF-α in hyperuricemic mice (B).

[0025] Figure 12 Effects of postbiotics on kidney damage in hyperuricemic mice (A); Effects of postbiotics on liver damage in hyperuricemic mice (B). Detailed Implementation

[0026] The materials used in this invention are as follows: MRS broth culture medium was purchased from Qingdao High-Tech Industrial Park Haibo Biotechnology Co., Ltd.; SPF-grade Kunming mice were purchased from Shandong Pengyue. A 3K15 medical centrifuge was purchased from SIGMA Centrifuge GmbH, Germany. A DHP-9162 electric thermostatic incubator was purchased from Zhongyi Guoke (Beijing) Technology Co., Ltd. A BKQ-B7511 pressure steam sterilizer was purchased from Shandong Boke Biotechnology Co., Ltd. Multiskan FC microplate reader and UV spectrophotometer were purchased from Thermo Fisher Scientific (Shanghai) Instruments Co., Ltd. An SW-CJ-2FD clean bench was purchased from Suzhou Antai Air Technology Co., Ltd.

[0027] Strains Isolation and Identification 1. Strains Isolation On April 25, 2023, at Gujiadao Wharf in Qingdao, sea urchin intestines were collected and placed in sterile MRS liquid medium, then shaken and incubated for 1-2 days. The medium was then serially diluted with physiological saline, with 200 μL of each diluted medium being spread onto solid medium. The medium was incubated upside down for 2-3 days. Colonies with different morphologies were picked and streaked, and after isolation and purification, a bacterial strain was obtained and named HYPostbio-Y1.

[0028] 2. Strain identification Morphological characteristics: Colony morphology of strain HYPostbio-Y1 on MRS agar medium (aerobic incubation at 37°C for 24 h), such as... Figure 1 and Figure 2 As shown, the colonies are off-white, small, round, with neat edges, smooth and moist surface, slightly raised, and opaque.

[0029] Biological characteristics: The Gram staining result is purple, indicating that the bacterium is a Gram-positive bacterium.

[0030] Genetic characterization: 16S rRNA gene analysis was performed. Genomic DNA was extracted from strain HYPostbio-Y1, and PCR amplification was performed using universal primers for the 16S rRNA gene. The resulting 1488 bp gene sequence is shown below: Based on the homology comparison of standard strain sequences in the EZBioCloud database, the 16S rRNA gene of strain HYPostbio-Y1 showed 99.26% homology with the 16S rRNA gene of standard strain Weizmannia coagulans DSM 1=ATCC7050. Combining its morphological characteristics, biological, physiological and biochemical characteristics and gene sequence characteristics, strain HYPostbio-Y1 was finally identified as Bacillus coagulans.

[0031] The bacterium was deposited on June 10, 2026, at the China General Microbiological Culture Collection Center (CGMCC, address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, 100101, China), with accession number CGMCC No. 39337.

[0032] Other materials used in this invention, unless otherwise stated, are commercially available. Other terms used in this invention, unless otherwise specified, generally have the meanings commonly understood by those skilled in the art. The invention is further described in detail below with reference to specific embodiments and data. The following embodiments are merely illustrative and not intended to limit the scope of the invention in any way.

[0033] Example 1

[0034] The steps for activating the bacterial strain are as follows: Bacillus coagulans was inoculated from 30% glycerol cryovials (2% inoculum) into MRS medium and incubated at 37 °C for 24 h. Activation was continued to the third generation at a 2% inoculum. The cultured bacterial suspension was centrifuged at 4 °C and 4000 r / min for 10 min, the supernatant was discarded, and the suspension was reconstituted with 5 mL of physiological saline. The total bacterial count was determined using a hemocytometer. The bacterial suspension was diluted with physiological saline to obtain the desired concentration: Bacillus coagulans (1×10⁻⁶) 9 (CFU / mL).

[0035] The steps for preparing Postbio-B are as follows: Inoculate Bacillus coagulans culture at a volume fraction of 3% (V / V) into MRS medium and incubate at 37 °C for 24 h until the bacterial count in the culture reaches 1.6 × 10⁻⁶. 9CFU / mL or higher, autoclave at 121 °C for 20 min, centrifuge at 4000×g at 4 °C for 10 min, collect the supernatant, and use a high-pressure tangential flow apparatus (GCM-S-20, Guochu Technology) with a 5KD molecular weight membrane core for ultrafiltration separation to obtain the cutoff component <5KD, which is Postbio-B, stored in a -20 °C refrigerator for later use.

[0036] I. Verification of the inhibitory effect of metagenin on xanthine oxidase As shown in Table 1, phosphate buffer, sample, and xanthine oxidase (0.2 U / mL) were added sequentially to 96-well microplates. The plates were incubated at 37°C for 4 min, and then xanthine substrate solution (1 mmol / L) was added to initiate the reaction. Absorbance was measured at 295 nm using a microplate reader, with measurements taken every 2 min for a total of 12 min. Since xanthine has absorbance at 295 nm, the absorbance values ​​without the sample and enzyme were subtracted. Each sample was tested in triplicate. The inhibition rate of xanthine oxidase by the measured sample was calculated using the following formula.

[0037]

[0038] Table 1. Xanthine oxidase inhibition assay system

[0039] The experimental results showed that Postbio-B had an inhibition rate of 82.24% against XOD.

[0040] II. Efficacy Verification of In Vitro Hyperuric Acid Cell Experiment 1. Determination of the cytotoxicity of metabiotics Count HK-2 cells in the logarithmic growth phase and adjust the cell suspension concentration to 1×10⁻⁶. 6 Cells were seeded at a density of 100 µL in 96-well plates and cultured at 37 ℃ for 24 h. Using DMEM complete medium as a control, the experimental groups were supplemented with medium containing 20–200 μL / mL of post-biotic. Each group was cultured in triplicate, and the cells were cultured for another 24 h at 37 ℃ and 5% CO2. After discarding the medium and washing, 100 μL of medium containing 10% CCK-8 was added to each well, and the cells were incubated at 37 ℃ for 1 h. The OD450 value was measured using a microplate reader, and cell viability was expressed as a percentage of the control group.

[0041]

[0042] The test results are as follows Figure 3 As shown: Compared with the blank control group, cell viability remained above 80% within the concentration range of 20–80 μL / mL, indicating that Postbio-B was essentially non-toxic to cells. However, the addition of 100 μL / mL Postbio-B resulted in cell viability below 80%, indicating that concentrations of Postbio-B at 100 μL / mL and above were cytotoxic and inhibited cell growth. Therefore, within the safe concentration range, three concentrations of 20, 40, and 80 μL / mL were selected for application in the cell model.

[0043] 2. Determination of uric acid content in hyperuricemic cells Collect HK-2 cells in logarithmic growth phase, count them, and adjust the concentration to 1×10⁻⁶. 6 Cells were seeded at 100 μL / mL in 24-well plates and incubated at 37°C for 24 h. The cells were washed twice with PBS. A control group, a model group with 2.5 mM adenosine, a positive control group with 100 μM allopurinol, and experimental groups with 20, 40, and 80 μL / mL of urobilinogen were set up. After each group was treated with the corresponding reagents and cultured, the uric acid content in the cell supernatant was detected using a uric acid kit.

[0044] The test results are as follows Figure 4 As shown: After adenosine stimulation, the uric acid content in cells was significantly higher than that in the control group (P<0.05). Compared with the model group, the Postbio-B treatment group had a significant inhibitory effect on uric acid (P<0.05), and the higher the concentration of postbiotic, the more obvious the inhibitory effect.

[0045] 3. Measurement of inflammatory factors in hyperuricemic cells Cells were arranged at a ratio of 1×10 6 Cells were seeded at a density of 1 / mL in 24-well plates for 24 h. Three different concentrations of postbiotics and adenosine were selected within the safe drug concentration range for co-stimulation for 24 h. Each group was divided into three replicates. The supernatant was collected and the levels of cellular inflammatory factors (IL-1β, IL-6, TNF-α) were determined according to the ELISA kit instructions.

[0046] The test results are as follows Figure 5 As shown: After adenosine treatment, cells expressed large amounts of pro-inflammatory factors IL-1β, IL-6, and TNF-α (P<0.05), indicating that adenosine induced an inflammatory response in the cells. The addition of Postbio-B significantly reduced the secretion level of IL-1β (P<0.05).

[0047] 4. Determination of key uric acid transport proteins in hyperuricemic cells Cells were loaded at a rate of 1×10 6Cells / mL were seeded in 24-well plates and cultured for 24 h. Three safe drug concentrations were selected and co-stimulated with adenosine for 24 h (3 replicates per group). After washing with PBS, Western blotting and IP lysis buffer were added to a freshly prepared lysis buffer of PMSF at a ratio of 9:1. After lysis at 4 °C for 30 min, the cells were centrifuged at 13400×g for 10 min. The supernatant was stored at -80 °C and the protein concentration was measured by the BCA method. After electrophoresis on a 10% separating gel and a 5% stacking gel at 140 V, the gel was equilibrated with methanol-activated PVDF membranes and assembled in the corresponding order. The membranes were then transferred at 200 mA for 1 h, blocked overnight with 1×TBST of 5% skim milk powder, incubated overnight at 4 °C with a 1:1000 dilution of primary antibody, washed with 1×TBST, incubated at room temperature for 1 h with a 1:3000 dilution of secondary antibody, and washed. Finally, the membranes were incubated with ECL working solution and then chemiluminescently imaged. The gray values ​​of the bands of uric acid transporters (URAT1, GLUT9, OAT1) were analyzed using Gelpro32 software.

[0048] The test results are as follows Figure 6 As shown: Compared with the control group (Ctrl), the relative expression levels of URAT1 and GLUT9 proteins in the Model group were significantly increased (P<0.05), indicating that hyperuricemia induction successfully upregulated the expression of URAT1 and GLUT9. After treatment with different concentrations of Postbio-B, the expression levels of URAT1 and GLUT9 were lower than those in the Model group. The experimental results show that the postbiotic Postbio-B can reduce uric acid reabsorption by downregulating URAT1 and GLUT9 expression, thereby exerting a uric acid-lowering effect. Compared with the control group (Ctrl), the relative expression level of OAT1 protein in the Model group was significantly decreased (P<0.05), indicating that hyperuricemia induction successfully downregulated OAT1 expression; different concentrations of postbiotics can significantly upregulate OAT1 protein expression in hyperuricemia-induced HK-2 cells, and Postbio-B promotes uric acid excretion by upregulating OAT1 expression, thereby exerting a uric acid-lowering effect.

[0049] II. Efficacy verification in in vitro hyperuricemia mouse experiments 1. Mouse breeding and grouping SPF-grade male Kunming mice, weighing 18–22 g, were used. All mice were kept at 50% ± 10% relative humidity and 23 ± 2 °C, with 12 hours of light and 12 hours of darkness daily. Mice were fed a standard diet and water to acclimatize. After 7 days of acclimatization, 32 male Kunming mice were randomly divided into 4 groups (n=8): blank control group (Ctrl), model group (Model), positive control group (All), and postbio-B group (Postbio-B).

[0050] Every morning, mice in the control group were administered 0.2 mL of 0.5% CMC-Na by gavage, while mice in the other groups were administered an equal volume of 250 mg / kg PO and 100 mg / kg adenine (prepared by grinding in a mortar and pestle and adding to 0.5% CMC-Na) by gavage daily to induce hyperuricemia (HUA). After 14 days of modeling, two mice were randomly selected to determine their serum uric acid concentration to confirm successful HUA model establishment. If successful, subsequent drug administration was initiated. After HUA model establishment, the control group continued to be treated with 0.5% CMC-Na by gavage, while the other groups continued to be treated with 250 mg / kg PO and 100 mg / kg adenine by gavage daily. Two hours after administration of the modeling drugs, mice in the control and model groups were administered 0.5% CMC-Na by gavage; mice in the positive control group were administered allopurinol (10 mg / kg) by gavage; and mice in the post-biotic group were administered Postbio-B by gavage. All medications were administered once daily at the same time for 14 consecutive days, with body weight recorded every 7 days. During the experiment, the mice were allowed free access to food and water.

[0051] The test results are as follows Figure 7 As shown: During the experiment, observations of the mice in each group revealed that the Ctrl group mice were in better spirits, had glossy fur, moved freely, and reacted quickly. Some HUA mice exhibited lethargy, sluggish movement, loss of appetite, and dull fur. In the HUA mouse model, during the modeling phase (0-14 days), the weight of the Ctrl group mice steadily increased, while the weight of the Model group and all intervention groups showed a decreasing trend. During the intervention phase (14-28 days), the weight of the Model group mice further decreased, while the weight of the All group and Postbio-B group showed varying degrees of recovery, effectively alleviating the weight loss in the model mice.

[0052] 2. Determination of uric acid content in serum and feces of hyperuricemic mice Blood was centrifuged at 3000 r / min for 10 min, and the clear supernatant was carefully collected for analysis. Mouse feces were accurately weighed and added to 9 volumes of 0.9% physiological saline at a mass (g) / volume (mL) ratio of 1:9. The mixture was mechanically homogenized under ice-water bath conditions to prepare a 10% homogenate. This homogenate was centrifuged at 3000 r / min for 10 min, and the supernatant was collected for analysis. The uric acid content in serum and feces was determined according to the kit instructions.

[0053] The test results are as follows Figure 8 As shown: Compared with the Ctrl group, the serum uric acid level in the Model group mice was significantly increased (P<0.05), indicating that the HUA mouse model was successfully established. After intervention with allopurinol and Postbio-B, the serum uric acid levels in both groups were significantly reduced (P<0.05). There was no significant difference in fecal uric acid between the Model group, All group and the Ctrl group (P>0.05), while the fecal uric acid level in the Postbio-B group was significantly increased, indicating that Postbio-B can accelerate the clearance of uric acid in the body by promoting intestinal uric acid excretion.

[0054] 3. Determination of xanthine oxidase and adenosine deaminase activities in serum and liver of hyperuricemic mice Accurately weigh mouse liver and add 9 volumes of 0.9% physiological saline at a mass (g) / volume (mL) ratio of 1:9. Homogenize mechanically under ice-water bath conditions to prepare a 10% homogenate. Centrifuge at 3000 r / min for 10 min and collect the supernatant for analysis. Determine the levels of XOD and ADA in serum and liver according to the kit instructions.

[0055] The test results are as follows Figure 9 As shown: Compared with the Ctrl group, serum XOD activity in the Model group mice was significantly increased (P<0.05). After intervention with allopurinol and the postbiotic Postbio-B, serum XOD activity significantly decreased (P<0.05), indicating that Postbio-B can directly reduce uric acid synthesis by inhibiting XOD activity. Compared with the Ctrl group, liver ADA activity in the Model group mice was significantly increased (P<0.05). Although liver ADA activity showed a decreasing trend after allopurinol intervention, it was not significantly different from that in the Model group (P>0.05); however, liver ADA activity in the Postbio-B group was significantly lower than that in the Model group (P<0.05), indicating that Postbio-B can effectively inhibit liver ADA activity.

[0056] 4. Determination of serum creatinine and blood urea nitrogen levels in hyperuricemic mice According to the kit instructions, the levels of CRE and BUN in serum were measured.

[0057] The test results are as follows Figure 10 As shown: Compared with the Ctrl group, the serum CRE and BUN levels in the Model group were significantly increased (P<0.05), indicating that HUA had caused significant damage to the renal filtration function of mice. After intervention with allopurinol and postbiotic Postbio-B, the serum CRE and BUN levels in both groups of mice were significantly lower than those in the Model group (P<0.05), indicating that Postbio-B can effectively alleviate hyperuricemia-induced renal filtration dysfunction in mice and exert a good renal protective effect.

[0058] 5. Determination of inflammatory factors in the serum of hyperuricemic mice Serum levels of IL-1β and TNF-α were measured using a commercial ELISA kit.

[0059] The test results are as follows Figure 11 As shown: Compared with the Ctrl group, the serum levels of IL-1β and TNF-α in the Model group mice were significantly increased (P<0.05), indicating that the hyperuricemic state activated the systemic inflammatory response. Allopurinol and Postbio-B intervention significantly reduced IL-1β levels (P<0.05), while TNF-α levels, although showing a decreasing trend, were not significantly different from those in the Model group (P>0.05). These results demonstrate that Postbio-B can inhibit the abnormal increase in pro-inflammatory cytokine levels and alleviate the inflammatory response in HUA mice.

[0060] 6. Liver and kidney tissue sections of hyperuricemic mice Mouse liver and kidney tissues fixed in 4% paraformaldehyde were rinsed overnight, dehydrated, embedded in paraffin, cut into 5 μm sections, baked, stained with hematoxylin and eosin (HE), and mounted. The morphological characteristics of the tissues were observed using a slide scanner.

[0061] The test results are as follows Figure 12 As shown: After HE staining, kidney tissue sections from mice in the Ctrl group showed uniform staining, normal glomerular morphology and structure, clear renal tubular structure, and tightly packed tubular epithelial cells with no obvious inflammation. Compared with the Ctrl group, mice in the Model group showed renal tubular dilation, flattening and atrophy of epithelial cells, renal interstitial fibrosis, and inflammatory cell infiltration. After intervention with allopurinol and Postbio-B, the pathological damage in both groups was alleviated, and the renal tubular structure, renal interstitial fibrosis, and inflammatory infiltration were improved, approaching the level of the Ctrl group. Liver damage and repair were similar to those in the kidneys. In conclusion, Postbio-B can alleviate potassium oxonate and adenine-induced liver and kidney damage in HUA mice.

[0062] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any other way. Any person skilled in the art may make changes or modifications to the above-disclosed technical content to create equivalent embodiments. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the protection scope of the present invention.

Claims

1. Application of Bacillus coagulans postbiotic in the preparation of formulations with xanthine oxidase inhibitory activity.

2. Application of Bacillus coagulans postbiotic in the preparation of preparations with uric acid-lowering effects.

3. Application of Bacillus coagulans postbiotics in the preparation of formulations that reduce inflammation caused by high uric acid.

4. Application of Bacillus coagulans postbiotics in the preparation of formulations for the prevention and treatment of hyperuricemia.

5. The application according to any one of claims 1 to 4, characterized in that, The Bacillus coagulans postbiotic was prepared by the following method: Inoculate Bacillus coagulans culture into MRS medium and incubate at 30–40°C for 12–48 h until the bacterial count in the culture reaches at least 1.6 × 10⁻⁶. 9 CFU / mL; then sterilize, centrifuge, collect the supernatant, ultrafilter, and obtain Bacillus coagulans postgenetic agent.

6. The application according to claim 5, characterized in that, The Bacillus coagulans is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC No. 39337.

7. The application according to claim 5, characterized in that, The bacterial count in the Bacillus coagulans culture is at least 1.0 × 10⁻⁶. 9 CFU / mL; the inoculum size is 1-5%, V / V.

8. The application according to claim 5, characterized in that, Ultrafiltration was performed using a 5KD molecular weight membrane core to obtain a cutoff component of <5KD.

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