Lactobacillus mucilaginosus and application thereof in preparation of uric acid reducing preparation

Abstract

The invention belongs to the technical field of microbial fermentation, and discloses lactobacillus mucilaginosus and application thereof in preparation of a uric acid reducing preparation. The survival rate of the fermented lactobacillus mucus in simulated gastric juice with the pH value of 2.0 for 3 hours is greater than 92%, the survival rate in simulated intestinal juice environment containing 0.5% of cholate is greater than 86%, and the viability of the fermented lactobacillus mucus is obviously higher than that of a conventional strain; and the tea has the effects of reducing uric acid, resisting oxidation and inhibiting pathogenic bacteria. The xanthine oxidase inhibition rate of the natural medicinal and edible compound potion obtained by mixing and co-fermenting the strain and existing bacterial powder is greater than 77% (45% or more higher than that of an unfermented group), and the oxidation resistance after fermentation can be improved by 50% at the same time. The beverage is suitable for being drunk as a daily functional food for a long time, and provides a natural and healthy collaborative diet relieving scheme for patients with hyperuricemia and gout.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of microbial fermentation, in particular to a strain of fermented Lactobacillus mucus and its application in preparation of a preparation for reducing uric acid. BACKGROUND

[0002] In recent years, with the change of dietary structure and lifestyle, the incidence of hyperuricemia has increased year by year, and has become an important health problem worldwide. Globally, the prevalence of hyperuricemia has also increased significantly. In addition, studies have shown that hyperuricemia is closely related to gout, chronic kidney disease, cardiovascular disease and other diseases. For example, about 90% of gout patients are accompanied by hyperuricemia, and the risk of chronic kidney disease in patients with hyperuricemia is 2-3 times higher than that in normal population. These data show that hyperuricemia is not only a metabolic disease, but also a major problem in global public health that needs to be solved urgently.

[0003] Hyperuricemia and the gout it causes are a chronic metabolic disease that needs long-term management. In addition to taking medication as prescribed, it is also necessary to maintain uric acid homeostasis by continuously adjusting dietary habits. However, traditional dietary interventions often have problems such as difficulty in implementation and slow results, and there is an urgent need for functional foods or drinks that have significant efficacy and long-term safety for consumption in the market. SUMMARY

[0004] The primary purpose of the present application is to overcome the shortcomings and deficiencies of the prior art, and to provide a strain of fermented Lactobacillus mucus.

[0005] Another purpose of the present application is to provide a food fermentation agent.

[0006] Still another purpose of the present application is to provide the use of the above-mentioned fermented Lactobacillus mucus or food fermentation agent in the preparation of a preparation for reducing uric acid.

[0007] Still another purpose of the present application is to provide a compound food and medicine homologous drink and a preparation method thereof.

[0008] The above-mentioned purposes of the present application are achieved by the following technical solutions: A strain of fermented Lactobacillus mucus, named Lactobacillus fermentum (Lactobacillus fermentum) Limosilactobacillus fermentum ) BW-3, with the accession number GDMCC No: 67568, preserved in the Guangdong Microbial Culture Collection Center of the Guangzhou Institute of Microbiology, Guangdong Academy of Sciences, located at No. 59, Building 5, Guangzhou Institute of Microbiology, Guangdong Academy of Sciences, 100, Martyrs' Road, Guangzhou, on December 29, 2025.

[0009] A food fermentation agent containing the above-mentioned fermented Lactobacillus mucus.

[0010] The food fermentation agent also contains at least one of Lactobacillus rhamnosus, Lactobacillus plantarum, Bifidobacterium animalis and Lactobacillus casei; preferably contains Lactobacillus rhamnosus, Lactobacillus plantarum, Bifidobacterium animalis and Lactobacillus casei complex.

[0011] The food fermentation agent preferably contains the mixed bacterial agent of the above-mentioned Lactobacillus muciaginosus, Lactobacillus rhamnosus, Lactobacillus plantarum, Bifidobacterium animalis and Lactobacillus casei in a cell number ratio of 4:1:1:1:1.

[0012] The above-mentioned Lactobacillus muciaginosus or the above-mentioned food fermentation agent is used for preparing a preparation for reducing uric acid or a food suitable for gout patients.

[0013] A preparation method of a compound food-drug homologous beverage, comprising the following steps: (1) decocting a traditional Chinese medicine composition with water to obtain an extract, sterilizing the extract to obtain a compound water extract to be fermented; wherein the traditional Chinese medicine composition comprises 15-25 parts of yiyi- ren, 20-40 parts of dried tangerine or orange peel, 15-25 parts of radix astragali, 5-15 parts of licorice, 30-50 parts of persicae semen, 15-25 parts of poria cocos, 5-15 parts of puerariae radix, 30-50 parts of hawthorn, 3-6 parts of alismatis rhizoma, and 3-6 parts of mori folium; (2) inoculating the compound water extract to be fermented with the above-mentioned Lactobacillus muciaginosus or the above-mentioned food fermentation agent, and fermenting to obtain a compound food-drug homologous beverage.

[0014] The traditional Chinese medicine composition in step (1) preferably comprises 20 parts of yiyi- ren, 30 parts of dried tangerine or orange peel, 20 parts of radix astragali, 10 parts of licorice, 40 parts of persicae semen, 20 parts of poria cocos, 10 parts of puerariae radix, 40 parts of hawthorn, 5 parts of alismatis rhizoma, and 5 parts of mori folium.

[0015] The decocting in step (1) is preferably performed as follows: first, 5-15 times the mass of water of the traditional Chinese medicine composition is added and decocted for 0.5-2 hours to collect the extract; then, 3-8 times the mass of water of the traditional Chinese medicine composition is added to the residue and decocted for 0.5-1.5 hours to collect the extract; and the extracts obtained in the two steps are combined; more preferably, 10 times the mass of water of the traditional Chinese medicine composition is first added and decocted for 1 hour to collect the extract; then, 5 times the mass of water of the traditional Chinese medicine composition is added to the residue and decocted for 0.5 hour to collect the extract; and the extracts obtained in the two steps are combined.

[0016] The sterilization in step (1) is preferably performed by pasteurization.

[0017] The pasteurization is preferably performed at 80-90°C for 10-20 minutes; more preferably, at 85°C for 15 minutes.

[0018] The Lactobacillus muciappilus in step (2) is added in the form of a seed solution.

[0019] The food fermentation agent in step (2) is added in the form of a seed solution.

[0020] The seed solution is obtained by inoculating the Lactobacillus muciappilus or the food fermentation agent described above into MRS medium and culturing at 35-38°C with shaking.

[0021] The concentration of the seed solution is preferably 10 8 CFU / mL.

[0022] The inoculation amount in step (2) is preferably 5-20% by volume; more preferably 10-15% by volume.

[0023] The fermentation temperature in step (2) is preferably 25-45°C; more preferably 30-40°C; most preferably 35-37°C.

[0024] The fermentation time in step (2) is preferably 12-96h; more preferably 24-72h; most preferably 36-48h.

[0025] A compound medicinal and edible homologous beverage is obtained by the preparation method described above.

[0026] The present application has the following advantages: (1) After isolation and purification, a new Lactobacillus fermentum (Lactobacillus fermentum BW-3) is screened and obtained. Limosilactobacillus fermentum The strain has good surface hydrophobicity, self-aggregation ability, lysozyme resistance, gastrointestinal fluid tolerance, and significant xanthine oxidase inhibition ability, pathogenic bacteria resistance and antioxidant capacity. These characteristics enable the strain to better reach the gastrointestinal tract and exert its probiotic function.

[0027] (2) The fermented compound medicinal and edible beverage prepared from the Lactobacillus fermentum obtained by the present application has excellent uric acid-lowering effect and antioxidant effect. Experimental results show that the xanthine oxidase inhibition rate of the medicinal and edible material composition prepared by the present application is 32.42%, and the xanthine oxidase inhibition rate of the mixed fermentation beverage is 77.62%, which is increased by 45.20%. Through mixed fermentation synergism, the uric acid-lowering effect and antioxidant effect of the medicinal and edible material are comprehensively improved, and the probiotic function is also possessed, which is suitable for developing into functional beverage, solid granules and other daily foods, and provides a safe, effective and easy-to-promote health solution for patients with hyperuricemia and gout. BRIEF DESCRIPTION OF DRAWINGS

[0028] Figure 1 It is a colony morphology photograph of Lactobacillus muciappilus BW-3.

[0029] Figure 2 Figure for hemolytic effect of L. fermentum BW-3.

[0030] Figure 3 Figure for effect of L. fermentum BW-3 alone, other bacteria mixed fermentation (not containing BW-3) and co-fermentation of BW-3 and other bacteria on the effect of reducing uric acid of compound medicinal food and beverage.

[0031] Figure 4 Figure for change of xanthine oxidase inhibition rate of water extract of compound medicinal food material fermented by mixed bacteria with fermentation time.

[0032] Figure 5 Figure for change of xanthine oxidase inhibition rate of water extract of compound medicinal food material fermented by mixed bacteria with fermentation temperature.

[0033] Figure 6 Figure for change of xanthine oxidase inhibition rate of water extract of compound medicinal food material fermented by mixed bacteria with inoculum amount. DETAILED DESCRIPTION

[0034] The application will be described in further detail below with reference to the embodiments and drawings, and the described embodiments are only part of the embodiments of the application, rather than all the embodiments. Based on the embodiments in the application, all other embodiments obtained by those of ordinary skill in the art without creative labor fall within the protection scope of the application. Example 1

[0035] Culture of L. fermentum BW-3 (1) Preparation of MRS medium The components of the MRS medium are as follows: 10.0 g / L of proteose peptone, 10.0 g / L of beef extract powder, 4.0 g / L of yeast extract powder, 2.0 g / L of ammonium citrate, 5.0 g / L of sodium acetate, 0.2 g / L of magnesium sulfate, 0.05 g / L of manganese sulfate, 2.0 g / L of potassium phosphate dibasic, 20.0 g / L of glucose and 1.0 g / L of Tween 80.

[0036] Preparation method: dissolve the above compounds with ultrapure water and make up to 1 L, adjust the pH to 5.7, and sterilize in a high-pressure sterilization pot at 121℃ for 20 min.

[0037] (2) Isolation and purification of fermentation strains Sample fish tea, meat tea and distiller's grains and inoculate into MRS liquid medium, 37℃, 180 rpm shaking culture for 24 h, then use inoculation ring to spread the culture liquid on the culture dish containing solid MRS medium, 37℃ culture for 48 h, then pick single colonies and streak for isolation and purification and screening.

[0038] (3) Screening of strains effectively inhibiting xanthine oxidase The strains were inoculated into MRS culture medium and cultured at 37°C for 24 h, and then 2 mL of the culture solution was centrifuged at 5,000 r for 5 min, and the supernatant was collected for determination of xanthine oxidase inhibition activity.

[0039] Determination of xanthine oxidase inhibition rate: 50 μL of the sample and 50 μL of a xanthine oxidase (XOD) solution (0.092 U / mL, solvent: 0.1 M, pH 7.4 PBS) were incubated at 37°C for 10 min, and then 100 μL of a xanthine solution (0.1 mM) was added to initiate the enzymatic reaction, and the reaction was performed for 4 min. The absorbance was read at 290 nm on an enzyme marker, and the absorbance was recorded.

[0040] The XOD inhibition rate was calculated as follows: inhibition rate (%) = [1-(B1-B0) / (A1-A0)]*100.

[0041] Table 1 Reaction system for determination of XOD inhibition rate

[0042] The test results are shown in Table 2, and the inhibition rates of the 9 isolated strains on xanthine oxidase are 7.41%-65.33%. Among them, the inhibition rate of strain BW-3 is relatively high, which is 65.33%, and strain BW-3 is selected for further study.

[0043] Table 2 Inhibition rate of isolated strains on xanthine oxidase

[0044] The BW-3 colony inoculated on the MRS solid culture medium plate is shown in Table 2, which is medium-sized, convex, wet, with a neat edge, and white round. Figure 1 After Gram staining, BW-3 is a Gram-positive bacterium.

[0045] (4) 16s rDNA identification of the strain The DNA of strain BW-3 was extracted using a Shenguo kit, and PCR amplification was performed using a universal primer and a downstream primer 1492R.

[0046] Upstream primer 27F: 5'-AGAGTTTGATCCTGGCTCAG-3'; Downstream primer 1492R: 5'-GGTTACCTTGTTACGACTT-3'.

[0047] The PCR reaction system consisted of: 10 µL 2×Taq Mix, 2 µL DNA template, 1 µL primer 27F (24.1 nmol concentration), 1 µL primer 1492R (22.8 nmol concentration), and 6 µL ddH2O.

[0048] The PCR amplification program was as follows: pre-denaturation at 94˚C for 5 minutes; denaturation at 95˚C for 30 seconds, annealing at 56˚C for 1 minute, extension at 72˚C for 30 seconds, for 35 cycles; and finally extension at 72˚C for 7 minutes.

[0049] The sequencing was performed by Suzhou Genewiz. The PCR amplification products were sequenced. Generally, the sequence quality at both ends of the sequencing is poor. The low-quality sequences at both ends were removed by quality cutting. The quality-controlled paired-end sequencing results were then assembled to obtain the 16S rDNA sequence shown in SEQ ID NO.1.

[0050] The 16S rDNA sequence is as follows:

[0051] The assembled 16S rDNA sequence was compared with the NCBI database. Based on the coverage and similarity of the comparison results, the sample with the highest comparison score was selected, confirming its species as *Lactobacillus fermentum*. *Lactobacillus fermentum* BW-3 was deposited at the Guangdong Provincial Center for Microbial Culture Collection on December 29, 2025, with accession number GDMCC No: 67568. Example 2

[0052] Probiotic characteristics of Lactobacillus fermentum BW-3 (surface hydrophobicity, self-aggregation, lysozyme resistance, gastrointestinal fluid tolerance, xanthine oxidase inhibition rate, antioxidant capacity, anti-pathogenic ability, hemolysis, and antibiotic sensitivity): (1) Surface hydrophobicity determination Lactobacillus fermentans BW-3 was inoculated into MRS medium and cultured at 37℃ and 180 rpm for 24 h. 30 mL of the 24-h culture was transferred to a 50 mL centrifuge tube and centrifuged at 4℃ and 6000 rpm for 10 min. The supernatant was discarded, and the culture was mixed with PBS buffer by pipetting. This washing process was repeated three times. The bacterial sludge was then resuspended in PBS buffer, and the OD was adjusted. 600 The result was adjusted to 0.6 ± 0.02, denoted as A0. 1 mL of xylene and 1 mL of chloroform were added to separate 10 mL centrifuge tubes, followed by the addition of 3 mL of OD. 600 The bacterial suspension was 0.6 ± 0.02 mg / L. After vortexing, it was allowed to stand at room temperature for 20 min. After 20 min, the aqueous phase was collected and the OD was measured. 600 The absorbance value at that point is denoted as A1.

[0053] The formula for calculating the surface hydrophobicity of a strain is as follows: hydrophobicity of strain (%) = (1-A1 / A0)×100.

[0054] Strains' surface hydrophobicity refers to their ability to detach from the aqueous phase and move towards other organic phases, and is related to their non-specific adhesion ability. Measurements showed that the surface hydrophobicity of *Lactobacillus myxobolus* BW-3 was 25.35%, indicating a certain degree of hydrophobicity.

[0055] (2) Determination of self-aggregation Inoculate *Lactobacillus fermentum* BW-3 into an anaerobic flask containing MRS broth and incubate at 37°C and 180 rpm for 24 h. Transfer 30 mL of the bacterial culture to a 50 mL centrifuge tube, centrifuge at 4°C and 5000 rpm for 10 min, then resuspend the bacterial culture in PBS buffer and adjust the OD. 600The value was recorded as B0, up to 0.6 ± 0.02. The suspension was allowed to stand at room temperature, and then 200 μL of the supernatant was collected at 3 h, 6 h, and 24 h to determine the OD value. 600 The absorbance at that point is denoted as B1.

[0056] The formula for calculating the self-aggregation ability of the strain is as follows: Self-aggregation (%) = (1-B1 / B0) × 100.

[0057] Strain self-aggregation refers to the ability of bacterial cells to combine and form cell aggregates, which is related to biofilm formation and helps the strain colonize intestinal epithelial cells. Measurements showed that the self-aggregation rate of *Lactobacillus fermentum* BW-3 gradually increased with storage time, reaching 92.27% at 24 hours. This high self-aggregation rate is beneficial for *Lactobacillus fermentum* to adhere to intestinal epithelial cells, helping them form a stable ecosystem in the intestine.

[0058] (3) Lysozyme resistance test To assess tolerance to lysozyme, *Lactobacillus fermentum* was inoculated and cultured for 24 h. The experiment consisted of a control group and an experimental group. In the control group, cultures were grown in MRS broth without lysozyme. In the experimental group, lysozyme was added to MRS broth at final concentrations of 1 mg / mL and 2 mg / mL. The different treatment groups were incubated at 37 °C for 24 h, and the OD of the cells was measured. 600 .

[0059] The test results showed that after culturing in MRS medium containing lysozyme (1 mg / mL and 2 mg / mL) for 24 h, the survival rate of Lactobacillus fermentum BW-3 reached 93.85% and 85.38%, respectively. This indicates that Lactobacillus fermentum BW-3 has good lysozyme tolerance, making it difficult to be decomposed by lysozyme in human saliva or gastric juice, and it can successfully pass through the digestive tract to colonize the intestine.

[0060] (4) Gastrointestinal fluid tolerance test Preparation of simulated gastric juice: Dissolve 0.62 g NaCl, 0.22 g KCl, 0.03 g CaCl2·2H2O, 0.027 g KH2PO4, 0.008 g MgCl2, and 0.112 g NaHCO3 in 200 mL of ultrapure water. Adjust the pH to 2.0 with 0.1 M HCl to obtain a gastric electrolyte solution. Take 150 mL of the above gastric electrolyte solution, add 0.036 g pepsin (activity ≥120 U / g) and 0.3 mL of 1 M sodium acetate solution (pH=5), and adjust the pH to 2.0 with 0.1 M HCl to obtain simulated gastric juice. Store at 4℃ for later use.

[0061] Preparation of simulated intestinal fluid: Dissolve 1.35 g NaCl, 0.163 g KCl, and 0.083 g CaCl2·2H2O in 250 mL of ultrapure water, and adjust the pH to 7.0 with 0.1 M NaHCO3 to obtain a small intestinal electrolyte solution. Take 200 mL of the above small intestinal electrolyte solution, and mix it with 200 mL of 7% w / v trypsin solution (i.e., 7 g / 100 mL, trypsin activity of 2000 U / g, solvent is 0.1 M, pH 7.4 PBS), 400 mL of 1% w / v bile salt solution (i.e., 1 g / 100 mL, solvent is 0.1 M, pH 7.4 PBS) and 26 mg of trypsin (130 U / mg). Adjust the pH to 7.0 with 0.1 M NaHCO3 to obtain simulated intestinal fluid.

[0062] Collect the bacterial cells from the culture medium of strain BW-3 with good growth condition by centrifugation, mix with PBS buffer by pipetting, wash the bacterial cells three times, and then resuspend them in simulated gastric and intestinal fluids. Incubate at 37°C and 150 rpm on a shaker. Samples of simulated gastric fluid and simulated intestinal fluid were taken at 0 h, 3 h and 6 h. The samples were serially diluted with PBS buffer and then plated on MRS solid agar plates for colony counting.

[0063] The formula for calculating the survival rate of bacterial strains is as follows: Survival rate of bacterial strains (%) = Nt / N0 × 100, where Nt is the number of colonies after 3 h or 6 h, and N0 is the number of colonies after 0 h.

[0064] Experimental results showed that the survival rates of *Lactobacillus mucinus* BW-3 in gastric juice were 92.28% and 78.49% after 3 hours and 6 hours, respectively; and in intestinal juice, the survival rates were 86.38% and 53.19%, respectively. This indicates that this *Lactobacillus* exhibits high tolerance to simulated gastrointestinal fluids and can stably exert its probiotic effects.

[0065] (5) Xanthine oxidase inhibition rate The testing method was the same as in Example 1. The results showed that Lactobacillus fermentum BW-3 exhibited a xanthine oxidase inhibition rate of 65.33%, indicating good potential for lowering uric acid.

[0066] (6) Antioxidant capacity determination 1) Determination of DPPH free radical scavenging ability: Take 500 μL of the supernatant obtained by centrifugation after 24 h of culture, add 500 μL of 0.2 mM DPPH solution, shake well, and let stand in the dark at room temperature for 30 min. Then centrifuge at 5000 r / min for 10 min, take the supernatant and measure the absorbance at 515 nm.

[0067] DPPH free radical scavenging rate (%) = 1 − (A 样品 -A 空白 ) / A 对照 ×100.

[0068] In the formula: A 空白 The absorbance value is for the blank group (using an equal volume of water to replace DPPH, adding only the supernatant of the bacterial solution to eliminate interference from the absorbance of the bacterial solution itself); A 样品 A is the absorbance value obtained from the supernatant of the bacterial culture to be tested; 对照 The absorbance value is for the control group (with water added, negative control).

[0069] 2) Determination of ABTS free radical scavenging ability: Take 50 μL of bacterial supernatant and add 400 μL of ABTS solution diluted with PBS (OD200). 734 =0.7), after thoroughly mixing, let it stand in the dark at room temperature for 10 min, and then measure the absorbance at 734 nm.

[0070] ABTS radical scavenging rate (Rdpph) = [A0 - A1 / A0] × 100% In the formula: A0 is the absorbance value of the blank sample (i.e., water of equal volume replacing the supernatant of the bacterial solution); A1 is the absorbance value of the sample.

[0071] Tests showed that Lactobacillus fermentum BW-3 had a DPPH free radical scavenging rate of 80.56% and an ABTS free radical scavenging rate of 91.67%.

[0072] (7) Determination of resistance to pathogens The antibacterial effect of cell-free supernatant (CFS) of isolated strains against two common enteric pathogens was evaluated using the perforation method. Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922 were selected as indicator bacteria in this experiment. First, Lactobacillus fermentum BW-3 was cultured in MRS broth at 37°C and 180 rpm for 24 h. Then, it was centrifuged at 4500 rpm for 10 min, and the supernatant was collected and filtered through a 0.22 μm sterile filter to obtain cell-free supernatant (CFS). Simultaneously, the two indicator bacteria were cultured in LB broth to the stationary phase and diluted to 10⁻⁶ liters each. 7 CFU / mL, take 100 μL and spread it evenly on LB solid medium. Make three wells (8 mm in diameter) on each medium, and then add 100 μL of CFS of fermented Lactobacillus mucinus BW-3. Use ordinary MRS liquid medium as a blank control. Place the medium in a 37°C incubator and incubate for 24 h, and measure and record the diameter of the inhibition zone.

[0073] The inhibition zone diameter of fermented Lactobacillus mucinus BW-3 against Staphylococcus aureus was 16.33±1.53 (mm), and the inhibition zone diameter of Escherichia coli was 18.67±1.15 (mm), demonstrating its good ability to inhibit pathogens and facilitating its probiotic effects in the gastrointestinal tract.

[0074] (8) Hemolytic assay Fermenting *Lactobacillus myxitis* BW-3 was inoculated onto Columbia blood agar plates and incubated at 37°C for 14–48 h. The presence or absence of hemolytic zones around the colonies was observed and photographed. Microbial hemolysis can be classified into three types: α-hemolysis, β-hemolysis, and γ-hemolysis. α-hemolysis refers to incomplete lysis of red blood cells, resulting in a surrounding green area; β-hemolysis refers to complete lysis of red blood cells, resulting in a surrounding clear area; and γ-hemolysis does not cause hemolysis.

[0075] result Figure 2 As shown, Lactobacillus fermentum BW-3 exhibits γ-hemolysis, meaning it does not cause hemolysis, thus meeting the safety requirements for probiotics.

[0076] (9) Antibiotic susceptibility testing Lactobacillus fermentum BW-3 was cultured in an anaerobic flask containing MRS medium for 24 hours. 100 μL of the bacterial culture was then transferred to solid MRS medium and spread evenly using a spreader. Antibiotic susceptibility testing discs (6 mm in diameter) were then gently placed on the plate, with three replicates per group. The size of the inhibition zone was determined according to the Clinical and Laboratory Standards Institute (CLST). The antibiotic susceptibility of the strain was assessed based on the size (diameter) of the inhibition zone.

[0077] Tests revealed that this fermenting *Lactobacillus mucinus* strain exhibits high sensitivity to most antibiotics, suggesting that it is generally safe in terms of antibiotic resistance. The table below shows the sensitivity test results for various antibiotics to this fermenting *Lactobacillus mucinus*.

[0078] Table 3. Susceptibility of strain (BW-3) to different types of antibiotics

[0079] Note: R indicates resistance; I indicates moderate sensitivity; S indicates sensitivity. Example 3

[0080] Effects of Lactobacillus mucin BW-3 fermentation alone, mixed fermentation with other bacteria (without BW-3), and co-fermentation of BW-3 with other bacteria on the uric acid-lowering effect of compound medicinal food and beverage preparations: (1) Preparation of fermentation strains: Group A (BW-3 single inoculum): BW-3 *Lactobacillus fermentum* was inoculated into MRS liquid medium and activated by culture at 37°C and 180 rpm for 24 h to obtain the bacterial culture. The OD was adjusted... 600 The value is 0.6, which means it is a single fermentation agent of Lactobacillus fermentum BW-3.

[0081] Group B (Other Bacterial Powder Mixtures): Four lyophilized bacterial powders were *Lactobacillus rhamnosus* HC1509 (Weikehaisi Shandong Bioengineering Co., Ltd., 10 billion CFU / g), *Lactobacillus plantarum* HC1775 (Weikehaisi Shandong Bioengineering Co., Ltd., 10 billion CFU / g), *Bifidobacterium animalis* HSLA-009 (Weikehaisi Shandong Bioengineering Co., Ltd., 10 billion CFU / g), and *Lactobacillus casei* HC1378 (Weikehaisi Shandong Bioengineering Co., Ltd., 10 billion CFU / g). Each bacterial powder was resuspended in MRS liquid medium (using the same amount and medium for each powder) according to the instructions and activated at 37°C for 30 minutes to 1 hour. The activated bacterial suspensions were then inoculated into MRS medium and cultured at 37°C and 180 rpm for 18-24 hours until the logarithmic growth phase (OD). 600 ≈0.6). Mix the bacterial solutions of the four strains in a volume ratio of 1:1:1:1 to obtain the mixed bacterial agent of other bacteria (excluding BW-3).

[0082] Group C (A+B mixed inoculum): Activated Lactobacillus fermentum BW-3 bacterial solution (OD) 600 Mix the bacterial culture (OD600≈0.6) prepared in group B with other bacteria (excluding BW-3) at a volume ratio of 1:1 until homogeneous. Adjust the OD of the mixed bacterial culture after mixing. 600 The concentration was 0.6, resulting in BW-3 as a co-fermentation agent with other bacteria.

[0083] (2) Preparation of water extracts from compound medicinal and edible materials: Weigh out 20 parts by weight of Coix seed, 30 parts by weight of dried tangerine peel, 20 parts by weight of Astragalus membranaceus, 10 parts by weight of licorice root, 40 parts by weight of peach kernel, 20 parts by weight of Poria cocos, 10 parts by weight of kudzu root, 40 parts by weight of hawthorn, 5 parts by weight of Alisma plantago-aquatica, and 5 parts by weight of mulberry leaf. Add 10 times the total weight of ultrapure water to the herbs and decoct for 1 hour, collecting the extract. Add 5 times the weight of ultrapure water to the residue and decoct for 0.5 hours, collecting and combining the two extracts. Perform pasteurization at 85℃ for 15 minutes to obtain the compound aqueous extract to be fermented. The XOD inhibition rate of the compound aqueous extract to be fermented was 32.42%.

[0084] (3) Inoculation and fermentation: The compound aqueous extract to be fermented was inoculated with the bacterial agents from groups A, B, and C respectively, at 35°C and an inoculum size of 10% (10 7Under the condition of CFU / mL, the fermentation was carried out at 180 rpm for 48 h with shaking, and the XOD inhibition rate of the fermentation broth was measured. The XOD inhibition rate was used to represent its urinary-reducing ability.

[0085] The results are as follows Figure 3 As shown, the XOD inhibition rate of the medicinal and edible compound fermented with Group A (BW-3 single agent) was 59.46% (an increase of 27.04% compared to before fermentation), the XOD inhibition rate of the medicinal and edible compound fermented with Group B (other mixed agents) was 42.02% (an increase of 9.6% compared to before fermentation), while the XOD inhibition rate of the medicinal and edible compound fermented with Group C (A+B mixed agents) was as high as 73.30% (an increase of 40.88% compared to before fermentation). The experiment found that compared with commercially available uric acid-lowering bacterial powders, adding the *Lactobacillus fermentata* BW-3 discovered in this invention for co-fermentation produces a synergistic effect, significantly improving the uric acid-lowering effect of the resulting medicinal and edible compound. Example 4

[0086] Changes in xanthine oxidase inhibition rate of mixed-culture fermented compound medicinal and edible material water extracts with fermentation time.

[0087] (1) Preparation of water extracts from compound medicinal and edible materials: Weigh out the following ingredients according to weight: 20 parts of Coix seed, 30 parts of dried tangerine peel, 20 parts of Astragalus membranaceus, 10 parts of licorice root, 40 parts of peach kernel, 20 parts of Poria cocos, 10 parts of kudzu root, 40 parts of hawthorn, 5 parts of Alisma plantago-aquatica, and 5 parts of mulberry leaf. Add 10 times the total weight of the herbs to ultrapure water, decoct and extract for 1 hour, and collect the extract. Then add 5 times the weight of ultrapure water to the residue, decoct and extract for 0.5 hours, collect and combine the two extracts, pasteurize at 85℃ for 15 minutes to obtain the compound aqueous extract to be fermented.

[0088] (2) Inoculation and fermentation: Inoculation Example 3 yielded a mixed bacterial inoculum (Lactobacillus fermentum BW-3, Lactobacillus rhamnosus, Lactobacillus plantarum, Bifidobacterium, and Lactobacillus casei bacterial powder) for Group C: At 35°C, 180 r / min, and an inoculum size of 10% (10 7 The aqueous extract of compound medicinal and edible materials was fermented for 96 hours under the condition of CFU / mL. Samples were taken at 0, 12, 24, 36, 48, 72 and 96 hours to determine the XOD inhibition rate of the fermentation broth.

[0089] The results are as follows Figure 4As shown, after 96 hours of fermentation culture of mixed bacteria and water extracts of compound medicinal and edible materials, the XOD inhibition rates of the fermentation broth were 32.42%, 46.06%, 51.67%, 60.10%, 71.32%, 62.76%, and 48.39%, respectively. The XOD inhibition rate of the fermentation broth first increased and then decreased with fermentation time, reaching a peak at 48 hours, indicating the best fermentation effect.

[0090] In addition, the medicinal and edible compound decoction fermented for 48 hours was tested according to the method in Example 1. Its DPPH free radical scavenging rate was 81.41% (51% higher than before fermentation) and its ABTS free radical scavenging rate was 84.58% (10% higher than before fermentation). Example 5

[0091] Example 3 shows the change in xanthine oxidase inhibition rate of the water extract of compound medicinal and edible materials fermented with mixed bacterial agents in group C as a function of fermentation temperature.

[0092] (1) Preparation of water extracts from compound medicinal and edible materials: Weigh out the following ingredients according to weight: 20 parts of Coix seed, 30 parts of dried tangerine peel, 20 parts of Astragalus membranaceus, 10 parts of licorice root, 40 parts of peach kernel, 20 parts of Poria cocos, 10 parts of kudzu root, 40 parts of hawthorn, 5 parts of Alisma plantago-aquatica, and 5 parts of mulberry leaf. Add 10 times the total weight of the herbs to ultrapure water, decoct and extract for 1 hour, collect the extract, add 5 times the weight of ultrapure water, decoct and extract for 0.5 hours, collect and combine the two extracts, pasteurize at 85℃ for 15 minutes to obtain the compound aqueous extract to be fermented.

[0093] (2) Inoculation and fermentation: At temperatures of 25°C, 30°C, 35°C, 40°C, and 45°C, at a speed of 180 r / min, and with an inoculum size of 10% (10 7 After fermenting the water extract of compound medicinal and edible materials for 48 hours under the condition of CFU / mL, the XOD inhibition rate of the fermentation broth was measured.

[0094] The results are as follows Figure 5 As shown, the XOD inhibition rate of the unfermented compound medicinal and edible material water extract was 32.42%. After 48 hours of fermentation with mixed cultures and the compound medicinal and edible material water extract, the XOD inhibition rates of the fermentation broth at temperatures of 25°C, 30°C, 35°C, 40°C, and 45°C were 41.47%, 59.41%, 69.08%, 62.27%, and 43.10%, respectively. The XOD inhibition rate of the fermentation broth first increased and then decreased with increasing fermentation temperature, reaching a peak at 35°C, which is the optimal fermentation temperature. Example 6

[0095] Example 3 shows the change in xanthine oxidase inhibition rate of the mixed bacterial agent fermented compound medicinal and edible material water extract in group C with the inoculum amount.

[0096] (1) Preparation of water extracts from compound medicinal and edible materials: Weigh out 20 parts by weight of coix seed, 30 parts by weight of dried tangerine peel, 20 parts by weight of astragalus root, 10 parts by weight of licorice root, 40 parts by weight of peach kernel, 20 parts by weight of poria cocos, 10 parts by weight of kudzu root, 40 parts by weight of hawthorn, 5 parts by weight of alisma rhizome, and 5 parts by weight of mulberry leaf. Add 10 times the weight of ultrapure water and decoct for 1 hour. Collect the extract and add 5 times the weight of ultrapure water. Decoction for 0.5 hours. Collect and combine the two extracts and pasteurize at 85°C for 15 minutes to obtain the compound water extract to be fermented.

[0097] (2) Inoculation and fermentation: Seed liquid viability is 10 8 The XOD inhibition rate of the fermentation broth was measured after fermentation of the water extract of compound medicinal and edible materials at 35℃ and 180 r / min for 48 h with inoculum amounts of 2%, 5%, 10%, 15% and 20%, respectively.

[0098] The results are as follows Figure 6 As shown, the XOD inhibition rates of the fermentation broth after fermentation with mixed bacteria and water extracts of compound medicinal and edible materials at 35°C for 48 h were 32.42%, 34.77%, 48.85%, 69.39%, 77.62%, and 53.42% at inoculum levels of 0%, 2%, 5%, 10%, 15%, and 20%, respectively. The XOD inhibition rate of the fermentation broth first increased and then decreased with increasing inoculum level, reaching a peak at 15%.

[0099] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A strain of fermenting *Lactobacillus mucilaginosus*, characterized by: The name of the fermenting mucus lactobacillus is Lactobacillus fermentum (Lactobacillus fermentum). Limosilactobacillus fermentum BW-3, with accession number GDMCC No: 67568, was deposited on December 29, 2025, at the Guangdong Provincial Microbial Culture Collection Center, Guangdong Academy of Sciences, located on the 5th floor of Building 59, Courtyard 100, Xianlie Middle Road, Guangzhou.

2. A food fermentation inoculant, characterized in that: Contains the fermented mucinous lactobacillus as described in claim 1.

3. The food fermentation agent according to claim 2, characterized in that: The food fermentation agent also contains at least one of Lactobacillus rhamnosus, Lactobacillus plantarum, Bifidobacterium animalis, and Lactobacillus casei.

4. The food fermentation agent according to claim 3, characterized in that: The food fermentation agent contains a mixed agent of Lactobacillus fermentans, Lactobacillus rhamnosus, Lactobacillus plantarum, Bifidobacterium animalis, and Lactobacillus casei, as described in claim 1, in a cell ratio of 4:1:1:1:

1.

5. The use of the fermenting Lactobacillus mucinus according to claim 1 or the food fermentation agent according to any one of claims 2 to 4 in the preparation of uric acid-lowering preparations or foods suitable for gout patients.

6. A method for preparing a compound medicinal and edible decoction, characterized in that... Includes the following steps: (1) The Chinese herbal medicine composition was decocted in water to obtain an extract that was sterilized, and a compound water extract to be fermented was obtained; wherein, the composition of the Chinese herbal medicine composition is as follows: 15-25 parts of coix seed, 20-40 parts of tangerine peel, 15-25 parts of astragalus, 5-15 parts of licorice, 30-50 parts of peach kernel, 15-25 parts of poria cocos, 5-15 parts of kudzu root, 30-50 parts of hawthorn, 3-6 parts of alisma, and 3-6 parts of mulberry leaf; (2) Inoculate the fermenting Lactobacillus mucinus according to claim 1 or the food fermentation agent according to any one of claims 2 to 4 into the compound water extract to be fermented, and ferment to obtain the compound medicine and food homology beverage.

7. The preparation method of the compound medicinal and edible decoction according to claim 6, characterized in that: The composition of the traditional Chinese medicine composition mentioned in step (1) is as follows: 20 parts of coix seed, 30 parts of tangerine peel, 20 parts of astragalus, 10 parts of licorice, 40 parts of peach kernel, 20 parts of poria cocos, 10 parts of kudzu root, 40 parts of hawthorn, 5 parts of alisma plantago-aquatica, and 5 parts of mulberry leaf.

8. The preparation method of the compound medicinal and edible decoction according to claim 6, characterized in that: The specific operation of the decoction in step (1) is as follows: First, add water of 5 to 15 times the mass of the Chinese herbal composition, decoct and extract for 0.5 to 2 hours, and collect the extract; then add water of 3 to 8 times the mass of the Chinese herbal composition to the dregs, decoct and extract for 0.5 to 1.5 hours, and collect the extract; combine the extracts obtained from the two decoctions. The sterilization method described in step (1) is pasteurization; The fermented *Lactobacillus mucinus* mentioned in step (2) is added in the form of a seed culture; The food fermentation agent mentioned in step (2) is added in the form of seed liquid; The amount of inoculation mentioned in step (2) is 5-20% by volume; The fermentation temperature described in step (2) is 25–45°C; The fermentation time described in step (2) is 12 to 96 hours.

9. The preparation method of the compound medicinal and edible decoction according to claim 8, characterized in that: The pasteurization conditions are sterilization at 80-90°C for 10-20 minutes; The concentration of the seed solution is 10. 8 CFU / mL; The amount of inoculation mentioned in step (2) is 10-15% by volume; The fermentation temperature described in step (2) is 35–37°C; The fermentation time described in step (2) is 24 to 72 hours.

10. A compound medicinal and edible beverage, characterized in that: It is obtained by the preparation method according to any one of claims 6 to 9.

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