Lactobacillus fermentum, a compound preparation thereof and application thereof

Abstract

The application provides a lactobacillus fermentum, a compound preparation thereof and application, and relates to the technical field of microorganisms.The lactobacillus fermentum HXJS4-1 disclosed in the application has better effect in nucleoside degradation than that of a strain in the prior art, can realize rapid degradation of nucleosides, has stronger inhibition on xanthine oxidase, effectively reduces the activity of xanthine oxidase, has obvious uric acid reducing effect in a mouse experiment, has certain anti-inflammatory effect, can be used in the prevention and treatment of hyperuricemia and gout, and provides a new probiotic or drug material for the treatment of hyperuricemia and gout.In addition, the strain has no obvious influence on the indexes of a sample body, proves that the strain has the characteristics of high safety, and can be widely applied to the fields of medicine and food.

Description

Technical Field

[0001] This application relates to the field of microbial technology, and in particular to a strain of Lactobacillus fermentum, its compound preparations, and its applications. Background Technology

[0002] Hyperuricemia (HUA) is a chronic clinical syndrome caused by purine metabolism disorders or abnormal uric acid excretion, resulting in a significantly higher than normal serum uric acid level. Generally, it is defined as a fasting serum uric acid level exceeding 420 μmol / L for men and 360 μmol / L for women on two separate occasions under normal purine dietary conditions. Gout is a crystalline arthritis caused by the deposition of monosodium urate crystals and is directly related to hyperuricemia due to purine metabolism disorders. In recent years, hyperuricemia in my country has shown a significant upward trend and a younger age of onset. Hyperuricemia is not only an early stage of gout but also an independent risk factor for hypertension, diabetes, coronary heart disease, and chronic kidney disease. Gout has become the "fourth high" after diabetes, hypertension, and hyperlipidemia, posing a serious threat to people's health.

[0003] Clinically, hyperuricemia is generally prevented and treated in three ways. The first is to reduce uric acid production using xanthine oxidase (XOD) inhibitors, such as allopurinol and febuxostat. The second is to increase uric acid excretion using URAT1 inhibitors, such as benzbromarone and ricinole. The third is to degrade uric acid using uricase drugs, such as raburicase and precancerous urate. However, xanthine oxidase inhibitors and URAT1 inhibitors are chemical drugs that can damage liver and kidney function and have strong toxic side effects. While uricase drugs are more effective, they have strong immunogenicity and are generally used as second-line drugs.

[0004] Under normal circumstances, the human body contains approximately 1200 mg of uric acid. About 600 mg is newly generated daily, while 600 mg is excreted, maintaining a balance. About 70% of the uric acid excreted is through the kidneys, with the remainder excreted in feces or further metabolized by intestinal flora. A small portion is excreted in the intestines or broken down by intestinal flora. In recent years, improving traditional metabolic diseases such as hyperuricemia based on gut microbiota has become a new research direction. Multiple studies have shown that regulating gut microbiota through probiotics and prebiotics plays an important role in regulating serum uric acid and its distribution in the body, and is helpful in the treatment of hyperuricemia and gout.

[0005] This demonstrates that current medications for hyperuricemia have numerous problems, exhibiting significant side effects and being unsuitable for long-term use. While a growing number of probiotics have been shown to be effective in preventing or alleviating hyperuricemia, their efficacy in the actual adjunctive treatment of hyperuricemia and related diseases urgently needs improvement. Summary of the Invention

[0006] To address the aforementioned issues, this invention aims to screen for probiotics and related preparations that can better reduce blood uric acid levels, alleviate hyperuricemia and gout, and are tolerant to carbon and / or nitrogen substrates.

[0007] On the one hand, this application provides a strain of Lactobacillus fermentum HXJS4-1, which is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC NO: 29677.

[0008] On the other hand, this application also provides a strain of Lactobacillus paracasei HXXJ-008, which is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC NO: 30483.

[0009] On the other hand, this application also provides a formulation comprising one or more of Lactobacillus fermentum, Lactobacillus paracasei, and / or Lactobacillus plantarum.

[0010] Preferably, the fermenting lactobacillus is Lactobacillus fermentum HXJS4-1.

[0011] More preferably, the *Lactobacillus paracasei* is *Lactobacillus paracasei* HXXJ-008.

[0012] Most preferably, the Lactobacillus plantarum is Lactobacillus plantarum HXJS3-1, which is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC NO: 29676.

[0013] In a preferred embodiment, the Lactobacillus fermentum is Lactobacillus fermentum HXJS4-1, which is deposited at the China General Microbiological Culture Collection Center with accession number CGMCC NO: 29677.

[0014] Preferably, the Lactobacillus fermentum may include Lactobacillus fermentum HXJS4-1, Lactobacillus fermentum HXJS4-1 inoculum, Lactobacillus fermentum HXJS4-1 dead bacterial suspension, Lactobacillus fermentum HXJS4-1 metabolites, and Lactobacillus fermentum HXJS4-1 extract.

[0015] In a preferred embodiment, the Lactobacillus paracasei is Lactobacillus paracasei HXXJ-008, which is deposited at the China General Microbiological Culture Collection Center with accession number CGMCC NO: 30483.

[0016] Preferably, the Lactobacillus paracasei may include Lactobacillus paracasei HXXJ-008, Lactobacillus paracasei HXXJ-008 bacterial agent, Lactobacillus paracasei HXXJ-008 dead bacterial suspension, Lactobacillus paracasei HXXJ-008 metabolites, and Lactobacillus paracasei HXXJ-008 extract.

[0017] Preferably, the *Lactobacillus plantarum* may include *Lactobacillus plantarum* HXJS3-1, *Lactobacillus plantarum* HXJS3-1 inoculum, *Lactobacillus plantarum* HXJS3-1 dead bacterial suspension, *Lactobacillus plantarum* HXJS3-1 metabolites, and *Lactobacillus plantarum* HXJS3-1 extract.

[0018] Those skilled in the art will understand that known excipients and solvents may be added to the formulation without affecting the activity and efficacy of the strain.

[0019] The excipients may be appropriate solvents, propellants, solubilizers, co-solvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, binding agents, penetration promoters, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, absorbents, diluents, flocculants and anti-flocculation agents, filter aids, release inhibitors, etc.

[0020] Furthermore, the formulation can be prepared using common methods, such as directly mixing the bacterial strain. One or more diluents or pharmaceutically acceptable carriers may also be added. In a preferred embodiment, the formulation can be prepared as an oral dosage form.

[0021] Furthermore, the total viable count of the *Lactobacillus fermentum*, *Lactobacillus paracasei*, and / or *Lactobacillus plantarum* is greater than 1 × 10⁻⁶. 8 CFU.

[0022] Preferably, 1×10 8 CFU-1×10 12 CFU.

[0023] The total number of viable bacteria of *Lactobacillus fermentum*, *Lactobacillus paracasei*, and / or *Lactobacillus plantarum* can be 1 × 10⁻⁶. 8 CFU, 1×10 9 CFU, 1×10 10 CFU, 1×10 11 CFU, 1×10 12 CFU and above.

[0024] In a preferred embodiment, the sum of the viable counts of the *Lactobacillus fermentum*, *Lactobacillus paracasei*, and / or *Lactobacillus plantarum* and their compound strains is greater than 1 × 10⁻⁶. 8 CFU.

[0025] Those skilled in the art will understand that the dosage of the bacterial strain can be adjusted according to the actual situation, as long as the sum of the total viable count of the three strains mixed randomly is greater than 1×10⁻⁶. 8 CFU is sufficient; this application only provides one example and does not impose specific limits on its upper limit.

[0026] This application is the first to use a combination of Lactobacillus fermentum HXJS4-1, Lactobacillus paracasei HXXJ-008, and Lactobacillus plantarum HXJS3-1 to achieve better results in lowering uric acid levels and reducing inflammatory factors in hyperuricemia, demonstrating a synergistic effect among the three strains, with effects reaching the level of the positive drug group.

[0027] In a preferred embodiment, the preparation may be a pharmaceutical product, probiotic yogurt, soy milk, compressed candy, and / or solid beverage.

[0028] On the other hand, this application also provides the use of the aforementioned Lactobacillus fermentum, Lactobacillus paracasei, or the aforementioned preparation in the degradation of nucleosides and / or the preparation of nucleoside-degrading products.

[0029] Preferably, the nucleoside includes one or more of inosine, guanosine, and adenosine; more preferably, the nucleoside includes inosine and / or guanosine.

[0030] The preferred strain is Lactobacillus fermentum HXJS4-1.

[0031] Preferably, the degradation is rapid degradation.

[0032] This application is the first to use *Lactobacillus fermentum* HXJS4-1 for nucleoside degradation. *Lactobacillus fermentum* HXJS4-1 can achieve rapid nucleoside degradation. In one specific embodiment, *Lactobacillus fermentum* HXJS4-1 OD... 600 When the value is 4.0, the inosine-guanosine degradation solution with a final concentration of approximately 1.26 mM can be completely degraded within 1 hour.

[0033] Preferably, the process of degrading nucleosides is not affected by carbon and / or nitrogen sources, that is, when sugars or proteins are present, the Lactobacillus fermentum or Lactobacillus paracasei described in this application can still maintain a high efficiency in degrading nucleosides.

[0034] Preferably, the carbon source can be selected from glucose, sucrose, starch, glycerol, etc., and the nitrogen source can be selected from yeast extract, peptone, yeast extract, beef extract, corn steep liquor, etc.

[0035] More preferably, the carbon source concentration can be 1-5 g / L, and the nitrogen source concentration can be 1-5 g / L.

[0036] The carbon source concentration can be any value among 1 g / L, 2 g / L, 3 g / L, 4 g / L, and 5 g / L, and the nitrogen source concentration can be any value among 1 g / L, 2 g / L, 3 g / L, 4 g / L, and 5 g / L.

[0037] In a preferred embodiment, the carbon source is sucrose and / or glucose, and the nitrogen source is yeast and its extracts.

[0038] On the other hand, this application also provides the use of the aforementioned Lactobacillus fermentum or Lactobacillus paracasei or the aforementioned preparation in inhibiting xanthine oxidase (XOD) activity and / or in the preparation of xanthine oxidase (XOD) inhibitory products.

[0039] This application is the first to verify the inhibitory ability of *Lactobacillus fermentum* HXJS4-1 and / or *Lactobacillus paracasei* HXXJ-008 on xanthine oxidase. *Lactobacillus fermentum* HXJS4-1 showed an inhibition rate of 71.5% on xanthine oxidase. Therefore, this application provides a novel microbial material for xanthine oxidase inhibitors and related products.

[0040] On the other hand, this application also provides the use of the aforementioned Lactobacillus fermentum or Lactobacillus paracasei or the aforementioned preparation for the purpose of inhibiting inflammation and / or preparing anti-inflammatory products for non-disease diagnosis and treatment purposes.

[0041] In this process, inflammation is suppressed by reducing the levels of inflammatory factors or increasing the levels of anti-inflammatory factors. These inflammatory factors include, but are not limited to, interleukin-1β (IL-1β), IL-6, TNF-α, IL-10, and IL-11.

[0042] This application is the first to use Lactobacillus fermentum HXJS4-1, Lactobacillus plantarum HXJS3-1 and / or Lactobacillus paracasei HXXJ-008 to inhibit inflammatory factors and reduce the number of inflammatory-related factors in the sample.

[0043] In a preferred embodiment, this application specifically verified the levels of inflammation-related factors in the liver and kidneys of samples, demonstrating that *Lactobacillus fermentum* HXJS4-1, *Lactobacillus plantarum* HXJS3-1, and *Lactobacillus paracasei* HXXJ-008, and combinations thereof, can effectively reduce the levels of inflammatory factors and increase the levels of anti-inflammatory factors in the liver and kidneys.

[0044] Its inhibition rates against IL-1β reached as high as 77.5% and 81.93%, respectively;

[0045] The highest inhibition rates against IL-6 were 84.7% and 71.78%, respectively;

[0046] The highest inhibition rates against TNF-α were 77.51% and 79.45%, respectively.

[0047] The highest recovery rates for IL-10 were 70.55% and 73.1%, respectively.

[0048] The highest inhibition rates against IL-11 were 77.77% and 83.19%, respectively.

[0049] Preferably, the sample is a hyperuricemia sample.

[0050] On the other hand, this application also provides the use of the aforementioned Lactobacillus fermentum, Lactobacillus paracasei, or the aforementioned preparation in the preparation of products that lower uric acid levels.

[0051] Preferably, the uric acid level can be the uric acid content in urine or the uric acid content in serum.

[0052] This application is the first to use Lactobacillus fermentum HXJS4-1, Lactobacillus plantarum HXJS3-1 and / or Lactobacillus paracasei HXXJ-008 to prepare products that lower uric acid levels and to use them to lower uric acid levels.

[0053] On the other hand, this application also provides the use of the aforementioned Lactobacillus fermentum or Lactobacillus paracasei or the aforementioned preparation in the preparation of products for the prevention and / or treatment of hyperuricemia.

[0054] Prevention and / or treatment of hyperuricemia can be achieved by inhibiting xanthine oxidase (XOD) activity, reducing the content of inflammatory factors while increasing the content of anti-inflammatory factors, and reducing uric acid levels.

[0055] On the other hand, this application also provides the use of the aforementioned Lactobacillus fermentum or Lactobacillus paracasei or the aforementioned preparation in the preparation of products for the prevention and / or treatment of gout.

[0056] The Lactobacillus fermentum HXJS4-1, Lactobacillus plantarum HXJS3-1 and / or Lactobacillus paracasei HXXJ-008 and their preparations described in this application can reduce the risk of hyperuricemia and effectively reduce the probability of hyperuricemia further developing into gout, thereby playing a role in preventing and / or treating gout.

[0057] Specifically, elevated serum uric acid levels lead to hyperuricemia, which in turn increases the concentration of inflammatory factors in the body, thus increasing the risk of gout. Furthermore, studies have shown that elevated inflammatory factor concentrations may lead to increased XOD activity, which can convert xanthine into uric acid, increasing uric acid production. Therefore, this application aims to reduce uric acid production by inhibiting XOD activity and simultaneously reduce the risk of gout by lowering inflammatory factor levels.

[0058] On the other hand, this application also provides the use of the aforementioned Lactobacillus fermentum and / or the aforementioned Lactobacillus paracasei and / or the aforementioned preparations in enhancing immunity.

[0059] On the other hand, this application also provides the use of the aforementioned Lactobacillus fermentum and / or the aforementioned Lactobacillus paracasei and / or the aforementioned formulation in the preparation of IL-11 inhibitors.

[0060] Furthermore, the IL-11 inhibitor can be used in oral and / or topical formulations for anti-aging and / or life extension.

[0061] Preferably, the IL-11 inhibitor can be used to prevent or treat a variety of age-related diseases selected from cardiovascular fibrosis, pulmonary fibrosis, cancer, diabetic complications, mitochondrial dysfunction, inflammation and cellular senescence, thrombocytopenia, muscle weakness and frailty, coronary artery disease, metabolic decline, coronary artery disease, hypertension, pulmonary hypertension, cerebrovascular disease, aortic disease, and other vascular diseases.

[0062] In addition, the Lactobacillus fermentum HXJS4-1, Lactobacillus plantarum HXJS3-1 and / or Lactobacillus paracasei HXXJ-008 described in this application have high safety and no significant effect on other indicators of the body after consumption, proving that they are safe and non-toxic.

[0063] The present invention has the following beneficial effects:

[0064] The *Lactobacillus fermentum* strain HXJS4-1 disclosed in this invention exhibits superior nucleoside degradation compared to existing strains, achieving rapid nucleoside degradation unaffected by carbon and / or nitrogen sources. This means that the *Lactobacillus fermentum* strain maintains its high nucleoside degradation capacity even when containing carbohydrates or proteins. Furthermore, it demonstrates stronger inhibition of xanthine oxidase, effectively reducing its activity. The strain also exhibits significant uric acid-lowering effects in hyperuricemic mice and possesses certain anti-inflammatory properties.

[0065] This invention also provides a compound preparation of Lactobacillus fermentum HXJS4-1, Lactobacillus plantarum HXJS3-1 and Lactobacillus paracasei HXXJ-008, demonstrating that the three strains have a synergistic effect and can be used in the prevention and treatment of hyperuricemia and gout, providing a new probiotic or pharmaceutical material for the treatment of hyperuricemia and gout.

[0066] This invention also provides the use of the aforementioned *Lactobacillus fermentum* and / or the aforementioned *Lactobacillus paracasei* and / or the aforementioned preparations in the preparation of IL-11 inhibitors. These can be used in oral and / or topical formulations for anti-aging and / or life-extending purposes, and can also be used for the prevention or treatment of various age-related diseases selected from cardiovascular fibrosis, pulmonary fibrosis, cancer, diabetic complications, mitochondrial dysfunction, inflammation and cellular senescence, thrombocytopenia, muscle weakness and frailty, coronary artery disease, metabolic decline, coronary artery disease, hypertension, pulmonary hypertension, cerebrovascular disease, aortic disease, and other vascular diseases.

[0067] In addition, consuming the above-mentioned strains had no significant impact on the body indicators of the samples, proving that they have high safety characteristics and can be widely used in the medical and food fields. Attached Figure Description

[0068] The accompanying drawings, which are included to provide a further understanding of this application and form part of this application, illustrate exemplary embodiments and are used to explain this application, but do not constitute an undue limitation of this application. In the drawings:

[0069] Figure 1 This is a morphological diagram of strain HXJS4-1;

[0070] Figure 2 This is a morphological diagram of strain HXXJ-008;

[0071] Figure 3 This is a statistical graph showing the IL-1β content in the liver and kidneys of mice in Example 8;

[0072] Figure 4 This is a statistical graph showing the IL-6 content in the liver and kidneys of mice in Example 8;

[0073] Figure 5 This is a statistical graph of XOD activity in mouse liver and serum from Example 9;

[0074] Figure 6 This is a statistical chart of serum uric acid levels in mice on days 0, 7, 14, and 21 in Example 9;

[0075] Figure 7 This is a statistical chart of uric acid content in mouse urine from Example 9;

[0076] Figure 8 This is a statistical graph showing the IL-1β content in the liver and kidneys of mice in Example 9;

[0077] Figure 9 This is a statistical graph showing the TNF-α content in the liver and kidneys of mice in Example 9;

[0078] Figure 10 This is a statistical graph showing the IL-6 content in the liver and kidneys of mice in Example 9;

[0079] Figure 11 This is a statistical chart of IL-11 content in the liver and kidneys of mice in Example 9;

[0080] Figure 12 This is a statistical chart of IL-10 content in the liver and kidneys of mice in Example 9;

[0081] Preservation of biological materials:

[0082] A strain of Lactobacillus fermentum HXJS4-1 was deposited on January 19, 2024, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO: 29677. The deposit address is No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, China, Institute of Microbiology, Chinese Academy of Sciences.

[0083] A strain of Lactobacillus plantarum HXJS3-1 was deposited on January 19, 2024, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO: 29676. The deposit address is No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, China, Institute of Microbiology, Chinese Academy of Sciences.

[0084] A strain of Lactobacillus paracasei HXXJ-008 was deposited on April 30, 2024, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO: 30483. The deposit address is No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, China, Institute of Microbiology, Chinese Academy of Sciences. Detailed Implementation

[0085] To more clearly illustrate the overall concept of this application, a detailed description is provided below with reference to the accompanying drawings and embodiments. Numerous specific details are set forth in the following description to provide a more thorough understanding of the invention. However, it will be apparent to those skilled in the art that the invention can be practiced without one or more of these details. In other instances, certain technical features well-known in the art have not been described to avoid confusion with the invention.

[0086] Unless otherwise specified in the examples, the conditions shall be performed according to the standard conditions or the conditions recommended by the manufacturer.

[0087] Unless otherwise specified, in the following embodiments, reagents or instruments whose manufacturers are not indicated are all conventional products that can be purchased commercially.

[0088] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in this invention all employ conventional techniques in the fields of microbiology, biochemistry, analytical chemistry, cell culture, and related areas.

[0089] The culture media and reagent components involved in the following examples are as follows:

[0090] MRS solid culture medium: 10g peptone, 5g beef meal, 20g glucose, 4g yeast extract, 5g sodium acetate, 2g dipotassium hydrogen phosphate, 0.2g magnesium sulfate, 2g triammonium citrate, 0.05g manganese sulfate, 1mL Tween 80, and 15g agar powder are added to 1000mL distilled water, stirred until dissolved, dispensed, and autoclaved at 121℃ for 15 minutes. This culture medium is mainly used for the isolation and cultivation of lactic acid bacteria.

[0091] MRS broth culture medium: 10g peptone, 5g beef meal, 20g glucose, 4g yeast extract, 5g sodium acetate, 2g dipotassium hydrogen phosphate, 0.2g magnesium sulfate, 2g triammonium citrate, 0.05g manganese sulfate, 1mL Tween 80. Add to 1000mL distilled water, heat to boiling to dissolve, dispense, and autoclave at 121℃ for 15min. This culture medium is mainly used for the cultivation of lactic acid bacteria.

[0092] PCA medium: 5g tryptone, 2g yeast extract, 1g glucose, 15g agar, dissolved in 1000mL distilled water, sterilized at 121℃ for 20min. This medium is mainly used for lactic acid bacteria colony counting.

[0093] Nucleoside degradation solution: 33.7 mg inosine and 35.7 mg guanosine were dissolved in 100 mM K3PO4 solution to prepare an inosine-guanosine degradation solution with a final concentration of approximately 1.26 mM.

[0094] HPLC mobile phase: Weigh 3.4g KH2PO4 and dissolve it in 950mL ultrapure water, then add 50mL methanol to prepare a 1L mobile phase.

[0095] Xanthine oxidase solution: Dissolve xanthine oxidase (100U) in 0.2mol / L phosphate buffer (pH 7.5) to a concentration of 0.5U / mL and store at 4℃.

[0096] 2mM xanthine substrate: Accurately weigh 30.422 mg of xanthine standard, dissolve it by sonication in 0.1 mol / L NaOH solution, and then bring the volume to 100 mL with 0.2 mol / L phosphate buffer (pH 7.5) to prepare a 2mM xanthine substrate solution.

[0097] 100mM K3PO4 buffer: Weigh 21.23g K3PO4, add 995mL of ultrapure water, dissolve, then add about 5mL of phosphate to adjust the pH to 7.0, and store at room temperature for later use.

[0098] In addition, the "water" mentioned in this invention includes any feasible water that can be used in the art, such as deionized water, distilled water, ion-exchanged water, double-distilled water, high-purity water, and purified water.

[0099] In the following examples, unless otherwise specified, % means wt%, i.e., weight percentage.

[0100] Example 1: Preliminary screening of Lactobacillus fermentation strains

[0101] Commercially available bacterial slurry solutions were diluted 10, 100, and 1000 times, respectively, and spread onto MRS solid medium containing 1% CaCO3, with three replicates per group. The samples were incubated at 37°C. After three days of incubation, 25 colonies of varying morphology were picked from single colonies with a clear clear zone on the plates and inoculated onto MRS broth medium. The colonies were incubated at 37°C and 220 rpm for 24 hours. They were then transferred to fresh MRS broth medium and incubated for 16 hours. Once the bacterial culture became turbid, the nucleoside degradation capacity was tested.

[0102] Example 2: Determination of Nucleoside Degradation Capacity

[0103] In this embodiment, the nucleoside degradation ability of the strain obtained in Example 1 was determined. Simultaneously, *Lactobacillus fermentum* BNCC138617, purchased from Beina Chuanglian Biotechnology Co., Ltd., was used as a control strain for testing.

[0104] The method for detecting nucleoside degradation capacity includes: using MRS broth medium to measure the OD of the fermentation broth from Example 1 or Lactobacillus plantarum BNCC336943. 600 Adjust the pH to 4.0. Centrifuge 1 mL of fermentation broth at 4℃ and 8000 rpm for 2 min, discard the supernatant, and collect the bacterial cells. Add 750 μL of physiological saline to the bacterial cells and wash them twice, collecting the washed bacterial cells. Add 750 μL of nucleoside degradation solution to the collected bacterial cells, using equal volumes of physiological saline and MRS broth as blank controls. Set up a heat inactivation control by inactivating the bacterial cells at 100℃ for 10 min. Incubate the prepared nucleoside degradation system at 37℃ and 120 rpm for 30 min, 60 min, 90 min, and 120 min, respectively. After the reaction, inactivate the system at 100℃ for 10 min and centrifuge at 12000 rpm for 2 min. Filter the supernatant through a 0.22 μm filter into a liquid chromatography vial for analysis.

[0105] Nucleoside content was determined by high-performance liquid chromatography (HPLC) with a mobile phase of 25 mM KH₂PO₄ containing 5% methanol. An ODS-3 (4.6 × 250 mm, 5 μm) column was used. Detection conditions: PDA detector, detection wavelength 254 nm, column temperature 35 °C, flow rate 1 mL / min, injection volume 20 μL, retention time 20 min.

[0106] The formula for calculating the nucleoside degradation rate is: (Initial nucleoside concentration - Remaining nucleoside concentration after reaction) / Initial nucleoside concentration × 100%

[0107] Finally, a lactic acid bacterium, HXJS4-1, with high nucleoside degradation ability was obtained through screening. Its nucleoside degradation ability is shown in Table 1.

[0108] Table 1. Degradation capacity of HXJS4-1 nucleoside

[0109]

[0110] As shown in Table 1, compared with the control strain BNCC138617, strain HXJS4-1 can rapidly degrade nucleosides, achieving almost complete nucleoside degradation within 1 hour.

[0111] Example 3: Identification of the strain

[0112] (1) Colony morphology characteristics

[0113] The strain HXJS4-1 obtained in Example 2 is a Gram-positive bacterium that grows well in MRS medium. The optimal growth temperature is 37°C, and the optimal pH is 6.5. It does not form spores, lacks flagella, and enters the stationary phase after 16 hours of shake-in culture. On MRS solid medium (plates), the bacterial plaques are milky white, round, with neat edges, opaque, and smooth. Morphology as shown... Figure 1 As shown.

[0114] (2) Molecular biological identification

[0115] In this embodiment, 16S rDNA sequencing was performed on strain HXJS4-1: A small amount of colony was picked from an MRS solid plate and added to 20 μL of prepared PCR reaction solution (the composition of which is shown in Table 2). PCR amplification was performed according to the pre-set program (as shown in Table 3). Afterwards, 4 μL of the solution was run for DNA gel electrophoresis for verification, and a band was found at approximately 1400 bp. The primer sequences involved in the amplification were: 27F: 5'-AGAGTTTGATCCTGGCTCAG-3'; 1492R: 5'-TACGG YTACCTTGTTACGACTT-3'.

[0116] The remaining PCR reaction solution was sent for sequencing, which was performed by Qingke Biotechnology Co., Ltd. The obtained 16S rDNA sequence (as shown in SEQ ID No. 1) was BLAST-aligned on NCBI and compared for homology with known strains to identify the classification of strain HXJS4-1. The strain was ultimately identified as *Lactobacillus fermentum*. This *Lactobacillus fermentum* HXJS4-1 was deposited on January 19, 2024, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC NO: 29677.

[0117] Table 2 PCR reaction system

[0118]

[0119] Table 3 PCR reaction procedure

[0120]

[0121] The 16S rDNA sequence of Lactobacillus fermentum HXJS4-1 (SEQ ID No. 1):

[0122]

[0123] Considering that human food may contain not only nucleosides but also many carbohydrates and proteins, this example further tested the degradation rate of nucleosides by *Lactobacillus fermentum* HXJS4-1 in the presence of different carbohydrate substrates. The specific substrates and detection methods are as follows:

[0124] Substrate 1: 1.26 mM inosine - 1.26 mM guanosine - 2 g / L sucrose - 2 g / L yeast extract;

[0125] Substrate 2: 1.26 mM inosine-1.26 mM guanosine-2 g / L glucose-2 g / L yeast extract;

[0126] Substrate 3: 1.26 mM inosine-1.26 mM guanosine.

[0127] Take OD 600 The Lactobacillus fermentum HXJS4-1 bacterial suspension with a pH of 4.0 was washed with physiological saline and then incubated with equal volumes of substrate solutions 1, 2, and 3 for 2 hours. Physiological saline and nucleoside degradation solution were used as blank controls. The determination method was the same as in Example 2. The nucleoside degradation ability of Lactobacillus fermentum HXJS4-1 under different conditions is shown in Table 4 below.

[0128] Table 4. Nucleoside degradation capacity of strains in the presence of carbon and nitrogen sources.

[0129]

[0130] The experimental results in Table 4 demonstrate that the presence of carbon and nitrogen sources does not affect the degradation of nucleosides by this strain.

[0131] Example 5: XOD Inhibition Ability Determination

[0132] Xanthine oxidase catalyzes the conversion of xanthine to uric acid, and the product uric acid has a characteristic absorption peak at 293 nm. The change in absorbance of the reaction system over a certain period of time, measured using an enzyme-linked immunosorbent assay (ELISA) reader, characterizes the amount of uric acid produced and can thus measure the change in xanthine oxidase activity.

[0133] In this embodiment, the xanthine oxidase (XOD) inhibition ability of Lactobacillus fermentum HXJS4-1 was determined. Lactobacillus fermentum BNCC138617 (purchased from the manufacturer) was used as a control. The specific determination method is as follows:

[0134] Take 0.75 mL of xanthine substrate solution and an equal volume of fermentation broth (OD200). 600Mix the Lactobacillus fermentum HXJS4-1 or Lactobacillus fermentum BNCC138617 bacterial suspensions with a value of 4.0, and finally add 0.5 mL of xanthine oxidase solution (0.5 U / mL) that has been incubated at 25 °C for 20 min. After reacting the reaction system at 25 °C for 30 min, measure the absorbance at a wavelength of 293 nm. Each group has three replicates.

[0135] The specific reaction groups and reaction systems are as follows:

[0136] Control sample ①: 0.75 mL xanthine + 0.5 mL xanthine oxidase + 0.75 mL PBS;

[0137] Control sample ②: 0.75 mL xanthine + 0.5 mL PBS + 0.75 mL fermentation broth;

[0138] Experimental sample ③: 0.75 mL xanthine + 0.5 mL xanthine oxidase + 0.75 mL fermentation broth;

[0139] Control sample ④: 0.75 mL xanthine + 1.25 mL PBS.

[0140] The XOD inhibition rate was calculated by detecting xanthine oxidase activity as follows, and the results are shown in Table 5 below.

[0141] XOD inhibition rate calculation formula: ((Absorbance value of control sample ① - Absorbance value of control sample ④) - (Absorbance value of test sample ③ - Absorbance value of control sample ②)) / (Control sample ① - Control sample ④) × 100%

[0142] Table 5 XOD Inhibition Rate

[0143] HXJS4-1 71.5％ BNCC138617 11.3％

[0144] As shown in Table 5, the *Lactobacillus fermentum* HXJS4-1 obtained in this application achieved an inhibition rate of 71.5% against xanthine oxidase, while the inhibition rate of xanthine oxidase in the control strain was only 11.3%.

[0145] Example 6: Screening of Lactobacillus paracasei HXXJ-008

[0146] Undamaged cow dung was collected by sample collectors from the Narat Grassland in Xinjiang. Wearing disposable sterile gloves, the sampling shovel and spoon were disinfected with 75% alcohol. 5-10g of the core portion was extracted from the cow dung, placed in a sterile sampling bag, sealed, and stored. Sample information was recorded, and the sample was kept in a cold storage box (with ice packs inside). It was then mailed to a Beijing laboratory via low-temperature cold chain express on the same day for experimental purposes. Weigh 1g of cow dung sample and place it in a sterile homogenizing cup. Add 10mL of sterile physiological saline and mix using a vortex mixer for 1-2 minutes to prepare a suspension. Filter the suspension through 4 layers of sterile gauze. Dilute the filtered suspension 10 times, 100 times, and 1000 times respectively and spread it onto MRS solid medium containing 1% CaCO3. After three days of incubation, pick 25 colonies with different morphologies from single colonies with obvious clear zones on the plates. Sequencing the 16S rDNA revealed a strain of Lactobacillus paracasei, named HXXJ-008.

[0147] Example 7: Identification of Lactobacillus paracasei HXXJ-008

[0148] (1) Colony morphology characteristics

[0149] The strain HXXJ-008 obtained in Example 6 is a Gram-positive bacterium that grows well in MRS medium. The optimal growth temperature is 37°C, and the optimal growth pH is 6.5. It does not form spores, lacks flagella, and enters the stationary phase after 16 hours of shake-in culture. On MRS solid plates, the bacterial plaques are milky white, round, with neat edges, opaque, and smooth. Figure 2 As shown.

[0150] (2) Molecular biological identification

[0151] 16S rDNA sequencing: A small amount of colony was picked from an MRS solid plate and added to 20 μL of prepared PCR reaction solution (see Table 6 for the composition). PCR amplification was performed according to the pre-set program (as shown in Table 7). Afterwards, 4 μL of the amplified DNA was run on a DNA gel electrophoresis plate for verification. A band was observed at approximately 1400 bp. Primer sequences involved in the amplification were: 27F: 5'-AGAGTTTGATCCTGGCTCAG-3'; 1492R: 5'-TACGGYTACCTTGTTACGACTT-3'.

[0152] The remaining PCR reaction solution was sent for sequencing, which was performed by Qingke Biotechnology Co., Ltd. The obtained 16S rDNA sequence (as shown in SEQ ID No. 2) was BLAST-aligned on NCBI and compared for homology with known strains to identify the strain. The strain was ultimately identified as *Lactobacillus paracasei*. This *Lactobacillus fermentum* HXXJ-008 was deposited on April 30, 2024, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, China, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCC NO: 30483.

[0153] Table 6 PCR reaction system

[0154]

[0155] Table 7 PCR reaction procedure

[0156]

[0157]

[0158] The 16S rDNA sequence of Lactaseibacillus paracasei HXXJ-008 (SEQ ID No. 2):

[0159] TTAGACGGCTCCGCTCCCTAAAAGGGTTACGCCACCGGCTTCGGGTGTTACAAACTCTC

[0160] ATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCGTGCTGAT

[0161] CCGCGATTACTAGCGATTCCGACTTCGTGTAGGCGAGTTGCAGCCTACAGTCCGAACT

[0162] GAGAATGGCTTTAAGAGATTAGCTTGACCTCGCGGTCTCGCAACTCGTTGTACCATCCA

[0163] TTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACC

[0164] TTCCTCCGGTTTGTCACCGGCAGTCTTACTAGAGGTGCCCAACTAAATGCTGGCAACTA

[0165] GTCATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGAC

[0166] GACAACCATGCACCACCTGTCATTTTGCCCCCGAAGGGGAAACCTGATCTCTCAGGTG

[0167] ATCAAAAAGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATG

[0168] CTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTTGAGTTTCAACCTTGCGGTCGTACT

[0169] CCCCAGGCGGAATGCTTAATGCGTTAGCTGCGGCACTGAAGGGCGGAAACCCCTCCAA

[0170] CACCTAGCATTCATCGTTTACGGCATGGACTACCAGGGTATCTAATCCTGTTCGCTACCC

[0171] ATGCTTTCGAGCTCAGCGTCAGTTACAGACCAGACAGCCGCCTTCGCCACTGGTGTT

[0172] CTTCCATATATCTACGCATTTCACCGCTACACATGGAGTTCCACTGTCCTCTTCTGCACT

[0173] CAAGTTTCCCAGTTTCCGATGCGCTTCCTCGGTTAAGCCGAGGGCTTTCACATCAGACT

[0174] TAAAAAACCGCCTGCGCTCGCTTTACGCCCAATAAATCCGGATAACGCTTGCCACCTAC

[0175] GTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTGGATACCGTCACG

[0176] CCGACAACAGTTACTCTGCCGACCATTCTTCTCCAACAACAGAGTTTTACGACCCGAA

[0177] AGCCTTCTTCACTCACGGCGGCGTTGCTCCATCAGACTTGCGTCCATTGTGGAAGATTCC

[0178] CTACTGCTGCCTCCCGTAGGAGTTTGGGCCGTGTCTCAGTCCCAATGTGGCCGATCAA

[0179] CCTCTCAGTTCGGCTACGTATCATCGCCTTGGTGAGCCATTACCTCACCAACTAGCTAAT

[0180] ACGCCGCGGGTCCATCCAAAAGCGATAGCTTACGCCATCTTTCAGCCAAGAACCATGC

[0181] GGTTCTTGGATCTATGCGGTATTAGCATCTGTTTCCAAATGTTATCCCCACTTAAGGGC

[0182] AGGTTACCCACGTGTTACTCACCCGTCCGCCACTCGTTCCATGTTGAATCTCGGTGCAA

[0183] GCACCGATCATCAACAAGAACTCGTTCGACTTGCA

[0184] Example 8: Lactobacillus paracasei HXXJ-008 mouse experiment

[0185] 1. Modeling and feeding of mice with hyperuricemia

[0186] Eighteen male mice, 4 weeks old and weighing 16-20g, were purchased and allowed to acclimatize for one week before the experiment. They were housed under standard laboratory conditions at 22℃±1℃ and 60% relative humidity, with a 12-hour light / 12-hour dark cycle (7:00-19:00 light). The mice were fed standard commercial mouse food, with a probiotic (Lactobacillus paracasei HXXJ-008) dosage of 1×10⁻⁶. 9 CFU / each. Divided into the following groups on average:

[0187] (1) Control group: fed basic feed and gavaged with 0.5 mL of sterile skim milk daily.

[0188] (2) Model group: fed a basal diet, and gavage with 300 mg / kg / d potassium oxonate (OP) and 10 g / kg / d yeast extract in sterile skim milk.

[0189] (3) HUA+HXXJ-008: Feed a basal diet containing 300 mg / kg / d potassium oxonate (OP) and 10 g / kg yeast extract powder, and also contains probiotic HXXJ-008 with a live count of 1×10⁻⁶. 9 The probiotics were dissolved in an equal volume and concentration of sterile skim milk and administered via gavage (CFU).

[0190] 2. Detection of inflammatory factor markers

[0191] On day 21 of feeding, liver and kidney samples were collected from mice. The samples were rinsed with physiological saline, diluted with 1 mL of PBS in pre-weighed test tubes, and stored at -20°C for use. Fresh liver and kidney samples were ground on ice using a hand grinder and then centrifuged at low speed (8000 rpm, 10 min) to obtain sample suspensions. The levels of interleukin-1β (IL-1β) and IL-6 in the liver and kidneys were measured according to the instructions provided by Beijing Bioscience.

[0192] Test results are shown Figure 3-4 It can be seen that the Lactobacillus paracasei HXXJ-008 treatment group has a good inhibitory effect on the inflammatory factors interleukin-1β (IL-1β) and IL-6 in the liver and kidney of hyperuricemic mice. Figure 3 It was found that *Lactobacillus paracasei* HXXJ-008 inhibited the inflammatory factor IL-1β in the liver and kidney of hyperuricemic mice by 42.82% and 64.09%, respectively. Figure 4 It can be seen that Lactobacillus paracasei HXXJ-008 inhibited the inflammatory factor IL-6 in the liver and kidney of hyperuricemic mice by 44.67% and 58.99%, respectively, and had a good inhibitory effect on inflammatory factors.

[0193] Example 9: Mouse Experiment with Compound Strains

[0194] 1. Modeling and feeding of mice with hyperuricemia

[0195] Sixty male mice, 4 weeks old and weighing 16-20g, were purchased and allowed to acclimatize for one week before the experiment. They were housed under standard laboratory conditions at 22℃±1℃ and 60% relative humidity, with a 12-hour light / 12-hour dark cycle (7:00-19:00 light). The mice were fed standard commercial mouse food and divided into the following groups:

[0196] (1) Control group: fed basic feed and gavaged with 0.5 mL of sterile skim milk daily.

[0197] (2) Model group: fed a basal diet, and gavage with 300 mg / kg / d potassium oxonate (OP) and 10 g / kg / d yeast extract in sterile skim milk.

[0198] (3) HUA+ treatment group 1: fed a basal diet containing 300 mg / kg / d potassium oxonate (OP) and 10 g / kg yeast extract powder, and also containing 1×10 live probiotics HXJS3-1. 9 The probiotics were dissolved in an equal volume and concentration of sterile skim milk and administered via gavage (CFU).

[0199] (4) HUA+ treatment group 2: fed a basal diet containing 300 mg / kg / d potassium oxonate (OP) and 10 g / kg yeast extract powder, and also containing 1×10 live probiotics HXJS4-1. 9 An equal volume and concentration of CFU of sterile skim milk was administered via gavage.

[0200] (5) HUA+ treatment group 3: fed a basal diet containing 300 mg / kg / d potassium oxonate (OP) and 10 g / kg yeast extract powder, and also containing probiotic HXXJ-008 with a live count of 1×10⁻⁶. 9 An equal volume and concentration of CFU of sterile skim milk was administered via gavage.

[0201] (6) HUA+ treatment group 4: fed a basal diet containing 300 mg / kg / d potassium oxonate (OP) and 10 g / kg yeast extract, along with equal amounts of probiotics HXJS3-1, HXJS4-1, and HXXJ-008, with a total live bacteria count of 1×10⁻⁶. 7 An equal volume and concentration of CFU of sterile skim milk was administered via gavage.

[0202] (7) HUA+ treatment group 5: fed a basal diet containing 300 mg / kg / d potassium oxonate (OP) and 10 g / kg yeast extract, along with equal amounts of probiotics HXJS3-1, HXJS4-1, and HXXJ-008, with a total live bacteria count of 1×10⁻⁶. 8 An equal volume and concentration of CFU of sterile skim milk was administered via gavage.

[0203] (8) HUA+ treatment group 6: fed a basal diet containing 300 mg / kg / d potassium oxonate (OP) and 10 g / kg yeast extract, along with equal amounts of probiotics HXJS3-1, HXJS4-1, and HXXJ-008, with a total live bacteria count of 1×10⁻⁶. 9An equal volume and concentration of CFU of sterile skim milk was administered via gavage.

[0204] (9) HUA+ treatment group 7: fed a basal diet containing 300 mg / kg / d potassium oxonate (OP) and 10 g / kg yeast extract, along with equal amounts of probiotics HXJS3-1, HXJS4-1, and HXXJ-008, with a total live bacteria count of 1×10⁻⁶. 11 An equal volume and concentration of CFU of sterile skim milk was administered via gavage.

[0205] (10) HUA+ positive drug group: The basal diet containing 300 mg / kg / d potassium oxychloride (OP) and 10 g / kg / d yeast extract powder, and an equal volume of sterile skim milk containing 5 mg / kg / d allopurinol was administered by gavage.

[0206] Feed continuously for 21 days according to the above standards.

[0207] 2. Detection indicators and results

[0208] (1) Uric acid content detection

[0209] A urine

[0210] During the 21-day feeding period, mice from each group were moved to clean, empty cages every three days for 1 hour to collect urine samples. The fresh urine was diluted 10 times with distilled water and placed in a 60°C water bath to ensure no urate crystals were deposited in the urine samples. The uric acid content of all samples was then tested according to the instructions provided with the UA test kit (Nanjing Jiancheng, China). Testing was performed immediately after sample collection.

[0211] The effects of each treatment group on the uric acid content in the urine of hyperuricemic mice are shown in the figure. Figure 7 It is evident that gavage administration of probiotics to mice significantly reduces uric acid levels in the urine of hyperuricemic mice. The HUA+ treatment groups 6 and 7 showed the most significant effects, comparable to the positive control group (allopurinol). The effect of HUA+ treatment group 4 was comparable to that of single-strain administration, while the effect of HUA+ treatment group 5 was significantly better than that of single-strain administration. This indicates that the uric acid-lowering effect is enhanced with increasing gavage count of probiotics when using a combination of three probiotics. A gavage count greater than 1×10⁻⁶ probiotics is considered optimal. 8 CFU / each can significantly reduce uric acid in urine.

[0212] B blood

[0213] Blood samples were collected from each group of mice on days 0, 7, 14, and 21 and allowed to stand for 30 minutes. The collected blood samples were then centrifuged at 4000 rpm for 20 minutes to obtain serum. The uric acid content was then measured according to the instructions provided by the UA test kit (Nanjing Jiancheng, China).

[0214] The effects of each treatment group on serum uric acid levels in hyperuricemic mice are shown in the table below. Figure 6 It was observed that all probiotic treatment groups effectively inhibited the uric acid content in the serum of hyperuricemic mice. The HUA+ treatment groups 6 and 7 showed the highest inhibition rates, with serum uric acid inhibition rates of 56.81% and 59.85%, respectively, comparable to the positive control group. The uric acid-lowering effect was enhanced with increasing gavage count of live bacteria. The combined use of three live bacteria resulted in a gavage count greater than 1×10⁻⁶. 8 CFU / each can significantly reduce uric acid in urine.

[0215] (2) XOD activity index detection

[0216] On day 21 of feeding, liver and blood samples were collected from mice. The liver samples were then rinsed with physiological saline, diluted with 1 mL of PBS in pre-weighed test tubes, and stored at -20°C for use. Fresh liver samples were ground on ice using a hand grinder and then centrifuged at low speed (8000 rpm, 10 min) to obtain a liver sample suspension. XOD was detected in the liver samples (according to the instructions provided with the Nanjing Jiancheng XOD Detection Kit). The collected blood samples were centrifuged at 4000 rpm for 20 min to obtain serum, and XOD activity in the serum was detected according to the instructions provided with the Nanjing Jiancheng XOD Detection Kit.

[0217] The inhibition of XOD activity in the liver and serum of hyperuricemic mice by each treatment group is shown in the figure. Figure 5 It was observed that all probiotic treatment groups had a certain inhibitory effect on XOD activity in the liver and serum of hyperuricemic mice. The HUA+ treatment groups 6 and 7 showed the highest inhibition rates, with HUA+ treatment group 6 achieving inhibition rates of 85.71% and 78.21%, respectively, and HUA+ treatment group 7 achieving inhibition rates of 86.93% and 78.69%, respectively. These results were comparable to the positive control group, effectively inhibiting the production of uric acid from purines. The uric acid-lowering effect was enhanced with increasing gavage count of live bacteria. The combined use of three live bacteria resulted in a gavage count greater than 1×10⁻⁶. 8 CFU / each can significantly and effectively inhibit the production of uric acid from purines.

[0218] (3) Detection of inflammatory factor markers

[0219] On day 21 of feeding, liver and kidney samples were collected from mice. The samples were rinsed with physiological saline, diluted with 1 mL of PBS in pre-weighed test tubes, and stored at -20°C for use. Fresh liver and kidney samples were ground on ice using a hand grinder and then centrifuged at low speed (8000 rpm, 10 min) to obtain sample suspensions. The levels of interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α), and IL-11 in the liver and kidneys were measured according to the instructions provided by Beijing Biosciences.

[0220] Test results are shown Figure 8-11 It was found that each probiotic treatment group had a good inhibitory effect on the inflammatory factors interleukin-1β (IL-1β), IL-6, tumor necrosis factor TNF-α, and IL-11 in the liver and kidneys of hyperuricemic mice. The uric acid-lowering effect was enhanced with the increase of the number of live bacteria administered by gavage. The number of live bacteria administered by gavage with the combination of three live bacteria was greater than 1×10⁻⁶. 8 CFU / unit can significantly inhibit inflammatory factors interleukin-1β (IL-1β), IL-6, tumor necrosis factor TNF-α, and IL-11. According to Figure 8 It was found that HUA+ treatment groups 6 and 7 showed the best inhibitory effect on the inflammatory factor IL-1β in the liver and kidneys of hyperuricemic mice. The inhibition rates of HUA+ treatment group 6 were 76.61% and 80.83%, respectively, while those of HUA+ treatment group 7 were 77.5% and 81.95%, respectively, comparable to the positive control group. Figure 9 It was found that HUA+ treatment groups 6 and 7 showed the best inhibitory effect on the inflammatory factor TNF-α in the liver and kidneys of hyperuricemic mice. The inhibition rates of HUA+ treatment group 6 were 76.92% and 77.43%, respectively, while those of HUA+ treatment group 7 were 77.51% and 79.45%, respectively, second only to the positive control group. Figure 10 It was found that HUA+ treatment groups 6 and 7 showed the best inhibitory effect on the inflammatory factor IL-6 in the liver of hyperuricemic mice. The inhibition rates of HUA+ treatment group 6 were 83.1% and 71.35%, respectively, while those of HUA+ treatment group 7 were 84.7% and 71.78%, respectively, second only to the positive control group. Figure 11 It can be seen that HUA+ treatment group 6 and HUA+ treatment group 7 have the best inhibitory effect on the inflammatory factor IL-11 in the liver and kidney of hyperuricemic mice. The inhibition rates of HUA+ treatment group 6 are 76.96% and 81.67%, respectively, and the inhibition rates of HUA+ treatment group 7 are 77.77% and 83.19%, respectively, which are second only to the positive drug group.

[0221] (4) Detection of anti-inflammatory factor IL-10

[0222] The anti-inflammatory factor IL-10 in the liver and kidneys of hyperuricemic mice was detected using a method for detecting inflammatory factor indicators.

[0223] Test results are shown Figure 12 It can be seen that HUA+ treatment groups 1-7 all showed good recovery effects on the anti-inflammatory factor IL-10 in the liver and kidneys of hyperuricemic mice. Among them, HUA+ treatment groups 5, 6 and 7 showed more significant recovery effects on the anti-inflammatory factor IL-10 in the liver and kidneys of hyperuricemic mice. The inhibition rates of HUA+ treatment group 6 were 66.7% and 71.56%, respectively, and the inhibition rates of HUA+ treatment group 7 were 70.55% and 73.1%, respectively, which were better than the positive drug group.

[0224] In summary, strains HXJS3-1, HXJS4-1, and HXXJ-008 significantly reduced serum uric acid, urinary uric acid, XOD activity in the liver and serum, and the levels of inflammatory factors (IL-1β, IL-6, TNF-α, and IL-11) in the liver and kidneys of hyperuricemic mice, while increasing the level of the anti-inflammatory factor IL-10. However, the combined use of these three strains was more effective. When the combined use of these three strains resulted in a gavage count of 1×10⁻⁶ live bacteria, the effect was significantly enhanced. 7 When the CFU / vial count is equal to that of a single bacterium, the effect is comparable. When the number of live bacteria administered via gavage is greater than 1×10⁻⁶. 8 When CFU / unit is used, it can better prevent and treat hyperuricemia, and its efficacy is similar to or comparable to that of the positive control drug allopurinol.

[0225] Example 10 Application Example

[0226] (1) Preparation of probiotic yogurt containing Lactobacillus fermentum HXJS4-1

[0227] Lactobacillus fermentum HXJS4-1 was activated in MRS culture medium, and OD was measured. 600 When the concentration is 0.8-1.2, take 1 mL of bacterial solution and centrifuge at 8000 rpm for 5 min in a low-temperature centrifuge. Wash it three times with sterile physiological saline. Add the cleaned bacterial strain to 100 mL of sterile milk and ferment at 42℃ for 8-10 h. Then inoculate Streptococcus thermophilus and Lactobacillus bulgaricus for secondary fermentation to form probiotic yogurt.

[0228] (2) Preparation of soy milk containing Lactobacillus fermentum HXJS4-1

[0229] Select high-quality soybeans, wash them three times with clean water, soak them until the soybean skin can be removed, add boiling water and grind them, then keep them warm at 80°C or above for 10 minutes, filter and centrifuge to obtain crude soy milk, add Lactobacillus fermentum HXJS4-1 when the temperature drops to 37°C, the amount of Lactobacillus fermentum HXJS4-1 added is the same as the amount added in Example 10 (1), and store it at 4°C to obtain soy milk containing Lactobacillus fermentum HXJS4-1.

[0230] (3) Preparation of compressed candies containing Lactobacillus fermentum HXJS4-1

[0231] Based on the weight proportions, 50 parts sorbitol, 15 parts microcrystalline cellulose, 10 parts Lactobacillus fermentum HXJS4-1, and 5 parts magnesium stearate are mixed and then granulated and compressed into tablets using a dry mixing method. The tablets are 0.5g / tablets with a live bacteria count of 10 billion CFU.

[0232] (4) Preparation of solid beverages containing Lactobacillus fermentum HXJS4-1

[0233] The solid beverage contains, by weight, 50 parts glucose, 15 parts maltodextrin, 10 parts Lactobacillus fermentum HXJS4-1, 10 parts soybean peptide powder, 10 parts resistant dextrin, and 8 parts isomaltooligosaccharide, which are thoroughly mixed and packaged.

[0234] (5) Preparation of solid beverages containing Lactobacillus fermentum HXJS4-1, Lactobacillus plantarum HXJS3-1, and Lactobacillus paracasei HXXJ-008

[0235] The solid beverage comprises, by weight, 50 parts glucose, 15 parts maltodextrin, 10 parts Lactobacillus fermentum HXJS4-1, 10 parts Lactobacillus plantarum HXJS3-1, 10 parts Lactobacillus paracasei HXXJ-008, 10 parts soybean peptide powder, 10 parts resistant dextrin, and 8 parts isomaltooligosaccharide, which are thoroughly mixed and packaged.

[0236] The above description is merely an embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principle of this application should be included within the scope of the claims of this application.

Claims

1. A strain of fermenting lactobacillus ( Lactobacillus fermentum HXJS4-1, characterized in that, It is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC NO: 29677.

2. A strain of Lactobacillus paracasei ( Lactobacillus paracasei HXXJ-008, characterized in that, It is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC NO: 30483.

3. A formulation, characterized in that, The formulation comprises one or more of *Lactobacillus fermentum* and / or *Lactobacillus paracasei*, wherein the *Lactobacillus fermentum* is the *Lactobacillus fermentum* as described in claim 1. Lactobacillus fermentum HXJS4-1; the Lactobacillus paracasei is the Lactobacillus paracasei described in claim 2 ( Lactobacillus paracasei HXXJ-008.

4. The formulation according to claim 3, wherein the formulation comprises *Lactobacillus plantarum*, and the *Lactobacillus plantarum* is *Lactobacillus plantarum* (…). Lactobacillus plantarum HXJS3-1 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO: 29676.

5. The formulation according to claim 3 or 4, characterized in that, The total number of viable counts of the Lactobacillus fermentum, Lactobacillus paracasei and / or Lactobacillus plantarum is greater than 1 x 10 8 CFU.

6. The use of the Lactobacillus fermentum as described in claim 1, or the Lactobacillus paracasei as described in claim 2, or the formulation as described in any one of claims 3-5, in the preparation of nucleoside-degrading products.

7. The use of the Lactobacillus fermentum as described in claim 1, or the Lactobacillus paracasei as described in claim 2, or the formulation as described in any one of claims 3-5, in the preparation of a product that lowers uric acid levels.

8. The use of the Lactobacillus fermentum as described in claim 1, or the Lactobacillus paracasei as described in claim 2, or the formulation as described in any one of claims 3-5, in the preparation of products for the prevention and / or treatment of hyperuricemia and / or gout.

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