Application of parabacteroides dielsii in preparation of product for treating hyperuricemia

Abstract

The invention discloses an application of parabacteroides dieldrii in preparation of a product for treating hyperuricemia. The activity of xanthine oxidase is inhibited through the parabacteroides dieldrii, uric acid related transporters are changed, and damaged organs are protected to play a uric acid reducing effect, so that chronic hyperuricemia is improved. In vitro, the parabacteroides dieldrii can inhibit the activity of xanthine oxidase. On the animal level, the parabacteroides dielsii can relieve liver and kidney injuries caused by hyperuricemia and can repair the injured intestinal barrier at the same time. Therefore, a design thought can be provided for an intestinal flora uric acid reducing preparation, so that a new intestinal flora target is provided for clinical treatment of hyperuricemia. According to the present invention, the activity of xanthine oxidase is inhibited in vitro, the uric acid transporter URAT1 in the kidney is reduced in vivo, the uric acid excretion transporter ABCG2 is increased, and the liver and kidney injury and the intestinal barrier damage caused by hyperuricemia are relieved so as to reduce the serum uric acid level and improve the hyperuricemia;

Description

TECHNICAL FIELD

[0001] The application relates to the field of biological medicine, and particularly relates to application of Parabacteroides distasonii in preparation of products for improving and treating hyperuricemia. BACKGROUND

[0002] Hyperuricemia (HUA) is a metabolic disease caused by excessive generation and / or excretion of uric acid (UA) due to abnormal purine metabolism, which leads to the blood uric acid exceeding the normal range. HUA not only causes gout, leading to joint deformity and dysfunction, but is also closely related to a variety of diseases. It has been confirmed that HUA is an independent risk factor for causing high blood pressure, high blood lipids, type II diabetes, kidney disease and cardiovascular events, and seriously threatens human health. At present, the drugs for treating HUA in the clinic are mainly divided into three categories: inhibiting the generation of uric acid, promoting the excretion of uric acid and uricase. Although the existing drugs have good effects, most of them take the liver and kidney as the action target, have larger adverse reactions, are difficult to balance the efficacy and toxicity, cannot be taken for a long time, and the organ damage problem is also an important factor affecting the research and development of new drugs. Therefore, it is particularly urgent to find a safe uric acid-lowering treatment strategy and target. The intestine is the largest site of UA excretion in the body except the kidney, and 1 / 3 of the UA is excreted through the intestine. The intestinal flora is currently considered a new and complex organ. Studies have shown that various chronic diseases in humans may be related to intestinal microecological disorders. The intestinal flora not only affects the metabolism of the host, but also plays an important role in maintaining the homeostasis of the intestinal environment, and greatly affects human health. The intestinal flora is also closely related to HUA, and to some extent, affects the metabolism and synthesis of uric acid.

[0003] Existing studies have shown that the intestinal flora of HUA and gout patients has phenomena such as reduced diversity, structural disorder, and changes in metabolic pathways and metabolic products, and other flora disorders. Anaerobic bacteria in the intestine can consume UA, and intestinal bacteria can partially compensate for the functional deficiency of HUA mice lacking uric acid oxidase, maintaining relatively stable UA levels. The destruction of intestinal flora increases the risk of gout, indicating the importance of intestinal flora in UA metabolism, which shows that regulating the intestinal flora is expected to become a new target for the treatment of HUA.

[0004] Parabacteroides distasonis (P. distasonis) is a gram-negative obligate anaerobic bacteria of the genus Parabacteroides, which is normal colonized in human intestinal tract, respiratory tract and oral cavity. Parabacteroides is a relatively new genus, and in recent years, P. distasonis has been considered as a new probiotic against metabolic syndrome. In recent years, the bacteria has shown good application prospect in metabolic diseases such as obesity and diabetes, which are closely related to HUA, and P. distasonis has not been associated with simple HUA. If it can be applied to improve simple hyperuricemia, it can obviously play a huge social benefit and produce significant economic value. SUMMARY

[0005] To solve the above technical problems, the present application is proposed.

[0006] The first aspect is the application of P. distasonis in the preparation of products for treating hyperuricemia. By inhibiting the activity of xanthine oxidase in vitro and reducing the kidney uric acid transporter URAT1 and increasing the uric acid excretion transporter ABCG2 in vivo, the serum uric acid level is reduced, and the hyperuricemia is treated, and the liver and kidney damage and intestinal barrier damage caused by high uric acid are relieved.

[0007] The second aspect is the application of the P. distasonis in the preparation of products for preventing and treating hyperuricemia, including but not limited to the application of the P. distasonis in the preparation of drugs for preventing and treating hyperuricemia, and the application in the preparation of health products or food for preventing and treating hyperuricemia.

[0008] The third aspect is the application of the P. distasonis in the preparation of products for inhibiting xanthine oxidase, including but not limited to the application of the P. distasonis in the preparation of drugs for inhibiting xanthine oxidase, and the application in the preparation of health products or food for inhibiting xanthine oxidase.

[0009] The P. distasonis is P. distasonis with the preservation number of GDMCC NO: 1.1564, and the sequence obtained by 16S bidirectional sequencing is shown in sequence 1.

[0010] The present application has the following advantages: by inhibiting xanthine oxidase activity, changing uric acid related transporters and protecting damaged organs, Parabacteroides distasonii can play a role in reducing uric acid and improve chronic hyperuricemia. In vitro, Parabacteroides distasonii can inhibit xanthine oxidase activity. At the animal level, Parabacteroides distasonii can relieve liver and kidney damage caused by hyperuricemia, and at the same time can repair the damaged intestinal barrier. Parabacteroides distasonii can protect organs such as liver and kidney while reducing uric acid. In summary, Parabacteroides distasonii can reduce uric acid levels and improve hyperuricemia through multiple targets and channels. This can provide design ideas for gut flora preparations for reducing uric acid, and provide new gut flora targets for clinical treatment of hyperuricemia. BRIEF DESCRIPTION OF DRAWINGS

[0011] Figure 1 The figure is the colony morphology of Parabacteroides distasonii. The colony morphology is silver-gray round, opaque, smooth surface and neat edge;

[0012] Figure 2 The figure is the result of Parabacteroides distasonii on chronic hyperuricemia mice. The mice were given high purine diet yeast extract by gavage and intraperitoneal injection of uricase inhibitor oxypotassium, and the mice were given drugs every day for 4 weeks to establish a chronic hyperuricemia mouse model. Blood was collected after 4 weeks, and serum was detected. Among them, A is the result of uric acid (UA), B is the result of serum creatinine (cr), C is the result of serum glutathione transaminase (ALT), and D is the result of serum glutathione transaminase (AST). Compared with the normal group, *** is p<0.001, **** is p<0.0001, compared with the model group, ## is p<0.01, #### is p<0.0001. The results show that the serum uric acid, serum creatinine (cr), serum liver enzymes AST and ALT of the mice after using Parabacteroides distasonii are significantly lower than those of the HUA model group, which shows that it can achieve the effect of reducing uric acid, and has a certain protective effect on the liver and kidney;

[0013] Figure 3 The figure is the protection effect of Parabacteroides distasonii on important organs of chronic hyperuricemia mice: from top to bottom are the staining results of kidney and liver, and from left to right are the normal group, the hyperuricemia model group and the Parabacteroides distasonii group. The HE pathological staining of liver and kidney shows that Parabacteroides distasonii can relieve liver and kidney damage caused by hyperuricemia. It is shown that Parabacteroides distasonii not only can play a role in reducing uric acid, but also can improve the pathological changes of organs caused by hyperuricemia;

[0014] Figure 4This image shows the protective effect of *Peribrobacter dysplasia* on the intestines – PAS glycogen staining of the colon. From top to bottom, the images show PAS glycogen staining of the colon in the normal group, the hyperuricemia model group, and the *Peribrobacter dysplasia* group. Compared with the normal group, the model group showed damaged intestinal villi and a reduction in goblet cells, indicating that *Peribrobacter dysplasia* can improve the aforementioned damage.

[0015] Figure 5 The image shows the effect of *Pseudomonas diffusa* on the expression level of ocludin protein in the intestine of hyperuricemic mice. Figure A is a Western blotting (WB) plot of ocludin protein expression levels, with the top row representing ocludin protein and the bottom row representing the internal control protein GAPDH. From left to right, the samples in each row represent the protein content of the normal group, the hyperuricemic model group, and the *Pseudomonas diffusa* group. Figure B is a statistical graph of ocludin protein expression levels from Figure A. Compared with the model group, **p<0.01 indicates that *Pseudomonas diffusa* can improve intestinal barrier damage caused by hyperuricemia.

[0016] Figure 6 This diagram illustrates the effects of *Pseudomonas diffusa* on relevant uric acid transporters. Image A shows the immunohistochemical staining of ABCG2 transporter protein in mouse colon tissue; Image B shows the immunohistochemical staining of URAT1 urate transporter in mouse kidney tissue. From left to right, the groups are: normal group, hyperuricemia model group, and *Pseudomonas diffusa* group. Compared to the normal group, the model group showed decreased intestinal ABCG2 protein and increased expression of transporter URAT1, indicating that *Pseudomonas diffusa* improves the levels of these uric acid transporters.

[0017] Figure 7 The diagram illustrates the in vitro inhibitory effect of *Pseudomonas diffusa* metabolites on xanthine oxidase: from left to right, the inhibition rates of xanthine oxidase in the allopurinol group, intracellular contents of *Pseudomonas diffusa*, and extracellular metabolites are shown. The results are as follows: when the average XOD inhibition rate of allopurinol was 96.19%, the XOD inhibition rate of *Pseudomonas diffusa* extracellular metabolites reached 75.83%, and the XOD inhibition rate of *Pseudomonas diffusa* intracellular contents reached 59.98%, indicating that *Pseudomonas diffusa* has a certain ability to inhibit uric acid production. That is, *Pseudomonas diffusa* has a certain inhibitory effect on xanthine oxidase, the rate-limiting enzyme in uric acid production, in vitro, and can reduce uric acid synthesis. Detailed Implementation

[0018] The following is combined Figures 1-7 The technical solutions of this invention have been clearly and completely described. However, the described embodiments are only a part of the embodiments of this invention, not all of them, and do not imply any limitation on the invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0019] Unless otherwise specified, the reagents, methods and equipment used in this invention are conventional reagents, methods and equipment in this technical field, and the reagents and materials used in the embodiments of this invention are all commercially available or can be prepared.

[0020] The reagents used are as follows: yeast extract; potassium oxonate; xanthine; xanthine oxidase. The *Parabacteroides distasonis* (P. distasonis) used in this embodiment was purchased from the Guangdong Provincial Microbial Culture Collection Center, accession number GDMCC NO: 1.1564. *Parabacteroides distasonis* was sequenced and identified by Beijing Qingke Biotechnology Co., Ltd., and the sequence obtained by 16S bidirectional sequencing is shown in Sequence 1.

[0021] The bacterial strain stored at -80℃ was activated on blood agar plates and incubated at 37℃ for 48 hours. Colony morphology was then observed. Results are as follows: Figure 1 As shown, the colonies are silvery-gray, round, opaque, with a smooth surface and neat edges.

[0022] Five-week-old SPF-grade male C57BL mice weighing 20±2g were randomly divided into three groups. All animals were housed at a temperature controlled between 24℃ and 26℃, with relative humidity controlled between 40% and 60%, and eight mice per cage. Experiments were conducted after one week of acclimatization. The relevant animal experiments were approved by the ethics review committee and obtained the necessary permits.

[0023] Specific implementation process:

[0024] (1) Resuscitation and identification of *Pseudomonas difficile*

[0025] Prepare BHI liquid culture medium at a ratio of 1L ultrapure water to 37g of the culture, sterilize at 121℃ for 30min, and store at 4℃ for later use; pick a small amount of *Pseudomonas difficile* powder into 5mL of the medium. In BHI medium, the culture was thoroughly mixed by pipetting and incubated in an anaerobic incubator at 37°C for 36 hours, after which the medium became turbid. A small amount of bacterial culture was taken and streaked onto a blood agar plate using the four-step streak method. The streaked blood agar plate was then inverted and incubated in an anaerobic incubator at 37°C for 48 hours. After removing the blood agar plate, silvery-gray, round colonies were observed to grow on the solid medium. Three larger, raised colonies were picked and placed in three 2mL liquid media, and then transferred to an anaerobic incubator at 37°C for 48 hours. After observing turbidity in the culture medium, 500μL of bacterial culture from each culture was sent to Beijing Qingke Biotechnology Co., Ltd. for 16S bidirectional sequencing. The obtained sequences were compared and identified with the NCBI database, and it was found that the sequence similarity with *Pseudomonas difficile* ATCC8503 was 99.3% (NCBI reference sequence: NR_074376.1). The remaining 1mL of bacterial culture was inoculated into a new BHI medium and cultured in an anaerobic environment for 36 hours before bacterial counting.

[0026] (2) Counting and collection of *Pseudomonas dilatatus*

[0027] The cultured bacterial stock solution was mixed with sterile culture medium at a ratio of 1×10⁻⁶. 6 and 1×10 8 The bacterial suspension was diluted according to the specified ratio. 200 μL of the diluted suspension was evenly spread onto blood agar plates using a sterile inoculation loop, and each plate was labeled with its corresponding dilution ratio. The plates were then incubated in an anaerobic incubator at 37°C for 48 hours. Simultaneously, 200 μL of the bacterial suspension was inoculated into a 96-well plate, and the OD value was measured at 600 nm. Calculations showed that a bacterial suspension concentration of approximately 2 × 10⁸ corresponds to an OD value of 0.8. 9 cfu / mL; the concentration of the bacterial culture was calculated based on the measured OD value after each day's culture and then used for subsequent experiments.

[0028] (3) Animal experimental protocol

[0029] Chronic hyperuricemia modeling protocol

[0030] 1) Grouping of experimental animals

[0031] Five-week-old SPF-grade male C57 mice weighing 20±2g were selected and divided into three groups according to their average weight, as shown in the table below. All animals were housed at a temperature controlled between 24℃ and 26℃, with relative humidity controlled between 40% and 60%, and eight mice per cage. Experiments were conducted after one week of acclimatization, as shown in Table 1 below. The relevant animal experiments were approved by the ethics review committee and obtained the necessary permits.

[0032] Table 1

[0033]

[0034] The drug preparations and concentrations for each group were as follows: 0.5 wt% sodium carboxymethyl cellulose was used as the drug solvent, along with potassium oxonate (36 mg / mL), yeast extract (1.8 g / mL), and *Pseudomonas difficile* (2 × 10⁻⁶). 8 (cfu / ml). All injections were administered at a volume of 100 μL / 10 g.

[0035] 2) Mice ate and drank normally during the experiment. Except for the control group, mice in each group were intraperitoneally injected with 300 mg / kg potassium oxonate solution and administered 15 mg / kg yeast extract solution (solvent: 0.5 wt% CMC-Na) by gavage at 9:30 am daily. The treatment groups were administered *Pseudomonas difficile* (2 × 10⁻⁶) by gavage half an hour after model establishment. 8(cfu / ml, 10mL / kg / d). The blank control group was given an equal volume of 0.5wt% sodium carboxymethyl cellulose by gavage and injection. The experiment lasted for 28 days. Blood was collected and mice were sacrificed on day 28 after the administration was completed. At the time of sacrifice, samples of kidney, liver, and intestinal tissue (duodenum, jejunum, ileum) were collected. Some samples were soaked in paraformaldehyde fixative, and others were stored at -80℃.

[0036] (4) Sample processing and determination

[0037] Sample processing and blood testing: After letting about 500 μL of blood stand at room temperature for 30 min, centrifuge at 3000 rpm for 15 min at 4 °C. Collect the serum and store it at -20 °C. Use a uric acid test kit, a creatinine test kit, and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) kits (Nanjing Jiancheng Bioengineering Institute) to measure the uric acid (UA), creatinine (cr), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in the serum, respectively.

[0038] Kidneys: After euthanizing the mice, the kidney tissue was quickly separated and the renal membrane was removed. The right kidney was placed in paraformaldehyde fixative for preservation and used for HE staining and immunohistochemical experiments.

[0039] Liver: After euthanizing the mice, the liver tissue was quickly separated, and the left lobe of each mouse was uniformly removed and preserved in paraformaldehyde fixative for HE staining.

[0040] Intestines: After euthanizing the mice, the duodenum, jejunum, and ileum were removed. Part of the intestines were preserved in paraformaldehyde fixative for HE staining, PAS glycogen staining, and immunohistochemical experiments, while the other part was stored at -80℃ for subsequent protein extraction.

[0041] (5) Periodic acid-Schiff (PAS) staining

[0042] Remove the periodic acid solution and equilibrate to room temperature. Add 100 μL of periodic acid solution to each intestinal tissue section and react in a humidified chamber in the dark for 10 minutes. Remove the periodic acid solution and soak the section in distilled water, washing on a shaker for 5 minutes. Then, add 100 μL of Schiff's reagent to each sample, place it in a humidified chamber, and stain in a 37°C oven in the dark for 30 minutes to 1 hour. After removing the staining solution, soak the section in distilled water and wash for 5 minutes. Then, add 100 μL of hematoxylin staining solution to each sample and stain for 30 seconds. Rinse three times with distilled water for 3 seconds each time to remove any excess stain. After drying in a 37°C oven, mount the sections with neutral resin and observe under a microscope.

[0043] (6) Western blot of proteins

[0044] Further investigation was conducted to examine the levels of uric acid transport and intestinal barrier-related proteins in the intestines of various experimental animals after gavage administration of *Pseudomonas difficile*. The main steps of protein immunoblotting are shown below:

[0045] 1) Extraction of total protein from intestinal tissue

[0046] First, remove the intestinal tissue from the -80℃ freezer and thaw it on ice. Place two magnetic beads in a 1.5ml EP tube. Weigh approximately 20mg of intestinal tissue and add 250μL of protein RIPA lysis buffer (add PMSF and phosphatase inhibitor at a 1:100 ratio before use). Place the EP tube in a homogenizer and homogenize for 5 seconds, pause for 5 seconds, and repeat for a total of 5 minutes. After homogenization, let it stand on ice for 10 minutes, then transfer the supernatant to a new 1.5mL centrifuge tube and centrifuge at 12000g for 15 minutes at 4℃. After centrifugation, transfer 200μL of the supernatant to a new 1.5mL centrifuge tube. Add the appropriate loading buffer and place the tube in a constant temperature metal bath at 100℃ for 5 minutes to denature the protein. Store at -80℃.

[0047] 2) Western blot assay for proteins

[0048] Separation was performed using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis at 200V for 40 minutes. Subsequently, proteins were transferred to a PVDF membrane. During wet transfer, a sandwich structure was constructed in the order of sponge-filter paper-gel-membrane-filter paper-sponge, gently rolling to remove air bubbles. The PVDF membrane was soaked in methanol for 1–5 seconds before use. After transfer, the membrane was blocked for 1 hour at room temperature with 5% skim milk prepared in TBST buffer, followed by dilution of the primary antibody with primary antibody dilution buffer and incubation overnight at 4°C. The next day, the primary antibody was discarded, and the membrane was washed 5 times with TBST buffer for 3 minutes each time. Subsequently, the membrane was incubated for 1 hour at room temperature with species-matched secondary antibody (1:5000, Jackson ImmunoResearch Laboratories, USA). The secondary antibody was discarded, and the membrane was washed 5 times with TBST buffer for 3 minutes each time. Protein bands were obtained by chemiluminescence exposure using the FDbio-Dura ECL kit, and grayscale values ​​were calculated using ImageJ. The experiment used GAPDH as an internal control and Occludin (1:5000) as the primary antibody.

[0049] (7) Immunohistochemical experiments

[0050] Kidney paraffin sections obtained from sampling were baked, dewaxed, and hydrated. They were then soaked in Tris-EDTA (TE 9.0) at pH 9.0 and heated in a microwave oven for 90 seconds, followed by antigen retrieval in a 65°C water bath for 1.5 hours. The sections were washed five times with PBS buffer for 3 minutes each time, and then reacted with 3% H2O2 for 10 minutes to inactivate endogenous peroxidase activity. Each tissue site was circled using a histochemical pen, washed with PBS buffer after 10 minutes, and then blocked with goat serum at room temperature for 1 hour. The sections were then incubated overnight at 4°C with primary antibodies URAT1 (1:2000) and ABCG2 (1:2000). The next day, the sections were warmed to room temperature for half an hour, washed with PBS buffer to remove the primary antibody, and then incubated with species-matched secondary antibody (1:200, Jackson Immuno Research Laboratories, USA) at room temperature for 1 hour. Finally, DAB staining was performed, and positive staining was observed under a microscope. Staining was then terminated by placing the sections in PBS buffer. Finally, the slides were stained with hematoxylin for 10 minutes, then blued again in PBS buffer, dehydrated, cleared, and mounted. Images were taken using an upright microscope.

[0051] (8) Determination of inhibition rate of *Pseudomonas difficile*

[0052] 1) Centrifuge the activated bacterial strain at 10,000 rpm for 10 min to collect the bacterial cells. Wash the bacterial cells 2-3 times with sterile PBS, then resuspend them in PBS and adjust the OD of the bacterial solution. 600 The OD value was 0.8, and the cells were incubated at 37℃ for 12 h. After centrifugation at 10000 rpm / min for 10 min, the supernatant was collected and filtered through a 0.22 μm microfiltration membrane to obtain cell metabolites. 600 The bacterial suspension with a concentration of 0.8 μL was disrupted by sonication at 200 W for 10 min under conditions of 5 s on and 5 s off. The disrupted material was centrifuged at 10,000 rpm for 10 min, and the supernatant was collected and filtered through a 0.22 μm microporous membrane to obtain the cell contents.

[0053] 2) According to Table 2, add phosphate buffer (pH 7.5), sample solution, and 0.5 U xanthine oxidase sequentially to a 96-well plate. Incubate at 37°C for 3 min, then add 100 μL of 0.5 mmol / L xanthine substrate. Record the absorbance value at 295 nm and read the absorbance (A) value after 5 min. The experiment included a sample group, an oxidase group, and a blank group. Allopurinol was used as a positive control. The inhibition rate (%) of the sample against xanthine oxidase was calculated using the following formula: Inhibition rate (%) = (1 - (A1 - A2) / (A2 - A3)) * 100, where A1 is the sample / positive experimental group, A2 is the negative control experimental group, and A3 is the negative control blank group.

[0054] Table 2

[0055]

[0056] The present invention has been described in detail above. The above description is only a preferred embodiment of the present invention and should not be construed as limiting the scope of the present invention. All equivalent changes and modifications made in accordance with the scope of this application should still fall within the scope of the present invention.

[0057]

Claims

1. The application of *Pseudomonas difficile* in the preparation of products for treating hyperuricemia, characterized by: By inhibiting xanthine oxidase activity in vitro, reducing the renal uric acid transporter URAT1 and increasing the uric acid excretion transporter ABCG2 in vivo, *Pseudomonas dilatatus* can lower serum uric acid levels and treat hyperuricemia, as well as alleviate liver and kidney damage and intestinal barrier impairment caused by hyperuricemia.

2. The application of *Pseudomonas difficile* according to claim 1 in the preparation of products for treating hyperuricemia, characterized in that: Application of *Pseudomonas dilatatus* in the preparation of products for the prevention and treatment of hyperuricemia.

3. The application of *Pseudomonas difficile* according to claim 1 in the preparation of products for treating hyperuricemia, characterized in that: Application of *Pseudomonas diffusa* in the preparation of products that inhibit xanthine oxidase.

4. The application of *Pseudomonas difficile* according to claim 2 in the preparation of products for treating hyperuricemia, characterized in that: Application of this *Pseudomonas dilatatus* in the preparation of drugs for the prevention and treatment of hyperuricemia.

5. The application of *Pseudomonas difficile* according to claim 2 in the preparation of products for treating hyperuricemia, characterized in that: Application of this *Pseudomonas dilataniae* in the preparation of health products or foods for the prevention and treatment of hyperuricemia.

6. The application of *Pseudomonas difficile* according to claim 3 in the preparation of products for treating hyperuricemia, characterized in that: Application of this *Pseudomonas diffusa* in the preparation of drugs that inhibit xanthine oxidase.

7. The application of *Pseudomonas difficile* according to claim 3 in the preparation of products for treating hyperuricemia, characterized in that: Application of this *Pseudomonas diffusa* in the preparation of health products or foods that inhibit xanthine oxidase.

8. The application of *Pseudomonas difficile* according to claim 1 in the preparation of products for treating hyperuricemia, characterized in that: The sequence obtained by 16S bidirectional sequencing of *Pseudomonas dilatatus* is shown in Sequence 1.

9. The application of *Pseudomonas difficile* according to claim 1 in the preparation of products for treating hyperuricemia, characterized in that: The *Pseudomonas dignitaria* is the strain with accession number GDMCC NO: 1.1564.

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