Probiotic abnormal Vickerhamomyces cerevisiae and application thereof in prevention of HUA

Abstract

The invention discloses a novel application of Wickerhamomyces anomalus YFJ252 (the preservation number is CGMCC (China General Microbiological Culture Collection Center) NO.28267) which is disclosed and preserved, in particular relates to an application of the strain and a metabolite of the strain in preparation of medicines, foods or health care products for preventing or treating hyperuricemia (HUA), and belongs to the technical field of microorganisms. The strain has excellent probiotic characteristics (acid and cholate resistance, high hydrophobicity, self-aggregation and co-aggregation capability, oxidation resistance, glycometabolism regulation, anti-inflammatory capability and the like) and a function of inhibiting the activity of xanthine oxidase (XO) in vitro. The serum uric acid, creatinine and urea nitrogen levels of a hyperuricemia mouse model induced by high fructose diet and oteracil potassium can be remarkably controlled, and a new way is provided for safely and effectively preventing HUA.

Description

Technical Field

[0001] The present invention belongs to the field of microbial technology and specifically relates to a strain of abnormal Wickham yeast ( Wickerhamomyces anomalus ) and its application in preventing hyperuricemia (HUA). Background Art

[0002] In recent years, the prevalence of hyperuricemia (HUA) has increased significantly due to changes in lifestyle and dietary habits, becoming a serious health concern for modern people. Hyperuricemia is a metabolic disease caused by abnormal purine metabolism or uric acid excretion, which can lead to gout, hypertension, atherosclerosis, and various cardiovascular diseases. Current treatments for HUA include allopurinol and benzbromarone, which can alleviate HUA by reducing uric acid (UA) synthesis or increasing UA excretion. However, medication use is often accompanied by side effects such as liver and kidney damage and toxic epidermal necrosis. Therefore, the development of safe and effective methods to alleviate hyperuricemia is urgently needed.

[0003] Xanthine oxidase (XO) is a key enzyme in uric acid synthesis. Under the action of XO, adenine and guanine are converted into hypoxanthine and xanthine, ultimately producing UA. Therefore, inhibiting XO activity and thereby reducing UA production is considered an effective approach to improving HUA. Currently, a number of natural product-based treatments have been developed to improve HUA by inhibiting XO activity, including Chinese herbal extracts, food-derived anti-uricosuric peptides, probiotics, and their metabolites. Probiotics are living microorganisms that are beneficial to the host and play an important role in promoting human health, particularly in regulating metabolic disorders.

[0004] Currently, the selection of probiotic strains for the treatment of HUA mainly focuses on lactic acid bacteria, and there are few reports on whether yeast has the function of alleviating or preventing HUA. Summary of the Invention

[0005] In view of the problems existing in the prior art, the present invention aims to provide a probiotic abnormal Wickham yeast and its application in preventing HUA. Wickerhamomyces anomalus , W.anomalus) were analyzed for their probiotic properties, their metabolite types were determined based on non-targeted metabolomics, and their uric acid-lowering ability in vitro and in a hyperuricemia mouse model was evaluated. The results showed that this strain not only has good acid and bile salt tolerance, hydrophobicity, self-aggregation, co-aggregation, antioxidant and hypoglycemic abilities, but can also significantly reduce uric acid levels, providing a new approach for the development of side-effect-free means of preventing HUA.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions.

[0007] The present invention discloses application of abnormal Wickham yeast YFJ252 (preservation number: CGMCC NO.28267) in preparing a product for preventing or treating hyperuricemia.

[0008] Furthermore, the product is a medicine, food or health product.

[0009] Furthermore, according to the above application, it is characterized in that the active ingredient of the product is the bacteria of abnormal Wickham yeast YFJ252 or its metabolites.

[0010] Furthermore, it is characterized in that the metabolites include substances that inhibit the activity of xanthine oxidase (XO).

[0011] The present invention also discloses a product for preventing or treating hyperuricemia, characterized in that it contains the bacteria of abnormal Wickham yeast YFJ252 or its metabolites as an active ingredient, and the strain preservation number is CGMCC NO.28267.

[0012] Furthermore, the product form includes oral preparations or probiotics.

[0013] The present invention also discloses the use of abnormal Wickham yeast YFJ252 (preservation number: CGMCC NO.28267) in preparing a preparation for inhibiting the activity of xanthine oxidase (XO).

[0014] Furthermore, the preparation can reduce serum uric acid, creatinine and urea nitrogen levels.

[0015] Compared with the prior art, the present invention has the following beneficial effects.

[0016] The present invention screened a yeast strain, Wickham's yeast YFJ252, with a high β-glucosidase production capacity from a plant-derived (fermented kimchi) sample. The strain was then analyzed for its safety (hemolytic activity, DNase production, and antibiotic sensitivity) and probiotic properties (acid and bile salt resistance, hydrophobicity, self-aggregation, co-aggregation, and antioxidant capacity). The results showed that the strain was non-hemolytic and did not produce DNase; it had good tolerance to acid and bile salts, and YFJ252 had hydrophobicity rates of 74.71%, 83.19%, and 37.21% for xylene, n-hexane, and ethyl acetate, respectively. It also exhibited good self-aggregation and co-aggregation abilities. It also showed strong resistance to DPPH free radicals and ABTS. + The free radical and hydroxyl radical scavenging rates reached 85.53%, 56.52%, and 63.73%, respectively. Metabolites in the fermentation broth of YFJ252 included 24 categories of metabolites, including organic heterocyclic compounds, organic acids and their derivatives, benzene ring compounds, alkaloids, lipids and lipid molecules, amino acids and peptides, fatty acids, shikimic acid and phenylpropanoids, oxygenated organic compounds, phenylpropanoids, and polyketides. These compounds are closely related to pathways such as lysine biosynthesis; alanine, aspartate, and glutamate metabolism; glyoxylate and dicarboxylic acid metabolism; arginine and proline metabolism; and pantothenic acid and coenzyme A biosynthesis. Furthermore, the strain exhibited inhibitory activities against α-amylase and α-glucosidase by 39.94% and 35.32%, respectively. Its ability to inhibit protein denaturation (inhibiting inflammation) reached 20.73%, and its inhibition rate against XO was 84.15%. A hyperuricemia mouse model was established by feeding mice a high-sugar diet (10 g fructose per 100 g feed) and continuously gavaging potassium oxonate (250 mg / kg) for 21 days. It was found that after the end of the 21-day experiment, the serum uric acid concentration (165.08 μmol / L) of mice in the potassium oxonate and high-sugar diet-induced model and YFJ252 group (HUA+YFJ252) was significantly lower than that of the model group (212.19 μmol / L) ( P <0.05). Comparison of creatinine and urea nitrogen revealed that serum creatinine and urea nitrogen concentrations in the model group were 27.84 μmol / L and 10.61 mmol / L, respectively. Simultaneous administration of YFJ252 during modeling reduced these concentrations to only 20.54 μmol / L and 9.88 mmol / L in the HUA + YFJ252 group. Therefore, we conclude that the abnormal Wickham yeast YFJ252 has the potential to prevent HUA. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 1 is a graph showing the results of the safety test of the strain of the present invention, wherein (a) hemolytic ability; (b) DNase production ability.

[0018] Figure 2 These are the results of the probiotic properties test of the strain of the present invention, including: (a) acid tolerance; (b) bile salt tolerance; (c) hydrophobicity; (d) self-aggregation and co-aggregation; and (e) antioxidant capacity.

[0019] Figure 3 These are the results of non-targeted metabolite analysis of the strain metabolites of the present invention, including: (a) classification and percentage circle diagram of YFJ252 metabolites; (b) PCA analysis diagram of YFJ252 metabolites; (c) KEGG pathway classification diagram of YFJ252 metabolites; and (d) bubble diagram of key metabolic pathways related to YFJ252 metabolites.

[0020] Figure 4 It is a graph showing the ability of the strain of the present invention to regulate sugar metabolism and inhibit protein denaturation.

[0021] Figure 5 The strain of the present invention (a) inhibits xanthine oxidase in vitro; (b) affects uric acid levels in mice; (c) affects serum creatinine levels in mice. * indicates P <0.05; *** indicates P <0.001; **** indicates P <0.0001. DETAILED DESCRIPTION

[0022] The present invention is further described in detail below with reference to specific embodiments. However, this should not be construed as limiting the scope of the present invention to the following embodiments, as all technologies implemented based on the present invention fall within the scope of the present invention.

[0023] Unless otherwise specified, all reagents and materials used in the present invention are commercially available.

[0024] Preparation of strain Abnormal Wickham yeast according to the disclosure of patent CN117625415A Wickerhamomyces anomalus YFJ252, and the following experiments were performed.

[0025] 1. Abnormal strain Wickham's yeast Wickerhamomyces anomalus Safety capabilities of YFJ252.

[0026] 1. Hemolytic.

[0027] The strain was streaked onto Columbia agar plates containing 5% sheep blood. After incubation at 28°C for 48 hours, the hemolysis on the plates was observed, with Staphylococcus aureus used as a positive control. The criteria for judgment were: the presence of a green area around the colony indicated that the strain exhibited α-hemolysis; the presence of a transparent area indicated that the strain exhibited β-hemolysis; and the absence of significant changes indicated that the strain exhibited γ-hemolysis or no hemolysis. The results showed that ( Figure 1 a) This strain is non-hemolytic.

[0028] 2. Produces DNase enzyme activity.

[0029] The strain's DNase production capacity was assessed by streaking YFJ252 onto DNase agar plates. After inoculation, the plates were incubated at 28°C for 48 hours. The presence of a clear zone on the plate indicated DNase production; the absence of a clear zone indicated a lack of DNase production. The results showed that ( Figure 1 b) There is no transparent zone around the bacterial colony, indicating that the strain does not produce DNase enzyme.

[0030] 3. Antibiotic sensitivity.

[0031] The antibiotic susceptibility of the strains was assessed using the disk diffusion method. Twelve antibiotics were selected: penicillin (10 μg), tetracycline (30 μg), ampicillin (10 μg), norfloxacin (10 μg), kanamycin (30 μg), clarithromycin (15 μg), gentamicin (10 μg), streptomycin (10 μg), erythromycin (15 μg), chloramphenicol (30 μg), clindamycin (2 μg), and vancomycin (30 μg). After incubation at 28°C for 24–48 h, the diameter of the inhibition zone was measured to assess the antibiotic susceptibility of the strains.

[0032] Table 1 shows the antibiotic susceptibility results. YFJ252 is insensitive, or resistant, to the listed antibiotics. The presence of antibiotics does not affect the growth of the strain, indicating that concurrent administration of this yeast during antibiotic treatment does not affect its survival or related functions. Furthermore, antibiotic resistance genes are not transferred between yeast and bacteria, reducing the possibility of resistance genes being transferred to intestinal bacteria and preventing the development of further bacterial resistance.

[0033] Table 1 Antibiotic sensitivity of YFJ252 Note: - stands for insensitive.

[0034] 2. Strains Wickerhamomyces anomalus The probiotic ability of YFJ252.

[0035] Further Wickerhamomyces anomalus The acid and bile salt tolerance, hydrophobicity, self-aggregation, co-aggregation and antioxidant capacity of YFJ252 were determined.

[0036] 1. Acid and bile salt resistant.

[0037] The activated strain was inoculated into YPD liquid medium containing different pH values (pH 2, pH 3, pH 4, pH 5) and different bile salt concentrations (0.1%, 0.2% and 0.3%) at a 2% inoculum volume and cultured at 28°C for 24 h. The OD values were adjusted every 2 h. 600 nm Measure the absorbance value.

[0038] The results are as follows Figure 2 a and Figure 2 As shown in Figure b, growth activity at pH 2 was lower than at other pH conditions, but overall the bacterium maintained a good growth trend. It grew well at pH 3, 4, and 5. In the first 24 hours, the presence of 0.1%, 0.2%, and 0.3% bile salts inhibited the growth of strain YFJ252. However, after 24 hours, the presence of different bile salt concentrations had little effect on its growth. Based on these results, we can conclude that strain YFJ252 has good acid and bile salt tolerance.

[0039] 2. Hydrophobicity.

[0040] The hydrophobicity of YFJ252 to xylene, n-hexane, and ethyl acetate was determined. The activated overnight culture was centrifuged at 8944 × g, 4°C for 10 min, the precipitate was collected, washed twice with 50 mM K2HPSO4 (pH 6.5), and resuspended to adjust the OD 560 nm Prepare the test bacterial solution at 0.8-1.0, which is recorded as OD1. Take 3 mL of the above bacterial solution and mix it with 1 mL of three organic solvents, namely xylene, n-hexane and ethyl acetate. Vortex the mixture for 2 minutes and then incubate it at 28℃ for 3 hours to separate the two phases. Take the aqueous phase at OD1. 560nm Measure its absorbance and record it as OD2. The formula for calculating hydrophobicity is: Where: OD1 is the initial absorbance of the bacterial suspension to be tested, and OD2 is the absorbance of the aqueous phase.

[0041] The results are as follows Figure 2 As shown in Figure c, the hydrophobicity of YFJ252 to xylene, n-hexane, and ethyl acetate reached 74.71%, 83.19%, and 37.21%, respectively. The hydrophobicity of probiotics is closely related to cell adhesion, indicating that this strain is relatively easy to colonize in the intestine.

[0042] 3. Self-aggregation and co-aggregation ability.

[0043] (1) Self-aggregation ability.

[0044] The activated strain was centrifuged at 8944 × g, 4 ° C for 10 min, the precipitate was collected, washed twice with PBS (pH 7.2) and resuspended to adjust the OD 600nm The above bacterial solution was cultured at 28℃, and the supernatant was taken every 2 hours at OD 600 nm Measure the absorbance value and record it as OD2. The formula for calculating self-aggregation is: Where: OD1 is the absorbance value of the test strain suspension at 0 h, and OD2 is the absorbance value of the test strain suspension after cultivation.

[0045] (2) Co-aggregation ability.

[0046] Escherichia coli ( Escherichia coli ) were co-cultured with YFJ252 as indicator bacteria for co-aggregation analysis. E. coli and YFJ252 suspensions were prepared according to the above method, and 2 mL of each suspension was mixed and vortexed for 10 seconds. The supernatant was collected every 2 hours and the OD was 0. 600 nm Determine the absorbance value and calculate the coaggregation formula: Where: OD1 and OD2 are YFJ252 and E. coli The absorbance of bacterial suspension at 0 h, OD3 for YFJ252 and E. coli Absorbance value of bacterial suspension after mixing.

[0047] The results are as follows Figure 2 As shown in Figure d, the aggregation rate of YFJ252 was only 37.53% at 2 hours, reaching 95.71% at 4 hours. Subsequently, as the self-aggregation time increased, the self-aggregation rate of YFJ252 reached a peak of 96.40% at 8 hours. The coaggregation rates of YFJ252 and E. coli showed no significant difference between 2 and 8 hours, ranging from 71.00% to 73.00%. This bacterium exhibited excellent self-aggregation ability, and the coaggregation rate with E. coli remained between 71.00% and 73.00%, indicating that it has a good ability to aggregate pathogens and could be used as a potential probiotic.

[0048] 4. Antioxidant capacity.

[0049] (1) DPPH free radical scavenging ability.

[0050] 100 μL of fermentation supernatant was mixed with 100 μL of 0.2 mmol / L DPPH ethanol solution, and 1 mL of PBS buffer (0.01 mol / L, pH 7.2) was used to replace the fermentation supernatant as a control group. The cells were cultured in the dark at room temperature for 30 min. 517nmThe absorbance was measured at 400 nm. A 1 mg / mL Vc solution was used as a positive control. The DPPH free radical scavenging ability was calculated using the following formula: Where: OD1 is the absorbance value of the sample, and OD2 is the absorbance value of the control group.

[0051] (2) Hydroxyl free radical scavenging ability.

[0052] 2 mL of PBS (pH 7.4), 1 mL of 0.25 mmol / mL o-phenanthroline solution, 1 mL of 0.75 mmol / L FeSO₄ solution, and 1 mL of fermentation supernatant were mixed, followed by the addition of 1 mL of 12% H₂O₂ and the reaction was carried out at 37°C for 60 min. A control was prepared by replacing the fermentation supernatant with the same volume of PBS solution. A blank control was prepared by replacing the fermentation supernatant and all other components in the reaction mixture with water. At OD 536nm The absorbance was measured at 400 nm. A 1 mg / mL Vc solution was used as a positive control. The hydroxyl radical scavenging capacity was calculated using the following formula: Where: OD0 is the absorbance value of the control group, OD1 is the absorbance value of the sample, and OD2 is the absorbance value of the blank group.

[0053] (3) ABTS + Free radical scavenging ability.

[0054] 7mM ABTS was mixed with 2.5mM potassium persulfate (1:1) and the mixture was reacted at room temperature in the dark for 24h to prepare radical cations and adjust the ABTS + The absorbance of the solution at 734 nm was 0.700 ± 0.02. Next, 500 μL of polysaccharide solutions of different concentrations and 1 mL of ABTS were added. + The solutions were mixed and reacted at room temperature for 10 minutes. The absorbance of the mixture was then measured at 734 nm. A 1 mg / mL Vc solution was used as a positive control. + Free radical scavenging rate calculation formula: Where: OD0 is the absorbance value of the blank control group, OD1 is the absorbance value of the sample, and OD2 is the absorbance value of the sample solution.

[0055] The results are as follows Figure 2 e shows: YFJ252's effects on DPPH free radical, hydroxyl free radical and ABTS +The free radical scavenging rates reached 85.53%, 63.73%, and 56.52%, respectively, compared to 96.40%, 98.88%, and 100%, respectively, for 1 mg / mL vitamin C. YFJ252 demonstrated comparable DPPH scavenging activity to 1 mg / mL vitamin C, and also exhibited strong scavenging abilities against the other two free radicals. These results demonstrate the bacteria's excellent antioxidant properties.

[0056] 3. Strains Wickerhamomyces anomalus Metabolite determination of YFJ252 fermentation broth.

[0057] The metabolite composition of the YFJ252 fermentation broth was analyzed using untargeted metabolomics using UHPLC-QTOF-MS. The following steps were performed: 100 μL of fermentation supernatant was transferred to an EP tube, and 400 μL of extraction buffer (methanol:acetonitrile = 1:1 (v / v)) containing an isotopically labeled internal standard was added. The sample was vortex-mixed for 30 s and sonicated for 10 min in an ice-water bath. The extract was allowed to stand at -40°C for 1 h. The sample was centrifuged at 13,800 × g for 15 min at 4°C. The supernatant was transferred to a vial and analyzed using a Vanquish (ThermoFisher Scientific) ultra-high performance liquid chromatograph. The target compounds were separated chromatographically on a Waters ACQUITY UPLC BEH Amide (2.1 mm × 50 mm, 1.7 μm) column. Phase A consisted of aqueous phase containing 25 mmol / L ammonium acetate and 25 mmol / L ammonia, and phase B consisted of acetonitrile. Sample plate temperature: 4°C, injection volume: 2 μL.

[0058] The experimental results detected 24 types of metabolic compounds, including organic heterocyclic compounds, organic acids and their derivatives, benzene ring compounds, alkaloids, lipids and lipid molecules, amino acids and peptides, fatty acids, shikimic acid and phenylpropanoids, oxygen-containing organic compounds, phenylpropanoids and polyketides. Figure 3 a).

[0059] The metabolite profiles of fermentation samples were analyzed using PCA ( Figure 3 b). PCA results showed that there was no significant difference in metabolites between different batches of YFJ252 fermentation broth, indicating that the metabolite results measured in the experiment were stable.

[0060] In order to further understand the corresponding metabolic pathways involved in the metabolites produced by YFJ252, the relevant data were imported into the KEGG database for metabolic pathway analysis. Figure 3c) It can be seen that the related metabolites mainly involve 15 metabolic pathways: phenylalanine metabolism, tyrosine metabolism, arginine and proline metabolism, lysine degradation, glycine, serine and threonine metabolism, alanine, aspartic acid and glutamate metabolism, arginine biosynthesis, glycerate and dicarboxylic acid metabolism, nucleotide metabolism, amino acid biosynthesis, 2-oxycarboxylic acid metabolism, carbon metabolism, ABC transporters, and pyrimidine metabolism.

[0061] Among them, the role of 5 metabolic pathways is more critical, namely lysine biosynthesis, alanine, aspartate and glutamate metabolism, glyoxylate and dicarboxylic acid metabolism, arginine and proline metabolism, and pantothenic acid and coenzyme A biosynthesis, especially the lysine biosynthesis pathway ( Figure 3 d).

[0062] 4. Strains Wickerhamomyces anomalus Functional characteristics of YFJ252.

[0063] 1. Regulate sugar metabolism ability.

[0064] (1) Inhibitory effect on α-amylase.

[0065] The sample (500 μL) and α-amylase (500 μL; 0.1 U / mL) were thoroughly mixed and incubated at 37°C for 10 minutes. Next, 500 μL of 1% soluble starch was added to the mixture and incubated under the same conditions. DNS (1 mL) was added to the mixture and boiled for 5 minutes to terminate the reaction. Finally, 10 mL of distilled water was added, and the absorbance was measured at 540 nm. α-amylase inhibition was calculated using the following formula: Where: OD0 is the absorbance value of the control group, OD1 is the absorbance value of the sample, and OD2 is the absorbance value of the sample blank.

[0066] (2) Inhibitory effect on α-glucosidase.

[0067] Thoroughly mix the sample solution (40 μL) with 40 μL of α-glucosidase solution (1 U / mL). After incubation at 37°C for 10 minutes, add 20 μL of p-nitrophenyl-α-D-glucopyranoside solution (16 mM). After incubation at 37°C for 15 minutes, terminate the reaction by adding 40 μL of Na₂CO₃ solution (0.2 M). Measure the absorbance of the mixture at 405 nm. Calculate the inhibition rate of α-glucosidase activity using the following formula: Where: OD1 is the absorbance value of the sample, OD2 is the absorbance value without enzyme, and OD3 is the absorbance value without sample.

[0068] The ability of YFJ252 to inhibit α-amylase and α-glucosidase was used to reflect its ability to lower blood sugar. Figure 4 As shown in the figure, the inhibitory ability of YFJ252 on α-amylase reached 39.94%, and the inhibitory ability on α-glucosidase reached 35.32%.

[0069] 2. Inhibit protein denaturation.

[0070] The anti-inflammatory potential of YFJ252 was indirectly reflected by analyzing its ability to inhibit protein denaturation. 100 μL of sample was mixed with 400 μL of 5% bovine serum albumin, and the pH was adjusted to 6.5 using 1 M HCl. The mixture was incubated at 37°C for 20 minutes, then at 51°C for 20 minutes. After cooling, 2.5 mL of PBS (0.01 M, pH 7.2) was added. Deionized water was used as a control instead of the sample. The absorbance of the mixture was measured at 416 nm. The formula for calculating the inhibition of albumin denaturation is: Where: OD1 is the absorbance value of the control group, and OD2 is the absorbance value of the sample group.

[0071] The results showed that ( Figure 4 ), YFJ252's ability to inhibit protein denaturation reached 20.73%, indicating that the strain has certain anti-inflammatory capabilities.

[0072] 5. Strains Wickerhamomyces anomalus Uric acid-lowering ability of YFJ252 in vitro and in vivo.

[0073] To explore Wickerhamomyces anomalus The ability of YFJ252 to reduce uric acid in vitro and in vivo was evaluated by measuring its ability to inhibit XO in vitro and the levels of uric acid, creatinine and urea nitrogen in hyperuricemic mice to alleviate or prevent HUA.

[0074] 1. Inhibit XO activity.

[0075] The overnight culture of the purified strain was centrifuged at 7104 × g for 10 min, the supernatant discarded, and the cells were washed twice with sterile PBS. The resulting cells were resuspended in PBS (0.1 mol / L, pH 7.0), incubated at 37°C for 12 h, and centrifuged at 7104 × g for 10 min. The supernatant was obtained and used to analyze XO inhibitory activity. The enzymatic reaction was divided into three groups: (1) blank group: 20 μL xanthine oxidase (0.1 U) and 20 μL 0.15 mM xanthine were added to 160 μL PBS; (2) sample group: 20 μL xanthine oxidase (0.1 U), 20 μL 0.15 mM xanthine, and 20 μL cell-free supernatant were added to 140 μL PBS; (3) positive control group: 20 μL xanthine oxidase (0.1 U), 20 μL 0.15 mM xanthine, and 20 μL allopurinol were added to 140 μL PBS. The absorbance values at 293 nm were measured at reaction times of 0 and 10 min. The inhibition rate of the strain on XO was calculated according to the following formula.

[0076] Where A s0 and A s A is the absorbance value of the sample group and the positive control group at 0 min and 10 min respectively. b0 and A b The absorbance values of the blank control at 0 min and 10 min, respectively.

[0077] Abnormal UA metabolism is one of the causes of HUA. UA is primarily synthesized in the liver, and XO is an important precursor for UA synthesis. Therefore, we aimed to evaluate the inhibitory effect of YFJ252 on XO to preliminarily determine whether this bacterium has the potential to treat HUA. The results, shown in Table 2, showed that YFJ252 had an inhibitory ability of 84.15% on XO, suggesting that this bacterium has the potential to treat or improve HUA.

[0078] Table 2 Inhibitory ability of YFJ252 on XO activity 2. Wickerhamomyces anomalus Uric acid-lowering ability of YFJ252 in a hyperuricemia mouse model.

[0079] To explore Wickerhamomyces anomalus The uric acid-lowering and HUA-preventive effects of YFJ252 were investigated using a high uric acid mouse model. Wickerhamomyces anomalusThe uric acid-lowering ability of YFJ252 bacterial suspension was determined. After overnight culture of YFJ252, the bacterial suspension was centrifuged at 7104×g for 10 min, the supernatant was discarded, and the suspension was washed twice with sterile PBS. Finally, the bacterial suspension was resuspended in PBS to a final colony count of 1×10 9 CFU / mL. Kunming mice aged 6 weeks and weighing 20-25 g were used in the animal experiment. Before the experiment, the mice were acclimated to a standard environment for one week with a temperature of 24±2℃ and a humidity of 50±5%. The mice were kept in a 12 / 12 h light-dark cycle and had free access to food and water. 60 mice were divided into 5 groups (n=12): (1) Control group: 21 days of continuous gavage with normal saline; (2) Model group: 21 days of continuous gavage with high-sugar diet (10 g fructose in 100 g feed) and 21 days of continuous gavage with potassium oxonate (250 mg / kg); (3) YFJ252 group: 21 days of continuous gavage with high-fat diet (10 g fructose in 100 g feed) and 250 mg / kg potassium oxonate on days 1-7, and 1×10 9 CFU / mL bacterial suspension; (4) APL group: given a high-fat diet (100 g of fructose in 100 g of feed) and gavage with potassium oxonate (250 mg / kg) on days 1 to 7, and gavage with 20 mg / kg of allopurinol on days 8 to 21; (5) HUA+YFJ252 group: given a high-fat diet (100 g of fructose in 100 g of feed) and gavage with potassium oxonate (250 mg / kg) and 1×10 9 CFU / mL bacterial suspension. After the experiment, blood was collected and the levels of serum uric acid, creatinine and urea nitrogen were measured.

[0080] The results are as follows Figure 5 As shown: When mice were gavaged with potassium oxonate and given YFJ252 at the same time, after 21 days of the experiment, the serum uric acid concentration of mice in the model group reached 212.19 μmol / L, while the serum uric acid concentration of mice in the HUA+YFJ252 group was only 165.08 μmol / L, which was significantly lower than that in the model group and the YFJ252 group and had no significant difference with the CON group ( Figure 5 a). Figure 5 b and Figure 5Figure c shows that serum creatinine and urea nitrogen concentrations in the model group were 27.84 μmol / L and 10.61 mmol / L, respectively. However, YFJ252 was able to reduce these concentrations, with concentrations in the HUA + YFJ252 group dropping to only 20.54 μmol / L and 9.88 mmol / L. This suggests that administering YFJ252 early in the modeling process can reduce serum creatinine and urea nitrogen levels in mice. Therefore, we conclude that the abnormal Wickham yeast YFJ252 has the potential to prevent HUA.

[0081] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the scope of the present invention. Those skilled in the art will readily appreciate that the present invention is susceptible to various modifications and variations. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention.

Claims

1. Abnormal Wickham yeast ( Wickerhamomycesanomalus ) Use of YFJ252 (Accession No.: CGMCC NO.28267) in the preparation of products for preventing or treating hyperuricemia.

2. The use according to claim 1, characterized in that The product is a medicine, food or health product.

3. The use according to claim 1 or 2, characterized in that The active ingredient of the product is the bacteria of abnormal Wickham yeast YFJ252 or its metabolites.

4. The use according to claim 3, characterized in that The metabolites include substances that inhibit xanthine oxidase (XO) activity.

5. A product for preventing or treating hyperuricemia, characterized in that: The invention comprises the bacteria of abnormal Wickham yeast YFJ252 or its metabolites as active ingredients, and the preservation number of the strain is CGMCC NO.28267.

6. The product according to claim 5, characterized in that The product forms include oral preparations or probiotics.

7. Use of the aberrant Wickham yeast YFJ252 (Accession No.: CGMCC NO. 28267) in the preparation of a preparation for inhibiting xanthine oxidase (XO) activity.

8. The use according to claim 7, characterized in that The formulation is able to reduce serum uric acid, creatinine and urea nitrogen levels.

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