Synergistic composition of pea polypeptide and plant-derived lactic acid bacteria and its application

Abstract

The present invention provides a cooperative composition of pea polypeptide and plant-derived lactic acid bacteria and its application. [Solution] Experiments have confirmed that the plant-derived lactic acid bacterium AR342 provided by the present invention can effectively inhibit xanthine oxidase activity and reduce serum uric acid levels. AR342 can also improve intestinal flora disorders caused by high uric acid, promote the proliferation of beneficial flora, and reduce the abundance of harmful intestinal bacteria. At the same time, it has been discovered that the pea polypeptide prepared by the present invention can also reduce uric acid content, and a cooperative composition of pea polypeptide and plant-derived lactic acid bacterium AR342 also has the activity of inhibiting uric acid production in the body, with an effect superior to that of treatment with plant-derived lactic acid bacterium AR342 or pea polypeptide alone. Based on the above, the present invention provides a new concept for the clinical treatment of hyperuricemia.

Description

[Technical Field]

[0001] The present invention relates to the field of biomedicine technology, in particular to a cooperative composition of pea polypeptide and plant-derived lactic acid bacteria and its application. [Background technology]

[0002] Uric acid is one of the end products of human metabolism. Normally, uric acid production and excretion are balanced to maintain a constant blood uric acid level. Excessive uric acid production or insufficient excretion leads to elevated blood uric acid levels, resulting in hyperuricemia. Hyperuricemia is a common metabolic disorder that can lead to complications such as gout, kidney stones, renal dysfunction, hypertension, and cardiovascular disease. Gout, a typical manifestation of hyperuricemia, is an inflammatory arthropathy caused by the deposition of uric acid salts in joints and soft tissues. Gout attacks typically involve severe pain, swelling, heat, and tenderness, severely impacting patients' quality of life.

[0003] Currently, drugs for treating hyperuricemia and gout are mainly divided into two categories. One is uric acid-lowering drugs, such as allopurinol and febuxostat, which reduce blood uric acid levels by inhibiting uric acid production. The other is uricosuric drugs, such as benzenesulfonic acid and probenecid, which increase uric acid excretion and thereby reduce uric acid accumulation in the body. However, the use of these drugs has certain limitations and side effects. For example, allopurinol can cause liver dysfunction, gastrointestinal disorders, and rashes; febuxostat can cause renal dysfunction, blood system disorders, and allergic reactions; benzenesulfonic acid can cause gastrointestinal irritation, edema, and hypertension; and probenecid can cause kidney stones, renal failure, and rashes. Therefore, the development of safe, effective, and side-effect-free uric acid-lowering and anti-gout drugs is urgently needed by patients with hyperuricemia and gout.

[0004] In recent years, numerous studies have demonstrated the close relationship between intestinal flora and uric acid metabolism. An imbalance in intestinal flora leads to increased uric acid production and absorption and decreased uric acid excretion, which can induce or worsen hyperuricemia and gout. Probiotics are active microorganisms that benefit host health. By regulating the balance of intestinal flora, they can improve intestinal function and affect various physiological activities, including host metabolism and immunity. Food-derived oligopeptides are a new type of nutritional supplement. They are a new protein hydrolyzed product made by enzymatic digestion of food proteins and have a molecular weight of less than 1000 Da. Small molecular peptides are more easily absorbed by the human body than proteins derived from the same source, and are characterized by good solubility, low viscosity, and high bioavailability. From a nutritional perspective, deep-sea fish are considered a high-purine diet and should be avoided by patients with hyperuricemia. However, food-derived oligopeptides extracted from the deep-sea fish bonito have been found to inhibit xanthine oxidase activity. Based on this, the present invention provides a cooperative composition of food-derived oligopeptides and plant-derived lactic acid bacteria, which exerts a synergistic effect to significantly inhibit the conversion of purines in the intestines, thereby lowering uric acid levels. Summary of the Invention [Problem to be solved by the invention]

[0005] The objective of the present invention is to solve the above-mentioned problems of the existing technology by providing a cooperative composition of pea polypeptide and plant-derived lactic acid bacteria and its application. The cooperative composition of pea polypeptide and plant-derived lactic acid bacteria provided by the present invention can significantly reduce uric acid levels, providing a new concept for the clinical treatment of hyperuricemia. [Means for solving the problem]

[0006] To achieve the above object, the present invention provides the following solutions.

[0007] The present invention provides a plant-based lactic acid bacterium (Lactobacillus plantarum) strain AR342, which has the storage number CGMCC No. 27368, the storage date of May 17, 2023, and the storage institution of the General Microbiology Center of the China Committee for the Conservation of Microorganisms, and is located at No. 3, Hall 1, Beichen West Road, Chaoyang District, Beijing.

[0008] The present invention also provides a cooperative composition of pea polypeptide and plant-derived lactic acid bacteria AR342, said cooperative composition comprising the pea polypeptide and the plant-derived lactic acid bacteria AR342 in a mass ratio of 1:1, The conservation number of the plant-derived lactic acid bacterium AR342 is CGMCC No. 27368, and the pea polypeptide is obtained by fermenting pea crude protein.

[0009] The present invention also provides a method for producing the cooperative composition, which comprises the step of mixing pea polypeptide and plant-derived lactic acid bacteria AR342.

[0010] Furthermore, the plant-derived lactic acid bacteria AR342 was mixed in the form of a bacterial suspension, and the concentration of the plant-derived lactic acid bacteria AR342 in the bacterial suspension was 1×10 9 cfu / mL.

[0011] Furthermore, said pea polypeptide may be produced by the following steps: Pea crude protein and sterilized water were mixed in a mass / volume ratio of (1-2):(1-3), and then inoculated with Aspergillus oryzae BR88 to carry out solid-state fermentation. After fermentation is complete, sterilized water is added, the mixture is homogenized, and the mixture is centrifuged to collect the supernatant. The supernatant is ultrafiltered and the filtrate is freeze-dried to obtain the pea polypeptide.

[0012] Furthermore, the storage number of the Aspergillus oryzae BR88 is CGMCC No. 10950, the storage date is June 3, 2015, the storage institution is the General Microbiology Center of the China Committee for the Conservation of Microorganism Species, the storage location is No. 3, Hall No. 1, Beichen West Road, Chaoyang District, Beijing, the inoculation amount is 5-15% v / m, and the fermentation conditions are 25°C for 30 hours.

[0013] Furthermore, the homogenization time is 5-10 minutes.

[0014] Furthermore, the ultrafiltration is carried out by passing the solution through 5000 Da and 1000 Da membranes in succession.

[0015] The present invention also provides the use of the cooperative composition in the manufacture of a drug for the prevention and / or treatment of hyperuricemia.

[0016] The present invention also provides a drug for preventing and treating hyperuricemia, which comprises the above cooperative composition. [Effects of the Invention]

[0017] The present invention discloses the following technical effects.

[0018] The plant-derived lactic acid bacterium AR342 provided by the present invention has excellent acid resistance and bile salt tolerance, and can effectively inhibit xanthine oxidase activity, thereby lowering serum uric acid levels. AR342 can significantly reduce renal inflammation levels and alleviate kidney cell damage caused by high uric acid. Furthermore, AR342 can improve intestinal flora disorders caused by high uric acid, promote the proliferation of beneficial flora, and reduce the abundance of harmful intestinal bacteria. The present invention also discovered that pea polypeptide can lower uric acid content, and that a cooperative composition of pea polypeptide and plant-derived lactic acid bacterium AR342 also has the activity of inhibiting uric acid production in the body, with a greater effect than treatment with plant-derived lactic acid bacterium AR342 or pea polypeptide alone. Based on these findings, the present invention provides a new concept for the clinical treatment of hyperuricemia. [Brief explanation of the drawings]

[0019] In order to more clearly explain the embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings used in the embodiments. It is obvious that the drawings in the following description are only some embodiments of the present invention, and those skilled in the art can obtain other drawings based on these drawings without paying creative labor. [Figure 1] This shows the effect of plant-derived lactic acid bacteria AR342 on uric acid content in Example 2. Allopurinol: positive control. [Figure 2] 1 shows the effect of plant-derived lactic acid bacteria AR342 on liver XOD activity in Example 2. [Figure 3] 1 shows the effect of plant-derived lactic acid bacteria AR342 on the kidney in Example 2. The scale bar is 100 μm. [Figure 4] This shows the effect of plant-derived lactic acid bacteria AR342 on the intestinal flora in Example 2. A: Composition of the intestinal flora of mice in each treatment group, B: Statistical graph of the abundance of Bifidobacterium, and C: Statistical graph of the abundance of Akkermansia muciniphila. Con: control group, Mod: model group. [Figure 5] This shows the effect of treatment with pea polypeptide alone on the uric acid content in Example 3. WPP: pea polypeptide, allopurinol: positive control. [Figure 6] 1 shows the effect of the combined treatment of pea polypeptide and plant-derived lactic acid bacteria AR342 on the uric acid content in Example 3. DETAILED DESCRIPTION OF THE INVENTION

[0020] While various exemplary embodiments of the present invention will be described in detail, this detailed description should not be construed as limiting the present invention, but rather as a more detailed description of certain aspects, features, and examples of the present invention.

[0021] It should be understood that the terms used herein are for the purpose of describing particular embodiments and are not intended to limit the present invention. Furthermore, with respect to numerical ranges in the present invention, it is understood that every intermediate value between the upper and lower limits of that range is also specifically disclosed. Any stated value or intermediate value within a range, as well as smaller ranges between other stated values ​​or intermediate values ​​within that range, are also included in the present invention. The upper and lower limits of these smaller ranges may or may not be included in the range.

[0022] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Only preferred methods and materials are described in the present invention; however, any methods and materials similar or equivalent to those described herein may be used in the practice or testing of the present invention. All publications mentioned herein are incorporated by reference to disclose and describe the methods and / or materials related to the publication. In the event of a conflict between an incorporated publication and the contents of this specification, the contents of this specification shall control.

[0023] Numerous modifications and variations can be made to the specific embodiments of the present specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments will also be apparent to those skilled in the art from the present specification. The present specification and examples are merely illustrative.

[0024] As used herein, the words "including," "including," "having," "containing," and the like are all open-ended terms, i.e., meaning "without being limited to."

[0025] The present invention has been achieved by screening for the plant-derived lactic acid bacterium (Lactobacillus plantarum) AR342 strain, which is capable of suppressing uric acid production. Experiments have shown that the pea polypeptide prepared in this invention can also reduce the amount of uric acid produced, and that a cooperative composition of the pea polypeptide and the plant-derived lactic acid bacterium AR342 exhibits a significantly superior effect than when either is used alone. Specific research is as follows:

[0026] Example 1 Isolation, cultivation and preservation of plant-derived lactic acid bacteria AR342

[0027] 1. Isolation and purification of the strain

[0028] Fermented vegetable samples were collected from the Ili region of Xinjiang, China. 2g of the sample was cut, added to 20mL of sterile water, and mixed uniformly. A gradient dilution was then performed to obtain a diluted solution. 200μL of the diluted solution was spread on MRS agar medium and anaerobically cultured at 37℃ for 48 hours. Strains with different colony morphologies were selected and streaked on MRS agar medium to obtain purified strains.

[0029] 2. Measurement of uric acid production rate of each strain

[0030] 2.1 Bacterial cell preparation

[0031] A single colony of each purified strain was picked and inoculated into MRS liquid medium, followed by anaerobically culturing at 37°C for 16 hours. The cells were then centrifuged at 6000 rpm for 5 minutes to obtain bacterial cells. The cells were then resuspended and washed twice in PBS phosphate buffer, and then centrifuged at 6000 rpm for 5 minutes to obtain the bacterial cells.

[0032] 2.2 Uric acid production rate measurement

[0033] Each bacterial cell was resuspended in PBS, the OD adjusted to 1.0, and 1 mL of the PBS-cell suspension was collected. 100 μL of xanthine oxidase (XOD) solution (0.2 U / mL) was added to 1 mL of the PBS-cell suspension and incubated at 37°C for 5 minutes. The mixture was then centrifuged at 6000 rpm for 5 minutes. 100 μL of the supernatant was dispensed into a 96-well plate, and 200 μL of 20 μg / mL xanthine solution was added. The mixture was incubated at 37°C for 3 minutes using a microplate reader. The absorbance was measured at 295 nm. The fermentation solution was replaced with PBS as a control. The uric acid production rate was calculated using the following formula:

number

[0034] As shown in Table 1, the measurement results showed that the uric acid production rate in the control group was 85.57%, and among the isolated strains, AR342 had the lowest uric acid production rate of 44.83%, followed by AR237, which had a uric acid production rate of 46.65%.

[0035] [Table 1]

[0036] 3. Strain identification

[0037] Strain AR342 was identified as having the lowest rate of uric acid production.

[0038] Chromosomal DNA of the purified AR342 strain was extracted and subjected to colony PCR using the 16s rDNA universal primer 27F / 1492R. After amplification, the 16s rDNA sequence was obtained by sequencing. High homology comparison of bacterial species was performed at NCBI, and the purified AR342 strain was identified as a plant-derived lactic acid bacterium.

[0039] Here, 27F is 5'-AGAGTTTGATCCTGGCTCAG-3' and 1492R is 5'-GGTTACCTTGTTACGACTT-3'.

[0040] The above-mentioned plant-derived lactic acid bacteria (Lactobacillus plantarum) AR342 strain was preserved at the General Microbiology Center of the China Committee for the Conservation of Microbial Species on May 17, 2023, with the preservation number CGMCC No. 27368, and is stored at No. 3, Hospital No. 1, Beichen West Road, Chaoyang District, Beijing.

[0041] Example 2 Plant-derived lactic acid bacteria AR342 can alleviate hyperuricemia in mice

[0042] 1. Preparation of hyperuricemia mouse model and experimental group assignment

[0043] 1.1 Preparation of solutions

[0044] 0.5% (m / v) carboxymethylcellulose sodium (CMC-Na) suspension: 0.5 g of carboxymethylcellulose sodium was accurately weighed into 100 mL of ultrapure water, and the mixture was sonicated, heated in a water bath, and thoroughly stirred until dissolved.

[0045] Oteracil potassium-adenine-carboxymethylcellulose sodium suspension: A suspension was prepared using 0.5% carboxymethylcellulose sodium, and the mass ratio of oteracil potassium to adenine in the suspension was 2.5:1.

[0046] Allopurinol-carboxymethylcellulose sodium suspension: Similarly, an allopurinol-carboxymethylcellulose sodium suspension was prepared using 0.5% carboxymethylcellulose sodium.

[0047] Plant-derived lactic acid bacteria AR342 suspension: AR342 cells collected by centrifugation were washed with saline twice, and then the cell concentration was adjusted to 1 × 10 with saline. 9 The concentration was adjusted to cfu / mL.

[0048] 1.2 Modeling and Grouping

[0049] The experiment took 28 days, with the adaptation period being days 1-7, the modeling period being days 8-14, and the intervention period being days 15-28.

[0050] Environmental Adaptation: The animals used in the experiment were 40 5-week-old C57BL / 6 mice weighing 18-25 g. After 7 days of environmental adaptation, the subsequent experiments were carried out.

[0051] Modeling: Hyperuricemia modeling: Mice were orally administered with oteracil potassium-adenine-carboxymethylcellulose sodium suspension at 9:00 every day, and a standard dose of 250 mg / kg oteracil potassium and 100 mg / kg adenine was orally administered according to the mouse body weight for 7 days to establish a hyperuricemia mouse model.

[0052] Forty mice were randomly divided into four groups: a control group (normal), a model group, a positive control group, and a treatment group. All mice in the model group, positive control group, and treatment group underwent hyperuricemia modeling, and the control group received oral administration of 0.2 mL of 0.5% CMC-Na suspension per mouse per day from days 8 to 21.

[0053] Intervention period: All mice were orally administered with 0.2 mL of 0.5% CMC-Na suspension per mouse per day during the intervention period. The model group, positive control group, and treatment group continued to receive an orally administered oteracil potassium-adenine-CMC-Na suspension during the intervention period. At the same time, the treatment group mice received 0.2 mL (1 × 10) of AR342 bacterial suspension. 9 cfu / mL) were orally administered to mice in the positive control group, and allopurinol-CMC-Na suspension was orally administered to mice in the positive control group according to the standard dose of 60 mg / kg of mouse body weight.

[0054] Sampling: After the intervention period, mice from each group were fasted for 12 hours, anesthetized, and blood collected by eyedroplet sampling. The blood was then transferred to sterile blood collection tubes and centrifuged at 3000 rpm for 10 minutes. The supernatant was then collected and temporarily stored in a 4°C refrigerator for use in measuring the uric acid content in mouse serum. After dissection, the mice were quickly excised and their livers, kidneys, and other tissues were rapidly frozen in liquid nitrogen to prevent autolysis. After aliquots, the tissues were stored in a -80°C freezer. The livers were used to measure mouse hepatic XOD activity, and the kidneys were used for H&E staining analysis.

[0055] Mouse feces were collected and fecal DNA was extracted according to the instructions of the EZNA Fecal DNA Kit (Omega Bio-tek, USA). The V3-V4 region of the bacterial 16S rRNA gene in the feces was amplified using universal primers. The primer sequences are as follows: 338F:5´-ACTCCTACGGGAGGCAGCA-3´ 806R:5´-GGACTACHVGGGTWTCTAAT-3´.

[0056] The amplified fragments were purified using a DNA extraction kit (Axygen Biosciences) and used to measure the intestinal flora.

[0057] 2. Serum uric acid measurement Mouse serum was taken and placed on an ice box. A 50 mg / L uric acid standard application solution was prepared immediately before use. 0.2 mL of mouse serum and 2 mL of tungstic acid protein precipitant were mixed uniformly and left to stand for 10 minutes, then centrifuged at 4°C and 3000 rpm / min for 5 minutes to obtain the supernatant. 1.6 mL of the supernatant was taken and 0.5 mL each of CUT reagent and phosphotungstic acid reagent was added, mixed thoroughly and uniformly in a centrifuge tube, and left to stand for 10 minutes. After the reaction was completed, 0.2 mL of the mixture was placed in a 96-well plate and the OD 690nm The absorbance of the uric acid standard application solution of 50 mg / L was measured and designated as measurement A. The measurement result was standard A. The measurement result was blank A. Ultrapure water was used instead of serum.

[0058] The formula for calculating uric acid content is as follows:

number

[0059] The results of uric acid content are shown in Figure 1. Compared with the control group, the serum uric acid level of mice in the model group was significantly elevated, and after oral administration of the AR342 bacterial suspension, the serum uric acid level of mice was significantly reduced, decreasing by 38.72%.

[0060] 3. Effect of AR342 on hepatic XOD activity in hyperuricemia mice

[0061] Liver XOD activity measurement: 0.1 g of mouse liver was accurately weighed and the blood was washed with saline. Nine times the mass of homogenization medium (0.9% saline) was added to the mouse liver, which was then mechanically homogenized and placed in an ice-water bath to prepare a 10% homogenate. The mixture was centrifuged at 3000 rpm / min at 4°C for 10 minutes in a refrigerated centrifuge, and the supernatant was transferred to a centrifuge tube and stored on an ice box. 20 μL of the liver homogenate was used to measure xanthine oxidase activity. Following the instructions in the xanthine oxidase activity kit, the reagents were added, mixed uniformly, and then heated in a water bath at 37°C for 20 minutes. A stop solution was then added and mixed uniformly. 200 μL was placed in a 96-well plate, zero-adjusted with distilled water, and the OD was measured. 530 XOD activity was measured in nm.

[0062] The results are shown in Figure 2. Compared with the control group, the liver XOD activity of mice in the model group was significantly increased, but after oral administration of the AR342 bacterial suspension, the liver XOD activity of mice was significantly decreased, decreasing by 32.67%.

[0063] Figure 3 shows H&E staining analysis of mouse kidneys. The results showed that the kidneys of the model mice exhibited severe kidney damage, with large numbers of inflammatory cells, large cell gaps, large intercellular cavitations, and a small number of necrotic and sloughed renal tubular epithelial cells. Some renal tubules were severely twisted, irregularly shaped, and had unclear borders. After oral administration of the AR342 bacterial suspension, the number of vacuoles and necrosis in renal tissues was significantly reduced, inflammation was alleviated, tissue borders were clear, and cell nuclei were distinct, demonstrating a clear alleviating effect on kidney injury.

[0064] 4. Effect of AR342 on intestinal flora in hyperuricemia mice

[0065] The purified and amplified mouse fecal DNA fragments were sent to Shanghai Meiji Biomedical Technology Co., Ltd. for sequencing and data analysis. The results are shown in Figure 4. The intestinal flora of the mice in the model group was significantly different from that of the control group. After oral administration of the AR342 bacterial suspension, the composition of the intestinal flora improved, and the abundance of beneficial bacterial genera such as Bifidobacterium and Akkermansia muciniphila was significantly increased.

[0066] Example 3: Pea peptides can alleviate hyperuricemia in mice

[0067] 1. Preparation of pea peptide (WPP)

[0068] 100 g of crude pea protein (a by-product of pea starch production, rich in protein and dietary fiber; the crude pea protein used in this study was commercially available with a crude protein content of ≥80%, purchased from Zhengzhou Guangwang Biotechnology Co., Ltd.) was mixed uniformly with sterilized water at a ratio of 1:1 (m / v). Then, Aspergillus oryzae BR88 (preserved at the General Microbiology Center of the China Microorganism Species Collection and Management Committee on June 3, 2015, CGMCC No. 10950) was inoculated into the crude pea protein, and solid-state fermentation was carried out. Fermentation conditions were as follows: Aspergillus oryzae inoculation rate 10% (v / m), fermentation at 25°C for 30 hours. After fermentation was completed, three times the mass of water was added and the mixture was homogenized for 5 minutes. After fermentation, the mixture was centrifuged at 8000 rpm and the supernatant was collected. This was then subjected to successive ultrafiltration using membranes with molecular weights of 5000 Da and 1000 Da. The filtrate after ultrafiltration was freeze-dried to obtain pea peptides.

[0069] 2. Treating hyperuricemia mice with WPP alone

[0070] The method described in Example 2 was used to model hyperuricemia mice. After successful modeling, the intervention period began. The groups were divided as follows: control group: normal mice were orally administered 0.2mL of 0.5% CMC-Na suspension per mouse per day; model group: hyperuricemia model mice were orally administered oteracil potassium-adenine-CMC-Na suspension, with a standard dose of 250mg / kg oteracil potassium and 100mg / kg adenine according to mouse body weight; WPP group: hyperuricemia model mice were orally administered WPP (200mg / kg body weight); positive control group: hyperuricemia model mice were orally administered allopurinol-CMC-Na suspension, with a standard dose of 60mg / kg mouse body weight.

[0071] The serum uric acid content of mice was measured according to the method described in Example 2. The results are shown in Figure 5. Compared with the control group, the serum uric acid level of mice in the model group was significantly elevated, but after oral administration of pea peptides, the serum uric acid level of mice was significantly reduced, decreasing by 17.56%.

[0072] Example 4 Synergistic treatment of hyperuricemia mice with pea peptides and plant-derived lactic acid bacteria AR342

[0073] 1. Preparation of a mixed suspension of WPP and AR342

[0074] Plant-derived lactic acid bacteria AR342 was redissolved in sterilized water to prepare a bacterial suspension (1 × 10 9 cfu / mL) was prepared, and then WPP powder was added to the bacterial suspension at a mass ratio of 1:1 with plant-derived lactic acid bacteria AR342, and the mixture was mixed uniformly.

[0075] 2. Synergistic treatment of hyperuricemia mice with WPP and AR342

[0076] Modeling of hyperuricemia mice was performed according to the method described in Example 2. After successful modeling, the intervention period began and the mice were divided into groups as follows: control group: normal mice were orally administered 0.2 mL of 0.5% CMC-Na suspension per mouse per day; model group: hyperuricemia model mice were orally administered oteracil potassium-adenine-CMC-Na suspension, and standard doses of 250 mg / kg oteracil potassium and 100 mg / kg adenine were orally administered according to the mouse body weight; WPP group: hyperuricemia model mice were orally administered WPP (200 mg / kg body weight); AR342 group: hyperuricemia model mice were orally administered 0.2 mL of AR342 bacterial suspension (1 × 10 9 cfu / mL); Co-intervention group: Hyperuricemia model mice were orally administered 0.2 mL of a mixed suspension of WPP and AR342; Positive control group: Hyperuricemia model mice were orally administered allopurinol-CMC-Na suspension, and oral administration was performed according to the standard dose of 60 mg / kg of mouse body weight.

[0077] The serum uric acid content of mice was measured according to the method of Example 2. The results are shown in Figure 6. Compared with the control group, the serum uric acid level of mice in the model group was significantly elevated. The combined therapeutic effect of pea peptide WPP and plant-derived lactic acid bacteria AR342 was the best, showing an effect equivalent to that of allopurinol, and the serum uric acid level of mice in the combined treatment group was significantly lower than that of mice treated with WPP or AR342 alone.

[0078] The above-mentioned embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Any modifications or improvements made by those skilled in the art to the technical solutions of the present invention without departing from the design spirit of the present invention should fall within the scope of protection defined by the claims of the present invention. [Accession number]

[0079] JPEG2026034870000005.jpg235168

[0080] JPEG2026034870000006.jpg235168

Claims

1. A plant-derived lactic acid bacterium (Lactobacillus plantarum) AR342 strain, characterized in that the plant-derived lactic acid bacterium AR342 has a storage number of CGMCC No. 27368.

2. a cooperative composition of pea polypeptide and plant-derived lactic acid bacterium AR342, said cooperative composition comprising pea polypeptide and plant-derived lactic acid bacterium AR342 in a mass ratio of 1:1, A cooperative composition of pea polypeptide and plant-derived lactic acid bacterium AR342, characterized in that the storage number of the plant-derived lactic acid bacterium AR342 is CGMCC No. 27368, and the pea polypeptide is obtained by fermenting pea crude protein.

3. 3. A method for producing a cooperative composition according to claim 2, comprising the step of mixing the pea polypeptide with the plant lactic acid bacterium AR342.

4. The plant-derived lactic acid bacteria AR342 was mixed in the form of a bacterial suspension, and the concentration of the plant-derived lactic acid bacteria AR342 in the bacterial suspension was 1×10 ９ The method of claim 3, wherein the concentration of the microbial cell culture medium is 10 ...

5. The pea polypeptide is produced by the following steps: Pea crude protein and sterilized water were mixed in a mass / volume ratio of (1-2):(1-3), and then the mixture was inoculated with Aspergillus oryzae BR88 and subjected to solid-state fermentation. After fermentation, the mixture is homogenized and centrifuged to collect the supernatant.

4. A method according to claim 3, characterized in that the supernatant is ultrafiltered and the filtrate is freeze-dried to obtain the pea polypeptide.

6. The stock number of the Aspergillus oryzae BR88 is CGMCC No. 10950, and the inoculation amount is 5-15% v / m.

6. The method according to claim 5, wherein the fermentation is carried out at 25° C. for 30 hours.

7. The method according to claim 5, wherein the homogenization time is 5 to 10 minutes.

8. 6. The method according to claim 5, wherein the ultrafiltration is carried out by passing the solution through membranes of 5000 Da and 1000 Da in succession.

9. Use of the cooperative composition according to claim 2 in the manufacture of a drug for the prevention and / or treatment of hyperuricemia.

10. A drug for preventing and treating hyperuricemia, comprising the cooperative composition of claim 2.

Images