Lactobacillus johnsonii 0225 and uses thereof
By screening and identifying Lactobacillus johnsonii strain 0225, a microbial agent was prepared for the production of uric acid-lowering drugs, which solved the problem of large side effects of existing drugs and achieved safe and effective uric acid-lowering effects and improved kidney function.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- HUAZHONG AGRI UNIV
- Filing Date
- 2026-02-26
- Publication Date
- 2026-06-02
Smart Images
Figure CN122128149A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of microbiology, specifically to Lactobacillus johnsonii O225 and its applications. Background Technology
[0002] Hyperuricemia (HUA), characterized by abnormally high serum uric acid (UA) levels, is a major risk factor for the progression of gout, kidney disease, and cardiovascular disease, and is on the rise in many countries. Currently, several approaches are available for treating hyperuricemia, including diet, medication, and biological therapies. Uric acid-lowering drugs such as xanthine oxidase (XOD) inhibitors (e.g., allopurinol) and benzbromarone are commonly used to treat hyperuricemia. However, these drugs often have serious side effects, including hepatotoxicity and potentially fatal allopurinol allergy syndrome.
[0003] With lifestyle changes, including dietary modifications targeting the gut microbiota, another treatment option may be emerging. The diverse microbial community in the gut contributes to the host's metabolism and is crucial for their physical and mental health. The gut and gut microbiota play important roles in regulating serum UA concentration and distribution. Probiotics can effectively modify and restore the host's gut microbiota. Increasingly, animal studies and clinical data demonstrate the ability of probiotic strains to reduce serum UA concentration and improve renal function, possibly by inhibiting the absorption of exogenous purines in the gut.
[0004] In recent years, the application of microorganisms to prevent hyperuricemia is still in its early stages of development. Therefore, it is necessary to find more efficient new microbial strains to deal with hyperuricemia, which is of great significance for the development of uric acid-lowering drugs and foods. Summary of the Invention
[0005] The purpose of this invention is to overcome the shortcomings of the prior art and provide a *Lactobacillus johnsonii* 0225 and its application. This invention uses MRS agar medium to isolate and screen microorganisms from the intestinal contents of mice previously treated with the natural product drug DIO in the laboratory, obtaining a strain. This strain is then isolated, purified, and subjected to morphological and molecular biological identification, confirming it as *Lactobacillus johnsonii*. Lactobacillus_johnsonii The applicant named the strain Lactobacillus johnsonii (Lactobacillus johnsonii). Lactobacillus_johnsonii Lactobacillus johnsonii 0225 has good uric acid-lowering activity and has the potential to be used to prepare uric acid-lowering drugs.
[0006] To achieve the above objectives, the technical solution designed by the present invention is as follows: This invention provides a Lactobacillus johnsonii ( Lactobacillus_johnsonii )0225, its accession number is CCTCC NO: M 2025549.
[0007] The above-mentioned Lactobacillus johnsonii ( Lactobacillus_johnsonii 0225 was deposited at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, Hubei Province, China, on March 20, 2025, with accession number CCTCC NO: M2025549.
[0008] The present invention also provides an application of the above-mentioned Lactobacillus johnsonii O225 in lowering uric acid.
[0009] The present invention also provides the application of the above-mentioned Lactobacillus johnsonii O225 in the preparation of uric acid-lowering drugs.
[0010] The present invention also provides a uric acid-lowering drug, wherein the drug contains the aforementioned Lactobacillus johnsonii O225.
[0011] Furthermore, the content of Lactobacillus johnsonii O225 in the drug is 1. 10 8~9 CFU / mL.
[0012] Furthermore, the content of Lactobacillus johnsonii O225 in the drug is 1. 10 9 CFU / mL.
[0013] The present invention also provides a method for preparing the uric acid-lowering drug according to claim 4, characterized in that it includes the following steps: 1) Activate Lactobacillus johnsonii O225; 2) The activated Lactobacillus johnsonii 0225 was fermented and cultured, and the bacterial cells were isolated from the cultured species and made into microbial agents.
[0014] Furthermore, in step 2), the culture medium used for fermentation is MRS broth medium, with the following formula: 10 g / L peptone, 8 g / L beef meal, 4 g / L yeast extract, 22 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium hydrogen citrate, 5 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate, and 1 g / L Tween 80, with a pH of 5.7 ± 0.2.
[0015] Furthermore, in step 2), the fermentation culture temperature is 37 ℃ and the time is 24-48 h.
[0016] The beneficial effects of this invention are: This invention enriches the variety of uric acid-lowering microorganisms, providing more strains for microbial degradation of uric acid. Furthermore, the *Lactobacillus johnsonii* provided by this invention has advantages such as good uric acid-lowering effect, good effect on improving renal function indicators, simple fermentation process, and low production cost, showing potential and broad prospects for further development of uric acid-lowering drugs. Attached Figure Description
[0017] Figure 1 The colony morphology of Lactobacillus johnsonii O225 on MRS agar medium.
[0018] Figure 2 This is a diagram of the fermentation cell suspension of Lactobacillus johnsonii O225.
[0019] Figure 3 The bar chart shows the in vivo index determination results of Lactobacillus johnsonii O225. Detailed Implementation
[0020] The present invention will now be described in further detail with reference to specific embodiments, so that those skilled in the art can understand it.
[0021] Example 1: Screening and identification of Lactobacillus johnsonii O225 1. Strain screening In a sterile operating room, weigh 0.5 g of mouse intestinal contents, add 100 mL of sterile physiological saline, mix thoroughly, and incubate at 37 ℃ and 180 r / min in a constant temperature shaker for 24 h. After enriching the bacterial culture, shake well and dilute to 10-fold using the 10-fold method. -1 10 -2 10 -3 10 -4 Dilute to 100 μL and transfer the bacterial culture to commercially available MRS agar plates. Allow colonies to form. Figure 1 ), observe its morphology, and purify and culture the isolated single colonies for further identification.
[0022] 2. Molecular biological identification The 16S rDNA gene of isolated single colonies was amplified using the following primers: 27 F: 5'- agatgatcatgg-3', 1492 R: 5'-tacggttaccttgttac-GACTT-3'; A 25 μL PCR reaction system was prepared: 0.5 μL template DNA, 0.5 μL each of forward and reverse primers (27F / 1492R), 12.5 μL 2×Taq enzyme, and ddH2O to a final volume of 25 μL. PCR amplification conditions were: 95 ℃ pre-denaturation for 5 min; 95 ℃ denaturation for 30 s, 55 ℃ annealing for 30 s, 72 ℃ extension for 1 min; 30 cycles; 72 ℃ extension for 10 min. 5 μL of PCR amplification product and DL2000 DNA Marker were taken, and the PCR amplification results were detected using a gel electrophoresis imaging analysis system. The PCR amplification product was sent to Beijing Qingke Biotechnology Co., Ltd., and the sequenced products were compared with the National Center for Biotechnology Information (NCBI) database or EzBioCloud (http: / / www.ezbiocloud.net) to identify the isolates. Lactobacillus_johnsonii The similarity was 100%, therefore the strain of this invention was identified as... Lactobacillus_ johnsonii The 16S rDNA sequence is as follows: TGTTCCAACCTTAGACGGCTGACTCCTATAAAGGTTATCCCACCGGCTTTGGGTGTTACAGACTCTCATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCGTGCTGATCCGCGATTACTAGCGATTCCAGCTTCGTGTAGGCGAGTTGCAGCCTACAGTCCGAACTGAGAACGGCTTTAAGAGATCCGCTTGCCTTCGCAGGTTCGCTTCTCGTTGTACCGTCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGACTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCATTAGAGTGCCCAACTTAATGATGGCAACTAATGACAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAGCCATGCACCACCTGTCTCAGCGTCCCCGAAGGGAACACCTAATCTCTTAGGTTTGCACTGGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAACCTTGCGGTCGTACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTGCAGCACTGAGAGGCGGAAACCTCCCAACACTTAGCACTCATCGTTTACGGCATGGACTACCAGGGTATCTAATCCTGTTCGCTACCCATGCTTTCGAGCCTCAGCGTCAGTTGCAGACCAGAGAGCCGCCTTCGCCACTGGTGTTCTTCCATATATCTACGCATTCCACCGCTACACATGGAGTTCCACTCTCCTCTTCTGCACTCAAGTTCAACAGTTTCTGATGCAATTCTCCGGTTGAGCCGAAGGCTTTCACATCAGACTTATTGAACCGCCTGCACTCGCTTTACGCCAATAAATCCGGACAACGCTTGCCCCTACGATTACCGCGGCTGCTGGCACGAAGTAGCCGTGATTTCTAA。
[0023] 3. Strain preservation The identified strains were cultured in fresh MRS broth and then stored in 50% glycerol at -80°C. Ultimately, the applicant named the screened strain *Lactobacillus johnsonii*. Lactobacillus_johnsonii Accession number 0225 was deposited at the China Center for Type Culture Collection (CCTCC), located at Wuhan University, Wuhan, Hubei Province, on March 20, 2025.
[0024] Example 2: Verification of the in vivo uric acid-lowering efficacy of Lactobacillus johnsonii O225 1. Preparation of Lactobacillus johnsonii O225 cell suspension Lactobacillus johnsonii 0225 was activated and inoculated into MRS medium, and cultured at 37 ℃ for 24-48 h to obtain a bacterial concentration of 1. 10 8 Centrifuge at 3000 r / min for 10 min at 4 ℃ to obtain bacterial cell precipitate. After washing with physiological saline, the bacterial cell precipitate is resuspended to a concentration of 1. 10 9 CFU / mL concentration, to obtain bacterial suspension ( Figure 2 Further verification of the efficacy of bacterial suspension in lowering uric acid in vivo.
[0025] 2. Evaluation of the uric acid-lowering ability of Lactobacillus johnsonii O225 Male KM mice (SPF grade), aged 6-8 weeks and weighing 30±5 g, were used in groups of 5. They were acclimatized for 3 days and fasted for 12 hours before modeling. The HUA mouse model was established by gavage administration of 200 mg / kg potassium oxonate, while the blank control group was administered an equal volume of 0.5% sodium carboxymethyl cellulose solution by gavage. One hour after daily gavage administration of potassium oxonate, mice were given 0.1 mL / 10 g bacterial suspension for 14 days.
[0026] After gavage, blood was collected via the eyeballs, left to stand for 3 hours, and then centrifuged at 6000 rpm for 5 minutes at room temperature. The supernatant serum was collected, and the levels of uric acid (UA), blood urea nitrogen (UREA), and creatinine (CREA) were measured.
[0027] The results showed that the levels of UA, UREA, and CREA were significantly increased in the model group mice. Lactobacillus_ johnsonii HZAU-0412 bacterial suspension significantly reduced the increase in serum uric acid in mice induced by potassium oxonate, and alleviated urea nitrogen and creatinine levels, bringing them closer to normal values. Figure 3 ).
[0028] Example 3 Preparation of uric acid-lowering microbial agents Lactobacillus johnsonii 0225 was activated and inoculated into MRS medium, and cultured at 37 °C for 24–48 h. The concentration of the bacterial suspension was determined by plate dilution colony counting. The bacterial cells were resuspended to a concentration of 1. 10 9 CFU / mL yields a microbial liquid preparation that can be used to degrade uric acid in vivo. This preparation contains live cells of Lactobacillus johnsonii O225 and has advantages such as high yield and low cost.
[0029] In summary, the present invention provides a microbial preparation for in vivo degradation of uric acid, which contains live cells of Lactobacillus johnsonii O225.
[0030] All other parts not described in detail are existing technologies. Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.
Claims
1. A type of Lactobacillus johnsonii ( Lactobacillus_johnsonii )0225, its accession number is CCTCC NO: M2025549.
2. The application of Lactobacillus johnsonii O225 as described in claim 1 in lowering uric acid.
3. The use of Lactobacillus johnsonii O225 as described in claim 1 in the preparation of uric acid-lowering drugs.
4. A uric acid-lowering drug, characterized in that: The drug contains Lactobacillus johnsonii O225 as described in claim 1.
5. The uric acid-lowering drug according to claim 4, characterized in that: The drug contains 1 part Lactobacillus johnsonii O225. 10 8~9 CFU / mL.
6. The uric acid-lowering drug according to claim 5, characterized in that: The drug contains 1 part Lactobacillus johnsonii O225. 10 9 CFU / mL.
7. A method for preparing the uric acid-lowering drug according to claim 4, characterized in that: Includes the following steps: 1) Activate Lactobacillus johnsonii O225; 2) The activated Lactobacillus johnsonii 0225 was fermented and cultured, and the bacterial cells were isolated from the cultured species and made into microbial agents.
8. The preparation method according to claim 7, characterized in that: In step 2), the culture medium used for fermentation is MRS broth medium, which has the following formula: 10 g / L peptone, 8 g / L beef meal, 4 g / L yeast powder, 22 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium hydrogen citrate, 5 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate, and 1 g / L Tween 80, with a pH of 5.7 ± 0.
2.
9. The preparation method according to claim 7, characterized in that: In step 2), the fermentation temperature is 37℃ and the time is 24-48 h.