US 12,391,913 B2
Lactiplantibacillus plantarum strain for alleviating hyperuricemia and combination and use thereof
Bailiang Li, Heilongjiang (CN); Zhongjiang Wang, Heilongjiang (CN); Zengbo Wang, Heilongjiang (CN); Zengwang Guo, Heilongjiang (CN); and Qingxue Chen, Heilongjiang (CN)
Assigned to Northeast Agricultural University, Heilongjiang (CN)
Appl. No. 18/717,423
Filed by Northeast Agricultural University, Heilongjiang (CN)
PCT Filed Dec. 5, 2023, PCT No. PCT/CN2023/136303
§ 371(c)(1), (2) Date Jun. 7, 2024,
PCT Pub. No. WO2024/193100, PCT Pub. Date Sep. 26, 2024.
Claims priority of application No. 202310288894.8 (CN), filed on Mar. 22, 2023.
Prior Publication US 2025/0109375 A1, Apr. 3, 2025
Int. Cl. C12N 1/20 (2006.01); A61K 35/00 (2006.01); A61K 35/741 (2015.01); A61P 19/06 (2006.01); C12R 1/01 (2006.01)
CPC C12N 1/205 (2021.05) [A61K 35/741 (2013.01); A61P 19/06 (2018.01); A61K 2035/115 (2013.01); C12R 2001/01 (2021.05)] 3 Claims
OG exemplary drawing
 
1. A method of preparing a combination for alleviating hyperuricemia, wherein the combination comprises a Lactiplantibacillus plantarum strain BL-17 at a viable bacterial cell concentration of higher than or equal to 1×106 CFU/mL or higher than or equal to 1×106 CFU/g, 0.01 wt % galacto-oligosaccharide, and 0.01 wt % adenylic acid in a 100 g/L sterilized skimmed milk aqueous solution, and the Lactiplantibacillus plantarum strain BL-17 is deposited in China Center for Type Culture Collection (CCTCC), Wuhan on Oct. 19, 2022, with an accession number of CCTCC NO: M20221596;
wherein the preparation method comprises the following steps:
step 1, streaking the Lactiplantibacillus plantarum strain BL-17 on an MRS solid medium on a plate, picking a single colony for Gram staining, and after a single pure colony of the Lactiplantibacillus plantarum strain BL-17 is observed microscopically, subculturing the Lactiplantibacillus plantarum strain BL-17 twice in an MRS liquid medium to ensure the Lactiplantibacillus plantarum strain BL-17 is viable, to obtain an MRS liquid culture comprising the Lactiplantibacillus plantarum strain BL-17, and mixing 50% (v/v) glycerin aqueous solution and the MRS liquid culture in a volume ratio of 2:3 to obtain a resulting mixture, and storing the resulting mixture at −80° C. for later use;
step 2, inoculating the cryopreserved Lactiplantibacillus plantarum strain BL-17 into 5 mL of the MRS liquid medium at an inoculum size of 4% to obtain a mixture, incubating the mixture at 37° C. for 20 hours, followed by subculturing the cryopreserved Lactiplantibacillus plantarum strain BL-17 for two passages to obtain an MRS liquid culture comprising a Lactiplantibacillus plantarum strain BL-17 at passage 2; and centrifuging the MRS liquid culture comprising the Lactiplantibacillus plantarum strain BL-17 at passage 2 at 4° C. and 6,000 r/min for 5 min to obtain a bacterial pellet, collecting the bacterial pellet, washing the bacterial pellet twice with sterile normal saline to obtain a washed bacterial pellet, and collecting the washed bacterial pellet for subsequent preparation of a bacterial suspension; and
step 3, resuspending the washed bacterial pellet in the 100 g/L sterilized skimmed milk aqueous solution and performing colony counting by a serial dilution and spread plate method to prepare a bacterial suspension with a concentration of 4.0×109 CFU/mL; and adding the galacto-oligosaccharide and the adenylic acid to the bacterial suspension contents such that the galacto-oligosaccharide and the adenylic acid are each at a concentration of 0.1 mg/mL; wherein
the combination is capable of attenuating an intestinal absorption of an inosine and inosine-related purine compounds, and an adenosine and adenosine-related purine compounds.