Condensaxtm weizmannia nj01 formulation degrading inosine and guanosine and uses thereof

Abstract

This invention belongs to the field of biotechnology and relates to a strain of *Weizmannii coagulans* NJ01 for the biodegradation of inosine and guanosine, which can produce enzymes that catalyze the degradation of inosine and guanosine. This invention also relates to a preparation of *Weizmannii coagulans* NJ01 for degrading inosine and guanosine, containing bacterial cells and / or crude enzymes of the aforementioned *Weizmannii coagulans* NJ01 strain. Research results show that the *Weizmannii coagulans* NJ01 strain and its produced enzymes provided by this invention are safe for human use and can efficiently biodegrade inosine and guanosine, demonstrating significant value and application prospects in the removal of inosine and guanosine for the prevention and treatment of hyperuricemia, gout, and related complications in humans.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology and relates to a Weizmannii coagulans NJ01 preparation that degrades inosine and guanosine and its application. Background Technology

[0002] Currently, there are approximately 197 million people worldwide suffering from hyperuricemia, and over 14.66 million suffering from gout. Early-stage hyperuricemia often presents with no obvious symptoms, but as it progresses, urate crystals deposit in the body, eventually leading to gout.

[0003] Inosine (also known as: inosine, molecular formula: C) 10 H 12 N 4O5 (molecular weight: 268.226) and guanosine (molecular formula: C 10 H 13 N 5O5 Inosine (molecular weight: 283.241) is a key compound in the purine metabolism pathway, generated by purine nucleotide salvage synthesis or de novo synthesis. Inosine and guanosine are precursors to uric acid formation, and their accumulation can lead to purine metabolism complications such as hyperuricemia and gout.

[0004] The number of people suffering from hyperuricemia and gout worldwide continues to rise, becoming a global public health problem. Traditional prevention and treatment methods for hyperuricemia mainly rely on two types: drug therapy and dietary intervention. However, drugs such as febuxostat and allopurinol have significant toxic side effects on the liver and kidneys, and dietary interventions vary greatly and are not very effective. Currently, there is still a lack of effective treatments for controlling hyperuricemia and gout in clinical practice. Therefore, it is necessary to conduct in-depth research on effective and safe drugs or methods for treating hyperuricemia complications. Summary of the Invention

[0005] One of the objectives of this invention is to address the problem of toxic side effects of existing drugs for the prevention and treatment of hyperuricemia and gout, and to provide a strain of Weizmannii coagulans NJ01 for the biodegradation of inosine and guanosine. The bacterial cells of this strain and the enzymes it produces can efficiently biodegrade inosine and guanosine, and have important application prospects in reducing uric acid production.

[0006] The second objective of this invention is to provide a *Weizmannii coagulans* NJ01 preparation for degrading inosine and guanosine and its application. The *Weizmannii coagulans* NJ01 preparation is made from the above-mentioned *Weizmannii coagulans* NJ01 strain used for biodegrading inosine and guanosine, and can efficiently biodegrade inosine and guanosine.

[0007] Therefore, the first aspect of the present invention provides a strain of Weizmannia coagulans NJ01 for the biodegradation of inosine and guanosine, which is capable of producing an enzyme that catalyzes the degradation of inosine and guanosine, and its accession number is CGMCC No.37022.

[0008] The second aspect of the present invention provides a *Weizmannii coagulans* NJ01 formulation for the biodegradation of inosine and guanosine, which contains bacterial cells and / or crude enzymes of the *Weizmannii coagulans* NJ01 strain as described in the first aspect of the present invention.

[0009] In some embodiments of the present invention, the *Wietzmannii* NJ01 preparation for degrading inosine and guanosine is a liquid preparation; preferably, in the liquid preparation for degrading inosine and guanosine, the concentration of *Wietzmannii* NJ01 strain cells is (1-10)×10⁻⁶. 8 / mL; and / or, in the liquid formulation for degrading inosine and guanosine, the protein concentration of the crude enzyme of *Weizmannia coagulans* strain NJ01 is 1-5 g / L.

[0010] In other embodiments of the present invention, the *Wietzmannii coagulans* NJ01 preparation for degrading inosine and guanosine is a solid powder preparation; preferably, in the solid powder preparation for degrading inosine and guanosine, the content of *Wietzmannii coagulans* NJ01 strain cells is (1-10)×10⁻⁶. 8 / g, more preferably (5-10)×10 8 / g; and / or, in the solid powder formulation of the degraded inosine and guanosine, the protein content of the crude enzyme of Weizmannia coagulans strain NJ01 is 1-5 g / kg, more preferably 3-5 g / kg.

[0011] A third aspect of the present invention provides a method for preparing a *Wietzmannii* NJ01 formulation that degrades inosine and guanosine as described in the second aspect of the present invention, comprising:

[0012] Step B: Inoculate the fermentation strain into the fermentation medium for fermentation culture to obtain the fermentation culture of Weizmannella coagulans strain NJ01.

[0013] Step C: Centrifuge the fermentation culture of Weizmannii NJ01 strain to harvest the bacterial cells of Weizmannii NJ01 strain.

[0014] The fermentation strain was obtained from the corresponding Weizmann's coagulans strain NJ01 through seed culture.

[0015] According to the present invention, the fermentation medium comprises, per 1L of water, the following components:

[0016] 5-10g of peptone; preferably 8-10g;

[0017] 5-10 g of yeast powder; preferably 8-10 g; and

[0018] 3-8 g of glucose; preferably 6-8 g;

[0019] Preferably, the pH value of the fermentation medium is 6-7.

[0020] More preferably, in step B, the fermentation culture temperature is 35-50℃, more preferably 35-40℃.

[0021] According to some embodiments of the present invention, the preparation method further includes:

[0022] Step K: The cell suspension of the *Weizmannia coagulans* strain NJ01 is subjected to cell disruption treatment under low temperature conditions to obtain a cell-free lysate of the *Weizmannia coagulans* strain NJ01.

[0023] Step L: Centrifuge the cell-free lysate of strain NJ01 of Weizmannia coagulans and take the supernatant cell-free extract as the crude enzyme of strain NJ01 of Weizmannia coagulans.

[0024] The low temperature is 0-4℃.

[0025] The fourth aspect of this invention provides the use of the *Wietzmannii coagulans* NJ01 preparation for degrading inosine and guanosine as described in the second aspect of this invention, or the *Wietzmannii coagulans* NJ01 preparation for degrading inosine and guanosine prepared by the preparation method described in the third aspect of this invention, in the preparation of inosine and guanosine-lowering agents, comprising:

[0026] Step D: Wash the cells of strain NJ01 of Weizmannia coagulans with physiological saline to obtain a pure cell product of strain NJ01 of Weizmannia coagulans.

[0027] Step E: In a physiological saline system, under low temperature conditions, ultrasonically disrupt the pure bacterial cells of strain NJ01 of Weizmannia coagulation. After centrifugation, take the supernatant to obtain cell-free extract as crude pure enzyme of strain NJ01 of Weizmannia coagulation.

[0028] Step F involves freeze-drying the bacterial cells and / or crude enzymes of *Weizmannia coagulans* strain NJ01, and then diluting the freeze-dried *Weizmannia coagulans* NJ01 preparation to produce inosine and guanosine preparations.

[0029] The low temperature is 0-4℃.

[0030] In some embodiments of the present invention, in step F, the freeze-dried *Weizmannii coagulans* NJ01 preparation is diluted with physiological saline to prepare liquid inosine and guanosine preparations; preferably, the liquid inosine and guanosine preparations are oral preparations.

[0031] In some other embodiments of the present invention, in step F, the freeze-dried *Weizmannii coagulans* NJ01 preparation is diluted with edible starch to prepare solid inosine and guanosine preparations; preferably, the solid inosine and guanosine preparations are oral preparations.

[0032] The inventors have discovered that the *Weizmannii coagulans* strain NJ01 and the enzymes it produces, which are used for the biodegradation of inosine and guanosine, are safe for human use and can efficiently biodegrade inosine and guanosine. They have significant value and application prospects in the efficient removal of inosine and guanosine for the prevention and treatment of hyperuricemia, gout and related complications in humans.

[0033] The above description is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention and to implement it in accordance with the contents of the specification, and in order to make the above and other objects, features and advantages of the present invention more apparent and understandable, specific embodiments of the present invention are described below. Attached Figure Description

[0034] To facilitate understanding of the present invention, a more detailed description will be provided below with reference to the accompanying drawings, during which the above and other objects, features, and advantages of the present invention will become more apparent. The accompanying drawings are provided to further illustrate the present invention and form part of the specification. They are used together with the embodiments of the present invention to explain the invention and do not constitute a limitation thereof.

[0035] Figure 1 A molecular phylogenetic tree based on 16S rRNA is shown for screening Weizmann's coagulans NJ01.

[0036] Figure 2 The figures show the kinetic curves of inosine and guanosine biodegradation by *Weizmannii coagulation* NJ01. The upper figure shows the kinetic curve of inosine biodegradation by *Weizmannii coagulation* NJ01, and the lower figure shows the kinetic curve of guanosine biodegradation by *Weizmannii coagulation* NJ01.

[0037] Figure 3 The figures show the kinetic curves of the degradation of inosine and guanosine catalyzed by the enzyme NJ01 of *Weizmannia coagulans*. The upper figure shows the kinetic curve of the degradation of inosine catalyzed by the enzyme NJ01 of *Weizmannia coagulans*, and the lower figure shows the kinetic curve of the degradation of guanosine catalyzed by the enzyme NJ01 of *Weizmannia coagulans*.

[0038] strain preservation

[0039] The strain provided in this invention is classified as *Weizmannia coagulans*, isolated and identified by Beijing Yiran Biotechnology Co., Ltd., and deposited at the China General Microbiological Culture Collection Center (CGMCC; address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing) on ​​December 11, 2025, with accession number CGMCC No. 37022. In this invention, this strain is named *Weizmannia coagulans* NJ01, also known as *Weizmannia coagulans* NJ01. Detailed Implementation

[0040] To facilitate understanding of the present invention, it will be described in detail below. However, before describing the present invention in detail, it should be understood that the present invention is not limited to the specific embodiments described. It should also be understood that the terminology used herein is for describing specific embodiments only and is not intended to be restrictive.

[0041] Unless otherwise defined, all terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While any methods and materials similar to or equivalent to those described herein may also be used in the practice or testing of this invention, preferred methods and materials are now described.

[0042] In this invention, the range of dosage concentration, temperature or other physical or chemical properties or characteristics, unless otherwise specified, covers or includes the upper and lower limits of that range.

[0043] I. Terminology

[0044] In this invention, the term "cell body" refers to the live and / or dead cells of *Weizmannii NJ01* coagulates.

[0045] In this invention, the term "crude enzyme" refers to the cell-free extract obtained by centrifuging and collecting the supernatant after the cells of *Weizmannii coagulans* NJ01 are broken.

[0046] The term "pure crude enzyme" used in this invention refers to a cell-free extract obtained by centrifuging and collecting the supernatant of pure cells of *Weizmannii NJ01* after they have been broken up, in contrast to crude enzyme.

[0047] The term "microbial preparation" as used in this invention refers to preparations of various forms made from microorganisms with medical research value as raw materials, using traditional or modern biotechnology, and used for the prevention (health care), treatment, and diagnosis of various physiological symptoms in the human body.

[0048] The term "edible starch" as used in this invention refers to starch that meets the "National Standard for Edible Starch" (GB31637-2016 National Food Safety Standard for Edible Starch).

[0049] The term "cell suspension" as used in this invention refers to the liquid obtained by adding approximately 50 mL of sterile physiological saline to a microbial fermentation culture after centrifugation to remove the supernatant, thereby suspending the precipitated microbial cells.

[0050] The "water" used in the invention for culture media or fermentation processes, unless otherwise specified, refers to sterile pure water obtained by filtration through a 0.22 μm filter membrane.

[0051] II. Implementation Plan

[0052] As mentioned earlier, approximately 197 million people worldwide suffer from hyperuricemia each year, and the number continues to rise, making it a global public health problem. Addressing the issues of toxic side effects from existing drug therapies and the inconsistent effectiveness of dietary control due to individual differences in constitution, the inventors of this invention have conducted a systematic and in-depth study on the utilization of microorganisms to degrade inosine and guanosine, based on the mechanism by which inosine and guanosine are precursors to uric acid formation and their accumulation leads to hyperuricemia and gout, among other purine metabolism complications.

[0053] The inventors have noted that while lactic acid bacteria can promote gut health, they also reduce the accumulation of uric acid to some extent through a series of metabolic regulation.

[0054] The inventors also noted that *Weizmannii coagulans* combines the advantages of both lactic acid bacteria and Bacillus, and is one of the Bacillus strains approved by the National Health Commission of my country for direct human consumption. Although *Weizmannii coagulans* can promote human gut health and improve human immune function, and has been widely used in food, animal feed, and health products, there are currently no research reports on its direct biodegradation of inosine and guanosine, thereby lowering uric acid levels.

[0055] Based on long-term research in microorganisms, the inventors have successfully screened a highly efficient pure strain of microorganism capable of biodegrading inosine and guanosine from enzymes produced through traditional natural fermentation in Weifang, Shandong, China. This strain and its produced enzymes are capable of efficiently biodegrading inosine and guanosine, possessing significant research value and promising application prospects in the efficient biodegradation of inosine and guanosine, thereby reducing uric acid production. This invention is thus derived.

[0056] Therefore, the first aspect of the present invention provides a strain of Weizmannia coagulans NJ01 for the biodegradation of inosine and guanosine. Studies have found that the Weizmannia coagulans NJ01 strain can produce one or more enzymes that can catalyze the degradation of inosine and guanosine.

[0057] Therefore, the Wickeman's strain NJ01, which relates to the first aspect of the present invention for the biodegradation of uric acid precursors inosine and guanosine, is capable of producing enzymes that catalyze the degradation of inosine and guanosine.

[0058] The inventors first successfully screened a strain of *Weizmannia coagulans* NJ01 from enzymes produced through traditional Chinese natural fermentation. Genomic DNA was extracted, and molecular identification, including PCR amplification and 16S rRNA sequencing, confirmed its identity as *Weizmannia coagulans*. Based on this, the strain was identified and named *Weizmannia coagulans* NJ01. This strain has been deposited at the China General Microbiological Culture Collection Center (CGMCC) under accession number CGMCC No. 37022.

[0059] The inventors have discovered that fermentation culture of Weizmannii coagulans strain NJ01 produces bacterial cells containing one or more enzymes capable of catalyzing the degradation of inosine and guanosine. In this invention, the mixture of these enzymes is referred to as crude enzyme, or Weizmannii coagulans NJ01 crude enzyme.

[0060] Further research revealed that centrifugation of the cell-free lysate after bacterial cell disruption, and the collection of the supernatant cell-free extract, served as the crude enzyme for *Weizmannii coagulans* NJ01. This readily explains why both the cells and the crude enzyme of *Weizmannii coagulans* strain NJ01 can catalyze the degradation of inosine and guanosine.

[0061] The results showed that the cells of the *Weizmannii coagulans* strain NJ01 were able to completely degrade and remove inosine or guanosine at an initial concentration of 500 mg / L within 12 hours.

[0062] The results also showed that the crude enzyme produced by the *Weizmannii coagulans* strain NJ01 was able to completely degrade and remove inosine or guanosine at an initial concentration of 500 mg / L within 12 hours at a protein concentration of 1.8 g / L.

[0063] Based on the above, the second to fourth aspects of the present invention further provide the use or application of the *Weizmannii coagulating* bacteria described in the first aspect of the present invention for the biodegradation of inosine and guanosine.

[0064] Specifically, the second aspect of the present invention provides a biodegradable inosine and guanosine preparation of Weizmannii coagulans NJ01, which belongs to a microbial preparation for biodegrading inosine and guanosine, and contains bacterial cells and / or crude enzymes of Weizmannii coagulans NJ01 strain as described in the first aspect of the present invention.

[0065] In some preferred embodiments of the present invention, the *Weizmannia coagulans* NJ01 preparation contains bacterial cells of the *Weizmannia coagulans* NJ01 strain as described in the first aspect of the present invention.

[0066] According to some embodiments of the present invention, the biodegradable inosine and guanosine-coagulating Weizmannii NJ01 preparation is a liquid preparation.

[0067] For example, in some embodiments of the present invention, the concentration of bacterial cells coagulating Weizmannii strain NJ01 in the liquid formulation of the biodegraded inosine and guanosine is (1-10)×10⁻⁶. 8 / mL.

[0068] For example, in other embodiments of the invention, the protein concentration of the crude enzyme of *Weizmannia coagulans* strain NJ01 is 1-5 g / L in the liquid formulation of the biodegradable inosine and guanosine.

[0069] According to other embodiments of the present invention, the biodegradable inosine and guanosine-coagulating Weizmannii NJ01 formulation is a solid powder formulation.

[0070] For example, in some embodiments of the present invention, the bacterial cell content of *Weizmannii* strain NJ01 in the solid powder formulation of the biodegraded inosine and guanosine is (1-10) × 10⁻⁶. 8 / g, preferably (5-10)×10 8 / g.

[0071] For example, in other embodiments of the present invention, in the solid powder formulation of the biodegraded inosine and guanosine, the protein content of the crude enzyme of Weizmannia coagulans strain NJ01 is 1-5 g / kg, preferably 3-5 g / kg.

[0072] A third aspect of the present invention provides a method for preparing a biodegradable inosine and guanosine-coagulating *Weizmannii* NJ01 formulation as described in the second aspect of the present invention, comprising:

[0073] Step B: Inoculate the fermentation strain into the fermentation medium and ferment at 35-50℃, preferably 35-40℃, at a shaking speed of 100-300 rpm for 2-3 days to obtain the fermentation culture of Weizmannella coagulans strain NJ01.

[0074] Step C: Centrifuge the fermentation culture of Weizmannii NJ01 strain to harvest the bacterial cells of Weizmannii NJ01 strain.

[0075] The fermentation strain was obtained from the corresponding Weizmann's coagulans strain NJ01 through seed culture.

[0076] As is known to those skilled in the art, 16S rRNA is commonly used internationally for molecular identification. Therefore, 16S rRNA can be used to compare similarities and obtain homology. Thus, the fermentation strain used in this invention is not limited to the field isolates used in this invention. 16S rRNA is the DNA sequence on the bacterial chromosome that encodes rRNA and exists in the genomes of all microorganisms. Figure 1 A molecular phylogenetic tree based on 16S rRNA is shown for the Weizmannella coagulans strain NJ01.

[0077] In step C above, the centrifugation process includes washing the precipitate (i.e., the cells of strain NJ01 of Weizmannii coagulans) obtained by centrifugation of the liquid fermentation culture with physiological saline, followed by centrifugation to obtain the cells of strain NJ01 of Weizmannii coagulans.

[0078] The present invention does not impose any particular restrictions on the centrifugal separation conditions in step C above. In some embodiments of the present invention, for example, the analyte can be centrifuged for 10 minutes at 8000-10000 r / min.

[0079] According to the method of the present invention, the fermentation culture is a shaker or fermenter fermentation culture of the microbial strain, and the fermentation strain is inoculated into the fermentation medium in the form of a seed liquid. The inoculation amount of the seed liquid is 0.1%-1% (v / v), preferably 0.2%-0.5% (v / v), and more preferably 0.5% (v / v).

[0080] In some specific embodiments of the present invention, the above-mentioned fermentation culture medium, based on 1L of water, includes the following components in 1L of water:

[0081] 5-10g of peptone;

[0082] 5-10 g of yeast powder; and

[0083] 3-8 g of glucose;

[0084] Preferably, the above fermentation medium, based on 1L of water, includes the following components in 1L of water:

[0085] 8-10 g of peptone;

[0086] 8-10 g of yeast powder; and

[0087] 6-8 g of glucose;

[0088] More preferably, the initial pH of the fermentation medium is adjusted to 6-7 using a 40% (wt / v) sodium hydroxide solution and a 36% (v / v) hydrochloric acid solution.

[0089] According to some embodiments of the present invention, the preparation method of the *Weizmannii coagulans* NJ01 preparation involved in the present invention further includes step A, which involves seed culture before step B: picking a single colony of the *Weizmannii coagulans* NJ01 strain provided by the present invention and inoculating it into 100 mL of fermentation liquid culture medium, and then culturing it in a shaker at 40°C and 200 r / min for 3 days to obtain the fermentation strain (seed liquid).

[0090] The inventors studied the effect of different temperatures on the growth of *Weizmannia coagulans* NJ01 and found that *Weizmannia coagulans* NJ01 grows rapidly at a temperature of 40℃.

[0091] According to some specific embodiments of the present invention, the preparation method further includes:

[0092] Step K: The cell suspension of the *Weizmannia coli* strain NJ01 is subjected to cell disruption treatment in an ice-water bath (i.e., an ice-water mixture, 0-4°C) to obtain a cell-free lysate of the *Weizmannia coli* strain NJ01.

[0093] Step L: The cell-free lysate of Weizmannia coagulans strain NJ01 is centrifuged, and the supernatant cell-free extract is used as the crude enzyme of Weizmannia coagulans strain NJ01.

[0094] The present invention does not impose any particular restrictions on the centrifugal separation conditions in step L above. In some embodiments of the present invention, for example, the analyte can be centrifuged at 15000-18000 r / min for 10-20 min.

[0095] The fourth aspect of this invention provides the use of the *W. coagulation* NJ01 preparation of biodegradable inosine and guanosine as described in the second aspect of this invention, or the *W. coagulation* NJ01 preparation of biodegradable inosine and guanosine prepared by the preparation method described in the third aspect of this invention, in the preparation of a drug that reduces inosine and guanosine and thereby reduces uric acid production, comprising:

[0096] Step D: Wash the cells of strain NJ01 of Weizmannia coagulans with physiological saline to obtain a pure cell product of strain NJ01 of Weizmannia coagulans.

[0097] Step E: In a physiological saline system, under low temperature conditions of 0-4℃, ultrasonically disrupt the pure bacterial cells of strain NJ01 of coagulated Weizmannii. After centrifugation, take the supernatant to obtain cell-free extract as crude enzyme pure of strain NJ01 of coagulated Weizmannii.

[0098] Step F involves freeze-drying the pure bacterial cells and / or crude enzyme of *Weizmannia coagulans* strain NJ01, and then diluting the freeze-dried *Weizmannia coagulans* NJ01 preparation to prepare a drug for inosine and guanosine.

[0099] In some embodiments of the present invention, in step F, the freeze-dried *Weizmannia coli* NJ01 preparation is diluted with physiological saline to prepare liquid isosorbide and guanosine preparations.

[0100] In some other embodiments of the present invention, in step F, the freeze-dried *Weizmannia coagulans* NJ01 preparation is diluted with edible starch to prepare solid inosine and guanosine preparations.

[0101] In some preferred embodiments of the present invention, the inosine and guanosine preparations are oral formulations.

[0102] III. Related materials and testing methods in this invention

[0103] 1. Materials

[0104] The enzyme samples of traditional Chinese fermented food involved in this invention were purchased in Shandong, China, and were licensed for use in scientific research activities.

[0105] 2. Detection Method

[0106] (1) The cell concentration in this invention is determined using the following method:

[0107] To determine the concentration of Weizmann's coagulans NJ01 cells, Weizmann's coagulans NJ01 culture was diluted with physiological saline and the cell concentration was directly measured using a flow cytometer (SYSMEX, Germany).

[0108] (2) The concentrations of inosine and guanosine in this invention are determined using the following method:

[0109] The method for determining the concentrations of inosine and guanosine involves dissolving the inosine and guanosine in pure aqueous solution according to the specified ratio, filtering the solution through a 0.22 μm filter membrane, and then determining the concentrations of inosine and guanosine in the liquid culture medium using a RID-20A high-performance liquid chromatograph (Shimadzu).

[0110] (3) The crude enzyme protein concentration in this invention is determined using the following method:

[0111] Cell-free extract of *Weizmannii coagulates* NJ01 was diluted with phosphate buffer solution and then Coomassie Brilliant Blue G-250 dye reagent was added in proportion and reacted for 10 minutes. The absorbance was measured at 595 nm using a 722S visible spectrophotometer (Shanghai Lingguang), and the protein concentration was calculated using the standard curve method.

[0112] IV. Examples

[0113] The present invention will be specifically described below through specific embodiments. Unless otherwise specified, the experimental methods described below are standard laboratory methods. Unless otherwise specified, the experimental materials described below are commercially available.

[0114] Example 1: Preparation of bacterial cells and crude enzymes of *Weizmannii coagulans* strain NJ01

[0115] (1) Prepare the growth medium for *Weizmannii coagulans* NJ01, with the following composition (per liter): 10.0 g peptone, 10.0 g yeast extract, and 5.0 g glucose. Add 100 mL of the prepared liquid culture medium to a 500 mL Erlenmeyer flask, sterilize under high temperature and high pressure (121 °C) for 20 minutes, and then sterilize again under ultraviolet irradiation in a clean workbench for 20 minutes.

[0116] (2) Under sterile conditions in a clean workbench, 0.5 mL of Weizmannia coagulans NJ01 bacterial suspension was inoculated into a triangular flask of liquid culture medium. After two days of batch culture at 40°C and 200 rpm, Weizmannia coagulans NJ01 cells were harvested by centrifugation (8000 rpm, 10 minutes) and discarding the supernatant.

[0117] Figure 1 The strain we screened was most closely related to *Weizmannia coagulans*, and was therefore named *Weizmannia coagulans* strain NJ01.

[0118] (3) Take 20 mL of the suspension of *Weizmannii coagulans* NJ01 cells and add it to a 50 mL glass tube. Then, insert the tube into ice water and use an ultrasonic cell disruptor to disrupt the *Weizmannii coagulans* NJ01 cells. The conditions are: ultrasonic power 400 W, interval of 2 seconds, ultrasonic oscillation for 10 seconds, and disruption time of 15 minutes (5 minutes each time). After the cells are disrupted, centrifuge the cell disruption solution at 15,000 rpm for 20 minutes, and then slowly pour out the supernatant as the cell-free extract (crude enzyme) of *Weizmannii coagulans* NJ01.

[0119] Example 2: Biodegradation of inosine and guanosine by *Weizmannii* strain NJ01 cells and crude enzymes

[0120] Based on different concentrations of inosine and guanosine, *Weizmannii coagulans* NJ01 cells and crude enzymes, prepared through culture, were added in a specific ratio as a rapid, safe, and efficient biocatalyst to achieve the rapid and efficient degradation and removal of inosine and guanosine. The results are as follows: Figure 2 and Figure 3 As shown.

[0121] Figure 2 This indicates that within 12 hours, the concentration of *Weizmannii* NJ01 cells decreased from 4.0 × 10⁻⁶. 6 / mL grew to 1.0×10 8 The strain can completely degrade inosine and guanosine at an initial concentration of 500 mg / L, indicating that *Weizmannii coagulans* NJ01 has a strong biodegradation ability for inosine and guanosine.

[0122] Figure 3 The results indicate that the cell-free extract (crude enzyme) of *Weizmannii coagulates* NJ01 can catalyze the degradation of inosine and guanosine. At a protein concentration of 1.8 g / L, it can completely degrade inosine and guanosine at an initial concentration of 500 mg / L in 12 hours.

[0123] It should be noted that the embodiments described above are merely preferred embodiments of the present invention, used to explain the present invention, and do not constitute any limitation on the present invention. The present invention has been described with reference to typical embodiments, but it should be understood that the terms used therein are descriptive and explanatory terms, not limiting terms. Modifications can be made to the present invention within the scope of the claims, and revisions can be made to the present invention without departing from the scope and spirit of the present invention. Although the present invention described herein relates to specific methods, materials, and embodiments, it does not mean that the present invention is limited to the specific examples disclosed herein; on the contrary, the present invention can be extended to all other methods and applications having the same function.

Claims

1. A strain of Weizmannia coagulans NJ01 for the biodegradation of inosine and guanosine, which produces an enzyme that catalyzes the degradation of inosine and guanosine, and its accession number is CGMCC No.37022.

2. A *Weizmannii coagulans* NJ01 formulation for the biodegradation of inosine and guanosine, comprising bacterial cells and / or crude enzymes of the *Weizmannii coagulans* NJ01 strain as described in claim 1.

3. The *Wietzmannii coagulans* NJ01 preparation according to claim 2, characterized in that, The *Wilhelminthes globulus* NJ01 formulation for degrading inosine and guanosine is a liquid formulation; in the liquid formulation for degrading inosine and guanosine, the cell concentration of *Wilhelminthes globulus* NJ01 strain is (1-10)×10⁻⁶. 8 / mL; and / or, in the liquid formulation for degrading inosine and guanosine, the protein concentration of the crude enzyme of *Weizmannia coagulans* strain NJ01 is 1-5 g / L.

4. The *Wietzmannii coagulans* NJ01 preparation according to claim 2, characterized in that, The *Wilhelminthes globulus* NJ01 formulation for degrading inosine and guanosine is a solid powder formulation; in the solid powder formulation for degrading inosine and guanosine, the cell concentration of *Wilhelminthes globulus* NJ01 strain is (1-10)×10⁻⁶. 8 / g; and / or, in the solid powder formulation of the degraded inosine and guanosine, the protein content of the crude enzyme of *Weizmannia coagulans* strain NJ01 is 1-5 g / kg.

5. The *Weizmannii coagulans* NJ01 preparation according to claim 4, characterized in that, In the solid powder formulation of degraded inosine and guanosine, the cell concentration of *Weizmannii* strain NJ01 is (5-10)×10⁻⁶. 8 / g; and / or, in the solid powder formulation of the degraded inosine and guanosine, the protein content of the crude enzyme of *Weizmannia coagulans* strain NJ01 is 3-5 g / kg.

6. A method for preparing a *Wietzmannii* NJ01 formulation that degrades inosine and guanosine as described in any one of claims 2-5, comprising: Step B: Inoculate the fermentation strain into the culture medium for fermentation culture to obtain the fermentation culture of Weizmannella coagulans strain NJ01. Step C: Centrifuge the fermentation culture of Weizmannii NJ01 strain to harvest the bacterial cells of Weizmannii NJ01 strain. The fermentation strain was obtained from the corresponding Weizmann's coagulans strain NJ01 through seed culture.

7. The preparation method according to claim 6, characterized in that, The fermentation medium, calculated per liter of water, comprises the following components per liter of water: 5-10g of peptone; preferably 8-10g; 5-10 g of yeast powder; preferably 8-10 g; as well as 3-8 g of glucose; preferably 6-8 g; The pH value of the fermentation medium is 6-7; In step B, the fermentation culture temperature is 35-50℃, preferably 35-40℃.

8. The preparation method according to any one of claims 6-7, characterized in that, The preparation method further includes: Step K: The cell suspension of the *Weizmannia coagulans* strain NJ01 is subjected to cell disruption treatment under low temperature conditions to obtain a cell-free lysate of the *Weizmannia coagulans* strain NJ01. Step L: Centrifuge the cell-free lysate of strain NJ01 of Weizmannia coagulans and take the supernatant cell-free extract as the crude enzyme of strain NJ01 of Weizmannia coagulans. The low temperature is 0-4℃.

9. The use of a *Weizmannii coagulans* NJ01 preparation for degrading inosine and guanosine as described in any one of claims 2-5, or a *Weizmannii coagulans* NJ01 preparation prepared by any one of claims 6-8, in the preparation of inosine and guanosine-lowering agents, comprising: Step D: Wash the cells of strain NJ01 of Weizmannia coagulans with physiological saline to obtain a pure cell product of strain NJ01 of Weizmannia coagulans. Step E: In a physiological saline system, under low temperature conditions, ultrasonically disrupt the pure bacterial cells of strain NJ01 of Weizmannia coagulation. After centrifugation, take the supernatant to obtain cell-free extract as crude pure enzyme of strain NJ01 of Weizmannia coagulation. Step F involves freeze-drying the pure bacterial cells and / or crude enzyme of the Weizmannii coagulans strain NJ01, and then diluting the freeze-dried Weizmannii coagulans NJ01 preparation to prepare a drug for degrading inosine and guanosine. The low temperature is 0-4℃.

10. The application according to claim 9, characterized in that, In step F, the freeze-dried *Weizmannii coagulans* NJ01 preparation is diluted with physiological saline to prepare liquid inosine and guanosine preparations; the prepared liquid inosine and guanosine preparations are oral formulations. Alternatively, in step F, the freeze-dried *Weizmannia coagulans* NJ01 preparation is diluted with edible starch to prepare solid inosine and guanosine preparations; the solid inosine and guanosine preparations are oral preparations.

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