CN119040210A

CN119040210A - Lactobacillus casei YWC-01 preparation for degrading uric acid and application thereof

Info

Publication number: CN119040210A
Application number: CN202411461982.4A
Authority: CN
Inventors: 闫海; 吴安情
Original assignee: Beijing Beike Yiran Biotechnology Co ltd
Current assignee: Beijing Beike Yiran Biotechnology Co ltd
Priority date: 2024-10-18
Filing date: 2024-10-18
Publication date: 2024-11-29

Abstract

The present invention belongs to the field of biotechnology, and relates to a Lactobacillus casei YWC-01 strain for degrading uric acid, which can produce an enzyme that catalyzes the degradation of uric acid. The present invention also relates to a Lactobacillus casei YWC-01 preparation for degrading uric acid, which contains bacterial cells and/or crude enzymes of Lactobacillus casei YWC-01 strains. The results of the study show that the Lactobacillus casei YWC-01 strain for biodegrading uric acid provided by the present invention and the enzyme produced therefrom are safe for the human body, and can efficiently biodegrade uric acid, and at an initial concentration of 8×10 ⁸ /mL of bacterial cells, uric acid with an initial concentration of 1g/L can be completely degraded and removed within 48 hours; at a crude enzyme protein concentration of 0.8g/L, uric acid with an initial concentration of 1g/L can be completely removed within 18 hours. Lactobacillus casei YWC-01 strain and the enzyme produced therefrom have important application prospects in the prevention and treatment of gout and corresponding complications.

Description

Lactobacillus casei YWC-01 preparation for degrading uric acid and application thereof

Technical Field

The invention belongs to the technical field of biology, and relates to a preparation of cheese bacillus YWC-01 for degrading uric acid, and a preparation method and application thereof.

Background

Hyperuricemia (Hyperuricemia, HUA) refers to the fact that under normal purine dietary conditions, fasting blood uric acid levels twice a day exceed the normal range, i.e., adult male blood uric acid levels are greater than 420. Mu. Mol/L, and female blood uric acid levels are greater than 360. Mu. Mol/L. According to the saturated solubility of uric acid in blood, above the above concentration, uric acid is liable to form crystals and deposit in tissues to form tophus, resulting in severe pain. At present, HUA patients in China are increased sharply, and serious threat is formed to the health of people.

In humans, intermediates in the purine metabolic pathway, such as inosine and guanosine, can be further metabolically converted to uric acid as the final product. Normally, about two-thirds of uric acid is excreted by the kidneys, with most of the uric acid being reabsorbed in the proximal tubules after filtration through the glomeruli, and then finally excreted by the urine through the process of secretion and reabsorption. Another about one third of uric acid is converted into allantoin in the intestine by the decomposition of intestinal bacteria and then discharged outside the body with feces. Since humans lack urate oxidase, uric acid cannot be degraded into more soluble allantoin, which hinders uric acid excretion, resulting in blood uric acid levels 3-10 times higher than in non-primate animals, which is also one of the important reasons for human susceptibility to HUA.

Currently, the treatment or improvement modes of HUA are mainly divided into medication and dietary intervention, wherein medication is a main mode, and commonly used drugs include allopurinol, febuxostat, benzbromarone, urate oxidase (labyrine) and the like, and mainly control blood uric acid level by inhibiting the activity of xanthine oxidase to reduce uric acid production or reducing uric acid reabsorption to increase uric acid excretion. The main mechanism of action of urate oxidase is to further degrade uric acid into more soluble allantoin, which is easier to excrete from the kidney. At present, although nucleoside substances such as inosine and the like are reported by the cheese bacillus, no research report exists in the aspect of the cheese bacillus for degrading uric acid. Although chemical drugs can play a role in controlling HUA development, a series of side effects such as anaphylaxis, liver injury, renal dysfunction, etc. are not neglected. Dietary intervention is mainly aimed at alleviating HUA by limiting the intake of high purine foods such as beer and fructose, and although blood uric acid levels can be controlled to some extent, the effect is not ideal. Therefore, research on a method for controlling HUA by removing uric acid more safely and efficiently has become a key scientific research problem to be solved urgently in the field of biological medicine at present.

Disclosure of Invention

Aiming at the problems of insignificant effect and side effects of medicines or foods for degrading and removing uric acid in the prior art, the invention provides a cheese bacillus YWC-01 strain for degrading uric acid, bacterial cells of the strain and enzymes produced by the bacterial cells are safe for human bodies, can efficiently biodegrade uric acid, and has important application prospects.

The invention also aims to provide a preparation of the cheese bacillus YWC-01 for degrading uric acid and application thereof, wherein the preparation of the cheese bacillus YWC-01 is prepared from the cheese bacillus YWC-01 strain for biodegrading uric acid, and can biodegrade uric acid with high efficiency.

To this end, a first aspect of the present invention provides a strain of uric acid degrading Lactobacillus casei (Lacticaseibacillus casei) YWC-01 capable of producing an enzyme for catalytic degradation of uric acid with the accession number CGMCC No.31534.

In some embodiments of the invention, the Lactobacillus casei YWC-01 strain is capable of degrading and removing uric acid at an initial concentration of 1g/L in 48 hours at an initial concentration of 8X 10 ⁸/mL of cells.

In other embodiments of the invention, the crude enzyme produced by the Lactobacillus casei YWC-01 strain is capable of completely removing uric acid at an initial concentration of 1g/L in 18 hours at a protein concentration of 0.8 g/L.

In a second aspect the present invention provides a preparation of uric acid degrading Lactobacillus casei YWC-01 comprising bacterial cells and/or crude enzyme of the strain of Lactobacillus casei YWC-01 as described in the first aspect of the invention, preferably the preparation of Lactobacillus casei YWC-01 comprises the strain of Lactobacillus casei YWC-01 as described in the first aspect of the invention.

In some embodiments of the invention, the uric acid-degrading preparation of Lactobacillus casei YWC-01 is a liquid preparation, preferably, the concentration of bacterial cells of Lactobacillus casei YWC-01 strain is (1-10) x 10 ⁸/mL in the uric acid-degrading liquid preparation, and/or the concentration of protein of crude enzyme of Lactobacillus casei YWC-01 strain is 0.1-1.0g/L in the uric acid-degrading liquid preparation.

In other embodiments of the present invention, the uric acid-degrading solid powder preparation of Lactobacillus casei YWC-01 is preferably one in which the content of cells of Lactobacillus casei YWC-01 strain is (1-10). Times.10 ⁸/g, more preferably (8-10). Times.10 ⁸/g, and/or in which the protein content of crude enzyme of Lactobacillus casei YWC-01 strain is 0.1-1.0g/kg, more preferably 8-10g/kg.

In a third aspect, the present invention provides a process for the preparation of a uric acid degrading preparation of Lactobacillus casei YWC-01 as defined in the second aspect of the invention, comprising:

Step B, inoculating a fermentation strain into a fermentation medium for fermentation culture to obtain a fermentation culture of a cheese bacillus YWC-01 strain;

Step C, carrying out centrifugal separation treatment on a fermentation culture of the Lactobacillus casei YWC-01 strain, and obtaining bacterial cells of the Lactobacillus casei YWC-01 strain;

wherein the fermentation strain is obtained by seed culture of corresponding cheese bacillus YWC-01 strain.

According to the invention, the fermentation medium comprises the following components in 1L of water, calculated as 1L of water:

Preferably, the pH of the fermentation medium is 7-8;

further preferably, in step B, the temperature of the fermentation culture is 30-40 ℃, preferably 38-40 ℃.

According to some embodiments of the invention, the method further comprises:

Step K, performing cell disruption treatment on the cell suspension of the Lactobacillus casei YWC-01 strain at a low temperature to obtain a cell-free disruption solution of the Lactobacillus casei YWC-01 strain;

step L, performing centrifugal separation on a cell-free disrupted solution of the Lactobacillus casei YWC-01 strain, and taking a supernatant cell-free extract as crude enzyme of the Lactobacillus casei YWC-01 strain;

Wherein the low temperature is 0-4 ℃.

According to a fourth aspect of the present invention there is provided the use of a preparation of uric acid degrading Lactobacillus casei YWC-01 as described in the second aspect of the present invention or a preparation of uric acid degrading Lactobacillus casei YWC-01 as described in the third aspect of the present invention in the manufacture of a uric acid lowering medicament comprising:

step D, washing bacterial cells of the cheese bacillus YWC-01 strain by using physiological saline to obtain a bacterial cell pure product of the cheese bacillus YWC-01 strain;

Step E, in a physiological saline system, under the low-temperature condition, adopting ultrasonic waves to crush a bacterial cell pure product of the cheese bacillus YWC-01 strain, and taking supernatant after centrifugal treatment to obtain a cell-free extract as a crude enzyme pure product of the cheese bacillus YWC-01 strain;

Step F, freeze-drying a bacterial cell pure product and/or a crude enzyme pure product of the Lactobacillus casei YWC-01 strain, and diluting a freeze-dried Lactobacillus casei YWC-01 preparation to prepare a medicament for degrading uric acid;

Wherein the low temperature is 0-4 ℃.

In some embodiments of the invention, in step F, the lyophilized preparation of Lactobacillus casei YWC-01 is diluted with physiological saline to form a liquid uric acid lowering agent.

In other embodiments of the present invention, in step F, the lyophilized preparation of Lactobacillus casei YWC-01 is diluted with an edible starch to form a solid uric acid lowering agent.

In some preferred embodiments of the invention, the uric acid lowering agent is an oral formulation.

The research shows that the cheese bacillus YWC-01 strain for biologically degrading uric acid and the enzyme produced by the strain are safe to human bodies, can biologically degrade uric acid, and have important application prospects in the aspect of efficiently removing uric acid for treating human HUA and corresponding complications.

Drawings

In order that the invention may be readily understood, the invention will be described with reference to the accompanying drawings.

FIG. 1 shows a molecular evolution tree based on 16S rDNA for screening of Lactobacillus casei YWC-01.

FIG. 2 is a kinetic profile of the biological degradation of uric acid by Lactobacillus casei YWC-01.

FIG. 3 is a kinetic profile of the enzymatic degradation of uric acid by Lactobacillus casei YWC-01.

Preservation of bacterial species

Cheese bacillus (Lacticaseibacillus casei) isolated and identified by Beijing Beike Yi-ran biotechnology limited company has been preserved in China general microbiological culture Collection center (CGMCC; address: china center for general microbiological study, no.3, xila 1, no.3, chaoyang area North Star, beijing, with a preservation date of 2024, 8 months, 2 days, and a preservation number of CGMCC No.31534. The strain of the present invention is designated as Lactobacillus casei YWC-01 strain (Lacticaseibacillus CASEI STRAIN YWC-01), which is also called Lactobacillus casei YWC-01.

Detailed Description

In order that the invention may be readily understood, the invention will be described in detail. Before the present invention is described in detail, it is to be understood that this invention is not limited to particular embodiments described. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

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

I. Terminology

The term "cell" as used herein refers to a living and/or dead cell of Lactobacillus casei.

The term "crude enzyme" as used herein refers to a cell-free extract obtained by disrupting cells of Lactobacillus casei and then centrifuging to obtain a supernatant.

The term "crude enzyme purified product" as used herein refers to a cell-free extract obtained by disrupting a bacterial cell purified product of Lactobacillus casei and then centrifuging to obtain a supernatant, relative to crude enzyme.

The term "microbial preparation" in the present invention refers to a preparation which is produced by using a microorganism having medical research value as a raw material and is used for preventing (health care), treating and diagnosing various physiological symptoms of human body by using a traditional technology or a modern biotechnology.

The term "edible starch" as used herein refers to starch which meets the national standard of edible starch (GB 31637-2016 national standard of food safety edible starch).

The term "water" used in the culture medium or the fermentation culture process of the invention refers to sterile pure water obtained by filtration through a 0.22. Mu. Filter, unless otherwise specified.

II. Description of the embodiments

As previously mentioned, existing dietetic and pharmaceutical therapies for the control of uric acid in humans have slow and insignificant efficacy and side effects. Accordingly, the present inventors have conducted extensive and intensive studies on the biodegradation of uric acid.

The inventors noted that although lactic acid bacteria promote intestinal health and reduce uric acid accumulation to some extent through a series of metabolic regulation, the strain used has not been reported to biodegrade uric acid directly and efficiently.

In recent years, research on human microorganisms is a research hotspot and a front edge of life science, and although domestic and foreign scholars screen microorganism strains capable of inhibiting uric acid synthesis and degrading uric acid precursors inosine, guanosine and the like from traditional foods, no evidence that cheese bacillus can directly and efficiently biodegrade uric acid is found.

The inventor also notices that the cheese bacillus (Lacticaseibacillus casei) belongs to prokaryotic microorganisms, is one of probiotic bacteria which can be directly eaten by people who pass the examination and approval of Wei Jian in China, has the functions of promoting the intestinal health of human beings and improving the immunity of human bodies, is widely applied to the fields of food, fermentation, beer and beverage production and the like, but has not been reported in research on the biodegradation of uric acid so far.

Based on the long-term microbial study, the inventor successfully screens a cheese bacillus from ferment (Lacticaseibacillus casei), and the inventor discovers that the strain cells and the generated enzyme can degrade uric acid with initial concentration of 1g/L in 48h and 18h respectively through research and exploration, which has very important research value and important application prospect in the aspect of high-efficiency biodegradation for removing uric acid. The present invention has been achieved thereby.

Thus, the Lactobacillus casei YWC-01 strain for biodegradation of uric acid according to the first aspect of the present invention is capable of producing an enzyme catalyzing the degradation of uric acid.

The inventors first successfully screened a strain of Lactobacillus casei YWC-01 from the ferments. The genomic DNA was extracted, and the resultant was amplified by PCR and identified as Lactobacillus casei (Lacticaseibacillus casei) by molecular identification based on 16S rDNA sequencing, and based on the above, the strain was identified and designated as Lactobacillus casei YWC-01 strain (Lacticaseibacillus CASEI STRAIN YWC 01). The strain is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.31534.

The present inventors have found that the fermentation culture of Lactobacillus casei YWC-01 strain produces yeast cells containing one or more enzymes capable of catalyzing the degradation of uric acid, and that the enzyme mixture is referred to as crude enzyme or Lactobacillus casei YWC-01 crude enzyme in the present invention.

Further studies have found that the cell-free disrupted solution after disruption of the cells of Lactobacillus casei YWC-01 is centrifuged and the supernatant cell-free extract is taken as crude enzyme of Lactobacillus casei YWC-01. It is thus readily understood that both the cells of the Lactobacillus casei YWC-01 strain and the crude enzyme are capable of catalyzing the degradation of uric acid.

The research result shows that the Lactobacillus casei YWC-01 strain can degrade and remove uric acid with the initial concentration of 1g/L completely within 48 hours under the initial concentration of 8X 10 ⁸/mL of bacterial cells.

The crude enzyme produced by the Lactobacillus casei YWC-01 strain can degrade uric acid with the initial concentration of 1g/L in 18 hours at the protein concentration of 0.8 g/L.

Based on the above, the second to fourth aspects of the present invention further provides the use or application of the cheese bacillus for biodegradation of uric acid according to the first aspect of the present invention.

In particular, the second aspect of the invention provides a preparation of Lactobacillus casei YWC-01 for biodegradation of uric acid, which belongs to a microbial preparation for biodegradation of uric acid, and comprises bacterial cells and/or crude enzymes of the Lactobacillus casei YWC-01 strain according to the first aspect of the invention.

In some preferred embodiments of the invention, the preparation of Lactobacillus casei YWC-01 comprises bacterial cells of the strain Lactobacillus casei YWC-01 as described in the first aspect of the invention.

According to some embodiments of the invention, the uric acid-biodegradable formulation of Lactobacillus casei YWC-01 is a liquid formulation.

In some embodiments of the invention, the concentration of bacterial cells of strain YWC-01 of Lactobacillus casei is (1-10). Times.10 ⁸/mL in the liquid formulation for biodegradation of uric acid.

In other embodiments of the invention, the crude enzyme of strain YWC-01 of Lactobacillus casei has a protein concentration of 0.1-1.0g/L in the liquid formulation for biodegradation of uric acid.

According to further embodiments of the present invention, the uric acid biodegradable formulation of Lactobacillus casei YWC-01 is a solid powder formulation.

In some embodiments of the present invention, the solid powder formulation for biodegradation of uric acid has a bacterial cell content of Lactobacillus casei YWC-01 strain of (1-10). Times.10 ₈/g, preferably (8-10). Times.10 ⁸/g.

In other embodiments of the present invention, the crude enzyme of Lactobacillus casei YWC-01 strain has a protein content of 0.1-1.0g/kg (mg/g), preferably 0.8-1.0g/kg, in the solid powder formulation for biodegradation of uric acid.

In a third aspect, the present invention provides a process for the preparation of a preparation of uric acid-biodegradable Lactobacillus casei YWC-01 as described in the second aspect of the invention, comprising:

step B, inoculating a fermentation strain into a fermentation medium, and standing and fermenting at 30-40 ℃, preferably 38-40 ℃ for 3-5 days to obtain a fermentation culture of the Lactobacillus casei YWC-01 strain;

As known to those skilled in the art, at present, 16S rRNA is commonly used internationally for molecular identification of prokaryotic microorganisms, and thus 16S rRNA can be used for comparison of similarity to obtain homology. The fermentation strain used in the present invention is not limited to the field isolate used in the present invention, and 16S rDNA is a DNA sequence corresponding to the encoded rRNA on the bacterial chromosome, and is present in the genome of all prokaryotic microorganisms. FIG. 1 shows the molecular evolution tree of Lactobacillus casei YWC-01 based on 16S rDNA.

In the step C, the centrifugal separation treatment comprises the steps of carrying out centrifugal separation treatment on the sediment (namely, bacterial cells of the Lactobacillus casei YWC-01 strain) obtained by the liquid fermentation culture, carrying out resuspension washing by using normal saline, and carrying out centrifugal separation treatment to obtain bacterial cells of the Lactobacillus casei YWC-01 strain.

The conditions for the centrifugation in the above step C are not particularly limited in the present invention, and in some embodiments of the present invention, for example, the to-be-separated substance may be centrifuged at 8000-10000r/min for 10min.

According to the method of the invention, the fermentation culture is standing of strain or fermentation culture in a fermentation tank, and the fermentation strain is inoculated into a fermentation culture medium in the form of seed liquid. The seed liquid is inoculated in an amount of 1 to 10% (v/v), preferably 5 to 10% (v/v), and more preferably 10% (v/v).

Specifically, the fermentation medium comprises the following components in 1L of water, based on 1L of water:

preferably, the fermentation medium comprises the following components in 1L of water, based on 1L of water:

in some embodiments of the invention, the initial pH of the fermentation medium is adjusted to 7-8 using 40% (wt/v) sodium hydroxide solution and 36% (v/v) hydrochloric acid solution.

According to some embodiments of the invention, the preparation method of the preparation of the cheese bacillus YWC-01 further comprises a step A of seed culture before the step B, wherein the step A is to pick up the single colony of the cheese bacillus YWC-01 provided by the invention, inoculate the single colony into 100mL of fermentation liquid culture medium, and perform stationary culture for 3 days at the temperature of 38 ℃ to obtain a fermentation strain (seed liquid).

The inventors studied the effect of different temperatures on the growth of Lactobacillus casei YWC-01 and found that Lactobacillus casei YWC-01 grows fast at a temperature of 38 ℃.

According to some embodiments of the invention, the method further comprises:

Step K, performing cell disruption treatment on the cell suspension of the Lactobacillus casei YWC-01 strain in an ice water bath (namely, an ice water mixture at 0-4 ℃) to obtain a cell-free disruption solution of the Lactobacillus casei YWC-01 strain;

And step L, carrying out centrifugal separation on the acellular disruption solution of the strain YWC-01 of the cheese bacillus, and taking the supernatant acellular extraction solution as crude enzyme of the strain YWC-01 of the cheese bacillus.

The conditions for the centrifugation in the above step L are not particularly limited in the present invention, and in some embodiments of the present invention, for example, the to-be-separated substance may be centrifuged at 15000-18000r/min for 10-20min.

According to a fourth aspect of the present invention there is provided the use of a preparation of uric acid-biodegradable Lactobacillus casei YWC-01 as described in the second aspect of the present invention or a preparation of uric acid-biodegradable Lactobacillus casei YWC-01 as described in the third aspect of the present invention in the preparation of a uric acid-lowering medicament comprising:

step E, in a physiological saline system, under the low temperature condition of 0-4 ℃, adopting ultrasonic waves to crush a bacterial cell pure product of the cheese bacillus YWC-01 strain, and taking a supernatant after centrifugal treatment to obtain a cell-free extract as a crude enzyme pure product of the cheese bacillus YWC-01 strain;

And F, freeze-drying the pure bacterial cell product and/or crude enzyme product of the Lactobacillus casei YWC-01 strain, and diluting the freeze-dried Lactobacillus casei YWC-01 preparation to prepare the uric acid reducing medicament.

III, related materials and detection methods in the present invention

1. Material

The ferment is prepared by taking apples as raw materials and fermenting the apples by natural strains.

2. Detection method

(1) The cell concentration in the invention is determined by the following method:

measurement of the cell concentration of Lactobacillus casei YWC-01 the culture of Lactobacillus casei YWC-01 was diluted with physiological saline to a multiple and the cell concentration was measured directly by flow cytometry (SYSMEX, germany).

(2) The uric acid concentration in the invention is measured by the following method:

the uric acid concentration is measured by a high performance liquid chromatograph (Shimadzu LC-20 AT), specifically a chromatographic column, namely Kromasil C18 (4.6X250 mm,5 mu particle diameter), a mobile phase, namely methanol, 0.5% acetic acid aqueous solution (10:90), the detection wavelength is 283nm, the flow rate is 1mL/min, the sample injection amount is 20 mu L, and the temperature is 35 ℃.

(3) The concentration of crude enzyme protein in the invention is determined by the following method:

Taking a cell-free extract of cheese bacillus YWC-01, diluting by a certain multiple by a phosphate buffer solution, adding a coomassie brilliant blue G-250 dye reagent proportionally for reacting for 10 minutes, measuring absorbance at 595nm by using a 722S visible spectrophotometer (Shanghai prismatic light), and calculating the protein concentration by using a standard curve method.

III, examples

The present invention will be specifically described below by way of specific examples. The experimental methods described below, unless otherwise specified, are all laboratory routine methods. The experimental materials described below, unless otherwise specified, are commercially available.

Example 1:

(1) A growth medium for Lactobacillus casei YWC-01 was prepared, which consisted of (per liter) peptone 10g, beef extract 10g yeast powder 10g, glucose 10g, and an initial pH of 7.5 was adjusted. 100 ml of the prepared liquid medium was added to a 500 ml Erlenmeyer flask, sterilized at high temperature and high pressure (115 ℃) for 30 minutes, and then sterilized under ultraviolet irradiation in a clean bench for 20 minutes.

(2) Under the aseptic condition in a clean bench, 10 milliliters of cheese bacillus YWC-01 bacteria liquid is inoculated into a triangular flask liquid culture medium, and after standing culture is carried out for 3 days at the temperature of 38 ℃, the cheese bacillus YWC-01 cells are obtained by a method of pouring out supernatant after centrifugation (8000 revolutions per minute, 10 minutes).

20ML of the Lactobacillus casei YWC-01 cell suspension is taken and added into a 50mL glass tube, and then the glass tube is inserted into ice water, and the Lactobacillus casei YWC-01 cells are crushed by an ultrasonic cell crusher under the conditions that the ultrasonic power is 400W, the interval is 2 seconds, the ultrasonic vibration is 10 seconds, and the crushing time is 15 minutes (5 minutes each time). After completion of cell disruption, the cell disruption solution was centrifuged at 15000 rpm for 20 minutes, and then the supernatant was slowly poured out as a cell-free extract (crude enzyme) of Lactobacillus casei YWC-01.

(3) According to different concentrations of uric acid, the cultured and prepared cheese bacillus YWC-01 cells and crude enzyme are used as a quick, safe and efficient biocatalyst, and are added according to a certain proportion, so that the aim of quickly and efficiently degrading and removing uric acid is fulfilled.

FIG. 1 shows that the strain we screened has recently related to Lactobacillus casei and is therefore designated Lactobacillus casei YWC-01 strain.

FIG. 2 shows that uric acid having an initial concentration of 1g/L can be completely degraded within 48 hours at an initial cell concentration of 8X 10 ⁸/mL of Lactobacillus casei YWC-01, indicating that Lactobacillus casei YWC-01 has a strong biodegradability for uric acid.

FIG. 3 shows that the cell-free extract (crude enzyme) of Lactobacillus casei YWC-01 is capable of catalyzing and degrading uric acid at a faster rate, and at a protein concentration of 0.8g/L, uric acid with an initial concentration of 1g/L can be completely degraded in 18 hours, and a higher uric acid degradation rate is achieved.

It should be noted that the above-mentioned embodiments are only preferred embodiments of the present invention, and are used for explaining the present invention, not to be construed as limiting the present invention. The invention has been described with reference to exemplary embodiments, but it is understood that the words which have been used are words of description and illustration, rather than words of limitation. Modifications may be made to the invention as defined in the appended claims, and the invention may be modified without departing from the scope and spirit of the invention. Although the invention is described herein with reference to particular means, materials and embodiments, the invention is not intended to be limited to the particulars disclosed herein, as the invention extends to all other means and applications which perform the same function.

Claims

1. A cheese bacillus YWC-01 strain for degrading uric acid can produce an enzyme for catalyzing and degrading uric acid, and the preservation number of the enzyme is CGMCC No.31534.

2. The strain of cheese bacillus YWC-01 according to claim 1, wherein said strain of cheese bacillus YWC-01 is capable of degrading and removing uric acid at an initial concentration of 1g/L in 48 hours at an initial concentration of 8 x 10 ⁸/mL of cells.

3. The strain of cheese bacillus YWC-01 according to claim 1, wherein the crude enzyme produced by the strain of cheese bacillus YWC-01 is capable of completely removing uric acid having an initial concentration of 1g/L in 18 hours at a protein concentration of 0.8 g/L.

4. A preparation of uric acid degrading lactobacillus casei YWC-01 comprising bacterial cells and/or crude enzyme of a lactobacillus casei YWC-01 strain as claimed in any of claims 1 to 3, preferably comprising a lactobacillus casei YWC-01 strain as claimed in any of claims 1 to 3.

5. The preparation of Lactobacillus casei YWC-01 as set forth in claim 4,

The preparation of the Lactobacillus casei YWC-01 for degrading uric acid is a liquid preparation, preferably, the concentration of bacterial cells of the Lactobacillus casei YWC-01 strain in the liquid preparation for degrading uric acid is (1-10) multiplied by 10 ⁸/mL, and/or the concentration of the crude enzyme protein of the Lactobacillus casei YWC-01 strain in the liquid preparation for degrading uric acid is 0.1-1.0g/L;

Or the uric acid-degrading solid powder preparation of Lactobacillus casei YWC-01, preferably wherein the content of cells of the strain of Lactobacillus casei YWC-01 is (1-10) x 10 ⁸/g, more preferably (8-10) x 10 ⁸/g, and/or the protein content of crude enzyme of the strain of Lactobacillus casei YWC-01 is 0.1-1.0g/kg, more preferably 0.8-1.0g/kg.

6. A process for the preparation of a uric acid degrading formulation of lactobacillus casei YWC-01 as defined in any of claims 4 or 5 comprising:

7. The method of claim 6, wherein the fermentation medium comprises the following components in 1L of water, based on 1L of water:

Preferably, the pH of the fermentation medium is 7-8;

8. The production method according to claim 6 or 7, characterized in that the production method further comprises:

Wherein the low temperature is 0-4 ℃.

9. Use of the uric acid degrading lactobacillus casei YWC-01 formulation as defined in claim 4 or 5 or the uric acid degrading lactobacillus casei YWC-01 formulation as defined in any of the claims 6-8 for the manufacture of a uric acid lowering medicament comprising:

Wherein the low temperature is 0-4 ℃.

10. The use according to claim 9, wherein,

In the step F, the lyophilized preparation of the cheese bacillus YWC-01 is diluted by using physiological saline to prepare a liquid uric acid reducing agent;

or in the step F, edible starch is used for diluting the freeze-dried preparation of the cheese bacillus YWC-01 to prepare a solid uric acid reducing medicament;

preferably, the uric acid lowering agent is an oral preparation.