KR20230064553A - Recombinant strain with uric acid resolution and use thereof - Google Patents

Abstract

The present invention relates to a recombinant strain with increased uric acid resolution and a use thereof. Since uric acid degradation activity is shown in a strain transformed by applying a recombinant expression vector containing pucL and pucM genes for encoding uricase and 5-hydroxyisourate hydrolases, a method for manufacturing a recombinant strain with increased uric acid resolution is usefully used. The recombinant strain manufactured by the manufacturing method is usefully used as a composition for preventing, treating, or improving a uric acid disease or a gouty disease.

Description

Recombinant strain with uric acid resolution and use thereof {Recombinant strain with uric acid resolution and use thereof}

The present invention relates to a recombinant strain having uric acid degrading ability and its use.

Uric acid is one of the decomposition products of purines, and excessive accumulation in the body can cause hyperuricemia or gout. When the serum uric acid level exceeds 7.0 mg/dL, it is called hyperuricemia, and due to this, uric acid crystallizes and is deposited in cartilage or joints, which is gout. In addition to gout, hyperuricemia causes arthritis, urinary stones, and kidney disease. Recently, the accumulation of uric acid in the body is increasing due to modern people's drinking habits and increased protein intake.

Since uric acid oxidase does not exist in the human body, uric acid becomes the final product in purine metabolism. However, other organisms, including microorganisms, have uric acid oxidase, which decomposes uric acid into allantoin. The solubility of uric acid is very low at 6mg/100mL, but the solubility of allantoin is 0.57g/100mL, which is less likely to accumulate in the body than uric acid. Thus, uric acid oxidase may be a way to prevent excessive accumulation of uric acid. On the other hand, two intermediate steps have been identified until uric acid is decomposed into allantoin. Uric acid is converted to 5-hydroxyisourate (HIU) by uric acid oxidase and 5-hydroxyisourate hydrolase (EC 3.5.2.17) by 5-hydroxyisourate hydrolase (EC 3.5.2.17). Isoleucic acid is converted back to 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline (OHCU). The conversion of OHCU to allantoin occurs rapidly. Therefore, if uric acid oxidase is provided into the human body through other organisms, including microorganisms, to promote uric acid decomposition, it will be helpful to improve diseases caused by the accumulation of uric acid.

1. Republic of Korea Patent Registration No. 10-1450511 (published on March 14, 2014)

An object of the present invention is to provide a method for preparing a recombinant strain having uric acid degrading ability.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating uric acid disease or gouty disease comprising the recombinant strain prepared by the above production method as an active ingredient.

Another object of the present invention is to provide a health functional food composition for preventing or improving uric acid disease or gouty disease, comprising the recombinant strain prepared by the above production method as an active ingredient.

In order to achieve the above object, the present invention comprises cloning a pucL or pucM gene, preparing a recombinant expression vector containing the cloned gene, and introducing the recombinant expression vector into a bacterial strain. It provides a method for producing a recombinant strain having uric acid degrading ability, comprising the step of transforming by.

In addition, the present invention provides a pharmaceutical composition for preventing or treating uric acid disease or gouty disease comprising the recombinant strain prepared by the above production method as an active ingredient.

In addition, the present invention provides a health functional food composition for preventing or improving uric acid disease or gouty disease comprising the recombinant strain prepared by the above production method as an active ingredient.

According to the present invention, uric acid degradation activity in a strain transformed by introducing a recombinant expression vector containing pucL and pucM genes encoding uric acid oxidase and 5-hydroxyisourate hydrolase By confirming that this appears, it can be usefully used as a method for preparing a recombinant strain having uric acid decomposition ability, and the recombinant strain prepared by the above method can be usefully used as a composition for preventing, treating or improving uric acid disease or gouty disease. It can be.

Figure 1 shows the uric acid decomposition process.
Figure 2 shows a vector map of pCB4170-p710-pucL (7.1 kb).
3 shows a vector map of pCB4170-p710-pucLM (7.4 kb).
4 is a transforming LC ( Leuconostoc It is the result of analyzing pucL and pucM expression in citrium ) strain through SDS-PAGE. M is a protein marker, 1 is soluble protein of wild-type (WT) LC strain, 2 is soluble protein of transformed LC strain, 3 is purified soluble protein of wild-type LC strain, 4 is purified of transformed LC strain It is a soluble protein.
Fig. 5 shows the results of uric acid clear zone analysis of the transformed LC strains on agar plates containing 0.3% (w/v) uric acid. Specifically, FIG. 5A is a Bs strain on an LB agar plate, and FIG. 5B is a transformed LC strain on an MRS agar plate. 1 is the positive control, 2 is the wild-type (WT) LC strain and 3-7 are the transgenic LC strains.
6 is an absorbance standard curve graph for each concentration of uric acid suspended in 0.1M boric acid buffer.
Figure 7 is the result of analyzing the change in absorbance over time of the transgenic LC strain (○) and the WT strain (●) at an absorbance of 293 nm.
Figure 8 is a result of comparing and analyzing the uric acid degrading enzyme (uricase) activity of the transformed strain and the WT strain.
Figure 9 is a result of comparing and analyzing the change in uric acid degrading enzyme (uricase) activity according to the incubation time of the transformed strain and the WT strain.

Hereinafter, the present invention will be described in more detail.

The present invention comprises the steps of cloning a pucL or pucM gene, preparing a recombinant expression vector containing the cloned gene, and integrating the recombinant expression vector into a bacterial strain to transform it. , It provides a method for producing a recombinant strain having uric acid degradation ability.

The pucL or pucM gene may encode uricase or 5-hydroxyisourate hydrolase.

The pucL gene may have a nucleotide sequence represented by SEQ ID NO: 1, and the pucM gene may have a nucleotide sequence represented by SEQ ID NO: 2.

In addition, the pucL or pucM gene may be derived from a Bacillus subtilis ssp. spizizenii strain, but is not limited thereto.

As shown in FIG. 1, uric acid is decomposed by uric acid oxidase and 5-hydroxyisouric acid hydrolase.

In addition, the present invention provides a recombinant strain with increased uric acid degradation ability prepared by the above production method.

The recombinant strain may exhibit uric acid decomposing activity.

In addition, the recombinant strain is Leuconostoc citrium ( Leuconostoc citrium ) strain, but is not limited thereto.

In addition, the present invention provides a pharmaceutical composition for preventing or treating uric acid disease or gouty disease comprising the recombinant strain prepared by the above production method as an active ingredient.

The uric acid disease or gouty disease is primary hyperuricemia, secondary hyperuricemia, primary gout, secondary gout, acute gouty arthritis, subcutaneous tophi, chronic gouty arthritis (chronic tophiarthritis), acute pain, chronic It may be at least one selected from the group consisting of pain, intractable pain, cancer pain, acute uric acid nephropathy, chronic uric acid nephropathy, and uric acid urolithiasis, but is not limited thereto.

The pharmaceutical composition of the present invention is prepared in a unit dose form or in a multi-dose container by formulating using a pharmaceutically acceptable carrier according to a method that can be easily performed by those skilled in the art. It can be prepared by incorporating into

The pharmaceutically acceptable carrier is one commonly used in formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like. The pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, and the like, in addition to the above components.

In the present invention, the content of the additives included in the pharmaceutical composition is not particularly limited and may be appropriately adjusted within the range of content used in conventional formulations.

The pharmaceutical composition may be formulated into an injectable formulation such as an aqueous solution, suspension, or emulsion, a pill, a capsule, a granule, a tablet, a cream, a gel, a patch, a spray, an ointment, a warning agent, a lotion, a liniment agent, a paste agent, and a cataplasma agent. It may be formulated in the form of one or more skin external preparations selected from the group consisting of, but is not limited thereto.

The pharmaceutical composition of the present invention may additionally contain pharmaceutically acceptable carriers and diluents for formulation. The pharmaceutically acceptable carriers and diluents include excipients such as starch, sugar and mannitol, fillers and extenders such as calcium phosphate, cellulose derivatives such as carboxymethylcellulose and hydroxypropylcellulose, gelatin, alginates, and polyvinyl pyrrolidone. binders such as talc, calcium stearate, lubricants such as hydrogenated castor oil and polyethylene glycol, disintegrants such as povidone and crospovidone, surfactants such as polysorbates, cetyl alcohol, glycerol and the like, but are not limited thereto. The pharmaceutically acceptable carrier and diluent may be biologically and physiologically compatible with the subject. Examples of diluents include, but are not limited to, saline, aqueous buffers, solvents and/or dispersion media.

The pharmaceutical composition of the present invention may be administered orally or parenterally (eg, intravenous, subcutaneous, intraperitoneal or topical application) depending on the desired method. For oral administration, it may be formulated into tablets, troches, lozenges, aqueous suspensions, oily suspensions, prepared powders, granules, emulsions, hard capsules, soft capsules, syrups, elixirs, and the like. In the case of parenteral administration, it may be formulated as an injection solution, suppository, powder for respiratory inhalation, aerosol for spray, ointment, powder for application, oil, cream, etc.

The dosage of the pharmaceutical composition of the present invention depends on the patient's condition, weight, age, sex, health condition, dietary constitution specificity, the nature of the preparation, the severity of the disease, the administration time of the composition, the administration method, the administration period or interval, the excretion rate and The range may vary depending on the type of drug, and may be appropriately selected by a person skilled in the art. For example, it may range from about 0.1 to 10,000 mg/kg, but is not limited thereto, and may be divided and administered once or several times a day.

The pharmaceutical composition may be administered orally or parenterally (eg, intravenous, subcutaneous, intraperitoneal or topical application) depending on the desired method. The pharmaceutically effective amount and effective dose of the pharmaceutical composition of the present invention may vary depending on the formulation method, administration method, administration time, administration route, etc. of the pharmaceutical composition, and those skilled in the art can achieve effective treatment for the desired treatment. The dosage can be easily determined and prescribed. Administration of the pharmaceutical composition of the present invention may be administered once a day, or may be divided and administered several times.

In addition, the present invention provides a health functional food composition for preventing or improving uric acid disease or gouty disease comprising the recombinant strain prepared by the above production method as an active ingredient.

The present invention can be generally used as a commonly used food.

The food composition of the present invention can be used as a health functional food. The above “functional health food” refers to food manufactured and processed using raw materials or ingredients having functional properties useful for the human body in accordance with the Health Functional Food Act, and “functional” refers to food that is not related to the structure and function of the human body. It refers to intake for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological functions.

The health functional food composition may contain conventional food additives, and the suitability as the “food additive” is determined in accordance with the General Rules of the Food Additive Code and General Test Methods approved by the Ministry of Food and Drug Safety, unless otherwise specified. It is judged according to the specifications and standards for the item.

Examples of the items listed in the “Food Additive Code” include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as dark pigment, licorice extract, crystalline cellulose, kaoliang pigment, guar gum, and mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.

The food composition of the present invention can be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills and the like. For example, among health functional foods in the form of capsules, hard capsules can be prepared by mixing and filling a composition according to the present invention with additives such as excipients in a conventional hard capsule, and soft capsules contain the composition according to the present invention. It can be prepared by mixing with additives such as excipients and filling in a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative, and the like, if necessary.

Definitions of terms for the excipients, binders, disintegrants, lubricants, corrigents, flavoring agents, etc. are described in literature known in the art, and include those having the same or similar functions. There is no particular limitation on the type of food, and it includes all health functional foods in a conventional sense.

In the present invention, the term "prevention" refers to all activities that suppress or delay uric acid disease or gouty disease by administering the composition according to the present invention.

In the present invention, the term "treatment" refers to all activities that improve or beneficially change uric acid disease or gouty disease by administering the composition according to the present invention.

In the present invention, the term "improvement" refers to all actions that improve the bad condition of uric acid disease or gouty disease by administration of the composition according to the present invention.

Hereinafter, examples will be described in detail to aid understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, but the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

[ Experimental example 1] Production of plasmids and transformed strains

E. coli TOP10 was used for cloning of all plasmids, and the transformed strain was Leuconostoc citrium ; Hereinafter referred to as LC) EFEL2700 strain (KACC 91348) was used. Uric acid oxidase and 5-hydroxyisourate hydrolase were cloned from Bacillus subtilis ( Bacillus subtilis ssp. spizizenii ; hereinafter referred to as Bs) strain (KACC 14741). Although 5-hydroxyisouric acid hydrolase has not been annotated in the Bacillus subtilis strain, a sequence showing homology when compared with the strain gene sequence through BLAST database analysis provided by NCBI was discovered and converted to pucM. Named and cloned.

Plasmids were prepared by PCR amplification of all DNA fragments using PrimeSTAR HS DNA polymerase. A backbone fragment was prepared from pCB4170-p710-Bgal. An insert fragment was prepared by cloning pucL from the genome of the Bs strain. Each fragment was ligated through Gibson Assembly to prepare pCB4170-p710-pucL as shown in FIG. 2 . A 6x his-tag was added to the C-terminus of the pucL gene to facilitate subsequent protein purification. A backbone fragment was again prepared from pCB4170-p710-pucL, pucM was cloned from the genome of the Bs strain, an insert fragment was prepared, and pCB4170-p710-pucLM was prepared as shown in FIG. 3 through Gibson Assembly. Similarly, 6x his-tag was added. pCB4170-p710-pucLM was transformed into an LC strain by electroporation. Electroporation was performed at 1.5 kV using a 0.1 cm electroporation cuvette. The transformed strain was cultured on MRS agar medium containing chloramphenicol at a concentration of 15 μg/mL, and then several colonies were cultured on MRS liquid medium at the same concentration of chloramphenicol. The base sequences of the pucL and pucM genes of the Bs strain are shown in Tables 1 and 2 below.

Sequence of pucL from Bacillus subtilis ssp. spizizenii KACC 14741 ATGTTCACAATGGATGACATGAACCAAATGGACATTCAAACACTGACAGACACGCTTGAATCTATTTTTGAACACTCTTCATGGATTGCGGAGAAAGCCGCGGCATTGCGGCCGTTTTCTTCCCTATCTGATCTTCACCACAAAATGGCCAGCATTGTAAAAGCCGCGGACCGCCAGACAGCTTGATTTAATCAACAAGCACCCCCCCACTCGGAACAAAGAATACAATGAGCGTCACCTCGG TAAGAGAGCAGCAAAACGCGGGACTCAGTAAACTTGAACAAGAGGAATACGAAGAATTTCTGAAGCTGAATGAACGCTATTATGAACGCTTCGGCTTTCCTTTTATTTTAGCGGTAAAGGGAAAGACGAAACAGGACATTTACCAAGCTCTGCTGGAAAGGCTTGAGAACGAACGAGAAACGGAGTTCCATCAGGCTCTTAAAGAAATTTACCGCATCGCCCGCTTTCGGCTGGCGGA CATCATAACTGAAAAAGGAGAGACGCAAATGAAAAGAACTATGTCTTATGGCAAAGGAAATGTATTTGCATACCGAACGTTTTTAAAGCCGCTCACAGGGGTACGGCAAATCCCTGAGTCTTCTTTTACAGGGAGAGACAATACTGTTGTCGGCATTGATGTGACGTGCGAAATTGGCGGAGATGCCTTTCTGACATCATTTATAGACGGAGATAATACACTCGTCGTGGCTACAGACTCAATGAAAAACT TTATTCAGAGCCATCTCGCTTCCTATGAAAGGGACGACAACCGAGGGATTCTTACACTATGTGGCACACCGATTTTTAGATACCTATTCTCATATGGACACTATCACTCTGACAGGCGAAGACATTCCGTTTGAGGCGATGCCTGCATACGAGGGGCAAGAACTCGGCACAAGCCACCTCGTCTTTAGAAGATCACGCAATGAACGCGCTCGGTCTGTGCTGAAAGCGGAACGAACCGG GGATACAATAACGATCACAGAGCAAGTACAGCGAAATCATGGATCTTCAGCTCGTCAAGGTGAGCGGCAACTCCTTCGTCGGCTTTATCCGGGACGAATATACGACTCTCCCGGAGGATGGCAACCGCCCTCTGTTTGTCTATTTAAACATCTGCTGGCAGTATGAAAACAGATGACGCACGCGCTTCTGATCCAGCCCGCTACGTCGCGGCTGAACAAGTCCGCGACTTGGCGAGCACC GTTTTTCACGAGCTCAAAACCCCTTCAATTCAAAATTTAATCTATCATATCGGCTGCAGAATATTAACGAGATTCCCGCAGCTCACTGATGTCAGCTTCCAATCTCAAAACCACACATGGGATACAGTCGTTGAAGAGATCCCAGGGTCCAAAGGAAAAGTCTACACCGAACCGCGCCCGCCATTCGGTTTCCAGCGCTTTACCGTGACAAGAGAAAGACGCCGAGAAAGAAAACAGAA AGCCGTCGAAAAATTAGGGAGCCTGAAAGCCTGA

Sequence of pucM from Bacillus subtilis ssp. spizizenii KACC 14741 ATGGGGAAGCTGACAACGCATATCTTGGATTTAACCTGCGGCAAACCCGCGGCGAACGTAAAAGTTGAATTGAAGAAGCTGGACGAAAGCATTATGAAAGAAGCATATACGAACAATGATGGGAGAGTCGATGTTCCTCTGCTGGCTGGTGAAGAGCTGGTGTCCGGAGAGTATGTGATGGAATTTCATGCAGGAGACTATTTCGCGAGTAAAAATGTGAACACGGCCGACCAGCCGT TTTTAACCATAGTCACCGTTCGTTTTCATCTTGCAGATCCGGAAGCTGATTATCACATCCCGCTTTTGCTGTCGCCGTTTGGATATCAGGTGTATAGAGGGAGTTAA

[ Example 1] Transformation strain uric acid resolution analyze

1-1. pucL and pucM Protein expression analysis

In order to confirm the pucL and pucM protein expression of the transformed strain, first, the supernatant of the cell lysate was purified using a Ni-NTA column. 2x Laemmli sample buffer was added to the supernatant of the cell lysate and the purified protein sample, heated at 98 ° C for 10 minutes, and a 15% acrylamide gel was used. Thereafter, the soluble protein extracted from the transformed strain and the protein sample purified using his-tag therefrom were analyzed through SDS-PAGE.

As a result, as shown in FIG. 4, uric acid oxidase and 5-hydroxyisouric acid hydrolase showed protein sizes of 56.5 kDa and 12.6 kDa, respectively, and it was confirmed that pucL and pucM expression appeared in the transformed strain. . This means that the transformed strain has the ability to degrade uric acid.

1-2. clear Clear zone analysis

In order to confirm the uric acid degradation ability of the transformed strain, the clear zone was analyzed. 5 μL of the transformed strain culture medium was dropped onto MRS agar medium supplemented with 0.3% (w/v) uric acid, and cultured for 24 hours. A wild-type (hereinafter referred to as WT) LC strain containing the CB4170 empty vector was used as a negative control.

As a result, as shown in Figure 5A, it was confirmed that the Bs strain has the ability to decompose uric acid. This means that pucL and pucM derived from the strain show activity. In addition, as shown in Figure 5B, in the case of the transformed LC strain designed to express both pucL and pucM derived from the above strain, growth inhibition appeared on the uric acid-containing plate, whereas the LC WT strain containing the pCB4170 empty vector did not grow. It was confirmed that the ring of inhibition did not appear. This means that the transformed strain developed uric acid degradation ability by transformation.

1-3. Absorbance analysis by uric acid concentration

Prior to the analysis of the uric acid degrading enzyme activity of the transformed strain, the absorbance of the standard solution for each concentration of uric acid was analyzed at a wavelength of 293 nm. After dissolving uric acid at 200 μM in boric acid buffer (pH 8.5), it was diluted through a serial dilution method, and a standard curve of uric acid at each concentration was drawn.

As a result, as shown in FIG. 6, the titer of the transformed LC strain was calculated to be 54 U/mg, and it was confirmed that the absorbance increased in proportion to the uric acid concentration as the uric acid concentration increased. This means that the conditions are well set to be suitable for measuring uric acid activity in the transformed strain.

1-4. Time-dependent absorbance analysis

In order to confirm the uric acid degradation ability of the transformed strain, the change in absorbance over time was analyzed. While the transformed strain was reacted at 30 ° C. for 30 minutes, the absorbance change over time was analyzed based on the initial absorbance of 293 nm. Leuconostoc citrium WT (wild type) strain containing the CB4170 empty vector was used as a negative control.

As a result, as shown in FIG. 7, it was confirmed that the absorbance value of the transformed strain decreased more significantly than that of the WT strain (control group). This means that the concentration of uric acid in the transformed strain decreases during the reaction, that is, the transformed strain has the ability to decompose uric acid.

1-5. uric acid degrading enzyme ( uricase ) activity assay

In order to confirm the uric acid degrading ability of the transformed strain, uric acid degrading enzyme (uricase) activity was analyzed. The culture solution was centrifuged to remove the supernatant, washed twice with 50 mM sodium phosphate buffer (pH 7.0), suspended in the same buffer, and disrupted with an ultrasonicator. The supernatant of the lysate was titered used in 40 μL of the supernatant of the cell lysate and 160 μL of boric acid buffer pH 8.5 in which 50 μM uric acid was dissolved were put into a 96-well microplate, and the change in absorbance based on the initial absorbance at 293 nm was analyzed at 30 ° C. 1 U of uric acid degrading enzyme was defined as the amount of enzyme that converts 1 μM of uric acid to allantoin in 1 minute at pH 8.5 and 30° C. Leuconostoc citrium WT strain containing the CB4170 empty vector was used as a negative control.

As a result, as shown in FIG. 8, it was confirmed that the uric acid lyase activity of 54 U/mg was shown in the transformed strain, whereas the uric acid lyase activity was not shown in the WT strain. This means that uric acid degrading enzyme activity was shown by transformation.

1-6. Analysis of uric acid degrading enzyme activity according to strain incubation time

In order to confirm the change in uric acid lyase activity according to the incubation time of the transformed strain, the transformed strain was cultured at 30 ° C. for 6, 12 and 18 hours, and then the uric acid lytic enzyme activity was analyzed over time. Leuconostoc citrium WT strain containing the CB4170 empty vector was used as a negative control.

As a result, as shown in FIG. 9, it was confirmed that the activity of uric acid degrading enzyme was the highest at 6 hours of incubation, and the activity decreased in the order of 12 and 18 hours. This means that when the transformed strain is commercialized in the future, side effects due to excessive activity of uric acid lyase in the human body after administration are less likely to occur.

Having described specific parts of the present invention in detail above, it is clear to those skilled in the art that these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. do. That is, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims (8)

cloning the pucL or pucM gene;
preparing a recombinant expression vector containing the cloned gene; and
A method for producing a recombinant strain having uric acid degrading ability, comprising the step of transforming by introducing the recombinant expression vector into a bacterial strain. The method according to claim 1, wherein the pucL or pucM gene encodes uricase or 5-hydroxyisourate hydrolase. The method according to claim 1, wherein the pucL gene has the nucleotide sequence represented by SEQ ID NO: 1. The method according to claim 1, wherein the pucM gene has the nucleotide sequence represented by SEQ ID NO: 2. The method according to claim 1, wherein the pucL or pucM gene is derived from a Bacillus subtilis ssp. spizizenii strain. A pharmaceutical composition for preventing or treating uric acid disease or gouty disease, comprising the recombinant strain prepared by the method of claim 1 as an active ingredient. The method according to claim 6, wherein the uric acid disease or gouty disease is primary hyperuricemia, secondary hyperuricemia, primary gout, secondary gout, acute gouty arthritis, subcutaneous tophi, chronic gouty arthritis (chronic tophiarthritis) A pharmaceutical composition, characterized in that at least one selected from the group consisting of acute pain, chronic pain, intractable pain, cancer pain, acute uric acid nephropathy, chronic uric acid nephropathy and uric acid urolithiasis. A health functional food composition for preventing or improving uric acid disease or gouty disease, comprising the recombinant strain prepared by the method of claim 1 as an active ingredient.

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