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CN-121991998-A - Regulation system for degrading uric acid to reduce uric acid level and application thereof

CN121991998ACN 121991998 ACN121991998 ACN 121991998ACN-121991998-A

Abstract

The application relates to the technical field of genetic engineering, in particular to a regulation system for degrading uric acid to reduce uric acid level and application thereof, wherein the system comprises a first functional gene, a second functional gene and a first terminator, wherein the first functional gene codes for a first functional protein capable of degrading uric acid, the first functional protein comprises one or more sequences shown as SEQ ID NO. 1-10 or at least 85% identical to the first functional protein, the second functional gene codes for a second functional protein capable of transporting uric acid, the second functional protein comprises one or more sequences shown as SEQ ID NO. 11-15 or at least 85% identical to the second functional protein, the first promoter is used for regulating the expression of the first functional gene and/or the second functional gene, and the first terminator is used for stopping the expression of the first functional gene and/or the second functional gene. The regulation and control system for degrading uric acid to reduce uric acid level provided by the embodiment of the application has high-efficiency uric acid degradation function.

Inventors

  • PAN LEI
  • FU XIAN
  • SHEN YUE

Assignees

  • 深圳华大生命科学研究院

Dates

Publication Date
20260508
Application Date
20241105

Claims (12)

  1. 1. A regulatory system for degrading uric acid to reduce uric acid levels, the regulatory system comprising: A first functional gene encoding a first functional protein capable of degrading uric acid, the first functional protein comprising one or more sequences as set forth in SEQ ID NOs 1-10 or having at least 85% identity thereto; A second functional gene encoding a second functional protein capable of transporting uric acid, the second functional protein comprising one or more sequences as set forth in SEQ ID NOS: 11-15 or having at least 85% identity thereto; a first promoter for regulating the expression of the first functional gene and/or the second functional gene, and A first terminator for terminating expression of the first functional gene and/or the second functional gene.
  2. 2. The regulation system of claim 1, wherein the regulation system further comprises: A second promoter for regulating the expression of the first functional gene and/or the second functional gene, and A second terminator for terminating the expression of the first functional gene and/or the second functional gene, Optionally, the second promoter and the first promoter are constitutive promoters.
  3. 3. The regulation system according to claim 1 or 2, characterized in that it comprises in particular: The first promoter is used for regulating and controlling the expression of the first functional gene; The first terminator is used for terminating the expression of the first functional gene; The second promoter for regulating the expression of the second functional gene, and The second terminator for terminating the expression of the second functional gene, Wherein the first promoter and the second promoter comprise any one of the sequences selected from SEQ ID NOS: 16-23 or any one of the sequences having at least 85% identity thereto, respectively, the first promoter preferably comprises any one of the sequences shown in SEQ ID NO:16 or any one of the sequences having at least 85% identity thereto, and the second promoter preferably comprises any one of the sequences shown in SEQ ID NOS: 17-23 or a sequence having at least 85% identity thereto; The first terminator and the second terminator respectively comprise any one sequence selected from SEQ ID NO. 24-26, the first terminator preferably comprises a sequence shown in SEQ ID NO. 24 or SEQ ID NO. 26, more preferably comprises a sequence shown in SEQ ID NO. 26, and the second terminator preferably comprises a sequence shown in SEQ ID NO. 25.
  4. 4. A regulatory system according to claim 3, wherein the regulatory system comprises: the first functional gene encoding the first functional protein comprising the sequence shown in SEQ ID NO.2 and any one of SEQ ID NO. 6-10, preferably comprising the sequence shown in SEQ ID NO. 2; the second functional gene encoding the second functional protein, wherein the second functional protein comprises a sequence shown in SEQ ID NO. 15; The first promoter comprises a sequence shown in SEQ ID NO. 16; The first terminator comprises a sequence shown in SEQ ID NO. 24 or SEQ ID NO. 26, preferably comprises a sequence shown in SEQ ID NO. 26; The second promoter comprising any one of the sequences shown in SEQ ID NO. 18-23, preferably comprising the sequence shown in SEQ ID NO. 18 or SEQ ID NO. 23, and The second terminator comprises a sequence shown in SEQ ID NO. 25.
  5. 5. The regulation and control system of claim 4, wherein the regulation and control system, The first functional protein comprises a sequence shown in SEQ ID NO. 2; The first terminator comprises the sequence shown in SEQ ID NO. 26, and The second promoter comprises the sequence shown as SEQ ID NO. 18 or SEQ ID NO. 23.
  6. 6. An expression vector comprising the regulatory system for degrading uric acid to reduce uric acid levels as defined in any one of claims 1 to 5, Optionally, the expression vector is a eukaryotic expression vector.
  7. 7. A recombinant microorganism comprising the regulatory system for degrading uric acid to reduce uric acid level as defined in any one of claims 1 to 5 or the expression vector as defined in claim 6, Alternatively, the microorganism is a Saccharomyces (Saccharomyces sp.) microorganism, preferably Saccharomyces cerevisiae (Saccharomyces cerevisiae), more preferably Saccharomyces cerevisiae (Saccharomyces cerevisiae var. Boulardii).
  8. 8. A composition for degrading uric acid to reduce uric acid levels comprising the recombinant microorganism and/or a metabolite thereof according to claim 7 and a pharmaceutically or dietetically acceptable adjuvant, Optionally, the microorganism in the composition retains activity, Alternatively, the composition is administered orally.
  9. 9. Use of the recombinant microorganism of claim 7 or the composition of claim 8 in the preparation of a food, health product, food additive or pharmaceutical product for degrading uric acid to reduce uric acid levels.
  10. 10. A pharmaceutical or health product for degrading uric acid to reduce uric acid levels comprising the recombinant microorganism according to claim 7 and/or a metabolite thereof or the composition according to claim 8, Optionally, the medicine or health product is in the form of tablet, capsule, granule, pill, powder, gel or solution.
  11. 11. A medicament for the treatment of hyperuricemia-related diseases comprising the recombinant microorganism and/or metabolite thereof according to claim 7 or the composition according to claim 8.
  12. 12. Use of a recombinant microorganism according to claim 7 or a composition according to claim 8 for the manufacture of a medicament for the treatment of hyperuricemia-related diseases, Alternatively, the hyperuricemia-related diseases include hyperuricemia, gout, uric acid stones, acute and chronic uric acid nephropathy.

Description

Regulation system for degrading uric acid to reduce uric acid level and application thereof Technical Field The application relates to the technical field of genetic engineering, in particular to a regulation system for degrading uric acid to reduce uric acid level and application thereof. Background Uric acid (Uric acid, UA) is an important intermediate of the purine (Purines) metabolic pathway. Clinically, normal levels of uric acid in humans are generally maintained in the range of 2.4-6.0mg/dL (female) and 3.4-7.0mg/dL (male). Genetic factors, medication for disease treatment, alcoholism, stay up, high purine dietary intake, etc. all increase the risk of unbalanced uric acid homeostasis in humans, leading to abnormally elevated blood uric acid levels, resulting in hyperuricemia (Hyperuricemia, HUA). Along with the improvement of social and economic level, the enrichment of material life and the acceleration of life rhythm, the incidence rate of hyperuricemia is greatly increased. Statistics in 2019 show that the prevalence rate of hyperuricemia in China is 13.3%, the overall prevalence rate of gout is 1.1%, and the trend of patient younger is shown. Recently, a series of applications research around the potential action site of uric acid excretion in the intestinal tract, and a scheme of treating hyperuricemia by using intestinal probiotics is developed, for example, functional elements such as urate oxidase and the like are introduced into the intestinal tract of a mouse so as to enhance the activity of uric acid metabolism in the intestinal tract and reduce the uric acid level of a human body. However, these intestinal probiotics are poor in quality and have the problems of insufficient uric acid degradation activity, need of being matched with or having potential cytotoxicity or easy to be absorbed by human bodies, and the like, so that the induction effect is drastically reduced. Furthermore, currently there are extremely limited chassis bacteria available for recombinant probiotic intervention routes. Thus, there is a need to develop beneficial strains that can efficiently and stably reduce uric acid levels to meet the increasing therapeutic demands for hyperuricemia and gout. Disclosure of Invention The present application solves at least one of the problems of the related art from the following aspects. To this end, embodiments of the present application provide a regulatory system for degrading uric acid to reduce uric acid levels comprising a first functional gene encoding a first functional protein capable of degrading uric acid comprising one or more sequences as shown in SEQ ID NOS: 1-10 or having at least 85% identity thereto, a second functional gene encoding a second functional protein capable of transporting uric acid comprising one or more sequences as shown in SEQ ID NOS: 11-15 or having at least 85% identity thereto, a first promoter for regulating the expression of the first functional gene and/or the second functional gene, and a first terminator for terminating the expression of the first functional gene and/or the second functional gene. In some embodiments, the regulatory system further comprises a second promoter for regulating expression of the first functional gene and/or the second functional gene, and a second terminator for terminating expression of the first functional gene and/or the second functional gene, optionally the second promoter and the first promoter are constitutive promoters. In some embodiments, the regulatory system specifically comprises the first promoter for regulating the expression of the first functional gene, the first terminator for terminating the expression of the first functional gene, the second promoter for regulating the expression of the second functional gene, and the second terminator for terminating the expression of the second functional gene, wherein the first and second promoters comprise any sequence selected from SEQ ID NO 16-23 or any sequence having at least 85% identity thereto, the first promoter preferably comprises any sequence selected from SEQ ID NO 16 or any sequence having at least 85% identity thereto, the second promoter preferably comprises any sequence selected from SEQ ID NO 17-23 or a sequence having at least 85% identity thereto, the first and second promoters comprise any sequence selected from SEQ ID NO 24-26, the first and second promoters preferably comprise any sequence selected from SEQ ID NO 17-23, the preferred sequence selected from SEQ ID NO 26 comprises any sequence selected from SEQ ID NO 26 and the preferred sequence selected from the group consisting of SEQ ID NO 26 and the preferred sequence comprises any sequence having at least 85% identity. In some embodiments, the regulatory system comprises the first functional gene encoding the first functional protein comprising any one of the sequences shown in SEQ ID NO. 2 and SEQ ID NO. 6-10, preferably comprising the sequence shown in SEQ ID NO. 2, the second