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Peer-reviewedCC BY-NC 4.0Microbial Biotechnology

Reprogramming Native Cryptic Plasmids Enables Antibiotic‐Selection‐Free Urate Degradation by Escherichia coli Nissle 1917

  • Yuan Chi1,2
  • Shiying Wu3
  • Fei Lin1,2
  • Tingwen Fan1,2
  • Hongwei Liu1,2
  • Huaiyi Yang1,2,4
4 affiliations
  1. 1State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology Chinese Academy of Sciences Beijing China
  2. 2University of Chinese Academy of Sciences Beijing China
  3. 3Key Laboratory of Plant Biotechnology of Liaoning Province School of Life Sciences, Liaoning Normal University Dalian Liaoning China
  4. 4Beijing Key Laboratory of Genetic Element Biosourcing & Intelligent Design for Biomanufacturing Beijing China
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Abstract

Hyperuricemia is associated with renal injury and disturbances in gut microbial ecology. Intestinal urate clearance by engineered probiotics is limited by antibiotic‐free functional maintenance and the oxygen requirement of urate oxidase (UOX). We reprogrammed the native cryptic plasmids of Escherichia coli Nissle 1917 (EcN) to construct M12U, a marker‐free dual‐plasmid strain. pMUT2 carried an HucR‐regulated urate‐responsive UOX module, while pMUT1 encoded a low‐oxygen‐responsive UOX module and Tat‐targeted Vitreoscilla haemoglobin (tat‐VHB). M12U degraded urate in vitro and retained activity under low‐oxygen conditions. In hyperuricemic mice (n = 5 per group), oral M12U reduced serum urate, creatinine, blood urea nitrogen, inflammatory cytokines and renal histopathological injury. 16S rRNA profiling showed increased microbial richness and a partial shift in community composition toward healthy controls. Shotgun metagenomics revealed enrichment of Lactobacillus johnsonii and Bacteroides acidifaciens , together with increased metabolic potential related to the pentose phosphate pathway and glutathione metabolism. Faecal metabolomics showed reduced uric acid abundance and increased thiamine monophosphate and argininosuccinic acid. These results support further evaluation of dual‐native‐plasmid architectures for gut‐directed urate clearance.

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Cite

BibTeXRIS
BibTeX
@article{chi2026reprogramming,
  title = {{Reprogramming Native Cryptic Plasmids Enables Antibiotic‐Selection‐Free Urate Degradation by Escherichia coli Nissle 1917}},
  author = {Yuan Chi and Shiying Wu and Fei Lin and Tingwen Fan and Hongwei Liu and Huaiyi Yang},
  journal = {Microbial Biotechnology},
  year = {2026},
  month = nov,
  doi = {10.1111/1751-7915.70469},
  url = {https://doi.org/10.1111/1751-7915.70469},
  note = {CC BY-NC 4.0},
}
RIS
TY  - JOUR
TI  - Reprogramming Native Cryptic Plasmids Enables Antibiotic‐Selection‐Free Urate Degradation by Escherichia coli Nissle 1917
AU  - Yuan Chi
AU  - Shiying Wu
AU  - Fei Lin
AU  - Tingwen Fan
AU  - Hongwei Liu
AU  - Huaiyi Yang
PY  - 2026
DA  - 2026/11/03
JO  - Microbial Biotechnology
DO  - 10.1111/1751-7915.70469
UR  - https://doi.org/10.1111/1751-7915.70469
AB  - Hyperuricemia is associated with renal injury and disturbances in gut microbial ecology. Intestinal urate clearance by engineered probiotics is limited by antibiotic‐free functional maintenance and the oxygen requirement of urate oxidase (UOX). We reprogrammed the native cryptic plasmids of Escherichia coli Nissle 1917 (EcN) to construct M12U, a marker‐free dual‐plasmid strain. pMUT2 carried an HucR‐regulated urate‐responsive UOX module, while pMUT1 encoded a low‐oxygen‐responsive UOX module and Tat‐targeted Vitreoscilla haemoglobin (tat‐VHB). M12U degraded urate in vitro and retained activity under low‐oxygen conditions. In hyperuricemic mice (n = 5 per group), oral M12U reduced serum urate, creatinine, blood urea nitrogen, inflammatory cytokines and renal histopathological injury. 16S rRNA profiling showed increased microbial richness and a partial shift in community composition toward healthy controls. Shotgun metagenomics revealed enrichment of Lactobacillus johnsonii and Bacteroides acidifaciens , together with increased metabolic potential related to the pentose phosphate pathway and glutathione metabolism. Faecal metabolomics showed reduced uric acid abundance and increased thiamine monophosphate and argininosuccinic acid. These results support further evaluation of dual‐native‐plasmid architectures for gut‐directed urate clearance.
SN  - 1751-7915
ER  -