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Background <p>Plasmid DNA (pDNA) is a key biomolecule for gene therapies, DNA vaccines, and mRNA-based applications. Although <i>Escherichia coli</i> is the established host for pDNA production, limitations such as moderate growth rates and endotoxin-associated challenges motivate the exploration of alternative production hosts. <i>Vibrio natriegens</i> is a fast-growing, non-pathogenic bacterium with a doubling time below 10&#xa0;min and high metabolic versatility. While it has been adopted for cloning and protein expression, its potential for pDNA production remains largely unexplored.</p> Results <p>We evaluated pDNA production by <i>V. natriegens</i> in comparison to <i>E. coli</i> in shake flask cultivations across four complex media (LB, BHI, 2xYT, TB) and one defined minimal medium (MSM). Under all tested conditions, <i>V. natriegens</i> exhibited faster biomass accumulation and enabled earlier pDNA isolation. Quantitative analysis of triplicate experiments revealed that <i>V. natriegens</i> achieved comparable or higher volumetric and specific pDNA yields in shorter cultivation times than <i>E. coli</i>. Agarose gel electrophoresis confirmed plasmid integrity, and functional validation using a cell-free expression system demonstrated efficient eGFP reporter expression from <i>V. natriegens</i>-derived plasmids.</p> Conclusions <p>This proof-of-concept study demonstrates that <i>V. natriegens</i> is a promising alternative host for pDNA production. Its combination of accelerated growth, robust pDNA yields, and functional plasmid quality highlights its potential for further development as a pDNA production platform.</p>

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Evaluation of Vibrio natriegens as a fast-growing alternative host for plasmid DNA production

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年5月15日

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  • Lara S. Möller,
  • Gabriel A. Monteiro,
  • Duarte M. F. Prazeres,
  • A. Rita Silva-Santos

摘要

Background

Plasmid DNA (pDNA) is a key biomolecule for gene therapies, DNA vaccines, and mRNA-based applications. Although Escherichia coli is the established host for pDNA production, limitations such as moderate growth rates and endotoxin-associated challenges motivate the exploration of alternative production hosts. Vibrio natriegens is a fast-growing, non-pathogenic bacterium with a doubling time below 10 min and high metabolic versatility. While it has been adopted for cloning and protein expression, its potential for pDNA production remains largely unexplored.

Results

We evaluated pDNA production by V. natriegens in comparison to E. coli in shake flask cultivations across four complex media (LB, BHI, 2xYT, TB) and one defined minimal medium (MSM). Under all tested conditions, V. natriegens exhibited faster biomass accumulation and enabled earlier pDNA isolation. Quantitative analysis of triplicate experiments revealed that V. natriegens achieved comparable or higher volumetric and specific pDNA yields in shorter cultivation times than E. coli. Agarose gel electrophoresis confirmed plasmid integrity, and functional validation using a cell-free expression system demonstrated efficient eGFP reporter expression from V. natriegens-derived plasmids.

Conclusions

This proof-of-concept study demonstrates that V. natriegens is a promising alternative host for pDNA production. Its combination of accelerated growth, robust pDNA yields, and functional plasmid quality highlights its potential for further development as a pDNA production platform.