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

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