Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Optimizing Conditions for the Production of Bacterial Extracellular Vesicles of Vibrio vulnificus and Analysis of the Inner Small RNA Compositions

  • Jeong Heon Park (Department of Life Sciences, Sogang University) ;
  • Suji Song (Department of Life Sciences, Sogang University) ;
  • Soyee Kim (Department of Life Sciences, Sogang University) ;
  • Minjeong Kim (Department of Life Sciences, Sogang University) ;
  • Kun-Soo Kim (Department of Life Sciences, Sogang University)
  • Received : 2023.10.04
  • Accepted : 2023.11.22
  • Published : 2024.01.28
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Abstract

Chemical and physical elements affecting the production of bacterial extracellular vesicles (BEVs) of the human pathogen Vibrio vulnificus were quantitatively assessed to optimize the conditions for the BEV production by using the western blot quantification for an outer membrane porin OmpU and by fluorescent dye FM4-64. When cells were cultured at 37℃ in an enriched medium (2 × Luria Bertani; 2 × LB) in the presence of EDTA, they produced about 70% more BEVs. BEVs were purified from the cells cultured in the established optimal conditions by the density gradient ultracentrifugation. The dynamic light scattering measurement of the purified BEVs showed that the diameter of them ranged from approximately 25 nm to 161 nm. We hypothesized that there may be some features in nucleotide sequences specific to RNAs packaged in BEVs compared to those in cellular RNA molecules. We compared the nucleotide sequences and abundance of sRNAs between in the cellular fraction and in BEVs through next-generation sequencing (NGS). While no distinct feature was observed in the nucleotide sequences of sRNAs between two groups, the length of sRNA fragments from BEVs were significantly shorter than those in cytoplasm.

Keywords

Acknowledgement

This work was supported by grants from the National Research Foundation (NRF) of Korea, funded by the Ministry of Science and ICT (2022R1A2C1008958), Republic of Korea.

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