[P002] Poster

1:00 PM - 1:50 PM JST(4:00 AM - 4:50 AM UTC)

[P01-116] Selective isolation of bacterial extracellular vesicle-producing bacteria from complex microbial communities by targeting high membrane curvature

○Hayato Kanki¹, Mizuki Kanno², Yusuke Sato³, Yosuke Tashiro^1,2 (1. Grad. Sch. Intgr. Sci. Tech. Shizuoka Univ. (Japan), 2. Grad. Sch. Sci. Tech. Shizuoka Univ. (Japan), 3. Grad. Sch. Intgr. Sci. Tech. Tohoku Univ. (Japan))

Keywords:

Bacterial extracellular vesicles (BEVs),Membrane curvature,Lipid packing deficiency,complex microbial communities

Bacterial extracellular vesicles (BEVs), nanosized particles (20–400 nm) released from bacterial membranes, play key roles in molecular transport and intercellular communication. However, the lack of methods to selectively isolate BEV-producing bacteria from complex microbial communities has limited our understanding of their ecological functions. We previously demonstrated that ApoC-NR, an amphipathic a-helical peptide probe that recognizes highly curved membrane regions associated with BEV formation, preferentially labels BEV-hyperproducing strains in a transposon mutant library of a model bacterium. In this study, we developed a strategy for the direct isolation of BEV-producing bacteria from complex microbial communities by integrating ApoC-NR labeling with magnetic separation. To evaluate its applicability, we applied this method to environmental aquatic samples. ApoC-NR successfully labeled indigenous bacteria in both freshwater and seawater, indicating active BEV production in situ. Furthermore, combining ApoC-NR labeling with magnetic bead-based separation enabled efficient isolation of native BEV-producing bacteria, including Vibrio and Enterobacter species. Magnetic separation significantly improved isolation efficiency compared to conventional approaches.This strategy enables rapid and selective isolation of BEV-producing bacteria from complex microbial communities and provides a practical tool for investigating BEV-mediated functions in natural environments.