Isolation and Characterization of Cyanobacterial Extracellular Vesicles

Cyanobacteria are a diverse group of photosynthetic, Gram-negative bacteria that play critical roles in global ecosystems and serve as essential biotechnology models. Recent work has demonstrated both marine freshwater cyanobacteria produce extracellular vesicles - small membrane-bound structures released from the outer surface microbes. While likely contribute to biological processes, their specific functional cyanobacterial biology remain largely unknown. To encourage advance research this area, detailed protocol is presented for isolating, concentrating, purifying vesicles. The current discusses methodologies have successfully isolated large cultures Prochlorococcus, Synechococcus, Synechocystis. Methods quantifying characterizing vesicle samples these strains presented. Approaches isolating aquatic field also described. Finally, typical challenges encountered with purification, methodological considerations different downstream applications, trade-offs between approaches discussed.