Extracellular vesicles produced by the G ram‐positive bacterium B acillus subtilis are disrupted by the lipopeptide surfactin

Summary Previously, extracellular vesicle production in G ram‐positive bacteria was dismissed due to the absence of an outer membrane, where G ram‐negative vesicles originate, and the difficulty in envisioning how such a process could occur through the cell wall. However, recent work has shown that G ram‐positive bacteria produce extracellular vesicles and that the vesicles are biologically active. In this study, we show that B acillus subtilis produces extracellular vesicles similar in size and morphology to other bacteria, characterized vesicles using a variety of techniques, provide evidence that these vesicles are actively produced by cells, show differences in vesicle production between strains, and identified a mechanism for such differences based on vesicle disruption. We found that in wild strains of B . subtilis , surfactin disrupted vesicles while in laboratory strains harbouring a mutation in the gene sfp , vesicles accumulated in the culture supernatant. Surfactin not only lysed B . subtilis vesicles, but also vesicles from B acillus anthracis , indicating a mechanism that crossed species boundaries. To our knowledge, this is the first time a gene and a mechanism has been identified in the active disruption of extracellular vesicles and subsequent release of vesicular cargo in G ram‐positive bacteria. We also identify a new mechanism of action for surfactin.