Many bacteria release membrane vesicles, which are nanoscale spheres consisting of a cellular membrane containing biomolecules. Membrane vesicles can transport DNA and proteins, and are involved in bacterial interactions. They have potential applications in nanotechnology and biomedicine, including cancer treatment. However, the formation of membrane vesicles by bacteria is currently not well understood. In particular, the release mechanism of membrane vesicles of Gram-positive bacteria, which have a very thick cell wall, has remained an enigma.