New method for Membrane Vesicle (MV) extraction fromGram-positive Bacteria, a nanoparticle useful in biomedicine andnanotechnology

Background and Aim : Bacterial Membrane vesicles (BMVs) play noticeably roles in variousbiological processes such as variation in the composition of the microbiota and facilitate hostmicrobenetwork, particularly applications in biomedicine and nanotechnology and take part indeveloping novel vaccines. We aimed to describe the novel, simplest and less expensive MVextraction protocol with available equipment.Methods : Bacillus subtilis ۶۰۵۱ was cultured in Brain Heart Infusion (BHI) medium for ۲۴h withshaking in ~۲۵۰×rpm then removed B. subtilis cells and cellular debris by several speedscentrifugation (۴۰۰۰۰×g) and times at ۴ °C. Adding detergent to MV sediment, vortexing at highspeedand striking by hand for ۲۰ to ۳۰ minutes. MVs were pelleted just by one step centrifugationat ۵۲۰۰۰×g for ۲h then washed with a phosphate buffered saline solution (PBS) and filtrated by a۰.۴۵ μm syringe filter.Results : MVs morphologic produced by Bacillus subtilis ۶۰۵۱ were seen through ScanningElectron Microscopy (SEM) and Transmission Electron Microscopy (TEM), and the sphericalshape was confirmed, also, diameters MVs ۱۵۰ nm was assessed.Conclusion : The result of this study demonstrated that the novel protocol for extraction the Grampositivebacteria’s MVs is a simple by using common equipment in laboratories and can beperformed as an alternative method with low-speed centrifuges available in most researchlaboratories.

کلیدواژه ها:

MV ، gram positive bacteria ، novel protocol ، detergent