Mastoparan M, a tetradecapeptide toxin (INKAIAALAKKLL-NH2) from hornet venom and its D-form mastoparan M were synthesized chemically. All D- and L-mastoparan M forms were found to adopt 28% alpha-helical structures in a 30% trifluroethanol solution as shown by the circular dichroism spectrum. All-D mastoparan M caused 3H-thymidine release from labeled bacterial cells after incubation for 1 h and complete cell lysis by 4 h. Both L- and D-mastoparan M showed strong activity against gram-positive and gram-negative bacteria. All-D mastoparan M showed 2-fold higher antibacterial activity than L-mastoparan M. The effects of all-D mastoparan M on the surface morphology of Staphylococcus aureus (ATCC29213) and Escherichia coli (ATCC25922) were studied by scanning-beam electron microscopy. Blast-like bleb extrusions on the surface of some S. aureus and swellings on the end of E. coli were seen after culture with all-D mastoparan M. These findings indicated the all-D mastoparan M could kill bacteria by disrupting cells.