Hystatin is a group of compounds resulted from the derivatization of aaptamine. A series of hystatin 2 derivatives (3, 5-15) have been synthesized and characterized using modern spectroscopic techniques, namely IR, 1H and 13C NMR, UV-Vis and Mass Spectrometer. The synthesized compounds were evaluated for their antibacterial activity against selected bacterial strains of both Gram positive and Gram negative groups, namely Bacillus cereus, Staphylococcus aureus, Micrococcus sp, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae using standard microbiological protocol of disc-diffusion method. The antibacterial activities of the compounds were assessed by the presence and absence of inhibition zones and minimum inhibition concentration (MIC) values. In general, most of the derivatives showed significant antibacterial activity towards Gram positive bacteria. It was observed that compound 6 exhibited bacterial inhibition activity against all tested bacteria. Nonetheless, compounds 5, 6 and 14 showed no antibacterial activities towards both Gram positive and Gram negative bacteria.