Gemini cationic surfactant is a promising generation of surfactants characterized by their unique surface properties and biological activities. Herein, a series of three Gemini cationic surfactants containing ester bond with different alkyl chain length and their corresponding monomeric structures have been synthesized. The monomeric surfactant were synthesized via simple esterification of different fatty acids (stearic, myrisitic, lauric acid) with 2-dimethyl amino ethanol, followed by quaternization with 1-bromohexane; while, and 1,6-dibromohexane was used to prepare the corresponding Gemini cationic surfactants. The structures of the synthesized compounds were elucidated using 1H-NMR, FTIR spectroscopy. The biological activities and surface properties have been assayed and it was found that the prepared Gemini surfactants have higher antimicrobial efficiencies than the corresponding monomeric structure.