Three Gemini surfactants using adamantane as spacer group with different carbon chain length, nP-Ad-Pn (n=12, 14, 16), were synthesized from 1,3-adamantanedicarboxylic acid. The success of synthesis was confirmed by FT-IR spectra, 1H NMR spectra, and ESI-MS spectra. The surface-active properties, thermodynamic parameters and micropolarity of the Gemini surfactants containing adamantane were investigated through surface tension, electrical conductivity measurement and steady-state fluorescence. The results show that the values of CMC are 6?0-4, 1?0-4 and 2.5?0-5 mol?L-1 respectively. the standard Gibbs free energy change (?G0m) and the standard enthalpy changes (?H0m) are negative, indicating that the micellization is spontaneous and exothermic. Further, the antimicrobial activity of the synthesized compounds against S. aureus, B. subtilis, E. coli, P. aeruginosa, V. Parahemolyticus was also investigated, and this study showed that the MIC of 12P-Ad-P12 was between 0.2 mg/L and 4.2 mg/L, which showed that this compounds have excellent antibacterial activity against five studied bacteria.