Two series of cationic surfactants based on glycine molecule as sulfonate salt of N,N-diethyl glycine fatty esters having a hydrocarbon chain length of C-12,C-16 and C-18 followed by quaternerization with ethyl iodide were synthesized. Their chemical structures were confirmed using IR,H-1-NMR, and mass spectroscopy. The surface activity of these amphiphiles has been determined by measurement of their surface tension. Parameters like surface tension (gamma), critical micelle concentration (cmc), effectiveness (Pi(cmc)), efficiency (pC(20)), maximum surface excess (Gamma(max)), minimum surface area (Amin) and Gibbs free energy of the micellization (Delta G(mic)) and adsorption (Delta G(ads)) have been determined from the measurements obtained by surface tension method. All these surfactants showed much lower CMC values compared to the similar hydrophobic chain length surfactant. All synthesized compounds have been evaluated for their biological activity against bacteria and fungi.