Amino acid surfactants have mild performance and are sourced from renewable biomass. Compared to classical surfactants, Gemini surfactants have superior properties. The amino acid Gemini surfactants are believed to be adopted more widely. The Gemini surfactant with tripeptide structure, sodium di(lauroyl glutamyl) lysine (DLGL), was prepared by amidation utilizing methyl laurate, glutamate and lysine and characterized by 1H NMR, 13C NMR and MS. Additionally, the pKa value, surface activities, aggregation, foaming properties and emulsifying attributes of the DLGL surfactant in aqueous solution with varied pH values were examined. The results indicate that the protonation‐deprotonation behavior of the DLGL surfactant is highly dependent on pH values. The surface tension, critical micelle concentration (cmc), foamability and foam stability exhibited superior performance at pH 6 and 7. Conversely, superior emulsifying ability was observed at pH 9 and 10. Moreover, the spherical vesicles were formed by the DLGL surfactant at pH 6, 7 or 8 while the micelles were generated at pH 9 or 10.