A series of single surfactant systems, i.e, quaternary ammonium-based gemini surfactants with different spacers and alkyl chain lengths (m-n-m; m = 12, n = 2, 3, 4, 6; n = 3, m = 12, 14, 16), halogen-free surface-active ionic liquid (HF-SAILs) with different symmetries ([Cnmim][C12H25SO4]; n = 6, 8, 10, 12), and single-chain cationic surfactants including 1-dodecyl-3-methylimidazolium bromide ([C12mim]Br) and dodecyltrimethylammonium bromide (DTAB), along with certain combinations of different surfactants (12-3-12/[C12mim]Br and 12-3-12/DTAB) were applied to an aqueous/liquid crystal interface (ALI). All the surfactants could induce an orientational transition of liquid crystals (LCs) from a planar to homeotropic state, which caused a bright-to-dark optical shift. It was proved that double-chain surfactants and the mixed surfactants inclined to adsorb at the ALI triggering the orientational transition. Inspiringly, a quicker and more sensitive dark-to-bright optical response was observed for mixed surfactant system-decorated interfaces in contact with proteins (such as bovine serum albumin (BSA), lysozyme, and trypsin) as opposed to the single surfactant systems. The ALI decorated by the 12-3-12/[C12mim]Br system was particularly efficient and exhibited the most sensitive optical response for BSA (0.01 ng mL−1). The order parameters (SCD) of surfactants tails at the interface and the free energy of proteins with 12-3-12 and [C12mim]Br were calculated, respectively. The results explain that the 12-3-12/[C12mim]Br-laden ALI shows a quicker and more sensitive optical response for BSA. This work inspired us to study mixed surfactant systems-decorated LC interfaces and further provides new insights for different chemical and biological applications.