The development of new strategies for thermal stability and storage of enzymes is very important, considering the nonretention of catalytic activity by enzymes under harsh conditions of temperature. Following this, herein, a new approach based on the interfacial adsorption of lysozyme (LYZ) at nanointerfaces of ionic liquid (IL)-based microemulsions, for enhanced thermal stability of LYZ, is reported. Microemulsions (MEs) composed of dialkyl imidazolium-based surface active ILs (SAILs) as surfactants, ILs as the nonpolar phase, and ethylene glycol (EG) as the polar phase, without any cosurfactants, have been prepared and characterized in detail. Various regions corresponding to polar-in-IL, bicontinuous, and IL-in-polar phases have been characterized using conductivity measurements. Dynamic light scattering (DLS) measurements have provided insights into the size distribution of microdroplets, whereas temperature-dependent DLS measurements established the thermal stability of the MEs. Nanointerfaces formed by SAILs with EG in thermally stable reverse MEs act as fluid scaffolds to adsorb and provide thermal stability, up to 120 °C, to LYZ. Thermally treated LYZ upon extraction into a buffer shows enzyme activity owing to negligible change in the active site of LYZ, as marked by retention of microenvironment of Trp residues present in the active site of LYZ. The present work is expected to establish a new platform for the development of novel nanointerfaces utilizing biobased components for other biomedical applications.