We present a new strategy for a one-step preparation of macroscopic peptide-inorganic hybrid supramolecular films with functionality. It has been found that photothermal induction is of key importance to mediate the disassembly of supramolecular structures in solution phase, enrichment and reorganization at the interfaces associated with water evaporation. Incorporation of functional nanoscale objects into the film is achieved, providing the potential of integration of multiple functions. Texture and thickness of the hybrid film assembled at interfaces can be tuned and controlled by alteration of experimental parameters such as concentration and irradiation time. The photothermal treatment approach developed in this work is simple, flexible and controllable for design of macroscale functional supramolecular films, directly interfacing with biological systems. The photothermally-induced transformation of supramolecular structures is attractive for preparation of macroscale materials or devices towards a broad range of applications in biology and in nanotechnology.