Developing sustainable, “green” synthetic routes for functional nanomaterials is essential for advancing biocompatible nanotechnology. In this study, we report the bottom-up biomineralization of gold–peptide supersphere composites (Au–HF SCs) using a biocompatible His–Phe (HF) dipeptide via a one-pot strategy without the addition of a separate reducing reagent. The HF peptide simultaneously functions as a reducing agent, self-assembling unit, capping agent, and stabilizer, enabling the formation of uniform composite nanostructures simply by mixing and heating with Au(III). By varying the peptide/Au(III) molar ratio from 0.67 to 17, the primary particle size of Au nanoparticles (AuNPs) and the morphology of secondary spherical assemblies were tunably controlled. The catalytic performance of the resulting Au–HF SCs was evaluated via the reduction of 4-nitrophenol (4-NP) as a model reaction. Characterization revealed hierarchical structures consisting of primary AuNPs (8–13 nm) supported on secondary peptide spheres (190–320 nm). The apparent rate constant () exhibited characteristic concentration dependences on 4-NP and , which were accurately described by the Langmuir–Hinshelwood model. Kinetic analysis provided adsorption and surface reaction rate constants, revealing that Au–HF SCs with a primary particle size of approximately 10 nm exhibited the highest catalytic activity. Furthermore, Eyring and Arrhenius analyses were conducted to determine the activation enthalpy, activation entropy, and activation energy, verifying the correlation between catalyst structure and kinetic parameters. Peptide-templated gold superspheres demonstrate that catalytic activity depends on particle size and molecular-level structural order, offering a versatile platform for designing advanced, multifunctional nanocatalysts.


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