Programmable Multiphasic Condensates Formed via Evaporation–Induced Phase Separation of Minimal Peptide Model

Biomolecular self-assembly is ubiquitous in nature, encompassing both ordered and disordered structures to create sophisticated superstructures essential for complex biological functions. Protein and peptide condensates formed via liquid-liquid phase separation (LLPS) are characterize by disordered assembly, gaining significant interest due to their crucial role in physiological events and potential applications from drug delivery to biosensing. Short peptides with ordered structures have been widely explored as building blocks for nanoarchitectured materials, but they lack the disordered features that endow biological systems with flexibility and adaptability. Here we introduce a minimalistic peptide sticker-and-spacer model that forms biomolecular condensates with core-shell structure through phase separation and spontaneous evaporation. The design allows to derive the guidelines for programming condensate's architecture from homogeneous to multiphasic state via the selection of sticker and spacer. Furthermore, we demonstrate control over compartmentalization driven by intrinsic redox chemistry and post-assembly modification. The condensates efficiently encapsulate and protect small-molecule payloads and function as microreactors. The evaporation-induced spontaneous phase separation results in solidified condensates enriched with redox-active tyrosine, which serve as novel nano-bioreactors, promoting selective biomineralization and formation of uniform metal-peptide nanohybrids. Therefore, our study provides a framework for the artificial design of protocells mimetic multicompartmental condensates endowed with on-demand functionality.