Precise Control Over Kinetics Assembly of Potent Antifreeze Peptide from Ice-Templated Strategy

Precise kinetic control over peptide assembly into functional nanostructures remains a major challenging. Herein, we report a novel ice-templated assembly strategy driven by programmed ice recrystallization, which enables ultrafast, kinetically tunable fabrication of peptides into well-defined nanofibers with precisely controllable dimensions. The nanostructure size can be quantitatively correlated with recrystallization parameters, including temperature and annealing time, providing a direct route to regulate assembly kinetics. The 4-mercaptobenzoic acid-modified threonine pentamer (MT5) exhibits exceptional ice recrystallization inhibition (IRI) activity, originating from the aligned threonine residues that afford enhanced ice-binding affinity. Importantly, MT5 nanofibers enabled reduced-DMSO cell cryopreservation, maintaining post-thaw cell viability with 5% (v/v) DMSO at a level comparable to the standard 10% (v/v) DMSO formulation. Molecular dynamics simulations revealed that the neighboring threonine on MT5 match the spacing of one ice lattice on the primary prism plane. This ice-templated method breaks through the bottlenecks of conventional peptide assembly, offers a versatile and programmable platform for rapid fabrication of functional nanostructures, and unlocks new routes for the rational design of high-performance antifreeze materials.