Growth suppression of ice crystal basal face in the presence of a moderate ice-binding protein does not confer hyperactivity.

Ice-binding proteins (IBPs) affect ice crystal growth by attaching to crystal faces. We present the effects on the growth of an ice single crystal caused by an ice-binding protein from the sea ice microalga (IBP) that is characterized by the widespread domain of unknown function 3494 (DUF3494) and known to cause a moderate freezing point depression (below 1 °C). By the application of interferometry, bright-field microscopy, and fluorescence microscopy, we observed that the IBP attaches to the basal faces of ice crystals, thereby inhibiting their growth in the direction and resulting in an increase in the effective supercooling with increasing IBP concentration. In addition, we observed that the IBP attaches to prism faces and inhibits their growth. In the event that the effective supercooling is small and crystals are faceted, this process causes an emergence of prism faces and suppresses crystal growth in the direction. When the effective supercooling is large and ice crystals have developed into a dendritic shape, the suppression of prism face growth results in thinner dendrite branches, and growth in the direction is accelerated due to enhanced latent heat dissipation. Our observations clearly indicate that the IBP occupies a separate position in the classification of IBPs due to the fact that it suppresses the growth of basal faces, despite its moderate freezing point depression.