Polyol-Induced 100-Fold Enhancement of Bacterial Ice Nucleation Efficiency.

Ice-nucleating proteins (INPs) from bacteria like are among the most effective ice nucleators known. However, large INP aggregates with maximum ice nucleation activity (at approximately -2 °C) typically account for less than 1% of the overall ice nucleation activity in bacterial samples. This study demonstrates that polyols significantly enhance the assembly of INPs into large aggregates, dramatically improving bacterial ice nucleation efficiency.

Functional aggregation of cell-free proteins enables fungal ice nucleation.

Biological ice nucleation plays a key role in the survival of cold-adapted organisms. Several species of bacteria, fungi, and insects produce ice nucleators (INs) that enable ice formation at temperatures above -10 °C. Bacteria and fungi produce particularly potent INs that can promote water crystallization above -5 °C.

Measurement of Ice Nucleation Activity of Biological Samples.

Experimentation with ice-nucleating biomolecules is needed to advance the fundamental understanding of biotic heterogeneous ice nucleation. Standard experimental procedures vary with sample type. Here we describe a generalized primary purification and analysis process to measure ice nucleation activity of biological samples using an advanced freezing droplet assay.

Toward Understanding Bacterial Ice Nucleation.

Bacterial ice nucleators (INs) are among the most effective ice nucleators known and are relevant for freezing processes in agriculture, the atmosphere, and the biosphere. Their ability to facilitate ice formation is due to specialized ice-nucleating proteins (INPs) anchored to the outer bacterial cell membrane, enabling the crystallization of water at temperatures up to -2 °C. In this Perspective, we highlight the importance of functional aggregation of INPs for the exceptionally high ice nucleation activity of bacterial ice nucleators.

Membranes Are Decisive for Maximum Freezing Efficiency of Bacterial Ice Nucleators.

Ice-nucleating proteins (INPs) from are among the most active ice nucleators known, enabling ice formation at temperatures close to the melting point of water. The working mechanisms of INPs remain elusive, but their ice nucleation activity has been proposed to depend on the ability to form large INP aggregates. Here, we provide experimental evidence that INPs alone are not sufficient to achieve maximum freezing efficiency and that intact membranes are critical.