Water never freezes in these lipid ‘traps’

Three-dimensional model of the novel lipid mesophase: This cubic motif repeats regularly in the material. (Credit: Peter Rüegg/ETH Zurich)

Scientists have discovered a way to prevent water from freezing by confining in nanometer-scale lipid channels.

Even at extreme sub-zero temperatures the water retains the amorphous characteristics of a liquid.

Making ice cubes is a simple process: you take a plastic ice-cube tray, fill it with water, and put it in the freezer. Before long, the water crystallizes and turns to ice.

If you were to analyze the structure of ice crystals, you’d see that the water molecules arrange in regular three-dimensional lattice structures. In water, by contrast, the molecules have no organization, which explains why water flows.

For the new study, which appears in Nature Nanotechnology, researchers at ETH Zurich designed and synthesized a new class of lipids (fat molecules) to create a new form of “soft” biological matter known as a lipidic mesophase.

In this material, the lipids spontaneously self-assemble and aggregate to form membranes, behaving similarly to natural fat molecules. The membranes then adopt a uniform arrangement to form a network of connected channels that measure less than one nanometer in diameter. Temperature and water content, as well as the novel structure of the designed lipid molecules, determine the structure that the lipidic mesophase takes.

What’s so special about this structure is that—unlike in an ice-cube tray—the narrow channels have no room for water to form ice crystals, so it remains disordered even at extreme sub-zero temperatures. The lipids don’t freeze either.

Using liquid helium, the researchers cooled a lipidic mesophase consisting of a chemically modified monoacylglycerol to a temperature as low as negative 263 degrees Celsius—a mere 10 degrees above the absolute zero temperature—and still no ice crystals formed.

At this temperature, the researchers demonstrated and confirmed in a simulation that the water became “glassy.”

The tricky part

“The key factor is the ratio of lipids to water,” says Raffaele Mezzenga, professor in the Laboratory of Food & Soft Materials at ETH Zurich.

Accordingly, it is the water content in the mixture that determines the temperatures at which the geometry of the mesophase changes. If, for example, the mixture contains 12 percent water by volume, the structure of the mesophase will transition at about minus 15 degrees Celsius from a cubic labyrinth to a lamellar (plate-like) structure.

“What makes developing these lipids so tricky is their synthesis and purification,” says Ehud Landau, professor of chemistry at the University of Zurich. This is because lipid molecules have two parts; one that is hydrophobic (repels water) and one that is hydrophilic (attracts water), he says. “This makes them extremely difficult to work with.”

The soft biomaterial formed from the lipid membranes and water has a complex structure that minimizes the water’s contact with the hydrophobic parts and maximizes its interface with the hydrophilic parts.

Biomimicry

The researchers modeled the new class of lipids on membranes of certain bacteria. These bacteria also produce a special class of self-assembling lipids that can naturally confine water in their interior, enabling the microorganisms to survive in very cold environments.

“The novelty of our lipids is the introduction of highly strained three-membered rings into specific positions within the hydrophobic parts of the molecules. These enable the necessary curvature to produce such tiny water channels and prevent lipids to crystallize,” Landau says.

Keeping membranes intact

These new lipidic mesophases will serve primarily as a tool for other researchers, who can use them to non-destructively isolate, preserve, and study large biomolecules in a membrane-mimicking environment, for instance by using cryogenic electron microscopy.

Biologists are increasingly turning to this method to determine the structures and functions of large biomolecules such as proteins or large molecular complexes.

“In the normal freezing process, when ice crystals form they usually damage and destroy membranes and crucial large biomolecules, which prevents us from determining their structure and function when they interact with lipid membranes,” Mezzenga says.

But not with the new mesophase, which is non-destructive and preserves such molecules in their original state and in presence of the other key building block of life, that is the lipids. “Our research is paving the way for future projects to determine how proteins might be preserved in their original form and interact with lipid membranes at very low temperatures,” Mezzenga says.

Scientists could also use the new class of soft matter in potential applications wherever they need to prevent water from freezing.

“But our work wasn’t aimed at exotic applications,” Mezzenga says: “Our main focus was to give researchers a new tool to facilitate the study of molecular structures at low temperature without ice-interfering crystals, and ultimately to understand how two main components of life, i.e. water and lipids, interact under extreme conditions of temperature and geometrical confinement.”

Source: ETH Zurich