Designing cryo-enzymatic reactions in subzero liquid water by lipidic mesophase nanoconfinement.
2021
Cryo-enzymology provides the possibility to develop unconventional biological reactions and detect intermediates in ultrafast enzymatic catalysis processes, but also illuminates the understanding of life principles in extremely cold environments. The scarcity of biological or biomimetic host systems that provide liquid water at subzero temperatures inhibits the prosperity of cryo-enzymology. Here we introduce cryo-enzymatic reactions in subzero water nanoconfined within lipid mesophases formed by conventional lipids. We show that the enzymatic reactions that ensue outperform the homologue catalytic processes run at standard temperatures. We use phytantriol-based lipidic mesophases (LMPs), within which water remains in the liquid state down to −120 °C, and combine crystallization and dynamic studies of the confined water to provide a fundamental understanding of the physical status of water at subzero temperatures, which sets the stage for cryo-enzymatic reactions in these environments. In the model horseradish peroxidase oxidization, the cation free-radical product is stabilized in LMPs at −20 °C, in contrast to the fast-consuming reactions at temperatures above 0 °C. Furthermore, the LMP system also supports the cascade reaction and lipase reaction at subzero temperatures, at which enzymatic reactions with both hydrophilic and hydrophobic substrates are successfully carried out. Our designed LMP system opens access to the nature of confined water in the biomimetic environment and provides a platform for low-temperature biomacromolecule reconstitution and the cryogenic control of enzymatic reactions in bionanotechnology. By employing the nanoconfinement in the mesophase formed by conventional lipids, cryo-enzymatic reactions are performed in subzero liquid water.
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