The biophysical effects of deuterium oxide on biomolecules and living cells through open notebook science.
Creators: name: Salvagno, Anthony
Publisher: name: figshare
Year: 2013
Subjects: Science Policy
Biophysics

description: This dissertation explores various effects of deuterium oxide (D2O also known as heavy water) in nature. Water is everywhere and interacts with just about everything. As such, it would be quite a daunting task to characterize every effect that water exhibits on everything in the universe. This research is a small piece of the puzzle, and provides some fundamental understanding of how water interacts with other molecules. This is done from two viewpoints: (1) the effects of heavy water on living cells and (2) the effects of heavy water on molecules. Varying concentrations of deuterium oxide were used as the growing solvent for four different organisms: S. cerevisiae, E. coli, A. thaliana, and N. tabacum. In each case growth rates and morphology was assessed and compared to the wild type. Organisms were surveyed for potential phenotypes exhibited in the presence of extremely low and high concentrations of D2O. In every organism, growth is increasingly inhibited in higher concentrations of D2O compared to lower concentrations of D2O. In the case of tobacco, a root hair phenotype was exhibited in the presence of deuterium depleted water (&lt;1ppm deuterium atoms). Roots also grew faster in 1% D2O and DDW, compared to natural water. For Arabidopsis, root germination is statistically indistinguishable between DI water and 33% D2O. Growth of the plant in 10% D2O is identical to that of natural water, and potentially healthier. Meanwhile, plants grown in 60% D2O exhibit slower growth and leaf discoloration. Tests on E. coli reveal inconsistent growth rates, but exhibit increased growth in DDW when adapted to D2O. Cellular and colonial morphology is also very distinguished from the wt. Cells appear to remain joined after cellular fission, while colonies exhibit brainy structures. Yeast morphology is quite different. Yeast cells remain joined after mitosis in 99% D2O, causing large cellular aggregates, while colonies become slightly asymmetric. Adaptation of yeast to D2O was not possible. Molecular effects were examined using a variety of tools including: dynamic light spectroscopy, Fourier transform-infrared spectroscopy, cavity ring-down spectroscopy, and optical tweezers. Heat induced protein aggregation was possible in H2O, but prevented in the presence of D2O and analyzed via DLS. Deuterium exchange and replacement was observed and quantified using both FT-IR and CRDS. With FT-IR it was possible to identify differences between solvents, while the time-scale of hydrogen-deuterium exchange was quantified for bulk water with CRDS. Using optical tweezers, DNA was overstretched in both H2O and D2O. The average force for DNA overstretching was found to be ~2.5pN higher in D2O compared to H2O. Deuterium oxide has a stabilizing force on biomolecules, which prevents protein denaturing and can affect the timing for cellular processes. It is because of this molecular property that D2O is observed to affect organisms grown with D2O instead of H2O. Despite this, there seems to be an optimal concentration of deuterium which is above the natural concentration of 155.6ppm. In the presence of deuterium depleted water, cells exhibit signs of stress, further demonstrating that deuterium isn’t merely tolerated in solution, but actually required as hypothesized by Gilbert N. Lewis in 1934.