Studying of isotopic effects of heavy water (D2O) in biological systems on example of prokaryotic and eukaryotic cells
Isotope effects of deuterium (D) in cells of various taxonomic groups of prokaryotic and eucaryotic microorganisms realizing methylotrophical, chemoheterotrophical, photoorganotrophical, and photosynthetic ways of assimilation of carbon substrates (methylotrophic bacteria, halobacteria, green microalgae) are investigated at growth on media with heavy water (D2О). The method of step by step adaptation technique of cells to D2О is developed, consisting in plating of ctlls on 2% agarose nutrient media containing increasing gradient of concentration of D2О (from 0 up to 98% D2О) and the subsequent selection of stable to D2О cells. Obtained from growth media with a low gradient of D2О concentration, cells were further transferred on growth media with higher gradient of D2О concentration, up to 98% D2О. In the result of that technique it were obtained adapted to maximum concentration of D2О cells, biological material of which instead of hydrogen contained deuterium with levels of enrichment 95% D. Usage of cells adapted to D2О and deuterated natural compounds (protein, amino acids, nucleosides, pigments, carboxydrates, lipids), extracted from them, can find further application in biophysical researches and biomedical technologies