Chromatographic Separation of Deuterium-Labeled Amino Acids, Proteins, Carbohydrates and Lipids Isolated From Bacterial Objects

Abstract

We carried out the chromatographic separation of ²H-labeled amino acids, proteins, carbohydrates and lipids isolated from biological samples of various taxonomic groups of prokaryotic and eucaryotic microorganisms realizing methylotrophic, chemoheterotrophic, photo-organotrophic, and photosynthetic pathways of assimilation of carbon substrates (methylotrophic bacteria Brevibacterium methylicum, chemoheterotrophic bacteria Bacillus subtilis, photo-organotrophic halobacteria Halobacterium halobium, green algae Chlorella vulgaris) at the growth on media with heavy water (²H₂O). For all investigated microorganisms are submitted data on growth and adaptation on growth media containing as sources of deuterated substrates ²H₂O, [²H]methanol and hydrolysates of deutero-biomass of methylotrophic bacteria B. methylicum, obtained after the multistage adaptation to ²H₂O. The qualitative and quantitative composition of intra- and endocellular ²H-labeled amino acids, proteins, carbohydrates and lipids in conditions of adaptation to ²H₂O is investigated. The chromatographic separation was implemented by the methods of RP-HPLC, ion exchange chromatography and electrophoresis.

Key words: amino acids, bacteria, carboxydrates, lipids, heavy water