.) The incorporation of stable isotopes into proteins, coupled with multi-dimensional NMR techniques enables a rapid determination of protein structure and protein-ligand interaction, thus, aiding in the design and development of new pharmaceuticals. The growth of microorganisms using stable-isotope-labeled glycerol as the sole carbon source results in the production of labeled proteins from those organisms. The uncertain availability and high cost of labeled glycerol limits the wide-spread application of these new techniques. The goal of this research is to produce 13C- and 2H-labeled compounds that can be used as cell culture nutrients to produce labeled proteins and ligands. This project will optimize the production of 13C-glycerol from the photosynthetic microalgae Dunaliella salina and 2H-glycerol by chemical synthesis, and recover 13C-labeled glucose and amino acids from the D. salina biomass. Recovery of glucose and amino acids will lower the cost of producing 13C-glycerol, and provide other 13C-labeled nutrients that are useful for mammalian cell culture. At the conclusion of this project, several stable-isotope-labeled cell culture nutrients will be available to aid researchers in studies of protein structure and protein-ligand interactions, thus providing new tools for rationale drug design.
