PRODUCTION OF STABLE ISOTOPICALLY LABELED GLYCEROL

THE PRESENCE OF ENRICHED STABLE ISOTOPE CONTENTS IN PROTEINS, COUPLED WITH SOPHISTICATED TWO- AND THREE-DIMENSIONAL NUCLEAR MAGNETIC RESONANCE TECHNIQUES, ENABLES A RAPID AND EASY DETERMINATION OF AMINO ACID SEQUENCE AND PROTEIN STRUCTURE. GLYCEROL IS OFTEN USED AS THE SOLE CARBON SOURCE FOR THE GROWTH OF GENETICALLY MODIFIED BACTERIA THAT OVERPRODUCE PARTICULAR PROTEINS; IN THE PRESENCE OF LABELED GLYCEROL, THE PROTEINS WILL BE LABELED. DEUTERATED GLYCEROL AND 13C-GLYCEROL ARE COMMERCIALLY AVAILABLE, BUT HIGH COST AND UNCERTAIN AVAILABILITY LIMIT THEIR WIDESPREAD USAGE. THE GOAL OF THIS RESEARCH IS TO STUDY THE PRODUCTION AND DETERMINE THE ECONOMIC FEASIBILITY OF PRODUCING 13C- AND 2H-GLYCEROL. IT IS PLANNED TO PRODUCE ISOTOPICALLY LABELED GLYCEROL BIOLOGICALLY FROM THE PHOTOSYNTHETIC MICROALGA, DUNALIELLA SALINA, AN ORGANISM WIDELY REPORTED TO PRODUCE UPTO 50 PERCENT OF ITS BIOMASS AS GLYCEROL. FOR THE PRODUCTION OF DEUTERATED GLYCEROL, D. SALINA WILL BE GROWN IN D20 (HEAVY WATER). IT IS ALSO PLANNED TO CHEMICALLY SYNTHESIZE DEUTERATED GLYCEROL USING INEXPENSIVE STARTING MATERIALS AND TO COMPARE THE ECONOMICS OF CHEMICALS SYNTHESIS TO THOSE OF BIOLOGICAL PRODUCTION. SUCCESS IN ACHIEVING THIS GOAL WILL LAY THE GROUNDWORK FOR THE DEVELOPMENT OF STABLE ISOTOPICALLY LABELED COMPOUNDS THAT WILL BE VITAL FOR THE ELUCIDATION OF PROTEIN STRUCTURES.