Phosphorus-31 nuclear magnetic resonance studies of wild-type and glycolytic pathway mutants of Saccharomyces cerevisiae

摘要: High-resolution phosphorus-31 nuclear magnetic resonance (31P NMR) spectra of wild-type and mutant strains Saccharomyces cerevisiae were observed at a frequency 145.7 MHz. Levels various phosphorus metabolites investigated upon addition glucose under both aerobic anaerobic conditions. Three isolated their biochemical defects characterized: pfk lacked phosphofructokinase activity; pgi phosphoglucose isomerase cif had no catabolite repression the fructose bisphosphatase activity. Each strain was found to accumulate characteristic sugar phosphates when added cell suspension. In case deficient mutant, appearance pentose shunt metabolite observed. 31P NMR peak assignments made by pH titration acid extract cells. Separate signals for terminal, penultimate, central atoms in intracellular polyphosphates allowed estimation average molecular weight. Signals glycero(3)phosphochline, glycero(3)phosphoserine, glycero(3) phosphoethanolamine as well three types nucleotide diphosphate sugars could be The resting cells range 6.5--6.8 level adenosine 5'-triphosphate (ATP) low. Upon introduction oxygen, ATP increased considerably reached value 7.2--7.3, irrespective external medium pH, indicating active proton transport these A new representing inorganic phosphate one cellular organelles, whose differed from cytoplasmic detected appropriate