The physiological function of periplasmic glucose oxidation in phosphate-limited chemostat cultures of Klebsiella pneumoniae NCTC 418.
Periplasmic oxidation of glucose into gluconate and 2-ketogluconate in Klebsiella pneumoniae occurs via glucose dehydrogenase (GDH) and gluconate dehydrogenase (GaDH), respectively. Since, as is shown here, in the presence of glucose, gluconate and 2-ketogluconate are not further metabolized intracellularly the physiological function of this periplasmic route was studied. It was found that periplasmic oxidation of glucose could function as an alternative production route of ATP equivalents. Instantaneous activation of either GDH or GaDH reduced the rate of degradation of glucose via glycolysis and the tricarboxylic acid (TCA) cycle in vivo. Furthermore, aerobic, magnesium- and phosphate-limited chemostat cultures with glucose as the carbon source showed high GDH plus GaDH activities in contrast to nitrogen- and sulphate-limited cultures. However, when fructose, which is not degraded by GDH, was the carbon source, specific oxygen consumption rates under these four conditions were essentially the same. The latter observation suggests that high transmembrane phosphate gradients which are supposedly present under phosphate-limited conditions do not cause high energetic demands due to futile cycling of phosphate ions. In addition, dissipation of the transmembrane phosphate gradient of phosphate-limited cells immediately increased the rate of intracellular glucose degradation. It is concluded that under phosphate-limited conditions (i) extensive futile cycling of phosphate ions is absent and (ii) low concentrations of phosphate ions limit intracellular degradation of glucose.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Microbiology, E. C. Slater Institute, University of Amsterdam, The Netherlands.
E T
ET
Buurman
G J
GJ
ten Voorde
M J
MJ
Teixeira de Mattos
D000255:Adenosine Triphosphate / Q000378:metabolism; D002237:Carbohydrate Dehydrogenases / Q000378:metabolism; D002245:Carbon Dioxide / Q000378:metabolism; D003470:Culture Media; D004789:Enzyme Activation; D005947:Glucose / Q000378:metabolism*; D042843:Glucose 1-Dehydrogenase; D005948:Glucose Dehydrogenases / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D007700:Kinetics; D007711:Klebsiella pneumoniae / Q000378:metabolism*; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D010710:Phosphates / Q000378:metabolism
D003470:Culture Media; D010710:Phosphates; D002245:Carbon Dioxide; D000255:Adenosine Triphosphate; D002237:Carbohydrate Dehydrogenases; D005948:Glucose Dehydrogenases; C020577:gluconate 2-dehydrogenase; D042843:Glucose 1-Dehydrogenase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005947:Glucose