Novel enzymic machinery for the metabolism of oxalacetate, phosphoenolpyruvate, and pyruvate in Pseudomonas citronellolis.
The metabolic pathways for the interconversion of oxalacetate, phosphoenolpyruvate, and pyruvate in Pseudomonas citronellolis form an interlocking system (Scheme 1) that would appear to require complex regulatory mechanisms to permit a proper flow of metabolites through the pathways and to prevent futile cycling. Oxalacetate decarboxylase (I in Scheme 1), P-enolpyruvate synthase (II), P-enolpyruvate carboxylase (III), and pyruvate kinase (V) are constitutive enzymes in this organism. Pyruvate carboxylase (VI) is inducible and has its highest activity in cells grown on glucose or lactate, moderate activity in cells grown on acetate, citrate, or glutamate, and virtually no activity in aspartate-grown cells. P-enolpyruvate carboxykinase (IV) was not detected. The presence of these five enzymes in a single cell has not been previously reported. In Scheme 1, three futile cycles are possible: the simultaneous operation of Reactions I and VI; of Reactions II and V; or of I, II, and III. An examination of the regulatory properties of the individual enzymes after partial purification offers support for the hypothesis of an intricate regulatory system. Oxalacetate decarboxylase (I) is inhibited by acetyl-CoA; phosphoenolpyruvate carboxylase (III) is activated by acetyl-CoA and ADP and inhibited by aspartate; phosphoenolpyruvate synthase (II) is inhibited by 5'-AMP and phosphoenolpyruvate; and pyruvate kinase (V) is activated by 5'-AMP and 2 keto, 3-deoxy,6-phosphogluconate and inhibited by ATP. The presence of metabolites with reciprocal but reinforcing functions is noteworthy. As an example, acetyl-CoA both inhibits the breakdown of oxalacetate and stimulates its formation. Only pyruvate carboxylase appears to be regulated by the carbon substrates of the growth medium.
D000105:Acetyl Coenzyme A / Q000378:metabolism; D000244:Adenosine Diphosphate / Q000378:metabolism; D001224:Aspartic Acid / Q000494:pharmacology; D002262:Carboxy-Lyases / Q000378:metabolism; D003470:Culture Media; D004789:Enzyme Activation / Q000187:drug effects; D005942:Gluconates / Q000494:pharmacology; D007700:Kinetics; D010071:Oxaloacetates / Q000378:metabolism*; D010728:Phosphoenolpyruvate / Q000378:metabolism*; D011549:Pseudomonas / Q000187:drug effects / Q000378:metabolism*; D011770:Pyruvate Kinase / Q000378:metabolism; D011773:Pyruvates / Q000378:metabolism*
D003470:Culture Media; D005942:Gluconates; D010071:Oxaloacetates; D011773:Pyruvates; D001224:Aspartic Acid; D000244:Adenosine Diphosphate; D000105:Acetyl Coenzyme A; D010728:Phosphoenolpyruvate; D011770:Pyruvate Kinase; D002262:Carboxy-Lyases