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Metabolic Pathway for Propionate Utilization by Phosphorus-Accumulating Organisms in Activated Sludge: ¹³C Labeling and In Vivo Nuclear Magnetic Resonance.
Paulo C. Lemos, 2003.In vivo ¹³C and ³¹P nuclear magnetic resonance techniques were used to study propionate metabolism by activated sludge in enhanced biological phosphorus removal systems . The fate of label supplied in [3-¹³C]propionate was monitored in living cells subjected to anaerobic/aerobic cycles . During the anaerobic phase, propionate was converted to polyhydroxyalkanoates (PHA) with the following monomer composition: hydroxyvalerate, 74.2%; hydroxymethylvalerate, 16.9%; hydroxymethylbutyrate, 8.6%; and hydroxybutyrate, 0.3% . The isotopic enrichment in the different carbon atoms of hydroxyvalerate (HV) produced during the first anaerobic stage was determined: HV₅, 59%; HV₄, 5.0%; HV₃, 1.1%; HV₂, 3.5%; and HV₁, 2.8% . A large proportion of the supplied label ended up on carbon C-5 of HV, directly derived from the pool of propionyl-coenzyme A (CoA), which is primarily labeled on C-3; useful information on the nature of operating metabolic pathways was provided by the extent of labeling on C-1, C-2, and C-4 . The labeling pattern on C-1 and C-2 was explained by the conversion of propionyl-CoA to acetyl-CoA via succinyl-CoA and the left branch of the tricarboxylic acid cycle, which involves scrambling of label between the inner carbons of succinate . This constitutes solid evidence for the operation of succinate dehydrogenase under anaerobic conditions . The labeling in HV₄ is explained by backflux from succinate to propionyl-CoA . The involvement of glycogen in the metabolism of propionate was also demonstrated; moreover, it was shown that the acetyl moiety to the synthesis of PHA was derived preferentially from glycogen . According to the proposed metabolic scheme, the decarboxylation of pyruvate is coupled to the production of hydrogen, and the missing reducing equivalents should be derived from a source other than glycogen metabolism .

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