Propionate oxidation in Escherichia coli: evidence for operation of a methylcitrate cycle in bacteria.
Escherichia coli grew in a minimal medium on propionate as the sole carbon and energy source. Initially a lag phase of 4-7 days was observed. Cells adapted to propionate still required 1-2 days before growth commenced. Incorporation of (2-13C), (3-13C) or (2H3)propionate into alanine revealed by NMR that propionate was oxidized to pyruvate without randomisation of the carbon skeleton and excluded pathways in which the methyl group was transiently converted to a methylene group. Extracts of propionate-grown cells contained a specific enzyme that catalyses the condensation of propionyl-CoA with oxaloacetate, most probably to methylcitrate. The enzyme was purified and identified as the already-known citrate synthase II. By 2-D gel electrophoresis, the formation of a second propionate-specific enzyme with sequence similarities to isocitrate lyases was detected. The genes of both enzymes were located in a putative operon with high identities (at least 76% on the protein level) with the very recently discovered prp operon from Salmonella typhimurium. The results indicate that E. coli oxidises propionate to pyruvate via the methylcitrate cycle known from yeast. The 13C patterns of aspartate and glutamate are consistent with the further oxidation of pyruvate to acetyl-CoA. Oxaloacetate is predominantly generated via the glyoxylate cycle rather than by carboxylation of phosphoenolpyruvate.
Textor
S
S
Laboratorium für Mikrobiologie, Fachbereich Biologie, Phillips-Universität, D-35032 Marburg, Germany.
Wendisch
V F
VF
De Graaf
A A
AA
Müller
U
U
Linder
M I
MI
Linder
D
D
Buckel
W
W
D000595:Amino Acid Sequence; D002950:Citrate (si)-Synthase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D008956:Models, Chemical; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009876:Operon / Q000235:genetics; D010084:Oxidation-Reduction; D011422:Propionates / Q000378:metabolism*; D019289:Pyruvic Acid / Q000378:metabolism; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid
D011422:Propionates; D019289:Pyruvic Acid; D002950:Citrate (si)-Synthase; C092884:methylcitrate synthase; C029658:propionic acid