Routes of flavodoxin and ferredoxin reduction in Escherichia coli. CoA-acylating pyruvate: flavodoxin and NADPH: flavodoxin oxidoreductases participating in the activation of pyruvate formate-lyase.
Flavodoxin and ferredoxin become reduced in Escherichia coli cells by oxidoreductase reactions which use pyruvate and NADPH as electron donor substrates. The two enzymes, which are minor proteins of this organism, were measured through the reduced flavodoxin-dependent activation of pyruvate formate-lyase. The NADPH-dependent enzyme, obtained homogeneously through Procion-red affinity chromatography, was identified as the flavoprotein 'component R' described previously by Fujii and Huennekens [J. Biol. Chem. 249, 6745-6753 (1974)]. The pyruvate-dependent enzyme was identified as CoA-acetylating pyruvate:flavodoxin (ferredoxin) oxidoreductase. Its catalytic properties in the forward, reverse, and the 14CO2-pyruvate exchange reaction are reported. The dihydro form of flavodoxin was characterized as the particular species involved in the activation of pyruvate formate-lyase. The activation process still occurs with 70% of maximal efficiency when the ratio [NADPH]/([NADP] + [NADPH]) is fixed at the intracellular 'anabolic reduction charge' value of 0.45, in conjunction with the NADPH-dependent enzyme. The [2Fe-2S] ferredoxin, though being readily used as electron acceptor of both oxidoreductases and having a redox potential similar to flavodoxin, proved incompetent in mediating the activation of pyruvate formate-lyase.
D000105:Acetyl Coenzyme A / Q000096:biosynthesis*; D000123:Acetyltransferases / Q000378:metabolism*; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology*; D005287:Ferredoxin-NADP Reductase / Q000378:metabolism*; D005418:Flavodoxin / Q000378:metabolism*; D005420:Flavoproteins / Q000378:metabolism*; D007658:Ketone Oxidoreductases / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D010084:Oxidation-Reduction; D050743:Pyruvate Synthase; D013379:Substrate Specificity
D005418:Flavodoxin; D005420:Flavoproteins; D000105:Acetyl Coenzyme A; D005287:Ferredoxin-NADP Reductase; D007658:Ketone Oxidoreductases; D050743:Pyruvate Synthase; D009247:NADH, NADPH Oxidoreductases; D000123:Acetyltransferases; C022560:formate C-acetyltransferase