Properties of a fructose-1,6-diphosphate-activated lactate dehydrogenase from Acholeplasma laidlawii type A.
Acholeplasma laidlawii A possesses a nicotinamide adenine dinucleotide (NAD)-dependent l(+)-lactate dehydrogenase (LDH) which is activated specifically by low concentrations of fructose-1, 6-diphosphate (FDP). Studies with partially purified enzyme show that the kinetic response to FDP is hyperbolic. The enzyme is inhibited by inorganic phosphate, adenosine triphosphate, and high concentrations of reduced NAD (NADH). Low activity is demonstrable in the absence of FDP at pH 6.0 to 7.2, but FDP is absolutely required in the region of pH 8. FDP causes an upward shift in the optimum pH of the enzyme, which is near 7.2 in tris (hydroxymethyl)aminomethane buffer. Activation of the enzyme by FDP is markedly affected by substrate concentration; FDP lowers the apparent K(m) for pyruvate and NADH. The affinity of the enzyme for pyruvate is also influenced by H(+) concentration. The pyruvate analogue alpha-ketobutyrate serves as an effective substrate for the enzyme; when it is utilized, the enzyme is still activated by FDP. Reversal of the pyruvate reduction reaction catalyzed by the enzyme can be demonstrated with the 3-acetylpyridine analogue of NAD. The catalytic properties of the A. laidlawii enzyme and the known FDP-activated LDHs which occur among lactic acid bacteria are discussed.
D000128:Acholeplasma laidlawii / Q000032:analysis / Q000201:enzymology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D002087:Butyrates / Q000494:pharmacology; D002130:Calcium Phosphates; D002384:Catalysis; D003067:Coenzymes / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D005636:Fructosephosphates / Q000032:analysis / Q000494:pharmacology*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007651:Keto Acids / Q000494:pharmacology; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D008024:Ligands / Q000378:metabolism; D009174:Mycoplasma / Q000201:enzymology*; D010710:Phosphates / Q000494:pharmacology; D011773:Pyruvates / Q000494:pharmacology; D013237:Stereoisomerism
D002087:Butyrates; D002130:Calcium Phosphates; D003067:Coenzymes; D005636:Fructosephosphates; D007651:Keto Acids; D008024:Ligands; D010710:Phosphates; D011773:Pyruvates; D000255:Adenosine Triphosphate; D007770:L-Lactate Dehydrogenase