Abstract
Conditions promoting maximal in vitro activity of the particulate NADH:fumarate reductase from Fibrobacter succinogenes were determined. This system showed a pH optimum of 6.0 in K+ MES buffer only when salt (NaCl or KCl) was present. Salt stimulated the activity eightfold at the optimal concentration of 150m M. This effect was due to stimulation of fumarate reductase activity as salt had little effect on NADH: decylubiquinone oxidoreductase (NADH dehydrogenase). The stimulation of fumarate reductase by salt at pH 6.0 was not due to removal of oxaloacetate from the enzyme. Kinetic parameters for several inhibitors were also measured. NADH dehydrogenase was inhibited by rotenone at a single site with a K i of 1 μM. 2-Heptyl-4-hydroxyquinonline-N-oxide (HOQNO) inhibited NADH: fumarate reductase with a K i of 0.006 μM, but NADH dehydrogenase exhibited two HOQNO inhibition constants of approximately 1 μM and 24 μM. Capsaicin and laurylgallate each inhibited NADH dehydrogenase by only 20% at 100 μM. NADH dehydrogenase gave K m values of 1 μM for NADH and 4 μM for reduced hypoxanthine adenine dinucleotide.
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Published with the approval of the Director of the Agricultural Experiment Station, North Dakota State University, as journal article no. 2201
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Meinhardt, S.W., Glass, T.L. Characterization of the NADH dehydrogenase and fumarate reductase of Fibrobacter succinogenes subsp. succinogenes S85. Arch. Microbiol. 162, 329–334 (1994). https://doi.org/10.1007/BF00263780
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DOI: https://doi.org/10.1007/BF00263780