title: Membrane-associated redox activities in Thermotoga neapolitana.
abstract: Elemental sulfur reduction by the hyperthermophilic bacterium Thermotoga neapolitana provides an alternative to hydrogen evolution during fermentation. Electrons are transferred from reduced cofactors (ferredoxin and NADH) to sulfur by a series of unknown steps. One enzyme that may be involved is an NADH:methyl viologen oxidoreductase (NMOR), an activity that in other fermenting organisms is associated with NADH:ferredoxin oxidoreductase. We found that 83% of NMOR activity was contained in the pellet fraction of cell extracts subjected to ultracentrifugation. This pellet fraction, presumably containing cell membranes, was required for electron transfer to NAD+ from ferredoxin-dependent pyruvate oxidation. However, the NMOR activity in this fraction used neither Thermotoga nor clostridial ferredoxins as substrates. NMOR activity was also detected in aerobically prepared vesicles. By comparison with ATPase activities, NMOR was found primarily on the cytoplasmic face of these vesicles. During these studies, an extracytoplasmic hydrogenase activity was discovered. In contrast to the soluble hydrogenase, this hydrogenase activity was completely inhibited when intact cells were treated with cupric chloride and was present on the extracytoplasmic face of vescides. In contrast to a soluble hydrogenase reported in Thermotoga maritima, this activity was air-stable and was inhibited by low concentrations of nitrite.
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269-3125 USA.
authors.forename: S A
authors.initials: SA
authors.lastname: Käslin
authors.forename: S E
authors.initials: SE
authors.lastname: Childers
authors.forename: K M
authors.initials: KM
authors.lastname: Noll
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D005288:Ferredoxins / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D010084:Oxidation-Reduction; D016999:Sulfur-Reducing Bacteria / Q000378:metabolism; D013824:Thermus / Q000378:metabolism*
chemical_list: D005288:Ferredoxins; D009247:NADH, NADPH Oxidoreductases; C048744:NADH-methyl viologen reductase; D000251:Adenosine Triphosphatases