PubMed · 5489063
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M A Riederer-Henderson, P W Wilson. 1970. Nitrogen fixation by sulphate-reducing bacteria.. https://doi.org/10.1099/00221287-61-1-27
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In NiFe hydrogenases, electrons are transferred from the active site to the redox partner via a chain of three Iron-Sulfur clusters, and the surface-exposed [4Fe4S] cluster has an unusual His(Cys)3 ligation. When this Histidine (H184 in Desulfovibrio fructosovorans) is changed into a cysteine or a glycine, a distal cubane is still assembled but the oxidative activity of the mutants is only 1.5 and 3% of that of the WT, respectively. We compared the activities of the WT and engineered enzymes for H2 oxidation, H+ reduction and H/D exchange, under various conditions: (i) either with the enzyme directly adsorbed onto an electrode or using soluble redox partners, and (ii) in the presence of exogenous ligands whose binding to the exposed Fe of H184G was expected to modulate the properties of the distal cluster. Protein film voltammetry proved particularly useful to unravel the effects of the mutations on inter and intramolecular electron transfer (ET). We demonstrate that changing the coordination of the distal cluster has no effect on cluster assembly, protein stability, active-site chemistry and proton transfer; however, it slows down the first-order rates of ET to and from the cluster. All-sulfur coordination is actually detrimental to ET, and intramolecular (uphill) ET is rate determining in the glycine variant. This demonstrates that although [4Fe4S] clusters are robust chemical constructs, the direct protein ligands play an essential role in imparting their ability to transfer electrons.
In this study, the occurrence and metabolic capacities of sulfate reducing bacteria (SRB) were studied in 36 water samples taken from cooling towers of 30 different buildings, such as hotels and business centres in Istanbul. SRB were present in 14 cooling towers out of 30 (46.6%) buildings and while the lowest concentration of SRB was 10 cells/mL, the highest concentration was determined as 10(4) cells/mL. After the distribution of SRB within cooling towers was determined, several strains of SRB were isolated and characterized metabolically. The isolated strains were composed of vibroid cells, growing anaerobically by using sulfate as electron acceptor and lactate or pyruvate as electron donor. They could be related to the genus Desulfovibrio. In addition, the recorded temperature of water samples was between 12 and 33 degrees C and a significant relationship between the number of SRB and the water temperatures was not found.
Seventeen human clinical isolates representing four species of Desulfovibrio were characterized using 16S rRNA gene sequences and tests for catalase, indole, nitrate, bile, urease, formate-fumarate stimulation, desulfoviridin, motility, and hydrogen sulfide production, plus susceptibility to antimicrobial agents. Eighty additional strains representing 10 phenotypically similar genera (Bilophila, Selenomonas, Capnocytophaga, Campylobacter, Bacteroides, Sutterella, Anaerobiospirillum, Dialister, Veillonella, and Mobiluncus) were included for comparison. All Desulfovibrio species produced H2S and were desulfoviridin positive, and all Desulfovibrio species except D. piger were motile. The four Desulfovibrio species could be distinguished from each other using tests for catalase, indole, nitrate, urease, and growth on bile, with the following results (positive [+], negative [-], growth [G], and no growth [NG]): for D. piger, -, -, -, -, and G, respectively; for D. fairfieldensis, +, -, +, -, and G, respectively; for D. desulfuricans, -, -, +, +, and NG, respectively; and for D. vulgaris, -, +, -, -, and G, respectively. Resistance to the 10-microg colistin disk separated the Desulfovibrio species from most of the other genera, which were usually susceptible. These simple tests were useful for characterizing the Desulfovibrio species and differentiating them from other phenotypically similar genera.