The DSR system presumably forms intracellular sulfite that is oxidized by an enzyme system consisting of Sat, Apr, and Qmo proteins (Rodriguez et al., 2011). The electron acceptors, cytochrome c, and menaquinone (Fig. 1) are ultimately oxidized

by the photosynthetic reaction center. In cultures of Cba. tepidum that contain both sulfide and thiosulfate, sulfide is oxidized preferentially while sulfur globules are formed (Chan et al., 2008; Azai et al., 2009; Holkenbrink et al., 2011). Following sulfide depletion, thiosulfate and sulfur globules are oxidized to sulfate. The molecular mechanism of Alpelisib mw this phenomenon is poorly understood. Sulfide possibly inhibits thiosulfate oxidation either by substrate competition between sulfide and thiosulfate (the SOX system oxidizes sulfide in vitro; Ogawa et al., 2010) or by saturation of the electron acceptor pool. Regulation of sulfur metabolism genes in GSB is poorly described, but it is known that SoxA is induced by thiosulfate in Chlorobaculum thiosulfatiphilum (Verté et al. 2002). find more In the purple sulfur bacterium, Allochromatium vinosum, sox and dsr genes are expressed at a low constitutive level in the absence of reduced sulfur substrates and are induced by thiosulfate and sulfide, respectively (Grimm et al., 2010, 2011). Chlorobaculum tepidum TLS was grown under incandescent illumination in CL medium (Frigaard et al., 2004). For experiments comparing early and late exponential growth phase, wild-type

cells were grown at 45 °C in 1-L flasks under a light intensity of 200 μmol photons m−2 s−1. For experiments comparing wild type and the dsrM mutant (Holkenbrink et al., 2011), cells were grown at 42 °C in 15-mL

tubes under a light intensity of 50 μmol photons m−2 s−1 and harvested in the late exponential growth phase. Cells were harvested by centrifugation and stored at −20 °C prior to analysis. Bacteriochlorophyll c was determined by extracting the cell pellet with acetone : methanol (7 : 2 by vol) (Frigaard et al., 1997). Sulfide was measured using the colorimetric methylene blue method (Cline 1969). Sulfate Ribonucleotide reductase and thiosulfate were measured by ion chromatography (Dionex, Hvidovre, Denmark) using a carbonate buffer as eluent. Samples for analysis of elemental sulfur were dissolved in methanol and analyzed as S8 by HPLC using a Sykam pump (S1100), UV–VIS detector (Sykam S3200), Zorbax ODS-column (125 × 4 mm, 5 μm; Knauer, Berlin, Germany) and methanol as the eluent at a flow rate of 1 mL min−1. Elemental sulfur was detected at 265 nm. Cell pellets were thawed and extracted in acetone : methanol (7 : 2 by vol) to remove pigments. The colorless cell pellets were solubilized in an SDS-containing buffer (Laemmli, 1970) supplemented with a complete protease inhibitor cocktail (Roche, Hvidovre, Denmark) at 95 °C for 3–5 min and cleared by centrifugation. Prior to digestion, proteins were reduced with dithiothreitol (1 mM) and alkylated with iodoacetamide (5 mM).