A genomic region required for phototrophic thiosulfate oxidation in the green sulfur bacterium Chlorobium tepidum (syn. Chlorobaculum tepidum)

College of Marine and Earth Studies and Delaware Biotechnology Institute, University of Delaware, Rm 127 DBI, 15 Innovation Way, Newark, DE 19711, USA Correspondence Thomas E. Hanson tehanson{at}udel.edu The specific enzymes employed by Chlorobium tepidum for the anaerobic oxidation of thiosulfate, sulfide and elemental sulfur during anoxygenic photosynthesis are not well defined. In particular, it is unclear how C. tepidum completely oxidizes thiosulfate. A C. tepidum genomic region, encoding a putative quinone-interacting membrane-bound oxidoreductase (Qmo) complex ( CT0866–0868 ), hypothetical proteins ( CT0869–0875 ) and a sulfide : quinone oxidoreductase (SQR) homologue ( CT0876 ), was analysed for its role in anaerobic sulfur oxidation. Transcripts of genes encoding the Qmo complex, which is similar to archaeal heterodisulfide reductases, were detected by RT-PCR only while sulfide or elemental sulfur were being oxidized, whereas the SQR homologue and CT0872 were expressed during thiosulfate oxidation and into early stationary phase. A mutant of C. tepidum was obtained in which the region between CT0868 and CT0876 was replaced by a transposon insertion resulting in the truncation or deletion of nine genes. This strain, C5, was completely defective for growth on thiosulfate as the sole electron donor in C. tepidum , but only slightly defective for growth on sulfide or thiosulfate plus sulfide. Strain C5 did not oxidize thiosulfate and also displayed a defect in acetate assimilation under all growth conditions. A gene of unknown function, CT0872 , deleted in strain C5 that is conserved in chemolithotrophic sulfur-oxidizing bacteria and archaea is the most likely candidate for the thiosulfate oxidation phenotype observed in this strain. The defect in acetate assimilation may be explained by deletion of CT0874 , which encodes a homologue of 3-oxoacyl acyl carrier protein synthase. Abbreviations: bChl, bacteriochlorophyll; BTP, 1,3-bis(tris(hydroxymethyl)methylamino)propane; Gm, gentamicin; GSB, green sulfur bacteria; IVTM, in vitro transposition mutagenesis; Qmo, quinone-interacting membrane-bound oxidoreductase; SQR, sulfide, quinone oxidoreductase Two supplementary tables are available with the online version of this paper.