A moderately halophilic, obligately chemolithoautotrophic, sulfur-oxidizing bacterium, designated strain HRh1^T, was obtained from mixed sediment samples from hypersaline chloride–sulfate lakes in the Kulunda Steppe, in south-western Siberia (Russia), using aerobic enrichment culture at 1 M NaCl with thiocyanate as substrate. Cells of the isolate were short, non-motile rods with a Gram-negative type of cell wall. The bacterium was an obligate aerobe capable of chemolithoautotrophic growth with thiocyanate and thiosulfate. With thiosulfate, it grew at NaCl concentrations of 0.2–3.0 M (optimum 0.5 M) and at pH 6.3–8.0 (optimum pH 7.3–7.5). During growth on thiocyanate, cyanate was identified as an intermediate. The dominant cellular fatty acids were C_{16 : 0}and C_{18 : 1}ω7. Phylogenetic analysis based on 16S rRNA gene sequencing placed the isolate in the classGammaproteobacteriaas an independent lineage, with an unclassified marine sulfur-oxidizing bacterium as the closest culturable relative (93 % sequence similarity). A singlecbbLgene (coding for the key enzyme of the Calvin–Benson cycle of autotrophic CO₂assimilation) with relatively low similarity to any homologous genes found in chemolithoautotrophs was detected in strain HRh1^T. On the basis of our phenotypic and phylogenetic analysis, the halophilic isolate is proposed to represent a new genus and novel species,Thiohalobacter thiocyanaticusgen. nov., sp. nov. The type strain ofThiohalobacter thiocyanaticusis HRh1^T(=DSM 21152^T=UNIQEM U249^T).