Isolation of the anaerobic thermoacidophilic crenarchaeote Acidilobus saccharovorans sp nov and proposal of Acidilobales ord. nov., including Acidilobaceae fam. nov and Caldisphaeraceae fam. nov.

An anaerobic acidophilic, hyperthermophilic archaeon, designated strain 345-15(T), was isolated from an acidic hot spring of Kamchatka (Russia). Cells of strain 345-15(T) were regular or irregular cocci, 1-2 mu m in diameter, with flagella. Strain 345-15(T) grew optimally at 80-85 degrees C and pH 3.5-4.0 and fermented a wide range of carbohydrates, including polysaccharides. Acetate, ethanol and lactate were the fermentation products. Growth was stimulated by elemental sulfur and thiosulfate, which were reduced to hydrogen sulfide. The G + C content of the DNA was 54.5 mol%. The 16S rRNA gene sequence analysis indicated that strain 345-15(T) belonged to the genus Acidilobus. The level of DNA-DNA hybridization between strain 345-15(T) and Acidilobus aceticus 1904(T) was 61%. Thus, strain 345-15(T) was considered as representing a novel species of the genus Acidilobus, with the name Acidilobus saccharovorans sp. nov. (type strain, 345-15(T) = DSM 16705(T) = VKM B-2471(T)), which shared the main morphological and physiological properties of the genus but differed by the presence of flagella and the spectrum of substrates utilized. Phylogenetic analysis showed that the genus Acidilobus, with its species Acidilobus aceticus, Acidilobus saccharovorans sp. nov. and 'Acidilobus sulfurireducens', and the genus Caldisphaera, represented by Caldisphaera lagunensis and 'Caldisphaera draconis', formed a separate cluster that adjoins the cluster formed by the species of the order Desulfurococcales. Members of the Acidilobus-Caldisphaera cluster are thermophilic, organotrophic anaerobic cocci that can be distinguished from all species of the order Desulfurococcales on the basis of acidophily. Based on these considerations, we propose a new family, Acidilobaceae fam. nov., to accommodate the subcluster of hyperthermophiles represented by the genus Acidilobus, a new family, Caldisphaeraceae fam. nov., for the subcluster of extreme thermophiles represented by the genus Caldisphaera, and a new order, Acidilobales ord. nov., to accommodate the two new families.