Thermococcus nautili sp nov., a hyperthermophilic archaeon isolated from a hydrothermal deep-sea vent

Thermococcus nautili, strain 30-1(T) (formerly reported as Thermococcus nautilus), was isolated from a hydrothermal chimney sample collected from the East Pacific Rise at a depth of 2633 m on the la chainette PP57' area Cells were motile, irregular cocci with a polar tuft of flagella (0.8-1.5 mu m) and divided by constriction. The micro-organism grew optimally at 87.5 degrees C (range 55-95 degrees C), at pH 7 (range pH 4-9) and with 2% NaCl (range 1-4%). Doubling time was 64 min in Zillig's broth medium under optimal conditions. Growth was strictly anaerobic. It grew preferentially in the presence of elemental sulfur or cystine, which are reduced to H2S, on complex organic substrates such as yeast extract, tryptone, peptone, Casamino acids and casein. Slow growth was observed on starch and pyruvate. Strain 30-1(T) was resistant to chloramphenicol and tetracyclin (at 100 mu g ml(-1)) but sensitive to kanamycin and rifampicin. The G+C content of the genomic DNA was 54 mol%. Strain 30-1(T) harboured three plasmids named pTN1, pTN2 and pTN3 and produced membrane vesicles that incorporate pTN1 and pTN3. As determined by 16S rRNA gene sequence analysis, strain 30-1(T) is related most closely to Thermococcus sp. AM4 (99.3% similarity) and Thermococcus gammatolerans DSM 15229(T) (99.2 %). DNA-DNA hybridization values (in silico) with these two closest relatives were below the threshold value of 70% (33 % with Thermococcus sp. AM4 and 32% with T. gammatolerans DSM 15229(T)) and confirmed that strain 30-1 represents a novel species. On the basis of the data presented, strain 30-1(T) is considered to represent a novel species of the genus Thermococcus, for which the name Thermococcus nautili sp. nov. is proposed. The type strain is 30-1(T) (=CNCM 4275=JCM 19601).