A hyperthermophilic and amylolytic prokaryote, designated Rt3, was isolated
from a thermal spring near Rotorua, New Zealand. The 16S rRNA gene of Rt3
was cloned and sequenced with the aim of determining its phylogenetic affil
iations. The phylogenetic analysis of this sequence, which included a selec
tion of archaebacterial and eubacterial 16S rRNA sequences, indicates that
Rt3 most likely belongs to the archaebacterial order Thermococcales. An amy
lase gene (amyA) from Rt3, encoding a highly thermostable amylase activity,
was cloned and its DNA sequence determined. Transcriptional signals typica
l of archaebacteria were evident in this sequence. The sequence is homologo
us to a broad range of enzymes from the AMY superfamily and contains a typi
cal N-terminal signal peptide. Phylogenetic analysis and comparison of stru
ctural features with other AMY superfamily enzymes reveals that, firstly, t
he closest homologues of the Rt3 amylase are members of the Bacillus and Pl
ant alpha-amylase groups; and secondly, that the Rt3 amylase is closely rel
ated to only one other currently known archaebacterial enzyme, i.e. an (AMY
superfamily) alpha-amylase from Natronococcus.