Molecular and comparative analysis of the hyperthermostable pyrococcus furiosus glutamate dehydrogenase and its gene

The gene for glutamate dehydrogenase (GDH) from Pyrococcus furiosus has been cloned, sequenced and expressed in Escherichia coli. Significant GDH activity could be detected in this host, allowing the further structure-function analysis of this hyperthermostable hexameric enzyme. The deduced primary sequence of the P. furiosus GDH was homologous to various bacterial, archaeal and eukaryal GDHs. Detailed comparative analysis of the primary sequences of these GDHs suggest that a decrease in Gly residues can be a general stabilizing feature of proteins functional under extreme conditions such as high temperatures or high salt concentrations whereas an increase in He residues and a decrease in Cys residues is typical for hyperthermostable enzymes.