Molecular characterization and heterologous expression of hypCD, the first two [NiFe] hydrogenase accessory genes of Thermococcus litoralis.
The hypCD genes, encoding the counterparts of mesophilic proteins involved in the maturation of [NiFe] hydrogenases, were isolated from the hyperthermophilic archaeon Thermococcus litoralis. The deduced gene products showed 30-40% identity to the corresponding mesophilic proteins. HypC and HypD were synthesized by the T7 expression system. Heterologous complementation experiments were done in Escherichia coli and Ralstonia eutropha strains lacking functionally active hypC and hypD genes. Only the cytoplasmic hydrogenase of R. eutropha could be processed by HypD from T. litoralis. This was the first demonstration of mesophilic hydrogenase processing using a hyperthermophilic archaeal accessory protein to produce an active enzyme.
Takács
M
M
Department of Biotechnology, University of Szeged, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, 6726 Szeged, Temesvári krt. 62, Hungary.
Rákhely
G
G
Kovács
K L
KL
D019843:Archaeal Proteins; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D020118:Cupriavidus necator / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D006864:Hydrogenase / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D011506:Proteins; D019712:Thermococcus / Q000201:enzymology / Q000235:genetics*; D014158:Transcription, Genetic
D019843:Archaeal Proteins; D001426:Bacterial Proteins; C491586:HypC protein, Thermococcus litoralis; C108875:HypD protein, Bacteria; D011506:Proteins; C043296:nickel-iron hydrogenase; D006864:Hydrogenase