title: Molecular study and partial characterization of iron-only hydrogenase in Desulfovibrio fructosovorans.
abstract: An iron-only hydrogenase was partially purified and characterized from Desulfovibrio fructosovorans wild-type strain. The enzyme exhibits a molecular mass of 56 kDa and is composed of two distinct subunits HydA and HydB (46 and 13 kDa, respectively). The N-terminal amino acid sequences of the two subunits of the enzyme were determined with the aim of designing degenerate oligonucleotides. Direct and inverse polymerase chain reaction techniques were used to clone the hydrogenase encoding genes. A 9-nucleotide region located 75 bp upstream from the translational start codon of the D. fructosovorans hydA gene was found to be highly conserved. The analysis of the deduced amino acid sequence of these genes showed the presence of a signal sequence located in the small subunit, exhibiting the consensus sequence which is likely to be involved in the specific export mechanism of hydrogenases. Two ferredoxin-like motives involved in the coordination of [4Fe-4S] clusters were identified in the N-terminal domain of the large subunit. The amino acid sequence of the [Fe] hydrogenase from D. fructosovorans was compared with the amino acid sequences from eight other hydrogenases (cytoplasmic and periplasmic). These enzymes share an overall 18% identity and 28% similarity. The identity reached 73% and 69% when the D. fructosovorans hydrogenase sequence was compared with the hydrogenase sequences from Desulfovibrio vulgaris Hildenborough and Desulfovibrio vulgaris oxamicus Monticello, respectively.
authors.affiliation: Laboratoire de Bioénergétique et Ingénierie des Protéines, UPR 9036-CNRS, Marseille, France.
authors.forename: L
authors.initials: L
authors.lastname: Casalot
authors.forename: C E
authors.initials: CE
authors.lastname: Hatchikian
authors.forename: N
authors.initials: N
authors.lastname: Forget
authors.forename: P
authors.initials: P
authors.lastname: de Philip
authors.forename: Z
authors.initials: Z
authors.lastname: Dermoun
authors.forename: J P
authors.initials: JP
authors.lastname: Bélaïch
authors.forename: M
authors.initials: M
authors.lastname: Rousset