Phylogenetic comparison of opine dehydrogenase sequences from marine invertebrates.

Three cDNA sequences encoding putative opine dehydrogenase (OpDH) enzymes from the mussel Mytilus galloprovincialis were obtained. The deduced amino acid sequences were clearly distinguishable from each other, showing that several OpDH transcripts could occur in the mussel tissues (p distance 0.46-0.55). When these sequences were aligned and compared with published databank proteins, the range of identity among the M. galloprovincialis OpDH and the strombine dehydrogenase from Ostrea edulis was 51-59 %, the best hit in the three comparisons, followed by OpDH enzymes from other marine invertebrates. Sequence alignment revealed structural motifs possibly related to the binding sites of the substrates. A phylogenetic analysis compared M. galloprovincialis OpDH and annotated sequences belonging to five phyla and seven taxonomic classes, including 19 species, representing the five OpDH protein family members. The phylogenetic tree clustered the OpDH enzymes according to the evolutionary relationships of the species and not to the biochemical reaction catalyzed.

Taxonomic study of Bathygadidae fishes (Gadiformes) from Atlantic Spanish waters combining morphological and molecular approaches.

From 2009 to 2011 eleven specimens belonging to four bathygadid species of the family Bathygadidae were captured in two different locations in the northern waters of Spain. The morphometric measurements and meristic characters of these specimens are given. The specimens were identified as belonging to the genera Gadomus Regan, 1903, and Bathygadus Günther, 1878, including the following species: Gadomus dispar (Vaillant, 1888), Gadomus longifilis (Goode & Bean, 1885), Gadomus arcuatus (Goode & Bean, 1886) and Bathygadus melanobranchus Vaillant, 1888. As a result, a new northern limit of distribution of G. arcuatus from the northeastern Atlantic is reported. The first molecular identification and genetic interrelationships of Bathygadidae species, based on the mitochondrial COI nucleotide sequences -DNA barcodes- is reported. Sequences corresponding to specimens from the same species were identical and the overall mean genetic diversity (uncorrected p-distance) was 0.096 ± 0.008. Based on a morphological and meristic examination of the specimens, as well as on the available literature, an updated key of the members of the family Bathygadidae from the north-eastern Atlantic Ocean is provided.

Identification of octopine dehydrogenase from Mytilus galloprovincialis.

A cDNA encoding the putative octopine dehydrogenase (OcDH) from the mussel Mytilus galloprovincialis was cloned and sequenced. The complete coding region was expressed in the bacteria Escherichia coli and the recombinant protein was purified. The M. galloprovincialis OcDH appears to have the highest affinity for the amino acid substrate L-arginine (88.22%), compared to L-alanine (9.04%) and glycine (2.74%). This enzyme showed no activity when taurine or ß-alanine was used as substrate. These data strongly support that this recombinant enzyme is octopine dehydrogenase and not another opine dehydrogenase such as alanopine or strombine dehydrogenases. The superimposition of the theoretical three-dimensional model of the M. galloprovincialis OcDH and the crystal structure of its homologous counterpart from the great scallop Pecten maximus showed interesting changes in the amino acid binding site which could explain the differences found in the substrate affinity between the two molluscs. A phylogenetic analysis was performed comparing M. galloprovincialis OcDH and annotated sequences representing the five opine dehydrogenase (OpDH) protein family members. The phylogenetic tree which was obtained clustered the OpDH enzymes according to the evolutionary relationships of the species and not to the biochemical reaction catalysed. Octopine dehydrogenase has been identified in the Mytilidae family for the first time, having previously only been established in one other marine invertebrate (P. maximus).

Dual function of fish hepcidin: response to experimental iron overload and bacterial infection in sea bass (Dicentrarchus labrax).

The role of hepcidin in iron metabolism regulation and bacterial infection has been the focus of recent attention. However, in spite of the growing number of hepcidin genes known from different organisms, little is known about its putative dual function in fish. The aim of this study was to characterize the sea bass hepcidin gene and to study its role in iron metabolism and infection. The novel sea bass hepcidin gene was found to be organized into two introns and three exons with several copies present in the genome. The transcript showed a constitutive low basal expression being mainly expressed in liver and encoding a putative 85 residues long peptide. Fish were submitted either to iron status modulation or bacterial infection and the hepcidin transcript levels were analysed along with a number of other parameters. Liver hepcidin expression was found to increase in both the iron-overloaded and infected fish, while in the iron-deficient fish no alteration in expression levels was detected. These results point to the evolutionary conservation of hepcidin's dual functions.