title: The Vanadium Iodoperoxidase from the marine flavobacteriaceae species Zobellia galactanivorans reveals novel molecular and evolutionary features of halide specificity in the vanadium haloperoxidase enzyme family.
abstract: Vanadium haloperoxidases (VHPO) are key enzymes that oxidize halides and are involved in the biosynthesis of organo-halogens. Until now, only chloroperoxidases (VCPO) and bromoperoxidases (VBPO) have been characterized structurally, mainly from eukaryotic species. Three putative VHPO genes were predicted in the genome of the flavobacterium Zobellia galactanivorans, a marine bacterium associated with macroalgae. In a phylogenetic analysis, these putative bacterial VHPO were closely related to other VHPO from diverse bacterial phyla but clustered independently from eukaryotic algal VBPO and fungal VCPO. Two of these bacterial VHPO, heterogeneously produced in Escherichia coli, were found to be strictly specific for iodide oxidation. The crystal structure of one of these vanadium-dependent iodoperoxidases, Zg-VIPO1, was solved by multiwavelength anomalous diffraction at 1.8 Å, revealing a monomeric structure mainly folded into α-helices. This three-dimensional structure is relatively similar to those of VCPO of the fungus Curvularia inaequalis and of Streptomyces sp. and is superimposable onto the dimeric structure of algal VBPO. Surprisingly, the vanadate binding site of Zg-VIPO1 is strictly conserved with the fungal VCPO active site. Using site-directed mutagenesis, we showed that specific amino acids and the associated hydrogen bonding network around the vanadate center are essential for the catalytic properties and also the iodide specificity of Zg-VIPO1. Altogether, phylogeny and structure-function data support the finding that iodoperoxidase activities evolved independently in bacterial and algal lineages, and this sheds light on the evolution of the VHPO enzyme family.
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France; CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France.
authors.forename: Jean-Baptiste
authors.initials: JB
authors.lastname: Fournier
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France; CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France.
authors.forename: Etienne
authors.initials: E
authors.lastname: Rebuffet
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France; CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Delage
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France; CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France.
authors.forename: Romain
authors.initials: R
authors.lastname: Grijol
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France; CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Meslet-Cladière
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France; CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France.
authors.forename: Justyna
authors.initials: J
authors.lastname: Rzonca
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France; CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Potin
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France; CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France.
authors.forename: Gurvan
authors.initials: G
authors.lastname: Michel
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France; CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France.
authors.forename: Mirjam
authors.initials: M
authors.lastname: Czjzek
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France; CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique, Roscoff, France catherine.leblanc@sb-roscoff.fr.
authors.forename: Catherine
authors.initials: C
authors.lastname: Leblanc
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D019143:Evolution, Molecular; D041965:Flavobacteriaceae / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification*; D007453:Iodide Peroxidase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D005810:Multigene Family; D009195:Peroxidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D012623:Seawater / Q000382:microbiology*; D016415:Sequence Alignment; D013379:Substrate Specificity; D014639:Vanadium / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D014639:Vanadium; C414928:vanadium iodoperoxidase; D009195:Peroxidase; D007453:Iodide Peroxidase