Open AccessIdentification and Characterization of a Novel Bacterial Sulfite Oxidase with No Heme Binding Domain from Deinococcus radiodurans
Author(s) -
Giovanni D’Errico,
Anna Di Salle,
Francesco La Cara,
Mosè Rossi,
Raffaele Cannio
Publication year - 2006
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.188.2.694-701.2006
Subject(s) - deinococcus radiodurans , biology , sulfite oxidase , deinococcus , microbiology and biotechnology , escherichia coli , biochemistry , gene , enzyme
An open reading frame (draSO ) encoding a putative sulfite oxidase (SO) was identified in the sequence of chromosome II ofDeinococcus radiodurans ; the predicted gene product showed significant amino acid sequence homology to several bacterial and eukaryotic SOs, such as the biochemically and structurally characterized enzyme fromArabidopsis thaliana . Cloning of theDeinococcus SO gene was performed by PCR amplification from the bacterial genomic DNA, and heterologous gene expression of a histidine-tagged polypeptide was obtained in a molybdopterin-overproducing strain ofEscherichia coli . The recombinant protein was purified to homogeneity by nickel chelating affinity chromatography, and its main kinetic and chemical physical parameters were determined. Northern blot and enzyme activity analyses indicated thatdraSO gene expression is constitutive inD. radiodurans and that there is no increase upon exposure to thiosulfate and/or molybdenum(II).