title: Expression and characterization of Cu/Zn superoxide dismutase from Wuchereria bancrofti.
abstract: The Cu/Zn superoxide dismutase gene from Wuchereria bancrofti (Cu/Zn WbSOD) was isolated by PCR using degeneracy primers. The complete Cu/Zn WbSOD consisted of 1,032 nucleotides containing 4 exons (477 nucleotides) and 3 introns. The molecular phylogenetic analysis of the Cu/Zn WbSOD gene in comparison with those from other organisms revealed that the gene was classified in the same clade to those of filarial Brugia malayi and Brugia pahangi (bootstrap value at 90). The nucleotide and deduced amino acid sequences of Cu/Zn WbSOD exhibited the similarity to those of intracellular Cu/Zn SOD of B. malayi and B. pahangi. The amino acid comparison of Cu/Zn WbSOD to others revealed that the binding sites and active sites were conserved. The expression of this gene yielded 16.366 kDa in size. After Ni-IDA column purification, the enzyme showed specific activity of 8.5 U/mg and 42.1% yield. The enzyme activity was inhibited when 6 mM KCN was added.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Sukhumvit 23, Bangkok 10110, Thailand.
authors.forename: Paisarn
authors.initials: P
authors.lastname: Khawsak
authors.forename: Pornpimon
authors.initials: P
authors.lastname: Kanjanavas
authors.forename: Piyapa
authors.initials: P
authors.lastname: Kiatsomchai
authors.forename: Kosum
authors.initials: K
authors.lastname: Chansiri
mesh_terms: D000818:Animals; D001665:Binding Sites; D020134:Catalytic Domain; D003001:Cloning, Molecular; D016000:Cluster Analysis; D017124:Conserved Sequence; D017931:DNA Primers / Q000235:genetics; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005091:Exons; D015870:Gene Expression; D007438:Introns; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011190:Potassium Cyanide / Q000494:pharmacology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D014958:Wuchereria bancrofti / Q000201:enzymology* / Q000235:genetics*
chemical_list: D017931:DNA Primers; D018509:DNA, Helminth; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D013482:Superoxide Dismutase; D011190:Potassium Cyanide