title: Characterization of a HMT2-like enzyme for sulfide oxidation from Pseudomonas putida.
abstract: The open reading frame pp0053, which has a high homology with the sequence of mitochondrial sulfide dehydrogenase (HMT2) conferring cadmium tolerance in fission yeast, was amplified from Pseudomonas putida KT2440 and expressed in Escherichia coli JM109(DE3). The isolated and purified PP0053-His showed absorption spectra typical of a flavin adenine dinucleotide (FAD)--binding protein. The PP0053-His catalyzed a transfer of sulfide-sulfur to the thiophilic acceptor, cyanide, which decreased the Km value of the enzyme for sulfide oxidation and elevated the sulfide-dependent quinone reduction. Reaction of the enzyme with cyanide elicited a dose-dependent formation of a charge transfer band, and the FAD-cyanide adduct was supposed to work for a sulfur transfer. The pp0053 deletion from P. putida KT2440 led to activity declines of the intracellular catalase and ubiquinone-H2 oxidase. The sulfide-quinone oxidoreductase activity in P. putida KT2440 was attributable to the presence of pp0053, and the activity showed a close relevance to enzymatic activities related to sulfur assimilation.
authors.affiliation: School of Agriculture, Meiji University, Higahimita 1-1-1, Kawasaki, 214-8571, Japan.
authors.forename: Hiroomi
authors.initials: H
authors.lastname: Shibata
authors.forename: Shigeki
authors.initials: S
authors.lastname: Kobayashi
mesh_terms: D000595:Amino Acid Sequence; D016227:Benzoquinones / Q000378:metabolism; D017353:Gene Deletion; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010084:Oxidation-Reduction; D016958:Pseudomonas putida / Q000201:enzymology* / Q000254:growth & development; D011808:Quinone Reductases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D013440:Sulfides / Q000378:metabolism; D013455:Sulfur / Q000378:metabolism*
chemical_list: D016227:Benzoquinones; D013440:Sulfides; C004532:quinone; D013455:Sulfur; D011808:Quinone Reductases; C073412:sulfide quinone reductase