| Title: | Purification and characterization of dimethylsulfide monooxygenase from Hyphomicrobium sulfonivorans |
| Author(s): | Boden R; Borodina E; Wood AP; Kelly DP; Murrell JC; Schafer H; |
| Address: | "School of Life Sciences, University of Warwick, Wellesbourne, UK" |
| ISSN/ISBN: | 1098-5530 (Electronic) 0021-9193 (Print) 0021-9193 (Linking) |
| Abstract: | "Dimethylsulfide (DMS) is a volatile organosulfur compound which has been implicated in the biogeochemical cycling of sulfur and in climate control. Microbial degradation is a major sink for DMS. DMS metabolism in some bacteria involves its oxidation by a DMS monooxygenase in the first step of the degradation pathway; however, this enzyme has remained uncharacterized until now. We have purified a DMS monooxygenase from Hyphomicrobium sulfonivorans, which was previously isolated from garden soil. The enzyme is a member of the flavin-linked monooxygenases of the luciferase family and is most closely related to nitrilotriacetate monooxygenases. It consists of two subunits: DmoA, a 53-kDa FMNH(2)-dependent monooxygenase, and DmoB, a 19-kDa NAD(P)H-dependent flavin oxidoreductase. Enzyme kinetics were investigated with a range of substrates and inhibitors. The enzyme had a K(m) of 17.2 (+/- 0.48) muM for DMS (k(cat) = 5.45 s(-)(1)) and a V(max) of 1.25 (+/- 0.01) mumol NADH oxidized min(-)(1) (mg protein(-)(1)). It was inhibited by umbelliferone, 8-anilinonaphthalenesulfonate, a range of metal-chelating agents, and Hg(2)(+), Cd(2)(+), and Pb(2)(+) ions. The purified enzyme had no activity with the substrates of related enzymes, including alkanesulfonates, aldehydes, nitrilotriacetate, or dibenzothiophenesulfone. The gene encoding the 53-kDa enzyme subunit has been cloned and matched to the enzyme subunit by mass spectrometry. DMS monooxygenase represents a new class of FMNH(2)-dependent monooxygenases, based on its specificity for dimethylsulfide and the molecular phylogeny of its predicted amino acid sequence. The gene encoding the large subunit of DMS monooxygenase is colocated with genes encoding putative flavin reductases, homologues of enzymes of inorganic and organic sulfur compound metabolism, and enzymes involved in riboflavin synthesis" |
| Keywords: | "Bacterial Proteins/genetics/*metabolism Cloning, Molecular Electrophoresis, Polyacrylamide Gel Gene Expression Regulation, Bacterial/physiology Hyphomicrobium/*enzymology/genetics/*metabolism Metals/metabolism Mixed Function Oxygenases/chemistry/genetics/;" |
| Notes: | "MedlineBoden, Rich Borodina, Elena Wood, Ann P Kelly, Donovan P Murrell, J Colin Schafer, Hendrik eng Research Support, Non-U.S. Gov't 2011/01/11 J Bacteriol. 2011 Mar; 193(5):1250-8. doi: 10.1128/JB.00977-10. Epub 2011 Jan 7" |
