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Literature summary for 1.8.5.3 extracted from
- Kinetic and spectroscopic characterization of tungsten-substituted DMSO reductase from Rhodobacter sphaeroides (2018), J. Biol. Inorg. Chem., 23, 295-301 .
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | pre-steady-state kinetics and enzyme-monitored turnover, and steady-state kinetics, kinetic analysis of molybdenum-DMSO reductase and a tungsten-substituted form of DMSO reductase, overview | Cereibacter sphaeroides | |
| 0.064 | - |
Dimethylsulfoxide | W-DMSOR, pH 6.0, temperature not specified in the publication | Cereibacter sphaeroides |  |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Molybdenum | required, Mo-DMSOR | Cereibacter sphaeroides | |
| additional information | kinetic and spectroscopic analysis of molybdenum-DMSO reductase and a tungsten-substituted form of DMSO reductase, overview. Partial reduction with sodium dithionite yields a well-resolved W(V) EPR signal of the so-called high-g split type that exhibits markedly greater g-anisotropy than the corresponding Mo(V) signal of the native form of the enzyme, with the g values shifted to higher magnetic field by as much as DELTAgave = 0.056. Deuteration of the enzyme confirms that the coupled proton is solvent-exchangeable, allowing to accurately simulate the tungsten hyperfine coupling. Global curve-fitting analysis of UV/vis absorption spectra observed in the course of the reaction of the tungsten-substituted enzyme with sodium dithionite affords a well-defined absorption spectrum for the W(V) species. The absorption spectrum for this species exhibits significantly larger molar extinction coefficients than either the reduced or the oxidized spectrum. This spectrum, in conjunction with those for fully oxidized W(VI) and fully reduced W(IV) enzyme, is used to deconvolute the absorption spectra seen in the course of turnover, in which the enzyme is reacted with sodium dithionite and DMSO, demonstrating that the W(V) is an authentic catalytic intermediate that accumulates to approximately 50% of the total enzyme in the steady state. At the end of turnover, in the presence of excess dithionite, the fully reduced W-DMSOR accumulates. That no W(IV)-DMSO intermediate is seen during turnover indicates that DMS does not rebind to the oxidized W-DMSOR | Cereibacter sphaeroides | |
| Tungsten | W-DMSOR, can substitute for molybdenum, the W-DMSOR shows 30fold higher activity than the molybdenum-containing enzyme, Mo-DMSOR. Rapid-scanning stopped-flow traces showing the enzyme monitored turnover of 0.032 mM W-DMSOR with 10 mM sodium dithionite and 0.25 mM DMSO performed in 50 mM KH2PO4, 0.6 mM EDTA, pH 6.0 at 25°C | Cereibacter sphaeroides | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| dimethylsulfoxide + menaquinol | Cereibacter sphaeroides | - |
dimethylsulfide + menaquinone + H2O | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Cereibacter sphaeroides | Q57366 | Rhodobacter sphaeroides strain 16 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| native Mo-DMSOR and engineered W-DMSOR, the latter from cell-free supernatant by ultracentrifugation at 150000 x g, ammonium sulfate fractionation, reduction with sodium dithionite followed by reoxidation with DMSO, hydrophobic interaction chromatography, and desalting gel filtration, followed by two different steps of anion exchange chromatography and ultrafiltration | Cereibacter sphaeroides |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| dimethylsulfoxide + menaquinol | - |
Cereibacter sphaeroides | dimethylsulfide + menaquinone + H2O | - |
? | |
| dimethylsulfoxide + methyl viologen | - |
Cereibacter sphaeroides | dimethylsulfide + oxidized methyl viologen + H2O | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| DMSO reductase | - |
Cereibacter sphaeroides |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1120 | - |
Dimethylsulfoxide | W-DMSOR, pH 6.0, temperature not specified in the publication | Cereibacter sphaeroides | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 7.5 | assay at | Cereibacter sphaeroides |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| methyl viologen | - |
Cereibacter sphaeroides | |
| molybdenum cofactor | Moco | Cereibacter sphaeroides | |
| additional information | kinetic and spectroscopic analysis of molybdenum-DMSO reductase and a tungsten-substituted form of DMSO reductase, overview. Partial reduction with sodium dithionite yields a well-resolved W(V) EPR signal of the so-called high-g split type that exhibits markedly greater g-anisotropy than the corresponding Mo(V) signal of the native form of the enzyme, with the g values shifted to higher magnetic field by as much as DELTAgave = 0.056. Deuteration of the enzyme confirms that the coupled proton is solvent-exchangeable, allowing to accurately simulate the tungsten hyperfine coupling. Global curve-fitting analysis of UV/vis absorption spectra observed in the course of the reaction of the tungsten-substituted enzyme with sodium dithionite affords a well-defined absorption spectrum for the W(V) species. The absorption spectrum for this species exhibits significantly larger molar extinction coefficients than either the reduced or the oxidized spectrum. This spectrum, in conjunction with those for fully oxidized W(VI) and fully reduced W(IV) enzyme, is used to deconvolute the absorption spectra seen in the course of turnover, in which the enzyme is reacted with sodium dithionite and DMSO, demonstrating that the W(V) is an authentic catalytic intermediate that accumulates to approximately 50% of the total enzyme in the steady state | Cereibacter sphaeroides |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 17500 | - |
Dimethylsulfoxide | W-DMSOR, pH 6.0, temperature not specified in the publication | Cereibacter sphaeroides | |
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