General Stability | Organism |
---|---|
Ni2+ stabilizes | Rhodococcus opacus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ni2+ | 0.5-1 mM, strong inhibition of artificial electron acceptor reduction | Rhodococcus opacus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05 | - |
methyl viologen | at pH 9.4 | Rhodococcus opacus | |
0.15 | - |
NAD+ | - |
Rhodococcus opacus | |
0.38 | - |
FAD | - |
Rhodococcus opacus | |
0.44 | - |
methylene blue | - |
Rhodococcus opacus | |
0.9 | - |
methyl viologen | at pH 8.0 | Rhodococcus opacus | |
4 | - |
ferricyanide | - |
Rhodococcus opacus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
soluble | - |
Rhodococcus opacus | - |
- |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | NAD+ reduction with H2 is completely dependent on the presence of divalent metal ions Ni2+, Co2+, Mg2+ or Mn2+ or of high salt concentrations between 500-1500 mM | Rhodococcus opacus | |
Mg2+ | NAD+ reduction with H2 is completely dependent on the presence of divalent metal ions Ni2+, Co2+, Mg2+ or Mn2+ or of high salt concentrations between 500-1500 mM | Rhodococcus opacus | |
Mn2+ | NAD+ reduction with H2 is completely dependent on the presence of divalent metal ions Ni2+, Co2+, Mg2+ or Mn2+, or of high salt concentrations of 500-1500 mM | Rhodococcus opacus | |
additional information | stimulation of activity by salt is greater the less chaotrophic the anion | Rhodococcus opacus | |
Ni2+ | NAD+ reduction with H2 is completely dependent on the presence of divalent metal ions Ni2+, Co2+, Mg2+ or Mn2+ or of high salt concentrations between 500-1500 mM | Rhodococcus opacus | |
Ni2+ | highest specific activity with NiCl2, optimal concentration: 1 mM | Rhodococcus opacus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
27000 | - |
1 * 64000 + 1 * 56000 + 1 * 31000 + 1 * 27000, native hydrogenase dissociates into two subunit dimers of 64000 Da and 1 * 31000 Da, and of 56000 Da and 27000 Da respectively, SDS-PAGE | Rhodococcus opacus |
31000 | - |
1 * 64000 + 1 * 56000 + 1 * 31000 + 1 * 27000, native hydrogenase dissociates into two subunit dimers of 64000 Da and 1 * 31000 Da, and of 56000 Da and 27000 Da respectively, SDS-PAGE | Rhodococcus opacus |
56000 | - |
1 * 64000 + 1 * 56000 + 1 * 31000 + 1 * 27000, native hydrogenase dissociates into two subunit dimers of 64000 Da and 1 * 31000 Da, and of 56000 Da and 27000 Da respectively, SDS-PAGE | Rhodococcus opacus |
64000 | - |
1 * 64000 + 1 * 56000 + 1 * 31000 + 1 * 27000, native hydrogenase dissociates into two subunit dimers of 64000 Da and 1 * 31000 Da, and of 56000 Da and 27000 Da respectively, SDS-PAGE | Rhodococcus opacus |
178000 | - |
sucrose density gradient centrifugation | Rhodococcus opacus |
200000 | - |
gel filtration | Rhodococcus opacus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus opacus | - |
- |
- |
Rhodococcus opacus 1B | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Rhodococcus opacus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
45 | - |
- |
Rhodococcus opacus |
Storage Stability | Organism |
---|---|
4°C, 500 mM potassium phosphate, pH 7.0, 3 d, 10% loss of activity | Rhodococcus opacus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
FAD + H2 | 99% activity compared to NAD+ | Rhodococcus opacus | FADH2 | - |
r | |
H2 + NAD+ | electron acceptor FAD | Rhodococcus opacus | H+ + NADH | - |
r | |
H2 + NAD+ | artificial electron acceptor benzyl viologen | Rhodococcus opacus | H+ + NADH | - |
r | |
H2 + NAD+ | artificial electron acceptor methyl viologen | Rhodococcus opacus | H+ + NADH | - |
r | |
H2 + NAD+ | electron acceptor NAD+ | Rhodococcus opacus | H+ + NADH | - |
r | |
H2 + NAD+ | artificial electron acceptor ferricyanide | Rhodococcus opacus | H+ + NADH | - |
r | |
H2 + NAD+ | artificial electron acceptor methylene blue | Rhodococcus opacus | H+ + NADH | - |
r | |
H2 + NAD+ | electron acceptor FAD | Rhodococcus opacus 1B | H+ + NADH | - |
r | |
H2 + NAD+ | artificial electron acceptor benzyl viologen | Rhodococcus opacus 1B | H+ + NADH | - |
r | |
H2 + NAD+ | artificial electron acceptor methyl viologen | Rhodococcus opacus 1B | H+ + NADH | - |
r | |
H2 + NAD+ | electron acceptor NAD+ | Rhodococcus opacus 1B | H+ + NADH | - |
r | |
H2 + oxidized benzyl viologen | 50fold higher specific activity compared to NAD+ reduction | Rhodococcus opacus | H+ + reduced benzyl viologen | - |
r | |
H2 + oxidized methyl viologen | - |
Rhodococcus opacus | H+ + reduced methyl viologen | - |
? | |
H2 + oxidized methyl viologen | - |
Rhodococcus opacus 1B | H+ + reduced methyl viologen | - |
? | |
H2 + oxidized methylene blue | - |
Rhodococcus opacus | H+ + reduced methylene blue | - |
r | |
oxidized methylene blue + H2 | 1058% activity compared to NAD+ | Rhodococcus opacus | reduced methylene blue + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
tetramer | 1 * 64000 + 1 * 56000 + 1 * 31000 + 1 * 27000, native hydrogenase dissociates into two subunit dimers of 64000 Da and 1 * 31000 Da, and of 56000 Da and 27000 Da respectively, SDS-PAGE | Rhodococcus opacus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
electron donor NADH | Rhodococcus opacus |
6.1 | - |
electron acceptor NAD+, no addition of Ni2+, Tris/MES buffer | Rhodococcus opacus |
7.8 | 8 | electron acceptor NAD+, in triethanolamine/HCl buffer containing NiCl2 | Rhodococcus opacus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Rhodococcus opacus |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
6 | - |
1 day: 9% loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 56% loss of activity in the absence of NiCl2 and MgCl2, 3 days: 44% loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 80% loss of activity in the absence of NiCl2 and MgCl2 | Rhodococcus opacus |
7 | - |
1 day: no loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 62% loss of activity in the absence of NiCl2 and MgCl2, 3 days: 21% loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 87% loss of activity in the absence of NiCl2 and MgCl2 | Rhodococcus opacus |
8 | - |
1 day: no loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 86% loss of activity in the absence of NiCl2 and MgCl2, 3 days: 27% loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, complete loss of activity in the absence of NiCl2 and MgCl2 | Rhodococcus opacus |