Activating Compound | Comment | Organism | Structure |
---|---|---|---|
glutathione | up to 0.1 mM: activation, above 0.1 mM: inactivation | Rhodococcus opacus | |
NH4+ | activates | Rhodococcus opacus | |
phosphate | increase in NAD-reduction rate | Rhodococcus opacus | |
SO42- | activates | Rhodococcus opacus |
General Stability | Organism |
---|---|
5 mM Mg2+ and 0.5 mM Ni2+ stabilizes | Rhodococcus opacus |
addition of 5 mM Mg2+ + 0.5 mM Ni2+ stabilizes | Rhodococcus opacus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
cyanide | 1 mM, 33% inhibition | Rhodococcus opacus | |
glutathione | above 0.1 mM: inactivation, up to 0.1 mM: activation | Rhodococcus opacus | |
iodoacetate | 100 mM, 36% inhibition | Rhodococcus opacus | |
Na+ | - |
Rhodococcus opacus | |
p-chloromercuribenzoate | 0.02 mM, complete inhibition, reversed by addition of glutathione | Rhodococcus opacus | |
Sulfide | 0.1 mM, 10% inhibition | Rhodococcus opacus | |
Triethanolamine buffer | at high concentration | Rhodococcus opacus | |
Tris-buffer | at high concentration | Rhodococcus opacus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.063 | - |
H2 | - |
Rhodococcus opacus | |
0.123 | - |
NAD+ | - |
Rhodococcus opacus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
K+ | activates | Rhodococcus opacus | |
Mg2+ | NAD-reduction: no linear kinetics in absence of metals, 0.5 mM Ni2+ and 5 mM Mg2+ required | Rhodococcus opacus | |
Ni2+ | NAD-reduction: no linear kinetics in absence of metals, 0.5 mM Ni2+ and 5 mM Mg2+ required | Rhodococcus opacus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus opacus | - |
- |
- |
Rhodococcus opacus 1B | - |
- |
- |
Purification (Comment) | Organism |
---|---|
protamine sulfate, ammonium sulfate, DEAE-cellulose, Sephadex G-200 | Rhodococcus opacus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
103 | - |
- |
Rhodococcus opacus |
Storage Stability | Organism |
---|---|
-20°C, 4 mg/ml protein concentration, pH 7.0 | Rhodococcus opacus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
H2 + NAD+ | artificial electron acceptor benzyl viologen | Rhodococcus opacus | H+ + NADH | - |
? | |
H2 + NAD+ | artificial electron acceptor methyl viologen | Rhodococcus opacus | H+ + NADH | - |
? | |
H2 + NAD+ | electron acceptor NAD+ | Rhodococcus opacus | H+ + NADH | - |
? | |
H2 + NAD+ | artificial electron acceptor methylene blue, only in presence of catalytic amounts of NAD+ | Rhodococcus opacus | H+ + NADH | - |
? | |
H2 + NAD+ | artificial electron acceptor methylene blue | Rhodococcus opacus | H+ + NADH | - |
? | |
H2 + NAD+ | artificial electron acceptor benzyl viologen | Rhodococcus opacus 1B | H+ + NADH | - |
? | |
H2 + NAD+ | artificial electron acceptor methyl viologen | Rhodococcus opacus 1B | H+ + NADH | - |
? | |
H2 + NAD+ | electron acceptor NAD+ | Rhodococcus opacus 1B | H+ + NADH | - |
? | |
H2 + NAD+ | artificial electron acceptor methylene blue, only in presence of catalytic amounts of NAD+ | Rhodococcus opacus 1B | H+ + NADH | - |
? | |
H2 + NAD+ | artificial electron acceptor methylene blue | Rhodococcus opacus 1B | H+ + NADH | - |
? |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
enzyme prepared in Tris or triethanolamine buffer: 90% loss of activity after 10 min, enzyme prepared in K-phosphate buffer: 30% loss of activity after 10 min | Rhodococcus opacus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
methylene blue reduction, catalytic amounts of NAD+ required | Rhodococcus opacus |
8 | 8.2 | electron acceptor NAD+ | Rhodococcus opacus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7 | 9.5 | triethanolamine buffer, Tris buffer pH 7.0: 40-45% activity, triethanolamine buffer, Tris buffer pH 9.5: 60-75% activity | Rhodococcus opacus |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
highest stability, 4°C, H2-atmosphere, 3 days | Rhodococcus opacus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
flavin | only when FMN or riboflavin are added in combination with Mg2+ or Ni2+, NAD-reducing system is activated | Rhodococcus opacus |