Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | slightly activating | Acinetobacter baylyi | |
DTT | slightly activating | Acinetobacter baylyi | |
EDTA | - |
Acinetobacter baylyi | |
iodoacetamide | slightly activating | Acinetobacter baylyi | |
PMSF | slightly activating | Acinetobacter baylyi | |
styrene oxide | slightly activating at 0.1 mM, inhibitory at 0.2 mM | Acinetobacter baylyi | |
Thiourea | slightly activating | Acinetobacter baylyi |
Cloned (Comment) | Organism |
---|---|
sequence comparisons and phylogenetic analysis, cloning in Escherichia coli strain DH5alpha, recombinant expression in Escherichia coli strain BL21(DE3) (pLysS) in inclusion bodies | Acinetobacter baylyi |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | complete inactivation at 0.1 mM, 38% activity remains in presence of 10 mM EDTA | Acinetobacter baylyi | |
additional information | poor inhibition by H2O2 | Acinetobacter baylyi | |
Styrene | - |
Acinetobacter baylyi | |
styrene oxide | slightly activating at 0.1 mM, inhibitory at 0.2 mM | Acinetobacter baylyi | |
thioanisole | - |
Acinetobacter baylyi |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Acinetobacter baylyi | |
0.0044 | - |
FAD | pH 7.5, 25°C, recombinant enzyme, with NADH | Acinetobacter baylyi | |
0.0055 | - |
FMN | pH 7.5, 25°C, recombinant enzyme, with NADH | Acinetobacter baylyi | |
0.011 | - |
riboflavin | pH 7.5, 25°C, recombinant enzyme, with NADH | Acinetobacter baylyi | |
0.024 | - |
NADH | pH 7.5, 25°C, recombinant enzyme, with FAD | Acinetobacter baylyi |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | no effect on enzyme activity by Cl- and NaN3 | Acinetobacter baylyi |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
FAD + NADH | Acinetobacter baylyi | - |
FADH2 + NAD+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Acinetobacter baylyi | Q6F936 | - |
- |
Purification (Comment) | Organism |
---|---|
refolded recombinant enzyme by anion exchange chromatography and gel filtration, or by nickel affinity chromatography and gel filtration, when a His10-tagged enzyme is expressed | Acinetobacter baylyi |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
reduced flavin + NAD+ = flavin + NADH + H+ | FAD reduction by recombinant His10-tagged SMOB-ADP1 follows a random sequential mechanism | Acinetobacter baylyi |
Renatured (Comment) | Organism |
---|---|
refolding of recombinant enzyme from Escherichia coli strain BL21(DE3) (pLysS) inclusion bodies, with Triton X100 used in the washing buffer, refolding from solubilization buffer containing 8 M urea, 10 mM DTT, 50 mM Tris-HCl, pH 8.0, per mg pellet at 50°C for 30 min, refolding in a 20fold volume of refolding buffer.containing 50 mM TrisHCl, pH 8.0, 240 mM NaCl, 10 mM KCl, 1 mM Na2EDTA, 1 mM reduced glutathione, 0.1 mM oxidized glutathione, and 0.05 % w/v PEG 4000, followed by ultrafiltration | Acinetobacter baylyi |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
FAD + NADH | - |
Acinetobacter baylyi | FADH2 + NAD+ | - |
r | |
FMN + NADH | - |
Acinetobacter baylyi | FMNH2 + NAD+ | - |
r | |
riboflavin + NADH | - |
Acinetobacter baylyi | reduced riboflavin + NAD+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 22665, His10-tagged enzyme, sequence calculation, 2 * 20000-25000, recombinant His10-tagged enzyme, SDS-PAGE, 2 * 18679, native enzyme, sequence calculation | Acinetobacter baylyi |
Synonyms | Comment | Organism |
---|---|---|
NAD(P)H:flavin oxidoreductase | - |
Acinetobacter baylyi |
NADH:flavin oxidoreductase | - |
Acinetobacter baylyi |
SMOB-ADP1 | - |
Acinetobacter baylyi |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 26 | assay at | Acinetobacter baylyi |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
7.8 | - |
FMN | pH 7.5, 25°C, recombinant enzyme, with NADH | Acinetobacter baylyi | |
21 | - |
riboflavin | pH 7.5, 25°C, recombinant enzyme, with NADH | Acinetobacter baylyi | |
56 | - |
FAD | pH 7.5, 25°C, recombinant enzyme, with NADH | Acinetobacter baylyi | |
64 | - |
NADH | pH 7.5, 25°C, recombinant enzyme, with FAD | Acinetobacter baylyi |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Acinetobacter baylyi |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.3 | 7.5 | activity irreversibly drops to under 5% at pH 5.3 indicating denaturation | Acinetobacter baylyi |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | no activity with NADPH | Acinetobacter baylyi | |
NADH | - |
Acinetobacter baylyi |
General Information | Comment | Organism |
---|---|---|
evolution | SMOB-ADP1 belongs to the flavin reductases of the HpaC-like subfamily. NAD(P)H:flavin oxidoreductase structure comparisons, overview | Acinetobacter baylyi |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0013 | - |
FAD | pH 7.5, 25°C, recombinant enzyme, with NADH | Acinetobacter baylyi | |
0.0014 | - |
FMN | pH 7.5, 25°C, recombinant enzyme, with NADH | Acinetobacter baylyi | |
0.0019 | - |
riboflavin | pH 7.5, 25°C, recombinant enzyme, with NADH | Acinetobacter baylyi | |
0.0027 | - |
NADH | pH 7.5, 25°C, recombinant enzyme, with FAD | Acinetobacter baylyi |