Activating Compound | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Bacillus subtilis | |
NADPH | about double activity compared to NADH | Bacillus subtilis |
Cloned (Comment) | Organism |
---|---|
frb gene inserted into plasmid pET21-a, introduced into Escherichia coli BL21 (DE3), cultivated at 30°C | Bacillus subtilis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5,5'-dithiobis(2-nitrobenzoic acid) | with 0.1 mM 28.8% activity | Bacillus subtilis | |
AgNO3 | with 1 mM 1.05% activity | Bacillus subtilis | |
CuCl2 | with 1 mM 11.2% activity | Bacillus subtilis | |
HgCl2 | with 1 mM 0.77% activity | Bacillus subtilis | |
additional information | with 1 mM ethylenediaminetetraacetic acid 99.8% activity, with 1 mM 2,2'-bipyridyl 81.8% activity | Bacillus subtilis | |
N-ethylmaleimide | with 1 mM 16.3% activity | Bacillus subtilis | |
p-chloromercuribenzoic acid | with 1 mM 15.6% activity | Bacillus subtilis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
CaCl2 | with 1 mM 84.5% activity | Bacillus subtilis | |
FeCl2 | with 1 mM 90.0% activity | Bacillus subtilis | |
MgCl2 | with 1 mM 92.6% activity | Bacillus subtilis | |
MnCl2 | with 1 mM 97.0% activity | Bacillus subtilis | |
NiCl2 | with 1 mM 86.6% activity | Bacillus subtilis | |
ZnCl2 | with 1 mM 96.8% activity | Bacillus subtilis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
20000 | - |
2 * 20000 | Bacillus subtilis |
40000 | - |
SDS-PAGE | Bacillus subtilis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
FMN + NADH + H+ | Bacillus subtilis | - |
FMNH2 + NAD+ | - |
? | |
FMN + NADH + H+ | Bacillus subtilis WU-S2B | - |
FMNH2 + NAD+ | - |
? | |
FMN + NADPH + H+ | Bacillus subtilis | 203% activity compared to NAD+ | FMNH2 + NADP+ | - |
? | |
FMN + NADPH + H+ | Bacillus subtilis WU-S2B | 203% activity compared to NAD+ | FMNH2 + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | - |
- |
- |
Bacillus subtilis WU-S2B | - |
- |
- |
Purification (Comment) | Organism |
---|---|
cells harvested by centrifugation, washing twice and suspended in 50 mM Tris-HCl buffer, pH 8.0, with 1 mM dithiothreitol and 10% glycerol, cells disrupted with ultraoscillator (20 kHz), debris removed by centrifugation, enzyme purified from cell-free extracts with ÄKTA explorer, applied to Toyopearl DEAE-650 M column, equilibrated with 50 mM Tris-HCl buffer, pH 8.0, washed, proteins eluted with linear gradient of 0-0.5 M KCl, active fractions (at 0.15-0.2 M KCl) combined and concentrated by ultrafiltration (membrane filter YM-30), enzyme purified with HiLoad 16/60 Superdex 200, equilibrated with 50 mM Tris-HCl buffer, pH 8.0 with 0.15 M NaCl, elution with same buffer | Bacillus subtilis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
42.1 | - |
FMN reduction with NADH, pH 6.0, 50°C | Bacillus subtilis |
85.5 | - |
FMN reduction with NADPH, pH 6.0, 50°C | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,3-dinitrobenzene + NADH + H+ | 10.6% activity compared to FMN | Bacillus subtilis | ? + NAD+ | - |
? | |
1,4-benzoquinone + NADH + H+ | 18.6% activity compared to FMN | Bacillus subtilis | 1,4-benzoquinol + NAD+ | - |
? | |
2 ferricyanide + NADH | 15.4% activity compared to FMN | Bacillus subtilis | 2 ferrocyanide + NAD+ + H+ | - |
? | |
2 ferricyanide + NADH | 15.4% activity compared to FMN | Bacillus subtilis WU-S2B | 2 ferrocyanide + NAD+ + H+ | - |
? | |
2 ferricytochrome c + NADH | 8.48% activity compared to FMN | Bacillus subtilis | 2 ferrocytochrome c + NAD+ + H+ | - |
? | |
2 ferricytochrome c + NADH | 8.48% activity compared to FMN | Bacillus subtilis WU-S2B | 2 ferrocytochrome c + NAD+ + H+ | - |
? | |
3-nitrophenol + NADH + H+ | 8.95% activity compared to FMN | Bacillus subtilis | ? + NAD+ | - |
? | |
4-nitroacetophenone + NADH + H+ | 17.9% activity compared to FMN | Bacillus subtilis | ? + NAD+ | - |
? | |
4-nitroaniline + NADH + H+ | 4.91% activity compared to FMN | Bacillus subtilis | ? + NAD+ | - |
? | |
4-nitrobenzoate + NADH + H+ | 21.2% activity compared to FMN | Bacillus subtilis | ? + NAD+ | - |
? | |
4-nitrophenol + NADH + H+ | 8.07% activity compared to FMN | Bacillus subtilis | ? + NAD+ | - |
? | |
4-nitrotoluene + NADH + H+ | 8.83% activity compared to FMN | Bacillus subtilis | ? + NAD+ | - |
? | |
FAD + NADH + H+ | 58.1% activity compared to FMN | Bacillus subtilis | FADH2 + NAD+ | - |
? | |
FMN + NADH + H+ | - |
Bacillus subtilis | FMNH2 + NAD+ | - |
? | |
FMN + NADH + H+ | - |
Bacillus subtilis WU-S2B | FMNH2 + NAD+ | - |
? | |
FMN + NADPH + H+ | 203% activity compared to NAD+ | Bacillus subtilis | FMNH2 + NADP+ | - |
? | |
FMN + NADPH + H+ | 203% activity compared to NAD+ | Bacillus subtilis WU-S2B | FMNH2 + NADP+ | - |
? | |
lumiflavin + NADH + H+ | 14.1% activity compared to FMN | Bacillus subtilis | reduced lumiflavin + NAD+ | - |
? | |
menadione + NADH + H+ | 19.0% activity compared to FMN | Bacillus subtilis | menadiol + NAD+ | - |
? | |
methyl-4-nitrobenzoate + NADH + H+ | 10.7% activity compared to FMN | Bacillus subtilis | ? + NAD+ | - |
? | |
methylene blue + NADH + H+ | 29.0% activity compared to FMN | Bacillus subtilis | reduced methylene blue + NAD+ | - |
? | |
NADH + H+ + oxidized 2,6-dichlorophenolindophenol | 23.0% activity compared to FMN | Bacillus subtilis | NAD+ + reduced 2,6-dichlorophenolindophenol | - |
? | |
NADH + H+ + oxidized 2,6-dichlorophenolindophenol | 23.0% activity compared to FMN | Bacillus subtilis WU-S2B | NAD+ + reduced 2,6-dichlorophenolindophenol | - |
? | |
nitrofurantoin + NADH + H+ | 10.1% activity compared to FMN | Bacillus subtilis | ? + NAD+ | - |
? | |
nitrofurazone + NADH + H+ | 13.5% activity compared to FMN | Bacillus subtilis | reduced nitrofurazone + NAD+ | - |
? | |
riboflavin + NADH + H+ | 29.0% activity compared to FMN | Bacillus subtilis | reduced riboflavin + NAD+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 20000 | Bacillus subtilis |
Synonyms | Comment | Organism |
---|---|---|
flavin reductase | - |
Bacillus subtilis |
Frb | - |
Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
- |
Bacillus subtilis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | 55 | maxiumum activity at 50°C, about 90% activity at 45°C, about 80% at 40 and 55°C, about 50-60% activity at 35 and 60°C, about 40% activity at 30°C, about 30% activity at 25°C, about 20% activity at 20 and 65°C, no activity at 70°C | Bacillus subtilis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | 70 | 30 min incubation at 20°C retains 100% activity, more than 90% activity is retained at 25-45°C, about 80% at 50°C, about 50% at 55°C, about 20% at 60°C, about 10% at 65°C, no activity at 70°C | Bacillus subtilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
- |
Bacillus subtilis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 10 | maximum activity at pH 6.0, more than 80% activity at pH 5.5, 6.5, and 7.0, about 75% activity at pH 7.5, about 30% activity at pH 8.5, less than 20% activity at pH 9-9.5, less than 10% activity at pH 4, 4.5, and 10, pH 4.0-6.0 citrate-phosphate buffer, pH 6.0-7.5 potassium-phosphate buffer, pH 7.5-8.5 Tris-HCl buffer, pH 8.5-10.0 glycine-NaOH buffer | Bacillus subtilis |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
2 | 12 | about 80% activity retained after 24 h incubation at pH 2.0-3.5, 90 to less than 100% at pH 4.0-5.5 and pH 8.0-12.0, 100% at pH 6.0-7.5, pH 4.0-6.0 citrate-phosphate buffer, pH 6.0-7.5 potassium-phosphate buffer, pH 7.5-8.5 Tris-HCl buffer, pH 8.5-10.0 glycine-NaOH buffer | Bacillus subtilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FMN | - |
Bacillus subtilis | |
NADH | - |
Bacillus subtilis | |
NADPH | - |
Bacillus subtilis |
General Information | Comment | Organism |
---|---|---|
metabolism | the flavin reductase couples with dibenzothiophene and dibenzothiophene sulfone monooxygenase in the thermophilic dibenzothiophene (DBT)-desulfurizing bacterium, the flavin reductase exhibits flavin reductase and nitroreductase activities | Bacillus subtilis |