Protein Variants | Comment | Organism |
---|---|---|
additional information | replacement of Tyr50 stacked on the si-face of the isoalloxazine ring of the flavin adenine dinucleotide prosthetic group modulates Bacillus subtilis ferredoxin-NADP+ oxidoreductase activity toward NADPH. The Y50G and Y50S mutations enhance the FAD fluorescence emission, whereas those of the wild type and Y50W mutant are quenched | Bacillus subtilis |
Y50G | site-directed mutagenesis, the mutant shows a blue shift of the FAD transition band and decreased thermal stability compared to wild-type. Using the diaphorase assay, the kcat values for the Y50G mutant in the presence of NADPH and ferricyanide is decreased to less than 5% of the wild-type activity | Bacillus subtilis |
Y50G | the mutation decreases thermal stability compared to the wild type enzyme | Bacillus subtilis |
Y50S | site-directed mutagenesis, the mutant shows a blue shift of the FAD transition band and decreased thermal stability compared to wild-type. Using the diaphorase assay, the kcat values for the Y50G mutant in the presence of NADPH and ferricyanide is decreased to less than 5% of the wild-type activity | Bacillus subtilis |
Y50S | the mutation decreases thermal stability compared to the wild type enzyme | Bacillus subtilis |
Y50W | site-directed mutagenesis, the mutant shows a blue shift of the FAD transition band and decreased thermal stability compared to wild-type. The mutant retains approximately 20 % reactivity in the diaphorase assay and BsFd-dependent cytochrome c reduction assay relative to wild-type | Bacillus subtilis |
Y50W | the mutation decreases thermal stability compared to the wild type enzyme, The mutant retains approximately 20% of wild type reactivity | Bacillus subtilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Bacillus subtilis | |
0.0019 | - |
NADPH | pH 7.0, 25°C, recombinant mutant Y50S | Bacillus subtilis | |
0.004 | - |
NADPH | pH 7.0, 25°C, recombinant mutant Y50W | Bacillus subtilis | |
0.0043 | - |
NADPH | pH 7.0, 25°C, recombinant mutant Y50G | Bacillus subtilis | |
0.0197 | - |
NADPH | pH 7.0, 25°C, recombinant wild-type enzyme | Bacillus subtilis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
40000 | - |
recombinant mutant enzymes, gel filtration | Bacillus subtilis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 oxidized ferredoxin + NADPH | Bacillus subtilis | - |
2 reduced ferredoxin + NADP+ + H+ | - |
r | |
2 oxidized ferredoxin + NADPH | Bacillus subtilis 168 | - |
2 reduced ferredoxin + NADP+ + H+ | - |
r | |
reduced ferredoxin + NADP+ + H+ | Bacillus subtilis | - |
oxidized ferredoxin + NADPH | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | - |
- |
- |
Bacillus subtilis | O05268 | - |
- |
Bacillus subtilis 168 | O05268 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2 reduced ferredoxin + NADP+ + H+ = 2 oxidized ferredoxin + NADPH | a hydride transfer reaction with NAD(P)H and two separate one-electron transfer reactions with ferredoxin are generally involved in te reaction process | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 oxidized ferredoxin + NADPH | - |
Bacillus subtilis | 2 reduced ferredoxin + NADP+ + H+ | - |
r | |
2 oxidized ferredoxin + NADPH | - |
Bacillus subtilis 168 | 2 reduced ferredoxin + NADP+ + H+ | - |
r | |
reduced cytochrome c + NADP+ | - |
Bacillus subtilis | oxidized cytochrome c + NADPH + H+ | - |
? | |
reduced ferredoxin + NADP+ + H+ | - |
Bacillus subtilis | oxidized ferredoxin + NADPH | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 40000, SDS-PAGE | Bacillus subtilis |
monomer | 1 * 40000, recombinant mutant enzymes, SDS-PAGE | Bacillus subtilis |
Synonyms | Comment | Organism |
---|---|---|
BsFNR | - |
Bacillus subtilis |
ferredoxin-NADP+ oxidoreductase | - |
Bacillus subtilis |
FNR | - |
Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Bacillus subtilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
16.2 | - |
NADPH | pH 7.0, 25°C, recombinant mutant Y50S | Bacillus subtilis | |
42.8 | - |
NADPH | pH 7.0, 25°C, recombinant mutant Y50G | Bacillus subtilis | |
183 | - |
NADPH | pH 7.0, 25°C, recombinant mutant Y50W | Bacillus subtilis | |
1012 | - |
NADPH | pH 7.0, 25°C, recombinant wild-type enzyme | Bacillus subtilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Bacillus subtilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Bacillus subtilis | |
FAD | flavin adenine dinucleotide prosthetic group, the FAD prosthetic group is noncovalently bound in the open conformation. The fluorescence intensity of the protein-bound FAD is influenced by its environment. The fluorescence emissions of enzyme mutants Y50G and Y50S enzymes increase six to sevenfold compared with that of free FAD, whereas the fluorescence emission of wild-type and Y50W enzymes is efficiently quenched to below 3% and 9% of that of free FAD. ENzyme residue Tyr50 plays an important role to stabilize the FAD prosthetic group | Bacillus subtilis | |
Ferredoxin | - |
Bacillus subtilis | |
NADP+ | - |
Bacillus subtilis | |
NADPH | - |
Bacillus subtilis |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a member of the flavoprotein superfamily | Bacillus subtilis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
8526 | - |
NADPH | pH 7.0, 25°C, recombinant mutant Y50S | Bacillus subtilis | |
9953 | - |
NADPH | pH 7.0, 25°C, recombinant mutant Y50G | Bacillus subtilis | |
45750 | - |
NADPH | pH 7.0, 25°C, recombinant mutant Y50W | Bacillus subtilis | |
51370 | - |
NADPH | pH 7.0, 25°C, recombinant wild-type enzyme | Bacillus subtilis |