Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3)pLysS cells | Vibrio harveyi |
Protein Variants | Comment | Organism |
---|---|---|
K167A | the mutant has apparently greatly increased Km and reduced kcat/Km for NADPH | Vibrio harveyi |
N134A | the mutant shows increased Km and reduced kcat/Km for NADPH | Vibrio harveyi |
R133A | the mutant shows increased Km and reduced kcat/Km for NADPH | Vibrio harveyi |
R15A | the mutant has apparently greatly increased Km and reduced kcat/Km for NADPH | Vibrio harveyi |
R225A | the mutant shows about wild type activity | Vibrio harveyi |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.009 | - |
NADPH | wild type enzyme, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi | |
0.01 | - |
NADPH | mutant enzyme R225A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi | |
0.016 | - |
NADPH | mutant enzyme R133A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi | |
0.034 | - |
NADPH | mutant enzyme N134A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Vibrio harveyi | - |
- |
- |
Purification (Comment) | Organism |
---|---|
HiTrap DEAE column chromatography, Mono Q column chromatography, and Superdex 75 gel filtration | Vibrio harveyi |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
FMN + NADPH + H+ | - |
Vibrio harveyi | FMNH2 + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
FRP | - |
Vibrio harveyi |
NADPH-flavin oxidoreductase | - |
Vibrio harveyi |
NADPH-FMN oxidoreductase | - |
Vibrio harveyi |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.2 | - |
NADPH | mutant enzyme N134A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi | |
9.2 | - |
NADPH | mutant enzyme R133A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi | |
19.2 | - |
NADPH | mutant enzyme R225A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi | |
23.8 | - |
NADPH | wild type enzyme, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Vibrio harveyi |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
12 | - |
NADPH | mutant enzyme R15A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi | |
25 | - |
NADPH | mutant enzyme K167A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi | |
150 | - |
NADPH | mutant enzyme N134A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi | |
570 | - |
NADPH | mutant enzyme R133A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi | |
2000 | - |
NADPH | mutant enzyme R225A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi | |
2670 | - |
NADPH | wild type enzyme, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C | Vibrio harveyi |