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Literature summary for 1.7.1.6 extracted from
- Functional role of Trp-105 of Enterococcus faecalis azoreductase (AzoA) as resolved by structural and mutational analysis (2008), Microbiology, 154, 2659-2667.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| into the vector pET-11a for expression in Escherichia coli BL21-Gold DE3 pLysS cells | Enterococcus faecalis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| W105A | mutant, complete loss of both affinity for FMN and enzyme activity | Enterococcus faecalis |
| W105F | mutant, lower Vmax value, decrease 30.6fold | Enterococcus faecalis |
| W105G | mutant, complete loss of both affinity for FMN and enzyme activity | Enterococcus faecalis |
| W105H | mutant, lower Vmax value, decrease 8.2fold | Enterococcus faecalis |
| W105Q | mutant, lower Vmax value, decrease 68.2fold | Enterococcus faecalis |
| W105Y | mutant, substitution does not significantly decrease the Vmax of the enzyme, 22% reduction | Enterococcus faecalis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0247 | - |
Methyl red | mutant W105F | Enterococcus faecalis |  |
| 0.0251 | - |
Methyl red | mutant W105H | Enterococcus faecalis | |
| 0.0254 | - |
Methyl red | wild-type enzyme | Enterococcus faecalis | |
| 0.0276 | - |
Methyl red | mutant W105Y | Enterococcus faecalis | |
| 0.0282 | - |
Methyl red | mutant W105Q | Enterococcus faecalis | |
| 0.158 | - |
NADH | wild-type enzyme | Enterococcus faecalis | |
| 0.166 | - |
NADH | mutant W105Y | Enterococcus faecalis | |
| 0.169 | - |
NADH | mutant W105Q | Enterococcus faecalis | |
| 0.171 | - |
NADH | mutant W105H | Enterococcus faecalis | |
| 0.175 | - |
NADH | mutant W105F | Enterococcus faecalis | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 23000 | - |
monomer, determined by SDS-PAGE | Enterococcus faecalis |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Enterococcus faecalis | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| by anion-exchange chromatography and by gel filtration | Enterococcus faecalis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| methyl red + NADH + H+ | - |
Enterococcus faecalis | ? + NAD+ | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homodimer | 2 * 23000 | Enterococcus faecalis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| AzoA | - |
Enterococcus faecalis |
| azoreductase | - |
Enterococcus faecalis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 23 | - |
activity assay at room temperature | Enterococcus faecalis |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.1 | - |
activity assay | Enterococcus faecalis |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FMN | - |
Enterococcus faecalis | |
| NADH | - |
Enterococcus faecalis | |
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