Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) pLysS cells | Pseudomonas aeruginosa |
Crystallization (Comment) | Organism |
---|---|
mutant enzyme Y131F in the presence of methyl red, sitting drop vapor diffusion method, using 0.1 M sodium acetate pH 4.5, 1.0 M diammonium hydrogen phosphate | Pseudomonas aeruginosa |
Protein Variants | Comment | Organism |
---|---|---|
Y131F | the mutant shows increased specific activity with methyl red and reduced specific activity with balsalazide compared to the wild type enzyme | Pseudomonas aeruginosa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0445 | - |
Methyl red | mutant enzyme Y131F, using NADPH as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
0.0927 | - |
Methyl red | wild type enzyme, using NADPH as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
0.0986 | - |
balsalazide | wild type enzyme, using NADPH as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
0.1029 | - |
balsalazide | mutant enzyme Y131F, using NADPH as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
0.535 | - |
NADPH | mutant enzyme Y131F, using methyl red as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
0.538 | - |
NADH | wild type enzyme, using methyl red as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
0.965 | - |
NADH | mutant enzyme Y131F, using methyl red as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
1.197 | - |
NADPH | wild type enzyme, using methyl red as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | Q9I5F3 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography | Pseudomonas aeruginosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-(4-aminophenylazo)benzenesulfonic acid + NADPH + H+ | about 25% specific activity compared to balsalazide | Pseudomonas aeruginosa | ? | - |
? | |
balsalazide + NADPH + H+ | 100% specific activity | Pseudomonas aeruginosa | ? | - |
? | |
methyl red + NADH + H+ | the enzyme exhibits lower activity with NADH compared to NADPH | Pseudomonas aeruginosa | ? | - |
? | |
methyl red + NADPH + H+ | about 50% specific activity compared to balsalazide | Pseudomonas aeruginosa | ? | - |
? | |
additional information | no activity with Amaranth, Tropaeolin, Orange II, Ponceau S, Ponceau BS, and Orange G | Pseudomonas aeruginosa | ? | - |
? | |
olsalazine + NADPH + H+ | less than 10% specific activity compared to balsalazide | Pseudomonas aeruginosa | ? | - |
? | |
sulfasalazine + NADPH + H+ | about 20% specific activity compared to balsalazide | Pseudomonas aeruginosa | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
AzoR1 | - |
Pseudomonas aeruginosa |
azoreductase 1 | - |
Pseudomonas aeruginosa |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
42 | 55 | whilst wild type AzoR1 loses 50% of its activity after incubation for 10 min at 55°C, mutant enzyme Y131F loses 50% of its activity after exposure to 42°C for 10 min, both enzymes are inactive after 10 min at 80°C | Pseudomonas aeruginosa |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
10 | - |
NADH | wild type enzyme, using methyl red as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
12.7 | - |
Methyl red | wild type enzyme, using NADPH as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
19.1 | - |
Methyl red | mutant enzyme Y131F, using NADPH as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
24.1 | - |
balsalazide | mutant enzyme Y131F, using NADPH as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
25.2 | - |
NADH | mutant enzyme Y131F, using methyl red as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
33.6 | - |
NADPH | wild type enzyme, using methyl red as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
35.5 | - |
NADPH | mutant enzyme Y131F, using methyl red as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa | |
36.8 | - |
balsalazide | wild type enzyme, using NADPH as cosubstrate, in 20 mM TrisHCl pH 8.0, temperature not specified in the publication | Pseudomonas aeruginosa |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FMN | dependent on | Pseudomonas aeruginosa | |
NADPH | - |
Pseudomonas aeruginosa |