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Literature summary for 1.7.1.6 extracted from

  • Wang, C.; Laurieri, N.; Abuhammad, A.; Lowe, E.; Westwood, I.; Ryan, A.; Sim, E.
    Role of tyrosine 131 in the active site of paAzoR1, an azoreductase with specificity for the inflammatory bowel disease prodrug balsalazide (2009), Acta Crystallogr. Sect. F, 66, 2-7.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.7.1.6

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) pLysS cells Pseudomonas aeruginosa

Crystallization (Commentary)

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

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 [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0445
-
 Methyl red mutant enzyme Y131F, using NADPH as cosubstrate, in 20 mM Tris–HCl 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 Tris–HCl pH 8.0, temperature not specified in the publication Pseudomonas aeruginosa
0.0986
-
 balsalazide wild type enzyme, using NADPH as cosubstrate, in 20 mM Tris–HCl pH 8.0, temperature not specified in the publication Pseudomonas aeruginosa
0.1029
-
 balsalazide mutant enzyme Y131F, using NADPH as cosubstrate, in 20 mM Tris–HCl 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 Tris–HCl 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 Tris–HCl 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 Tris–HCl 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 Tris–HCl pH 8.0, temperature not specified in the publication Pseudomonas aeruginosa

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa Q9I5F3
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Pseudomonas aeruginosa

Substrates and Products (Substrate)

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

Synonyms Comment Organism
AzoR1
-
 Pseudomonas aeruginosa
azoreductase 1
-
 Pseudomonas aeruginosa

Temperature Stability [°C]

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 [1/s]

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 Tris–HCl 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 Tris–HCl 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 Tris–HCl pH 8.0, temperature not specified in the publication Pseudomonas aeruginosa
24.1
-
 balsalazide mutant enzyme Y131F, using NADPH as cosubstrate, in 20 mM Tris–HCl 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 Tris–HCl 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 Tris–HCl 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 Tris–HCl pH 8.0, temperature not specified in the publication Pseudomonas aeruginosa
36.8
-
 balsalazide wild type enzyme, using NADPH as cosubstrate, in 20 mM Tris–HCl pH 8.0, temperature not specified in the publication Pseudomonas aeruginosa

Cofactor

Cofactor Comment Organism Structure
FMN dependent on Pseudomonas aeruginosa
NADPH
-
 Pseudomonas aeruginosa