Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Paenarthrobacter nitroguajacolicus |
sequence comparisons, overexpression of mutant N-terminally His6-tagged HODs in Escherichia coli strain M15 | Paenarthrobacter nitroguajacolicus |
Protein Variants | Comment | Organism |
---|---|---|
C69S | stable in its monomeric form | Paenarthrobacter nitroguajacolicus |
C69S | site-directed mutagenesis, the mutant has catalytic properties that are identical to those of wild-type HOD | Paenarthrobacter nitroguajacolicus |
E224A | decreased Km for O2 | Paenarthrobacter nitroguajacolicus |
E224A/C69S | site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme | Paenarthrobacter nitroguajacolicus |
F219Y | does not affect the kinetic parameters of the enzyme | Paenarthrobacter nitroguajacolicus |
F219Y/C69S | site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme | Paenarthrobacter nitroguajacolicus |
H102Q | Km for the heteroaromatic substrate is increased only 2.8fold, and kcat is reduced 2.3fold | Paenarthrobacter nitroguajacolicus |
H102Q/C69S | site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme | Paenarthrobacter nitroguajacolicus |
S101A | about 21fold increase in the Km for the organic substrate | Paenarthrobacter nitroguajacolicus |
S101A/C69S | site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme | Paenarthrobacter nitroguajacolicus |
S220N | does not drastically influence the kinetic parameters of the enzyme for the organic substrate, but it causes a 3.5fold decrease in Km for O2 and a 6.2fold decrease in kcat for O2 | Paenarthrobacter nitroguajacolicus |
S220N/C69S | site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme | Paenarthrobacter nitroguajacolicus |
Y196A | increased Km value | Paenarthrobacter nitroguajacolicus |
Y196A/C69S | site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme | Paenarthrobacter nitroguajacolicus |
Y196K | increased Km value | Paenarthrobacter nitroguajacolicus |
Y196K/C69S | site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme | Paenarthrobacter nitroguajacolicus |
Y196R | increased Km value | Paenarthrobacter nitroguajacolicus |
Y196R/C69S | site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme | Paenarthrobacter nitroguajacolicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | pH/pD dependence of apparent kcatO2/KmO2 of Hod C69S, overview | Paenarthrobacter nitroguajacolicus | |
0.0092 | - |
1H-3-Hydroxy-4-oxoquinaldine | wild type enzyme, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.0092 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant C69S | Paenarthrobacter nitroguajacolicus | |
0.0095 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant F219Y, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.0095 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant F219Y/C69S | Paenarthrobacter nitroguajacolicus | |
0.0154 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant E224A, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.0154 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant S220N, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.0154 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant E224A/C69S | Paenarthrobacter nitroguajacolicus | |
0.0154 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant S220N/C69S | Paenarthrobacter nitroguajacolicus | |
0.0257 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant H102Q, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.0257 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant H102Q/C69S | Paenarthrobacter nitroguajacolicus | |
0.0399 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant Y196K, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.0399 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant Y196K/C69S | Paenarthrobacter nitroguajacolicus | |
0.0836 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant Y196R, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.0836 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant Y196R/C69S | Paenarthrobacter nitroguajacolicus | |
0.0995 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant Y196A, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.0995 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant Y196A/C69S | Paenarthrobacter nitroguajacolicus | |
0.1907 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant S101A, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.1907 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant S101A/C69S | Paenarthrobacter nitroguajacolicus | |
0.325 | - |
O2 | mutant H102Q, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.325 | - |
O2 | pH 8.0, 30°C, recombinant mutant H102Q/C69S | Paenarthrobacter nitroguajacolicus | |
0.357 | - |
O2 | mutant S220N, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.357 | - |
O2 | pH 8.0, 30°C, recombinant mutant S220N/C69S | Paenarthrobacter nitroguajacolicus | |
0.711 | - |
O2 | mutant E224A, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.711 | - |
O2 | pH 8.0, 30°C, recombinant mutant E224A/C69S | Paenarthrobacter nitroguajacolicus | |
1.233 | - |
O2 | wild type enzyme, at 30°C | Paenarthrobacter nitroguajacolicus | |
1.233 | - |
O2 | pH 8.0, 30°C, recombinant mutant C69S | Paenarthrobacter nitroguajacolicus | |
1.27 | - |
O2 | mutant F219Y, at 30°C | Paenarthrobacter nitroguajacolicus | |
1.27 | - |
O2 | pH 8.0, 30°C, recombinant mutant F219Y/C69S | Paenarthrobacter nitroguajacolicus | |
1.37 | - |
O2 | mutant Y196R, at 30°C | Paenarthrobacter nitroguajacolicus | |
1.37 | - |
O2 | pH 8.0, 30°C, recombinant mutant Y196R/C69S | Paenarthrobacter nitroguajacolicus | |
1.382 | - |
O2 | mutant S101A, at 30°C | Paenarthrobacter nitroguajacolicus | |
1.382 | - |
O2 | pH 8.0, 30°C, recombinant mutant S101A/C69S | Paenarthrobacter nitroguajacolicus | |
1.6 | - |
O2 | mutant Y196K, at 30°C | Paenarthrobacter nitroguajacolicus | |
1.6 | - |
O2 | pH 8.0, 30°C, recombinant mutant Y196K/C69S | Paenarthrobacter nitroguajacolicus | |
1.646 | - |
O2 | mutant Y196A, at 30°C | Paenarthrobacter nitroguajacolicus | |
1.646 | - |
O2 | pH 8.0, 30°C, recombinant mutant Y196A/C69S | Paenarthrobacter nitroguajacolicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
1H-3-Hydroxy-4-oxoquinaldine + O2 | Paenarthrobacter nitroguajacolicus | - |
N-Acetylanthranilate + CO | - |
? | |
1H-3-Hydroxy-4-oxoquinaldine + O2 | Paenarthrobacter nitroguajacolicus Rü61a | - |
N-Acetylanthranilate + CO | - |
? | |
1H-3-Hydroxy-4-oxoquinaldine + O2 | Paenarthrobacter nitroguajacolicus R-61a | - |
N-Acetylanthranilate + CO | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paenarthrobacter nitroguajacolicus | Q7WSQ7 | - |
- |
Paenarthrobacter nitroguajacolicus | Q7WSQ7 | formerly Arthrobacter ilicis R-61a | - |
Paenarthrobacter nitroguajacolicus R-61a | Q7WSQ7 | formerly Arthrobacter ilicis R-61a | - |
Paenarthrobacter nitroguajacolicus Rü61a | Q7WSQ7 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Paenarthrobacter nitroguajacolicus |
recombinant mutant N-terminally His6-tagged HODs from Escherichia coli strain M15 by nickel affinity and anion exchange chromatography | Paenarthrobacter nitroguajacolicus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
3-hydroxy-2-methyl-1H-quinolin-4-one + O2 = N-acetylanthranilate + CO | catalytic mechanism, overview. H251 acts as a general base to abstract a proton from the organic substrate. Residue S101, which corresponds to the nucleophile of the catalytic triad of alpha/beta-hydrolases, presumably participates in binding the heteroaromatic substrate. H102 and residues located in the topological region of the triad's acidic residue appear to influence O2 binding and reactivity. Role of Y196 of Hod as hydrogen-bondforming residue to oxygen atoms or even proton donor to negative charges of catalytic intermediates | Paenarthrobacter nitroguajacolicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1H-3-Hydroxy-4-oxoquinaldine + O2 | - |
Paenarthrobacter nitroguajacolicus | N-Acetylanthranilate + CO | - |
? | |
1H-3-Hydroxy-4-oxoquinaldine + O2 | Hod has adapted active-site residues of the alpha/beta hydrolase fold for the dioxygenolytic reaction | Paenarthrobacter nitroguajacolicus | N-Acetylanthranilate + CO | - |
? | |
1H-3-Hydroxy-4-oxoquinaldine + O2 | - |
Paenarthrobacter nitroguajacolicus Rü61a | N-Acetylanthranilate + CO | - |
? | |
1H-3-Hydroxy-4-oxoquinaldine + O2 | Hod has adapted active-site residues of the alpha/beta hydrolase fold for the dioxygenolytic reaction | Paenarthrobacter nitroguajacolicus Rü61a | N-Acetylanthranilate + CO | - |
? | |
1H-3-Hydroxy-4-oxoquinaldine + O2 | - |
Paenarthrobacter nitroguajacolicus R-61a | N-Acetylanthranilate + CO | - |
? | |
1H-3-Hydroxy-4-oxoquinaldine + O2 | Hod has adapted active-site residues of the alpha/beta hydrolase fold for the dioxygenolytic reaction | Paenarthrobacter nitroguajacolicus R-61a | N-Acetylanthranilate + CO | - |
? | |
1H-3-hydroxy-4-oxoquinaldine + O2 | - |
Paenarthrobacter nitroguajacolicus | N-acetylanthranilic acid + CO | - |
? | |
1H-3-hydroxy-4-oxoquinaldine + O2 | - |
Paenarthrobacter nitroguajacolicus Rü61a | N-acetylanthranilic acid + CO | - |
? | |
1H-3-hydroxy-4-oxoquinaldine + O2 | - |
Paenarthrobacter nitroguajacolicus R-61a | N-acetylanthranilic acid + CO | - |
? | |
additional information | absence of viscosity effects and kinetic solvent isotope effects suggests that turnover of the ternary complex, rather than substrate binding, product release, or proton movements, involves the rate-determining step in the reaction catalyzed by Hod | Paenarthrobacter nitroguajacolicus | ? | - |
? | |
additional information | absence of viscosity effects and kinetic solvent isotope effects suggests that turnover of the ternary complex, rather than substrate binding, product release, or proton movements, involves the rate-determining step in the reaction catalyzed by Hod | Paenarthrobacter nitroguajacolicus Rü61a | ? | - |
? | |
additional information | absence of viscosity effects and kinetic solvent isotope effects suggests that turnover of the ternary complex, rather than substrate binding, product release, or proton movements, involves the rate-determining step in the reaction catalyzed by Hod | Paenarthrobacter nitroguajacolicus R-61a | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
1H-3-Hydroxy-4-oxoquinaldine 2,4-dioxygenase | - |
Paenarthrobacter nitroguajacolicus |
HOD | - |
Paenarthrobacter nitroguajacolicus |
HodC | - |
Paenarthrobacter nitroguajacolicus |
More | HOD belongs to the alpha/beta-hydrolase-fold superfamily of enzymes | Paenarthrobacter nitroguajacolicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Paenarthrobacter nitroguajacolicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.037 | 0.23 | O2 | mutant E224A, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.7 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant Y196K, at 30°C | Paenarthrobacter nitroguajacolicus | |
0.7 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant Y196K/C69S | Paenarthrobacter nitroguajacolicus | |
1 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant Y196R, at 30°C | Paenarthrobacter nitroguajacolicus | |
1 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant Y196R/C69S | Paenarthrobacter nitroguajacolicus | |
1.3 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant Y196A, at 30°C | Paenarthrobacter nitroguajacolicus | |
1.3 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant Y196A/C69S | Paenarthrobacter nitroguajacolicus | |
2.3 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant S101A, at 30°C | Paenarthrobacter nitroguajacolicus | |
2.3 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant S101A/C69S | Paenarthrobacter nitroguajacolicus | |
3.4 | - |
O2 | mutant Y196K, at 30°C | Paenarthrobacter nitroguajacolicus | |
3.4 | - |
O2 | mutant Y196R, at 30°C | Paenarthrobacter nitroguajacolicus | |
3.4 | - |
O2 | pH 8.0, 30°C, recombinant mutant Y196K/C69S | Paenarthrobacter nitroguajacolicus | |
3.4 | - |
O2 | pH 8.0, 30°C, recombinant mutant Y196R/C69S | Paenarthrobacter nitroguajacolicus | |
6.4 | - |
O2 | mutant Y196A, at 30°C | Paenarthrobacter nitroguajacolicus | |
6.4 | - |
O2 | pH 8.0, 30°C, recombinant mutant Y196A/C69S | Paenarthrobacter nitroguajacolicus | |
10 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant H102Q, at 30°C | Paenarthrobacter nitroguajacolicus | |
10 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant H102Q/C69S | Paenarthrobacter nitroguajacolicus | |
12.2 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant S220N, at 30°C | Paenarthrobacter nitroguajacolicus | |
12.2 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant S220N/C69S | Paenarthrobacter nitroguajacolicus | |
14.8 | - |
O2 | mutant H102Q, at 30°C | Paenarthrobacter nitroguajacolicus | |
14.8 | - |
O2 | pH 8.0, 30°C, recombinant mutant H102Q/C69S | Paenarthrobacter nitroguajacolicus | |
18.3 | - |
O2 | mutant S220N, at 30°C | Paenarthrobacter nitroguajacolicus | |
18.3 | - |
O2 | pH 8.0, 30°C, recombinant mutant S220N/C69S | Paenarthrobacter nitroguajacolicus | |
21 | - |
O2 | mutant F219Y, at 30°C | Paenarthrobacter nitroguajacolicus | |
22.8 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant F219Y, at 30°C | Paenarthrobacter nitroguajacolicus | |
22.8 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant F219Y/C69S | Paenarthrobacter nitroguajacolicus | |
22.9 | - |
1H-3-Hydroxy-4-oxoquinaldine | wild type enzyme, at 30°C | Paenarthrobacter nitroguajacolicus | |
22.9 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant C69S | Paenarthrobacter nitroguajacolicus | |
28.2 | - |
1H-3-Hydroxy-4-oxoquinaldine | mutant E224A, at 30°C | Paenarthrobacter nitroguajacolicus | |
28.2 | - |
1H-3-Hydroxy-4-oxoquinaldine | pH 8.0, 30°C, recombinant mutant E224A/C69S | Paenarthrobacter nitroguajacolicus | |
38.8 | - |
O2 | mutant S101A, at 30°C | Paenarthrobacter nitroguajacolicus | |
38.8 | - |
O2 | pH 8.0, 30°C, recombinant mutant S101A/C69S | Paenarthrobacter nitroguajacolicus | |
42 | - |
O2 | wild type enzyme, at 30°C | Paenarthrobacter nitroguajacolicus | |
79.1 | - |
O2 | mutant E224A, at 30°C | Paenarthrobacter nitroguajacolicus | |
79.1 | - |
O2 | pH 8.0, 30°C, recombinant mutant E224A/C69S | Paenarthrobacter nitroguajacolicus | |
110.9 | - |
O2 | mutant F219Y, at 30°C | Paenarthrobacter nitroguajacolicus | |
110.9 | - |
O2 | pH 8.0, 30°C, recombinant mutant F219Y/C69S | Paenarthrobacter nitroguajacolicus | |
113.7 | - |
O2 | wild type enzyme, at 30°C | Paenarthrobacter nitroguajacolicus | |
113.7 | - |
O2 | pH 8.0, 30°C, recombinant mutant C69S | Paenarthrobacter nitroguajacolicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Paenarthrobacter nitroguajacolicus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5.1 | 9.1 | pH profile, C59S Hod, overview | Paenarthrobacter nitroguajacolicus |
5.5 | 11 | - |
Paenarthrobacter nitroguajacolicus |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
5.5 | 11 | incubation for 5 min at pH between 5.5 and 11.0 does not result in significant loss of activity | Paenarthrobacter nitroguajacolicus |