Any feedback?
Literature summary for 3.5.2.2 extracted from
- Inhibition of a putative dihydropyrimidinase from Pseudomonas aeruginosa PAO1 by flavonoids and substrates of cyclic amidohydrolases (2015), PLoS ONE, 10, e0127634 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| recombinant expression of wild-type and mutant His-tagged enzymes | Pseudomonas aeruginosa |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D316A | site-directed mutagenesis, inactive mutant | Pseudomonas aeruginosa |
| H183A | site-directed mutagenesis, inactive mutant | Pseudomonas aeruginosa |
| H239A | site-directed mutagenesis, inactive mutant | Pseudomonas aeruginosa |
| H59A | site-directed mutagenesis, inactive mutant | Pseudomonas aeruginosa |
| H61A | site-directed mutagenesis, inactive mutant | Pseudomonas aeruginosa |
| K150A | site-directed mutagenesis, inactive mutant | Pseudomonas aeruginosa |
| N337A | site-directed mutagenesis, the mutant enzyme is active, but its activity is about 20fold less than that of the wild-type dihydropyrimidinase | Pseudomonas aeruginosa |
| S289A | site-directed mutagenesis, inactive mutant | Pseudomonas aeruginosa |
| Y155A | site-directed mutagenesis, inactive mutant | Pseudomonas aeruginosa |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 3-Amino-1,2,4-triazole | - |
Pseudomonas aeruginosa |  |
| 5-hydantoin acetic acid | - |
Pseudomonas aeruginosa | |
| Acetohydroxamate | - |
Pseudomonas aeruginosa | |
| allantoin | slight inhibition | Pseudomonas aeruginosa | |
| dihydromyricetin | competitive inhibitor, dihydromyricetin is docked in the active site pocket of dihydropyrimidinase, binding kinetics. Dihydromyricetin forms a stable complex with dihydropyrimidinase with the Kd value of 0.0226 mM | Pseudomonas aeruginosa | |
| dihydroorotate | slight inhibition | Pseudomonas aeruginosa | |
| galangin | - |
Pseudomonas aeruginosa | |
| kaempferol | - |
Pseudomonas aeruginosa | |
| additional information | myricetin and dihydromyricetin are computationally docked into the three-dimensional model of dihydropyrimidinase by using PatchDock. The dihydrouracil-complexed structure model of Pseudomonas aeruginosa dihydropyrimidinase is directly constructed by superimposing the crystal structure of the dihydrouracil-yeast dihydropyrimidinase complex (PDB ID 2FVK). Structure-inhibition relationship with dihydropyrimidinase, overview | Pseudomonas aeruginosa | |
| myricetin | - |
Pseudomonas aeruginosa | |
| Myricitrin | - |
Pseudomonas aeruginosa | |
| Orotic acid | - |
Pseudomonas aeruginosa | |
| quercetin | - |
Pseudomonas aeruginosa | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics | Pseudomonas aeruginosa |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | activates at 1 mM | Pseudomonas aeruginosa | |
| Mn2+ | activates at 1 mM | Pseudomonas aeruginosa | |
| additional information | no effects on enzyme activity by Cd2+, Ni2+, Mg2+, and Ca2+ at 1 mM | Pseudomonas aeruginosa | |
| Zn2+ | activates at 1 mM | Pseudomonas aeruginosa | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 5,6-dihydrouracil + H2O | Pseudomonas aeruginosa | - |
3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | Pseudomonas aeruginosa ATCC 15692 | - |
3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | Pseudomonas aeruginosa 1C | - |
3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | Pseudomonas aeruginosa PRS 101 | - |
3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | Pseudomonas aeruginosa DSM 22644 | - |
3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | Pseudomonas aeruginosa CIP 104116 | - |
3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | Pseudomonas aeruginosa LMG 12228 | - |
3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | Pseudomonas aeruginosa JCM 14847 | - |
3-ureidopropanoate | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas aeruginosa | Q9I676 | - |
- |
| Pseudomonas aeruginosa 1C | Q9I676 | - |
- |
| Pseudomonas aeruginosa ATCC 15692 | Q9I676 | - |
- |
| Pseudomonas aeruginosa CIP 104116 | Q9I676 | - |
- |
| Pseudomonas aeruginosa DSM 22644 | Q9I676 | - |
- |
| Pseudomonas aeruginosa JCM 14847 | Q9I676 | - |
- |
| Pseudomonas aeruginosa LMG 12228 | Q9I676 | - |
- |
| Pseudomonas aeruginosa PRS 101 | Q9I676 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant wild-type and mutant His-tagged enzymes by nickel affinity chromatography and dialysis to over 97% purity | Pseudomonas aeruginosa |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.74 | - |
substrate 5,6-dihydrouracil, pH 8.0, 25°C, purified recombinant enzyme in presence of 1 mM Cd2+ | Pseudomonas aeruginosa |
| 0.82 | - |
substrate 5,6-dihydrouracil, pH 8.0, 25°C, purified recombinant enzyme in absence of metal ion | Pseudomonas aeruginosa |
| 0.82 | - |
substrate 5,6-dihydrouracil, pH 8.0, 25°C, purified recombinant enzyme in presence of 1 mM Ca2+ | Pseudomonas aeruginosa |
| 0.82 | - |
substrate 5,6-dihydrouracil, pH 8.0, 25°C, purified recombinant enzyme in presence of 1 mM Mg2+ | Pseudomonas aeruginosa |
| 0.91 | - |
substrate 5,6-dihydrouracil, pH 8.0, 25°C, purified recombinant enzyme in presence of 1 mM Ni2+ | Pseudomonas aeruginosa |
| 1.72 | - |
substrate 5,6-dihydrouracil, pH 8.0, 25°C, purified recombinant enzyme in presence of 1 mM Mn2+ | Pseudomonas aeruginosa |
| 2.9 | - |
substrate 5,6-dihydrouracil, pH 8.0, 25°C, purified recombinant enzyme in presence of 1 mM Zn2+ | Pseudomonas aeruginosa |
| 5.8 | 5.9 | substrate 5,6-dihydrouracil, pH 8.0, 25°C, purified recombinant enzyme in presence of 1 mM Co2+ | Pseudomonas aeruginosa |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 5,6-dihydrouracil + H2O | - |
Pseudomonas aeruginosa | 3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | - |
Pseudomonas aeruginosa ATCC 15692 | 3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | - |
Pseudomonas aeruginosa 1C | 3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | - |
Pseudomonas aeruginosa PRS 101 | 3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | - |
Pseudomonas aeruginosa DSM 22644 | 3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | - |
Pseudomonas aeruginosa CIP 104116 | 3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | - |
Pseudomonas aeruginosa LMG 12228 | 3-ureidopropanoate | - |
r | |
| 5,6-dihydrouracil + H2O | - |
Pseudomonas aeruginosa JCM 14847 | 3-ureidopropanoate | - |
r | |
| 5-propylhydantoin + H2O | low activity | Pseudomonas aeruginosa | N-carbamoyl-D-valine | - |
? | |
| 5-propylhydantoin + H2O | low activity | Pseudomonas aeruginosa ATCC 15692 | N-carbamoyl-D-valine | - |
? | |
| 5-propylhydantoin + H2O | low activity | Pseudomonas aeruginosa 1C | N-carbamoyl-D-valine | - |
? | |
| 5-propylhydantoin + H2O | low activity | Pseudomonas aeruginosa PRS 101 | N-carbamoyl-D-valine | - |
? | |
| 5-propylhydantoin + H2O | low activity | Pseudomonas aeruginosa DSM 22644 | N-carbamoyl-D-valine | - |
? | |
| 5-propylhydantoin + H2O | low activity | Pseudomonas aeruginosa CIP 104116 | N-carbamoyl-D-valine | - |
? | |
| 5-propylhydantoin + H2O | low activity | Pseudomonas aeruginosa LMG 12228 | N-carbamoyl-D-valine | - |
? | |
| 5-propylhydantoin + H2O | low activity | Pseudomonas aeruginosa JCM 14847 | N-carbamoyl-D-valine | - |
? | |
| additional information | the catalytic efficiency of the enzyme toward dihydrouracil is higher than that toward phthalimide (fourfold) and 5-propylhydantoin (100fold). Therefore, this bacterial enzyme is suitably identified as a dihydropyrimidinase, not a hydantoinase. Allantoin and dihydroorotate are no substrates | Pseudomonas aeruginosa | ? | - |
- |
|
| additional information | the catalytic efficiency of the enzyme toward dihydrouracil is higher than that toward phthalimide (fourfold) and 5-propylhydantoin (100fold). Therefore, this bacterial enzyme is suitably identified as a dihydropyrimidinase, not a hydantoinase. Allantoin and dihydroorotate are no substrates | Pseudomonas aeruginosa ATCC 15692 | ? | - |
- |
|
| additional information | the catalytic efficiency of the enzyme toward dihydrouracil is higher than that toward phthalimide (fourfold) and 5-propylhydantoin (100fold). Therefore, this bacterial enzyme is suitably identified as a dihydropyrimidinase, not a hydantoinase. Allantoin and dihydroorotate are no substrates | Pseudomonas aeruginosa 1C | ? | - |
- |
|
| additional information | the catalytic efficiency of the enzyme toward dihydrouracil is higher than that toward phthalimide (fourfold) and 5-propylhydantoin (100fold). Therefore, this bacterial enzyme is suitably identified as a dihydropyrimidinase, not a hydantoinase. Allantoin and dihydroorotate are no substrates | Pseudomonas aeruginosa PRS 101 | ? | - |
- |
|
| additional information | the catalytic efficiency of the enzyme toward dihydrouracil is higher than that toward phthalimide (fourfold) and 5-propylhydantoin (100fold). Therefore, this bacterial enzyme is suitably identified as a dihydropyrimidinase, not a hydantoinase. Allantoin and dihydroorotate are no substrates | Pseudomonas aeruginosa DSM 22644 | ? | - |
- |
|
| additional information | the catalytic efficiency of the enzyme toward dihydrouracil is higher than that toward phthalimide (fourfold) and 5-propylhydantoin (100fold). Therefore, this bacterial enzyme is suitably identified as a dihydropyrimidinase, not a hydantoinase. Allantoin and dihydroorotate are no substrates | Pseudomonas aeruginosa CIP 104116 | ? | - |
- |
|
| additional information | the catalytic efficiency of the enzyme toward dihydrouracil is higher than that toward phthalimide (fourfold) and 5-propylhydantoin (100fold). Therefore, this bacterial enzyme is suitably identified as a dihydropyrimidinase, not a hydantoinase. Allantoin and dihydroorotate are no substrates | Pseudomonas aeruginosa LMG 12228 | ? | - |
- |
|
| additional information | the catalytic efficiency of the enzyme toward dihydrouracil is higher than that toward phthalimide (fourfold) and 5-propylhydantoin (100fold). Therefore, this bacterial enzyme is suitably identified as a dihydropyrimidinase, not a hydantoinase. Allantoin and dihydroorotate are no substrates | Pseudomonas aeruginosa JCM 14847 | ? | - |
- |
|
| phthalimide + H2O | - |
Pseudomonas aeruginosa | ? | - |
? | |
| phthalimide + H2O | - |
Pseudomonas aeruginosa ATCC 15692 | ? | - |
? | |
| phthalimide + H2O | - |
Pseudomonas aeruginosa 1C | ? | - |
? | |
| phthalimide + H2O | - |
Pseudomonas aeruginosa PRS 101 | ? | - |
? | |
| phthalimide + H2O | - |
Pseudomonas aeruginosa DSM 22644 | ? | - |
? | |
| phthalimide + H2O | - |
Pseudomonas aeruginosa CIP 104116 | ? | - |
? | |
| phthalimide + H2O | - |
Pseudomonas aeruginosa LMG 12228 | ? | - |
? | |
| phthalimide + H2O | - |
Pseudomonas aeruginosa JCM 14847 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| More | enzyme structure homology modeling | Pseudomonas aeruginosa |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| dht | - |
Pseudomonas aeruginosa |
| PA0441 | - |
Pseudomonas aeruginosa |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Pseudomonas aeruginosa |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
assay at | Pseudomonas aeruginosa |
IC50 Value
| IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
|---|---|---|---|---|---|
| 0.04 | - |
versus substrate 5-propylhydantoin, pH 8.0, 25°C | Pseudomonas aeruginosa | dihydromyricetin | |
| 0.048 | - |
versus substrate dihydrouracil, pH 8.0, 25°C | Pseudomonas aeruginosa | dihydromyricetin | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | dihydropyrimidinase is a member of the cyclic amidohydrolase family, which also includes allantoinase, dihydroorotase, hydantoinase, and imidase. These metalloenzymes possess very similar active sites and may use a similar mechanism for catalysis | Pseudomonas aeruginosa |
| additional information | enzyme structure homology modeling, determination and analysis of active site residues of dihydropyrimidinase, overview | Pseudomonas aeruginosa |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
