Any feedback?
Literature summary for 4.3.1.16 extracted from
- Gene cloning and expression of pyridoxal 5-phosphate-dependent L-threo-3-hydroxyaspartate dehydratase from Pseudomonas sp. T62, and characterization of the recombinant enzyme (2009), J. Biochem., 145, 661-668.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| ADP | addition of 10 mM increases enzyme activity to 106% of the control | Pseudomonas sp. T62 |  |
| AMP | addition of 10 mM increases enzyme activity to 144% of the control | Pseudomonas sp. T62 | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| the gene encoding L-THA DH is cloned and expressed in Escherichia coli | Pseudomonas sp. T62 |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| K53A | mutant enzyme is produced in Escherichia coli JM109 cells | Pseudomonas sp. T62 |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| ATP | addition of 10 mM decreases enzyme activity to 89% of the control | Pseudomonas sp. T62 | |
| CoCl2 | 1 mM causes inhibition enzyme activity (81% relative activity) | Pseudomonas sp. T62 | |
| CuCl2 | 1 mM causes inhibition enzyme activity (92% relative activity) | Pseudomonas sp. T62 | |
| D-serine | 5 mM decrease the enzyme activity to 27% | Pseudomonas sp. T62 | |
| EDTA | strong inhibition by EDTA (69% inhibition), suggesting that metal ions are involved in the enzyme reaction | Pseudomonas sp. T62 | |
| GDP | addition of 10 mM decreases enzyme activity to 73% of the control | Pseudomonas sp. T62 | |
| L-erythro-3-hydroxyaspartate | 5 mM, decrease the enzyme activity to 15% | Pseudomonas sp. T62 | |
| SnCl2 | 1 mM causes inhibition enzyme activity (70% relative activity) | Pseudomonas sp. T62 | |
| ZnCl2 | 1 mM causes inhibition enzyme activity (56% relative activity) | Pseudomonas sp. T62 | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | none of the other 3-hydroxy amino acids tested (DL-erythro- and D-threo-3-hydroxyaspartate, D-threonine, L-threonine, DL-allo-threonine, D-serine, L-serine, and DL-phenylserine) is a substrate for this enzyme, either at 5 mM or 50 mM, as in the case of the enzyme from the original strain | Pseudomonas sp. T62 | |
| 0.54 | - |
L-threo-3-hydroxyaspartate | exhibits dehydratase activity specific only to L-threo-3-hydroxyaspartate | Pseudomonas sp. T62 | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | By addition of 1 mM to the reaction mixture, relative enzyme activity increased to 159% of control activity. | Pseudomonas sp. T62 | |
| Mg2+ | By addition of 1 mM to the reaction mixture, relative enzyme activity increased to 196% of control activity. | Pseudomonas sp. T62 | |
| Mn2+ | By addition of 1 mM to the reaction mixture, relative enzyme activity increased to 151% of control activity. | Pseudomonas sp. T62 | |
| Mn2+ | effect of divalent cations are measured using the enzyme as control dialyzed against the buffer minus MnCl2, showing a decrease of the specific activity of the enzyme to approximately 36% of the initial activity after overnight dialysis | Pseudomonas sp. T62 | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 36000 | - |
determined by using a MALDI-TOF mass spectrometer, measured value is lower than that determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (40 kDa), and that of the enzyme purified from the original strain (approximately 39 kDa by SDS-PAGE). The discrepancy may be due to the surface charge of the protein or some unknown factors such as the conformation of the enzyme | Pseudomonas sp. T62 |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-threo-3-hydroxyaspartate | Pseudomonas sp. T62 | - |
oxaloacetate + NH3 | - |
r | |
| additional information | Pseudomonas sp. T62 | none of the other 3-hydroxy amino acids tested (DL-erythro- and D-threo-3-hydroxyaspartate, D-threonine, L-threonine, DL-allo-threonine, D-serine, L-serine, and DL-phenylserine) is a substrate for this enzyme, either at 5 mM or 50 mM, as in the case of the enzyme from the original strain | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas sp. T62 | A4F2N8 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| after centrifugation of cultivated cell culture with the recombinant enzyme, resulting supernatant is applied to a Ni-sepharose column connected to a fast protein liquid chromatography system. The column is equilibrated with buffer supplemented with 20 mM imidazole. The enzyme is eluted with imidazole gradient. | Pseudomonas sp. T62 |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
exhibits no detectable serine/aspartate racemase activity | Pseudomonas sp. T62 |
| additional information | - |
K53A mutant enzyme shows no detectable activity | Pseudomonas sp. T62 |
| 39 | - |
recombinant enzyme | Pseudomonas sp. T62 |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-threo-3-hydroxyaspartate | - |
Pseudomonas sp. T62 | oxaloacetate + NH3 | - |
r | |
| additional information | none of the other 3-hydroxy amino acids tested (DL-erythro- and D-threo-3-hydroxyaspartate, D-threonine, L-threonine, DL-allo-threonine, D-serine, L-serine, and DL-phenylserine) is a substrate for this enzyme, either at 5 mM or 50 mM, as in the case of the enzyme from the original strain | Pseudomonas sp. T62 | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| L-THA DH | - |
Pseudomonas sp. T62 |
| L-threo-3-hydroxyaspartate dehydratase | pyridoxal 5'-phosphate-dependent | Pseudomonas sp. T62 |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 35 | - |
- |
Pseudomonas sp. T62 |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 9 | - |
- |
Pseudomonas sp. T62 |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| pyridoxal 5'-phosphate | - |
Pseudomonas sp. T62 | |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | DL-aspartate, L-serine, DL-threonine, DL-allo-threonine, DL-phenylserine, or malonic acid did not cause significant inhibition of the enzyme reaction at 5 mM | Pseudomonas sp. T62 | |
| 0.2 | - |
L-erythro-3-hydroxyaspartate | strong competitive inhibition | Pseudomonas sp. T62 | |
| 22.8 | - |
D-serine | non-competitive inhibition | Pseudomonas sp. T62 | |
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
