Cloned (Comment) | Organism |
---|---|
gene OCC_11152, sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain Rosetta (DE3) | Thermococcus litoralis |
Protein Variants | Comment | Organism |
---|---|---|
C194A | site-directed mutagenesis, inactive mutant | Thermococcus litoralis |
C83A | site-directed mutagenesis, inactive mutant | Thermococcus litoralis |
C83A/C194A | site-directed mutagenesis, inactive mutant | Thermococcus litoralis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5,5'-dithiobis(2-nitrobenzoic acid) | 35% inhibition at 1 mM | Thermococcus litoralis | |
CoSO4 | 36% inhibition at 1 mM | Thermococcus litoralis | |
CuSO4 | 15% inhibition at 1 mM | Thermococcus litoralis | |
EDTA | 7% inhibition at 1 mM | Thermococcus litoralis | |
FeSO4 | 43% inhibition at 1 mM | Thermococcus litoralis | |
iodoacetic acid | complete inhibition at 1 mM | Thermococcus litoralis | |
MgSO4 | 48% inhibition at 1 mM | Thermococcus litoralis | |
MnSO4 | 18% inhibition at 1 mM | Thermococcus litoralis | |
N-ethylmaleimide | 62% inhibition at 1 mM | Thermococcus litoralis | |
NiSO4 | 31% inhibition at 1 mM | Thermococcus litoralis | |
p-chloromercuribenzoic acid | 36% inhibition at 1 mM | Thermococcus litoralis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | the activation energy of Tl-AspR is calculated to be 51.8 kJ/mol by Arrhenius plots | Thermococcus litoralis | |
30.2 | - |
L-aspartate | recombinant enzyme, pH 8.0, 90°C | Thermococcus litoralis | |
136 | - |
D-Aspartate | recombinant enzyme, pH 8.0, 90°C | Thermococcus litoralis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
CaCl2 | activates to 120% activity at 1 mM | Thermococcus litoralis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
56000 | - |
recombinant enzyme, gel filtration | Thermococcus litoralis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-aspartate | Thermococcus litoralis | - |
D-aspartate | - |
r | |
L-aspartate | Thermococcus litoralis ATCC 51850 / DSM 5473 / JCM 8560 / NS-C | - |
D-aspartate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermococcus litoralis | H3ZNI2 | isolated from a hot spring of a shallow sea in Naples, Italy | - |
Thermococcus litoralis ATCC 51850 / DSM 5473 / JCM 8560 / NS-C | H3ZNI2 | isolated from a hot spring of a shallow sea in Naples, Italy | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain Rosetta (DE3) by heat treatment of cell-free extract at 70°C for 30 min, centrifugation, dialysis, and anion exchange chromatography, dialysis, hydrophobic interaction chromatography, and another different anion exchange chromatography, followed by dialysis | Thermococcus litoralis |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | the organism is able to grow at 55-98°C in pH 4.0-8.0, optimally at 88°C and pH 6.0 | Thermococcus litoralis | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1550 | - |
crude recombinant, pH 8.0, 90°C | Thermococcus litoralis |
1590 | - |
purified recombinant enzyme after complete purification procedure, pH 8.0, 90°C | Thermococcus litoralis |
3870 | - |
partially purified recombinant enzyme, after the first anion exchange chromatographic step, pH 8.0, 90°C | Thermococcus litoralis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-aspartate | D-Asp is the preferred substrate | Thermococcus litoralis | L-aspartate | - |
r | |
D-aspartate | D-Asp is the preferred substrate | Thermococcus litoralis ATCC 51850 / DSM 5473 / JCM 8560 / NS-C | L-aspartate | - |
r | |
L-aspartate | - |
Thermococcus litoralis | D-aspartate | - |
r | |
L-aspartate | 56.4% of the activity in the reverse reaction direction | Thermococcus litoralis | D-aspartate | - |
r | |
L-aspartate | - |
Thermococcus litoralis ATCC 51850 / DSM 5473 / JCM 8560 / NS-C | D-aspartate | - |
r | |
L-aspartate | 56.4% of the activity in the reverse reaction direction | Thermococcus litoralis ATCC 51850 / DSM 5473 / JCM 8560 / NS-C | D-aspartate | - |
r | |
additional information | the enzyme also acts on L-cysteic acid and L-cysteine sulfinic acid (with below 30% activity compared to D-aspartate) in addition to D- and L-aspartic acids. No activity with D-glutamate, D-asparagine, D-glutamine, D-alanine, D-leucine, D-valine, D-proline, D-serine, D-threonine, and D-arginine | Thermococcus litoralis | ? | - |
? | |
additional information | the enzyme also acts on L-cysteic acid and L-cysteine sulfinic acid (with below 30% activity compared to D-aspartate) in addition to D- and L-aspartic acids. No activity with D-glutamate, D-asparagine, D-glutamine, D-alanine, D-leucine, D-valine, D-proline, D-serine, D-threonine, and D-arginine | Thermococcus litoralis ATCC 51850 / DSM 5473 / JCM 8560 / NS-C | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 25000-30000, recombinant enzyme, SDS-PAGE | Thermococcus litoralis |
Synonyms | Comment | Organism |
---|---|---|
OCC_11152 | - |
Thermococcus litoralis |
PLP-independent AspR | - |
Thermococcus litoralis |
pyridoxal 5'-phosphate independent aspartate racemase | - |
Thermococcus litoralis |
Tl-AspR | - |
Thermococcus litoralis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
95 | - |
the activity of Tl-AspR increases gradually and shows a maximum activity at 95°C | Thermococcus litoralis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
90 | - |
purified enzyme, approximately 50% of initial activity remains after incubation for 11 h | Thermococcus litoralis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
118000 | - |
L-aspartate | recombinant enzyme, pH 8.0, 90°C | Thermococcus litoralis | |
610000 | - |
D-Aspartate | recombinant enzyme, pH 8.0, 90°C | Thermococcus litoralis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Thermococcus litoralis |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
6 | 8 | purified enzyme, min. 50% activity remaining within this range | Thermococcus litoralis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | the enzyme does not require pyridoxal 5'-phosphate as a coenzyme | Thermococcus litoralis |
General Information | Comment | Organism |
---|---|---|
evolution | the essential cysteine residues are conserved as Cys83 and Cys194 | Thermococcus litoralis |
additional information | amino acid analysis of cell-free extract, overview | Thermococcus litoralis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3907.3 | - |
L-aspartate | recombinant enzyme, pH 8.0, 90°C | Thermococcus litoralis | |
4485.3 | - |
D-Aspartate | recombinant enzyme, pH 8.0, 90°C | Thermococcus litoralis |