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Literature summary extracted from
- Biochemical characterization of an extremely thermostable l-asparaginase from Thermococcus gammatolerans EJ3 (2014), J. Mol. Catal. B, 109, 122-129.
No PubMed abstract available
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.5.1.1 | expressed in Escherichia coli BL21(DE3) cells | Thermococcus gammatolerans |
General Stability
| EC Number | General Stability | Organism |
|---|---|---|
| 3.5.1.1 | after incubation with various concentration of urea for 6 h, the purified enzyme samples almost do not exhibit any change in enzyme activity even by incubation with 8 M of urea | Thermococcus gammatolerans |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.5.1.1 | 10 | - |
L-asparagine | at pH 8.5 and 85°C | Thermococcus gammatolerans |  |
| 3.5.1.1 | 24.1 | - |
D-asparagine | at pH 8.5 and 85°C | Thermococcus gammatolerans | |
| 3.5.1.1 | 39.5 | - |
L-glutamine | at pH 8.5 and 85°C | Thermococcus gammatolerans | |
| 3.5.1.1 | 77.6 | - |
D-glutamine | at pH 8.5 and 85°C | Thermococcus gammatolerans | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.5.1.1 | Mg2+ | marginal increase in enzyme activity in the presence of 1 mM Mg2+ (less than 15%) | Thermococcus gammatolerans | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 3.5.1.1 | 35000 | - |
2 * 35000, SDS-PAGE | Thermococcus gammatolerans |
| 3.5.1.1 | 36500 | - |
2 * 36500, predicted from amino acid sequence | Thermococcus gammatolerans |
| 3.5.1.1 | 74000 | - |
gel filtration | Thermococcus gammatolerans |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.5.1.1 | L-asparagine + H2O | Thermococcus gammatolerans | best substrate | L-aspartate + NH3 | - |
? | |
| 3.5.1.1 | L-asparagine + H2O | Thermococcus gammatolerans EJ3 | best substrate | L-aspartate + NH3 | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.5.1.1 | Thermococcus gammatolerans | C5A6Q4 | - |
- |
| 3.5.1.1 | Thermococcus gammatolerans EJ3 | C5A6Q4 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.5.1.1 | Ni-Sepharose column chromatography | Thermococcus gammatolerans |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 3.5.1.1 | 0.925 | - |
with D-glutamine as substrate, at pH 8.5 and 85°C | Thermococcus gammatolerans |
| 3.5.1.1 | 2.926 | - |
with L-glutamine as substrate, at pH 8.5 and 85°C | Thermococcus gammatolerans |
| 3.5.1.1 | 3.753 | - |
with D-asparagine as substrate, at pH 8.5 and 85°C | Thermococcus gammatolerans |
| 3.5.1.1 | 7.622 | - |
with L-asparagine as substrate, at pH 8.5 and 85°C | Thermococcus gammatolerans |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.5.1.1 | D-asparagine + H2O | - |
Thermococcus gammatolerans | D-aspartate + NH3 | - |
? | |
| 3.5.1.1 | D-asparagine + H2O | - |
Thermococcus gammatolerans EJ3 | D-aspartate + NH3 | - |
? | |
| 3.5.1.1 | D-glutamine + H2O | - |
Thermococcus gammatolerans | D-glutamate + NH3 | - |
? | |
| 3.5.1.1 | D-glutamine + H2O | - |
Thermococcus gammatolerans EJ3 | D-glutamate + NH3 | - |
? | |
| 3.5.1.1 | L-asparagine + H2O | best substrate | Thermococcus gammatolerans | L-aspartate + NH3 | - |
? | |
| 3.5.1.1 | L-asparagine + H2O | best substrate | Thermococcus gammatolerans EJ3 | L-aspartate + NH3 | - |
? | |
| 3.5.1.1 | L-glutamine + H2O | - |
Thermococcus gammatolerans | L-glutamate + NH3 | - |
? | |
| 3.5.1.1 | L-glutamine + H2O | - |
Thermococcus gammatolerans EJ3 | L-glutamate + NH3 | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 3.5.1.1 | homodimer | 2 * 35000, SDS-PAGE | Thermococcus gammatolerans |
| 3.5.1.1 | homodimer | 2 * 36500, predicted from amino acid sequence | Thermococcus gammatolerans |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.5.1.1 | L-asparaginase | - |
Thermococcus gammatolerans |
| 3.5.1.1 | L-asparagine amidohydrolase | - |
Thermococcus gammatolerans |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.5.1.1 | 85 | - |
- |
Thermococcus gammatolerans |
Temperature Range [°C]
| EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.5.1.1 | 70 | 95 | the enzyme shows more than 75% of relative activity in the range of 70-95°C | Thermococcus gammatolerans |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.5.1.1 | 70 | 95 | the enzyme retains more than 60% of its initial activityafter 30 min at 7095°C, and retains 66%, 60%, 54%, 53%, and 43% of its initial activity after 2 h of exposure at 70, 80, 85, 90, and 95°C, respectively | Thermococcus gammatolerans |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.5.1.1 | 1362 | - |
D-glutamine | at pH 8.5 and 85°C | Thermococcus gammatolerans | |
| 3.5.1.1 | 2048 | - |
L-glutamine | at pH 8.5 and 85°C | Thermococcus gammatolerans | |
| 3.5.1.1 | 3378 | - |
D-asparagine | at pH 8.5 and 85°C | Thermococcus gammatolerans | |
| 3.5.1.1 | 5721 | - |
L-asparagine | at pH 8.5 and 85°C | Thermococcus gammatolerans | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.5.1.1 | 8.5 | - |
- |
Thermococcus gammatolerans |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.5.1.1 | 8 | 10 | the enzyme shows more than 85% of relative activity at the range from pH 8.0-10.0 | Thermococcus gammatolerans |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.5.1.1 | 17.6 | - |
D-glutamine | at pH 8.5 and 85°C | Thermococcus gammatolerans | |
| 3.5.1.1 | 51.9 | - |
L-glutamine | at pH 8.5 and 85°C | Thermococcus gammatolerans | |
| 3.5.1.1 | 140.2 | - |
D-asparagine | at pH 8.5 and 85°C | Thermococcus gammatolerans | |
| 3.5.1.1 | 572.1 | - |
L-asparagine | at pH 8.5 and 85°C | Thermococcus gammatolerans | |
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