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Literature summary for 3.5.1.3 extracted from
- Purification and properties of dicarboxylate omega-amidase from Bacillus subtilis 168 and Thermus aquaticus YT-1 (1972), Arch. Biochem. Biophys., 153, 95-104.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| additional information | no activation by ATP or Coenzyme A | Thermus aquaticus | |
| additional information | no activation by ATP or Coenzyme A | Bacillus subtilis |
General Stability
| General Stability | Organism |
|---|---|
| does not require mercaptoethanol or succinate for stability | Thermus aquaticus |
| does not require mercaptoethanol or succinate for stability | Bacillus subtilis |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| additional information | amide and acyl transfer activity: no inhibition by high concentrations of hydroxylamine; no inhibition by EDTA | Bacillus subtilis | |
| additional information | amide and acyl transfer activity: no inhibition by high concentrations of hydroxylamine; no inhibition by EDTA | Thermus aquaticus | |
| NH4+ | biosynthesis of omega-amidase is repressed by free ammonium ions | Bacillus subtilis |  |
| NH4+ | synthesis of omega-amidase is repressed by about 50% | Thermus aquaticus | |
| phosphate | phosphate buffer: slightly inhibitory | Bacillus subtilis | |
| phosphate | phosphate buffer: slightly inhibitory | Thermus aquaticus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.3 | - |
succinamate | - |
Bacillus subtilis | |
| 1.34 | - |
succinamate | - |
Thermus aquaticus | |
| 30 | - |
succinate | - |
Thermus aquaticus | |
| 50 | - |
hydroxylamine | with substrate succinamate | Bacillus subtilis | |
| 63 | - |
succinate | - |
Bacillus subtilis | |
| 100 | - |
hydroxylamine | with substrate succinamate | Thermus aquaticus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| additional information | no activation by Mg2+ | Thermus aquaticus | |
| additional information | no activation by Mg2+ | Bacillus subtilis | |
| additional information | no requirement for any metal ions for hydroxamate formation | Thermus aquaticus | |
| additional information | no requirement for any metal ions for hydroxamate formation | Bacillus subtilis | |
| additional information | no Mg2+ requirement for omega-amidase activity | Thermus aquaticus | |
| additional information | no Mg2+ requirement for omega-amidase activity | Bacillus subtilis |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 36000 | 38500 | gel filtration | Thermus aquaticus |
| 120000 | - |
gel filtration | Bacillus subtilis |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2-oxoglutaramate + H2O | Thermus aquaticus | alpha-ketoglutaramate | 2-oxoglutarate + NH3 | - |
? | |
| 2-oxoglutaramate + H2O | Bacillus subtilis | alpha-ketoglutaramate | 2-oxoglutarate + NH3 | - |
? | |
| 2-oxoglutaramate + H2O | Thermus aquaticus | physiological function of omega-amidase might be hydrolysis or transfer of amide group of alpha-ketoglutarate | 2-oxoglutarate + NH3 | - |
? | |
| 2-oxoglutaramate + H2O | Bacillus subtilis | physiological function of omega-amidase might be hydrolysis or transfer of amide group of alpha-ketoglutarate | 2-oxoglutarate + NH3 | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bacillus subtilis | - |
168, wild-type | - |
| Bacillus subtilis 168 | - |
168, wild-type | - |
| Thermus aquaticus | - |
YT-1 | - |
| Thermus aquaticus | - |
gram-negative extreme thermophilic bacterium | - |
| Thermus aquaticus YT-1 | - |
YT-1 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Thermus aquaticus |
- |
Bacillus subtilis |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 10.9 | - |
- |
Bacillus subtilis |
| 66 | - |
- |
Thermus aquaticus |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -20°C, protein concentration above 0.1 mg/ml, 0.05 M potassium phosphate, pH 7.2, 0.05 M KCl, 3 months, quite stable | Bacillus subtilis |
| -20°C, protein concentration above 0.1 mg/ml, 3 months, quite stable | Thermus aquaticus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2-oxoglutaramate + H2O | alpha-ketoglutaramate | Thermus aquaticus | 2-oxoglutarate + NH3 | - |
? | |
| 2-oxoglutaramate + H2O | alpha-ketoglutaramate | Bacillus subtilis | 2-oxoglutarate + NH3 | - |
? | |
| 2-oxoglutaramate + H2O | physiological function of omega-amidase might be hydrolysis or transfer of amide group of alpha-ketoglutarate | Thermus aquaticus | 2-oxoglutarate + NH3 | - |
? | |
| 2-oxoglutaramate + H2O | physiological function of omega-amidase might be hydrolysis or transfer of amide group of alpha-ketoglutarate | Bacillus subtilis | 2-oxoglutarate + NH3 | - |
? | |
| asparagine + H2O | - |
Bacillus subtilis | aspartate + NH3 | - |
? | |
| fumarate + H2O | - |
Thermus aquaticus | ? | - |
? | |
| fumarate + hydroxylamine | acyl transfer reaction | Thermus aquaticus | fumaryl hydroxamate + H2O | - |
? | |
| L-asparagine + hydroxylamine | amide transfer reaction | Bacillus subtilis | L-asparaginyl hydroxamate + H2O | - |
? | |
| malate + H2O | - |
Thermus aquaticus | ? | - |
? | |
| malate + H2O | - |
Bacillus subtilis | ? | - |
? | |
| malate + hydroxylamine | acyl transfer reaction | Thermus aquaticus | malyl hydroxamate + H2O | - |
? | |
| malate + hydroxylamine | acyl transfer reaction | Bacillus subtilis | malyl hydroxamate + H2O | - |
? | |
| additional information | substrate specificity | Thermus aquaticus | ? | - |
? | |
| additional information | substrate specificity | Bacillus subtilis | ? | - |
? | |
| additional information | no reaction with monocarboxylic acids | Thermus aquaticus | ? | - |
? | |
| additional information | no reaction with monocarboxylic acids | Bacillus subtilis | ? | - |
? | |
| additional information | catalyzes several enzymatic reactions: amide transfer, acyl transfer, and amide hydrolysis, are consistent with acyl enzyme type of intermediate | Thermus aquaticus | ? | - |
? | |
| additional information | catalyzes several enzymatic reactions: amide transfer, acyl transfer, and amide hydrolysis, are consistent with acyl enzyme type of intermediate | Bacillus subtilis | ? | - |
? | |
| additional information | glutamine, acetamide, acetate, propionate, aspartate and glutamate not or very poor substrates | Thermus aquaticus | ? | - |
? | |
| additional information | glutamine, acetamide, acetate, propionate, aspartate and glutamate not or very poor substrates | Bacillus subtilis | ? | - |
? | |
| additional information | catalyzes hydrolysis of dicarboxylic acid amides and the formation of hydroxamates from amides, amide transfer, of the corresponding acids, acyl transfer | Thermus aquaticus | ? | - |
? | |
| additional information | catalyzes hydrolysis of dicarboxylic acid amides and the formation of hydroxamates from amides, amide transfer, of the corresponding acids, acyl transfer | Bacillus subtilis | ? | - |
? | |
| additional information | substrate specificity | Bacillus subtilis 168 | ? | - |
? | |
| additional information | no reaction with monocarboxylic acids | Bacillus subtilis 168 | ? | - |
? | |
| additional information | catalyzes several enzymatic reactions: amide transfer, acyl transfer, and amide hydrolysis, are consistent with acyl enzyme type of intermediate | Bacillus subtilis 168 | ? | - |
? | |
| additional information | glutamine, acetamide, acetate, propionate, aspartate and glutamate not or very poor substrates | Bacillus subtilis 168 | ? | - |
? | |
| additional information | catalyzes hydrolysis of dicarboxylic acid amides and the formation of hydroxamates from amides, amide transfer, of the corresponding acids, acyl transfer | Bacillus subtilis 168 | ? | - |
? | |
| additional information | substrate specificity | Thermus aquaticus YT-1 | ? | - |
? | |
| additional information | no reaction with monocarboxylic acids | Thermus aquaticus YT-1 | ? | - |
? | |
| additional information | catalyzes several enzymatic reactions: amide transfer, acyl transfer, and amide hydrolysis, are consistent with acyl enzyme type of intermediate | Thermus aquaticus YT-1 | ? | - |
? | |
| additional information | glutamine, acetamide, acetate, propionate, aspartate and glutamate not or very poor substrates | Thermus aquaticus YT-1 | ? | - |
? | |
| additional information | catalyzes hydrolysis of dicarboxylic acid amides and the formation of hydroxamates from amides, amide transfer, of the corresponding acids, acyl transfer | Thermus aquaticus YT-1 | ? | - |
? | |
| succinamate + H2O | amidase activity, hydrolysis proceeds at a slower rate than formation of succinyl hydroxamate in the presence of hydroxylamine | Thermus aquaticus | succinic acid + NH3 | - |
? | |
| succinamate + H2O | amidase activity, hydrolysis proceeds at a slower rate than formation of succinyl hydroxamate in the presence of hydroxylamine | Bacillus subtilis | succinic acid + NH3 | - |
? | |
| succinamate + hydroxylamine | amide transfer reaction | Thermus aquaticus | succinyl hydroxamate + NH3 | - |
r | |
| succinamate + hydroxylamine | amide transfer reaction | Bacillus subtilis | succinyl hydroxamate + NH3 | - |
r | |
| succinamide + H2O | - |
Thermus aquaticus | succinate + NH3 | - |
? | |
| succinamide + H2O | - |
Bacillus subtilis | succinate + NH3 | - |
? | |
| succinamide + hydroxylamine | amide transfer reaction | Thermus aquaticus | succinyl hydroxamate + ? | - |
r | |
| succinamide + hydroxylamine | amide transfer reaction | Bacillus subtilis | succinyl hydroxamate + ? | - |
r | |
| succinate + hydroxylamine | very high substrate concentration required for acyl transfer reaction | Thermus aquaticus | succinyl hydroxamate + H2O | - |
r | |
| succinate + hydroxylamine | very high substrate concentration required for acyl transfer reaction | Bacillus subtilis | succinyl hydroxamate + H2O | - |
r | |
| succinate + hydroxylamine | acyl transfer reaction | Thermus aquaticus | succinyl hydroxamate + H2O | - |
r | |
| succinate + hydroxylamine | acyl transfer reaction | Bacillus subtilis | succinyl hydroxamate + H2O | - |
r | |
| succinimide + hydroxylamine | amide transfer reaction | Thermus aquaticus | ? | - |
? | |
| succinimide + hydroxylamine | amide transfer reaction | Bacillus subtilis | ? | - |
? | |
| succinyl hydroxamate + H2O | - |
Thermus aquaticus | succinate + hydroxylamine | - |
r | |
| succinyl hydroxamate + H2O | - |
Bacillus subtilis | succinate + hydroxylamine | - |
r | |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Bacillus subtilis |
| 45 | 56 | broad temperature optimum | Bacillus subtilis |
| 70 | - |
assay at | Thermus aquaticus |
| 80 | - |
- |
Thermus aquaticus |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | 63 | about 50% of maximum activity | Bacillus subtilis |
| 63 | 87 | about 50% of maximum activity | Thermus aquaticus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| additional information | - |
- |
Thermus aquaticus |
| additional information | - |
- |
Bacillus subtilis |
| 6.5 | - |
acyl transferase activity, sharp optimum | Thermus aquaticus |
| 6.5 | - |
acyl transferase activity, sharp optimum | Bacillus subtilis |
| 7 | 9 | amide transferase activity, broad optimum | Thermus aquaticus |
| 7 | 9 | amide transferase activity, broad optimum | Bacillus subtilis |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| additional information | no requirement for any cofactors for hydroxamate formation | Thermus aquaticus | |
| additional information | no requirement for any cofactors for hydroxamate formation | Bacillus subtilis |
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