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Literature summary for 6.3.1.15 extracted from
- Cloning, overexpression, and purification of novobiocic acid synthetase from Streptomyces spheroides NCIMB 11891 (2000), J. Biol. Chem., 275, 21754-21760.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| overexpressed as a His6 fusion protein in Escherichia coli | Streptomyces niveus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| EDTA | 0.5 mM, reduces the enzyme activity to 1.4% of the maximal activity | Streptomyces niveus |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.019 | - |
3-dimethylallyl-4-hydroxybenzoate | pH 8.0, 30°C | Streptomyces niveus | |
| 0.131 | - |
3-amino-4,7-dihydroxy-8-methylcoumarin | pH 8.0, 30°C | Streptomyces niveus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | the reaction is dependent on the presence of Mg2+ or Mn2+ | Streptomyces niveus | |
| Mn2+ | the reaction is dependent on the presence of Mg2+ or Mn2+ | Streptomyces niveus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 56885 | - |
1 * 56885, calculated from sequence | Streptomyces niveus |
| 57000 | 58000 | gel filtration | Streptomyces niveus |
| 60000 | - |
1 * 60000, His-tagged protein, calculated from sequence | Streptomyces niveus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Streptomyces niveus | Q9L9F6 | - |
- |
| Streptomyces niveus NCIMB 11891 | Q9L9F6 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Streptomyces niveus |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 3.036 | - |
pH 8.0, 30°C | Streptomyces niveus |
Storage Stability
| Storage Stability | Organism |
|---|---|
| 4°C, purified enzyme loses 22% of its activity after 2 days | Streptomyces niveus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + 3-dimethylallyl-4-hydroxybenzoate + 3-amino-4,7-dihydroxy-8-methylcoumarin | the formation of an acyl adenylate from ring A and ATP is demonstrated by an ATP-PPi exchange assay. The purified nzyme exhibits both activation and transferase activity, i.e. it catalyzed both the activation of ring A as acyl adenylate and the subsequent transfer of the acyl group to the amino group of ring B. The reaction is specific for ATP as nucleotide triphosphate | Streptomyces niveus | AMP + diphosphate + novobiocic acid | - |
? | |
| ATP + 3-dimethylallyl-4-hydroxybenzoate + 3-amino-4,7-dihydroxy-8-methylcoumarin | the formation of an acyl adenylate from ring A and ATP is demonstrated by an ATP-PPi exchange assay. The purified nzyme exhibits both activation and transferase activity, i.e. it catalyzed both the activation of ring A as acyl adenylate and the subsequent transfer of the acyl group to the amino group of ring B. The reaction is specific for ATP as nucleotide triphosphate | Streptomyces niveus NCIMB 11891 | AMP + diphosphate + novobiocic acid | - |
? | |
| ATP + 3-geranyl-4-hydroxybenzoate + 3-amino-4,7-dihydroxy-8-methylcoumarin | - |
Streptomyces niveus | AMP + diphosphate + N-(2,7-dihydroxy-8-methyl-4-oxo-4H-chromen-3-yl)-4-hydroxy-3-[(2E)-6-methylhepta-2,5-dien-1-yl]benzamide | - |
? | |
| ATP + 3-geranyl-4-hydroxybenzoate + 3-amino-4,7-dihydroxy-8-methylcoumarin | - |
Streptomyces niveus NCIMB 11891 | AMP + diphosphate + N-(2,7-dihydroxy-8-methyl-4-oxo-4H-chromen-3-yl)-4-hydroxy-3-[(2E)-6-methylhepta-2,5-dien-1-yl]benzamide | - |
? | |
| ATP + 4-hydroxy-3-methylbenzoate + 3-amino-4,7-dihydroxy-8-methylcoumarin | - |
Streptomyces niveus | AMP + diphosphate + 3-[(2E)-but-2-en-1-yl]-N-(2,7-dihydroxy-8-methyl-4-oxo-4H-chromen-3-yl)-4-hydroxybenzylamide | - |
? | |
| ATP + 4-hydroxy-3-methylbenzoate + 3-amino-4,7-dihydroxy-8-methylcoumarin | - |
Streptomyces niveus NCIMB 11891 | AMP + diphosphate + 3-[(2E)-but-2-en-1-yl]-N-(2,7-dihydroxy-8-methyl-4-oxo-4H-chromen-3-yl)-4-hydroxybenzylamide | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 56885, calculated from sequence | Streptomyces niveus |
| monomer | 1 * 60000, His-tagged protein, calculated from sequence | Streptomyces niveus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| NovL | - |
Streptomyces niveus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Streptomyces niveus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
phosphate buffer | Streptomyces niveus |
| 8 | - |
Tris-HCl buffer | Streptomyces niveus |
General Information
| General Information | Comment | Organism |
|---|---|---|
| physiological function | the enzyme is involved in the biosynthesis of the aminocoumarin antibiotic novobiocin | Streptomyces niveus |
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