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Literature summary extracted from

  • Steffensky, M.; Li, S.M.; Heide, L.
    Cloning, overexpression, and purification of novobiocic acid synthetase from Streptomyces spheroides NCIMB 11891 (2000), J. Biol. Chem., 275, 21754-21760.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.1.15 overexpressed as a His6 fusion protein in Escherichia coli Streptomyces niveus

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.1.15 EDTA 0.5 mM, reduces the enzyme activity to 1.4% of the maximal activity Streptomyces niveus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.1.15 0.019
-
 3-dimethylallyl-4-hydroxybenzoate pH 8.0, 30°C Streptomyces niveus
6.3.1.15 0.131
-
 3-amino-4,7-dihydroxy-8-methylcoumarin pH 8.0, 30°C Streptomyces niveus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.1.15 Mg2+ the reaction is dependent on the presence of Mg2+ or Mn2+ Streptomyces niveus
6.3.1.15 Mn2+ the reaction is dependent on the presence of Mg2+ or Mn2+ Streptomyces niveus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.3.1.15 56885
-
 1 * 56885, calculated from sequence Streptomyces niveus
6.3.1.15 57000 58000 gel filtration Streptomyces niveus
6.3.1.15 60000
-
 1 * 60000, His-tagged protein, calculated from sequence Streptomyces niveus

Organism

EC Number Organism UniProt Comment Textmining
6.3.1.15 Streptomyces niveus Q9L9F6
-
-
6.3.1.15 Streptomyces niveus NCIMB 11891 Q9L9F6
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.1.15
-
 Streptomyces niveus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.3.1.15 3.036
-
 pH 8.0, 30°C Streptomyces niveus

Storage Stability

EC Number Storage Stability Organism
6.3.1.15 4°C, purified enzyme loses 22% of its activity after 2 days Streptomyces niveus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.1.15 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
-
 ?
6.3.1.15 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
-
 ?
6.3.1.15 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
-
 ?
6.3.1.15 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
-
 ?
6.3.1.15 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
-
 ?
6.3.1.15 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

EC Number Subunits Comment Organism
6.3.1.15 monomer 1 * 56885, calculated from sequence Streptomyces niveus
6.3.1.15 monomer 1 * 60000, His-tagged protein, calculated from sequence Streptomyces niveus

Synonyms

EC Number Synonyms Comment Organism
6.3.1.15 NovL
-
 Streptomyces niveus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.3.1.15 30
-
 assay at Streptomyces niveus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.1.15 7
-
 phosphate buffer Streptomyces niveus
6.3.1.15 8
-
 Tris-HCl buffer Streptomyces niveus

General Information

EC Number General Information Comment Organism
6.3.1.15 physiological function the enzyme is involved in the biosynthesis of the aminocoumarin antibiotic novobiocin Streptomyces niveus