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Literature summary extracted from

  • Rusnak, F.; Faraci, W.S.; Walsh, C.T.
    Subcloning, expression, and purification of the enterobactin biosynthetic enzyme 2,3-dihydroxybenzoate-AMP ligase: demonstration of enzyme-bound (2,3-dihydroxybenzoyl)adenylate product (1989), Biochemistry, 28, 6827-6835.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
6.2.1.71 additional information no activation by DTT Escherichia coli

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.2.1.71 overexpression in Escherichia coli JM105 Escherichia coli
6.2.1.71 structural gene entE subcloned into multi-copy plasmid pKK223-3 under control of tac-promoter, DNA sequence determination Escherichia coli

General Stability

EC Number General Stability Organism
6.2.1.71 glycerol, 50%, stabilizes dilute enzyme solutions Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.2.1.71 0.0027
-
 2,3-Dihydroxybenzoic acid pH 9.0, 37°C Escherichia coli
6.2.1.71 0.091
-
 2-hydroxybenzoic acid pH 9.0, 37°C Escherichia coli
6.2.1.71 0.093
-
 2,3,4-trihydroxybenzoic acid pH 9.0, 37°C Escherichia coli
6.2.1.71 0.242
-
 2,4-dihydroxybenzoic acid pH 9.0, 37°C Escherichia coli
6.2.1.71 0.552
-
 2,5-dihydroxybenzoic acid pH 9.0, 37°C Escherichia coli
6.2.1.71 1.12
-
 ATP pH 9.0, 37°C Escherichia coli

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.2.1.71 Mg2+ required Escherichia coli

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.2.1.71 59000
-
 2 * 59000, SDS-PAGE Escherichia coli
6.2.1.71 115000
-
 gel filtration Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.2.1.71 ATP + 2,3-dihydroxybenzoate Escherichia coli activation of 2,3-dihydroxybenzoic acid in the biosynthesis of siderophore enterobactin diphosphate + (2,3-dihydroxybenzoyl)adenylate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.2.1.71 Escherichia coli
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.2.1.71 4.8fold Escherichia coli

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.2.1.71 8.33
-
 partially purified enzyme Escherichia coli

Storage Stability

EC Number Storage Stability Organism
6.2.1.71 -70°C to 2°C, 25 mM Tris-HCl buffer, pH 8.0, 5 mM DTT, 10 mM MgCl2, 50% glycerol, stable for several months Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.2.1.71 ATP + 2,3,4-trihydroxybenzoic acid
-
 Escherichia coli diphosphate + (2,3,4-trihydroxybenzoyl)adenylate
-
 ?
6.2.1.71 ATP + 2,3-dihydroxybenzoate
-
 Escherichia coli diphosphate + (2,3-dihydroxybenzoyl)adenylate product remains enzyme-bound for further reaction in overall biosynthesis of enterobactin ?
6.2.1.71 ATP + 2,3-dihydroxybenzoate activation of 2,3-dihydroxybenzoic acid in the biosynthesis of siderophore enterobactin Escherichia coli diphosphate + (2,3-dihydroxybenzoyl)adenylate
-
 ?
6.2.1.71 ATP + 2,4-dihydroxybenzoic acid
-
 Escherichia coli diphosphate + (2,4-dihydroxybenzoyl)adenylate
-
 ?
6.2.1.71 ATP + 2,5-dihydroxybenzoic acid i.e. gentisic acid Escherichia coli diphosphate + (2,5-dihydroxybenzoyl)adenylate
-
 ?
6.2.1.71 ATP + 2-hydroxybenzoic acid i.e. salicylic acid, not 3- or 4-derivative Escherichia coli diphosphate + (2-hydroxybenzoyl)adenylate
-
 ?
6.2.1.71 additional information no substrates are benzoic acid, 2,6-dihydroxybenzoic acid, 2,4,6-trihydroxybenzoic acid, anthranilic acid, thiosalicylic acid Escherichia coli ?
-
 ?

Subunits

EC Number Subunits Comment Organism
6.2.1.71 dimer 2 * 59000, SDS-PAGE Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.2.1.71 37
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.2.1.71 0.45
-
 2,3,4-Trihydroxybenzoate pH 9.0, 37°C Escherichia coli
6.2.1.71 2.5
-
 2-hydroxybenzoic acid pH 9.0, 37°C Escherichia coli
6.2.1.71 3.15
-
 2,5-dihydroxybenzoic acid pH 9.0, 37°C Escherichia coli
6.2.1.71 3.23
-
 2,4-Dihydroxybenzoate pH 9.0, 37°C Escherichia coli
6.2.1.71 5.5
-
 2,3-Dihydroxybenzoate pH 9.0, 37°C Escherichia coli
6.2.1.71 5.83
-
 ATP pH 9.0, 37°C Escherichia coli