Literature summary extracted from

  • Sikora, A.L.; Wilson, D.J.; Aldrich, C.C.; Blanchard, J.S.
    Kinetic and inhibition studies of dihydroxybenzoate-AMP ligase from Escherichia coli (2010), Biochemistry, 49, 3648-3657.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.2.14 gene entE, expression in Escherichia coli strain BL21(DE3) Escherichia coli
6.3.2.14 gene entE, expression of His-tagged EntE in Escherichia coli strain BL21(DE3) Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.2.14 5'-O-[N-(2,3-dihydroxybenzoyl)sulfamoyl]adenosine
-
Escherichia coli
6.3.2.14 5'-O-[N-(salicyl)sulfamoyl]adenosine
-
Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.2.14 additional information
-
additional information two possible kinetic mechanisms can explain nonlinear kinetics: one-step slow association and two-step isomerization, bi-uni-uni-bi ping-pong kinetic mechanism, kinetic analysis, overview Escherichia coli
6.3.2.14 0.07
-
salicylic acid pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 0.07
-
3-hydroxybenzoate pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 0.0025
-
2,3-Dihydroxybenzoate pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 0.43
-
ATP pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 3.1
-
4-aminosalicylic acid pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 0.0029
-
phosphopantetheinylated EntB pH 7.8, 25°C, recombinant EntE Escherichia coli

Metals/Ions

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

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.2.14 6 ATP + 3 2,3-dihydroxybenzoate + 3 L-serine Escherichia coli overall reaction. EntB, -D, -E, and -F are then required to catalyze the ATP-dependent assembly of enterobactin from three molecules each of 2,3-dihydroxybenzoate and L-serine. EntD, a phosphopantetheinyl transferase, uses coenzyme A to phosphopantetheinylate S245 of the aryl carrier protein domain of EntB. Next, EntE catalyzes the transfer of 2,3-dihydroxybenzoate onto the phosphopantetheinylated (holo) EntB to yield the covalently arylated EntB. Finally, arylated EntB, ATP, and L-serine are used as substrates for the reaction catalyzed by EntF to generate enterobactin enterobactin + 6 AMP + 6 diphosphate
-
?
6.3.2.14 6 ATP + 3 2,3-dihydroxybenzoate + 3 L-serine Escherichia coli overall reaction. EntB, -D, -E, and -F are then required to catalyze the ATP-dependent assembly of enterobactin from three molecules each of 2,3-dihydroxybenzoate and L-serine. EntD, a phosphopantetheinyl transferase, uses coenzyme A to phosphopantetheinylate S245 of the aryl carrier protein domain of EntB. Next, EntE catalyzes the transfer of 2,3-dihydroxybenzoate onto the phosphopantetheinylated (holo)EntB to yield the covalently arylated EntB. Finally, arylated EntB, ATP, and L-serine are used as substrates for the reaction catalyzed by EntF to generate enterobactin enterobactin + 6 AMP + 6 diphosphate
-
?
6.3.2.14 ATP + 2,3-dihydroxybenzoate + phosphopantetheinylated EntB Escherichia coli reaction of EntE arylated EntB + AMP + diphosphate
-
?
6.3.2.14 ATP + arylated EntB + L-serine Escherichia coli reaction of EntF enterobactin + AMP + diphosphate
-
?

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.14 Escherichia coli P0ADI4 gene entB; gene entB
-
6.3.2.14 Escherichia coli P10378 gene entE; gene entE
-
6.3.2.14 Escherichia coli P11454 gene entF; gene entF
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.2.14 recombinant His-tagged EntE from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.2.14 additional information no activity with 4-aminobenzoic acid Escherichia coli ?
-
?
6.3.2.14 6 ATP + 3 2,3-dihydroxybenzoate + 3 L-serine overall reaction. EntB, -D, -E, and -F are then required to catalyze the ATP-dependent assembly of enterobactin from three molecules each of 2,3-dihydroxybenzoate and L-serine. EntD, a phosphopantetheinyl transferase, uses coenzyme A to phosphopantetheinylate S245 of the aryl carrier protein domain of EntB. Next, EntE catalyzes the transfer of 2,3-dihydroxybenzoate onto the phosphopantetheinylated (holo) EntB to yield the covalently arylated EntB. Finally, arylated EntB, ATP, and L-serine are used as substrates for the reaction catalyzed by EntF to generate enterobactin Escherichia coli enterobactin + 6 AMP + 6 diphosphate
-
?
6.3.2.14 6 ATP + 3 2,3-dihydroxybenzoate + 3 L-serine overall reaction. EntB, -D, -E, and -F are then required to catalyze the ATP-dependent assembly of enterobactin from three molecules each of 2,3-dihydroxybenzoate and L-serine. EntD, a phosphopantetheinyl transferase, uses coenzyme A to phosphopantetheinylate S245 of the aryl carrier protein domain of EntB. Next, EntE catalyzes the transfer of 2,3-dihydroxybenzoate onto the phosphopantetheinylated (holo)EntB to yield the covalently arylated EntB. Finally, arylated EntB, ATP, and L-serine are used as substrates for the reaction catalyzed by EntF to generate enterobactin Escherichia coli enterobactin + 6 AMP + 6 diphosphate
-
?
6.3.2.14 ATP + 2,3-dihydroxybenzoate + phosphopantetheinylated EntB reaction of EntE Escherichia coli arylated EntB + AMP + diphosphate
-
?
6.3.2.14 ATP + arylated EntB + L-serine reaction of EntF Escherichia coli enterobactin + AMP + diphosphate
-
?
6.3.2.14 ATP + 3-hydroxybenzoate + phosphopantetheinylated EntB reaction of EntE Escherichia coli arylated EntB + AMP + diphosphate
-
?
6.3.2.14 ATP + 4-aminosalicylic acid + phosphopantetheinylated EntB reaction of EntE Escherichia coli arylated EntB + AMP + diphosphate
-
?
6.3.2.14 ATP + salicylic acid + phosphopantetheinylated EntB reaction of EntE Escherichia coli arylated EntB + AMP + diphosphate
-
?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.3.2.14 25
-
assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.3.2.14 0.3
-
3-hydroxybenzoate pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 0.8
-
salicylic acid pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 2.8
-
ATP pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 2.8
-
2,3-Dihydroxybenzoate pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 2.8
-
phosphopantetheinylated EntB pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 4.4
-
4-aminosalicylic acid pH 7.8, 25°C, recombinant EntE Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.2.14 7.8
-
assay at Escherichia coli

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.2.14 ATP
-
Escherichia coli

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
6.3.2.14 additional information
-
additional information thermodynamics and inhibition kinetics, stopped-flow measurements Escherichia coli

General Information

EC Number General Information Comment Organism
6.3.2.14 metabolism chorismate is converted to 2,3-dihydroxybenzoate via the sequential catalytic activities of EntC, -B, and -A Escherichia coli

KCat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.3.2.14 11
-
salicylic acid pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 4.6
-
3-hydroxybenzoate pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 15
-
4-aminosalicylic acid pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 390
-
ATP pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 880
-
2,3-Dihydroxybenzoate pH 7.8, 25°C, recombinant EntE Escherichia coli
6.3.2.14 980
-
phosphopantetheinylated EntB pH 7.8, 25°C, recombinant EntE Escherichia coli