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Literature summary for 6.2.1.71 extracted from
- 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.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| additional information | no activation by DTT | Escherichia coli |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| overexpression in Escherichia coli JM105 | Escherichia coli |
| structural gene entE subcloned into multi-copy plasmid pKK223-3 under control of tac-promoter, DNA sequence determination | Escherichia coli |
General Stability
| General Stability | Organism |
|---|---|
| glycerol, 50%, stabilizes dilute enzyme solutions | Escherichia coli |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0027 | - |
2,3-Dihydroxybenzoic acid | pH 9.0, 37°C | Escherichia coli |  |
| 0.091 | - |
2-hydroxybenzoic acid | pH 9.0, 37°C | Escherichia coli | |
| 0.093 | - |
2,3,4-trihydroxybenzoic acid | pH 9.0, 37°C | Escherichia coli | |
| 0.242 | - |
2,4-dihydroxybenzoic acid | pH 9.0, 37°C | Escherichia coli | |
| 0.552 | - |
2,5-dihydroxybenzoic acid | pH 9.0, 37°C | Escherichia coli | |
| 1.12 | - |
ATP | pH 9.0, 37°C | Escherichia coli | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | required | Escherichia coli | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 59000 | - |
2 * 59000, SDS-PAGE | Escherichia coli |
| 115000 | - |
gel filtration | Escherichia coli |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + 2,3-dihydroxybenzoate | Escherichia coli | activation of 2,3-dihydroxybenzoic acid in the biosynthesis of siderophore enterobactin | diphosphate + (2,3-dihydroxybenzoyl)adenylate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| 4.8fold | Escherichia coli |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 8.33 | - |
partially purified enzyme | Escherichia coli |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -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)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + 2,3,4-trihydroxybenzoic acid | - |
Escherichia coli | diphosphate + (2,3,4-trihydroxybenzoyl)adenylate | - |
? | |
| ATP + 2,3-dihydroxybenzoate | - |
Escherichia coli | diphosphate + (2,3-dihydroxybenzoyl)adenylate | product remains enzyme-bound for further reaction in overall biosynthesis of enterobactin | ? | |
| ATP + 2,3-dihydroxybenzoate | activation of 2,3-dihydroxybenzoic acid in the biosynthesis of siderophore enterobactin | Escherichia coli | diphosphate + (2,3-dihydroxybenzoyl)adenylate | - |
? | |
| ATP + 2,4-dihydroxybenzoic acid | - |
Escherichia coli | diphosphate + (2,4-dihydroxybenzoyl)adenylate | - |
? | |
| ATP + 2,5-dihydroxybenzoic acid | i.e. gentisic acid | Escherichia coli | diphosphate + (2,5-dihydroxybenzoyl)adenylate | - |
? | |
| ATP + 2-hydroxybenzoic acid | i.e. salicylic acid, not 3- or 4-derivative | Escherichia coli | diphosphate + (2-hydroxybenzoyl)adenylate | - |
? | |
| additional information | no substrates are benzoic acid, 2,6-dihydroxybenzoic acid, 2,4,6-trihydroxybenzoic acid, anthranilic acid, thiosalicylic acid | Escherichia coli | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 59000, SDS-PAGE | Escherichia coli |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Escherichia coli |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.45 | - |
2,3,4-Trihydroxybenzoate | pH 9.0, 37°C | Escherichia coli | |
| 2.5 | - |
2-hydroxybenzoic acid | pH 9.0, 37°C | Escherichia coli | |
| 3.15 | - |
2,5-dihydroxybenzoic acid | pH 9.0, 37°C | Escherichia coli | |
| 3.23 | - |
2,4-Dihydroxybenzoate | pH 9.0, 37°C | Escherichia coli | |
| 5.5 | - |
2,3-Dihydroxybenzoate | pH 9.0, 37°C | Escherichia coli | |
| 5.83 | - |
ATP | pH 9.0, 37°C | Escherichia coli | |
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