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Literature summary extracted from
- Structure of chorismate mutase-like domain of DAHPS from Bacillus subtilis complexed with novel inhibitor reveals conformational plasticity of active site (2017), Sci. Rep., 7, 6364 .
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 5.4.99.5 | recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Bacillus subtilis |
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 2.5.1.54 | structure of N-terminal domain AroQ in complex with citrate and chlorogenic acid at 1.9 A and 1.8 A resolution, respectively. Helix H2' undergoes uncoiling at the first turn and increases the mobility of loop L1'. The side chains of Arg45, Phe46, Arg52 and Lys76 undergo conformational changes, which may play an important role in DAHPS regulation by the formation of the domain-domain interface. Chlorogenic acid binds with a higher affinity than chorismate | Bacillus subtilis |
| 5.4.99.5 | purified recombinant enzyme AroQ (BsCM_2) in complex with citrate and chlorogenic acid, sitting drop vapor diffusion method, mixing of 0.001 ml of 18 mg/ml protein in 25 mM Tris-HCl, pH 7.5, and 50 mM NaCl, with 0.001 ml of reservoir solution containing 1 M ammonium sulphate, 0.1 M potassium sodium tartrate, and 0.1 M sodium citrate, pH 5.8, 20°C, 15 days, X-ray diffraction structure determination and analysis at 1.9 A and 1.8 A resolution, respectively, molecular replacement using the structure of the N-terminal CM domain of bifunctional DAHPS from Listeria monocytogens (PDB ID 3NVT) as template | Bacillus subtilis |
| 5.4.99.5 | structure of N-terminal domain AroQ in complex with citrate and chlorogenic acid at 1.9 A and 1.8 A resolution, respectively. Helix H2' undergoes uncoiling at the first turn and increases the mobility of loop L1'. The side chains of Arg45, Phe46, Arg52 and Lys76 undergo conformational changes, which may play an important role in DAHPS regulation by the formation of the domain-domain interface. Chlorogenic acid binds with a higher affinity than chorismate | Bacillus subtilis |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.4.99.5 | chlorogenic acid | CGA, a structural analogue of chorismic acid, is an inhibitor of chorismate mutase, type II regulatory domain (BsCM_2). It binds to BsCM_2 with a higher affinity than chorismate. Similar to BsCM_2, in BsAroH, the chlorogenic acid's position is shifted from the transition state analogue position. The chlorogenic acid interacts with residues Arg63, Val73, Thr74 from one chain and Arg7, Arg90, Val114, Leu115, and Arg116 from the adjacent chain; CGA, a structural analogue of chorismic acid, is an inhibitor of chorismate mutase, type II regulatory domain (BsCM_2). It binds to BsCM_2 with a higher affinity than chorismate. The BsCM_2-CGA structure has several residues in alternate conformations. His73 exists as alternative conformation in both the chains. At active site S1, the chlorogenic acid makes hydrogen bonds with the side chain of Arg27, Lys38, Gln86, and the main chain atoms of Arg45, Asp47, and Phe79 of chain B. The ligand molecule also interacts with Lys38, Arg50, and Lys80 of chain B, and Arg10 of chain A through water bridge formation. However, at active site S2, along with the above interactions, the ligand forms a direct hydrogen bond with Lys80 and an additional water bridge-mediated hydrogen bond with Gln86 of chain A | Bacillus subtilis |  |
| 5.4.99.5 | additional information | the similarity of chlorogenic acid's interaction with both monofunctional chorismate mutases BsAroH and BsCM_2 may result in similar binding to both proteins; the similarity of chlorogenic acid's interaction with both monofunctional chorismate mutases BsAroH and BsCM_2 may result in similar binding to both proteins | Bacillus subtilis |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.4.99.5 | additional information | - |
additional information | Michaelis-Menten kinetics | Bacillus subtilis | |
| 5.4.99.5 | 0.067 | - |
chorismate | pH 7.5, 37°C, recombinant AroH | Bacillus subtilis | |
| 5.4.99.5 | 1.514 | - |
chorismate | pH 7.5, 37°C, recombinant BsCM_2 | Bacillus subtilis | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.4.99.5 | Chorismate | Bacillus subtilis | - |
Prephenate | - |
? | |
| 5.4.99.5 | Chorismate | Bacillus subtilis 168 | - |
Prephenate | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 2.5.1.54 | Bacillus subtilis | P39912 | bifunctional enzyme, catalyzes reactions of EC 2.5.1.54 and EC 5.4.99.5 | - |
| 2.5.1.54 | Bacillus subtilis 168 | P39912 | bifunctional enzyme, catalyzes reactions of EC 2.5.1.54 and EC 5.4.99.5 | - |
| 5.4.99.5 | Bacillus subtilis | P19080 | - |
- |
| 5.4.99.5 | Bacillus subtilis | P39912 | - |
- |
| 5.4.99.5 | Bacillus subtilis | P39912 | bifunctional enzyme, catalyzes reactions of EC 2.5.1.54 and EC 5.4.99.5 | - |
| 5.4.99.5 | Bacillus subtilis 168 | P19080 | - |
- |
| 5.4.99.5 | Bacillus subtilis 168 | P39912 | - |
- |
| 5.4.99.5 | Bacillus subtilis 168 | P39912 | bifunctional enzyme, catalyzes reactions of EC 2.5.1.54 and EC 5.4.99.5 | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 5.4.99.5 | recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, gel filtration, and ultrafiltration | Bacillus subtilis |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.4.99.5 | Chorismate | - |
Bacillus subtilis | Prephenate | - |
? | |
| 5.4.99.5 | Chorismate | - |
Bacillus subtilis 168 | Prephenate | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 5.4.99.5 | dimer | in the BsCM_2-chlorogenic acid structure, two active sites, S1 and S2, are located at the interface of two monomers | Bacillus subtilis |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 2.5.1.54 | AroA | - |
Bacillus subtilis |
| 5.4.99.5 | AroA | - |
Bacillus subtilis |
| 5.4.99.5 | AroH | - |
Bacillus subtilis |
| 5.4.99.5 | AroQ | - |
Bacillus subtilis |
| 5.4.99.5 | BsAroH | - |
Bacillus subtilis |
| 5.4.99.5 | BsCM_2 | - |
Bacillus subtilis |
| 5.4.99.5 | CM type 2 | - |
Bacillus subtilis |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 5.4.99.5 | 37 | - |
assay at | Bacillus subtilis |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.4.99.5 | 0.78 | - |
chorismate | pH 7.5, 37°C, recombinant BsCM_2 | Bacillus subtilis | |
| 5.4.99.5 | 46 | - |
chorismate | pH 7.5, 37°C, recombinant AroH | Bacillus subtilis | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 5.4.99.5 | 7.5 | - |
assay at | Bacillus subtilis |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 5.4.99.5 | evolution | the N-terminal domain of DAHPS from Bacillus subtilis is homologous to the AroQ class of chorismate mutase, type II. Bacillus subtilis also contains a monofunctional AroH class of chorismate mutase situated downstream of the shikimate pathway | Bacillus subtilis |
| 5.4.99.5 | additional information | AroH molecular docking, using crystal structure of BsAroH, PDB ID 2CHT, overview | Bacillus subtilis |
| 5.4.99.5 | additional information | structural basis of ligand binding into the active site of AroQ class of chorismate mutase from crystal structure analysis, conformational flexibility of active site loop, overview. Molecular dynamics results show that helix H2' undergoes uncoiling at the first turn and increases the mobility of loop L1'. The side chains of Arg45, Phe46, Arg52 and Lys76 undergo conformational changes, which may play an important role in DAHPS regulation by the formation of the domain-domain interface. BsCM_2 active site architecture and its regulatory role, molecular dynamics simulation, overview | Bacillus subtilis |
| 5.4.99.5 | physiological function | in Bacillus subtilis, the N-terminal domain of the bifunctional 3-deoxy-D-arabino-heptulosonate-7-phosphate-synthase (DAHPS), the first enzyme of the shikimate pathway, belongs to an AroQ class of chorismate mutase and is functionally homologous to the downstream AroH class chorismate mutase. BsCM_2 has a regulatory function in the bifunctional DAHPS enzyme, regulation of DAHPS enzyme activity by the CM2 domain, overview | Bacillus subtilis |
| 5.4.99.5 | physiological function | in Bacillus subtilis, the N-terminal domain of the bifunctional 3-deoxy-D-arabino-heptulosonate-7-phosphate-synthase (DAHPS), the first enzyme of the shikimate pathway, belongs to an AroQ class of chorismate mutase and is functionally homologous to the downstream AroH class chorismate mutase. BsCM_2 may also have a regulatory function in the bifunctional DAHPS enzyme | Bacillus subtilis |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.4.99.5 | 1.94 | - |
chorismate | pH 7.5, 37°C, recombinant BsCM_2 | Bacillus subtilis | |
| 5.4.99.5 | 686.6 | - |
chorismate | pH 7.5, 37°C, recombinant AroH | Bacillus subtilis | |
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