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Literature summary for 2.5.1.19 extracted from
- A T42M substitution in bacterial 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) generates enzymes with increased resistance to glyphosate (2003), Biosci. Biotechnol. Biochem., 67, 1405-1409.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of the chimeric mutant aroA-M1, comprising N-terminal part of the Escherichia coli enzyme and the C-terminal part of the enzyme from Salmonella typhimurium, and of T42M-aroA mutant in Escherichia coli strain BL21(DE3) | Salmonella enterica subsp. enterica serovar Typhimurium |
| expression of the wild-type enzyme, of the chimeric mutant aroA-M1, comprising N-terminal part of the Escherichia coli enzyme and the C-terminal part of the enzyme from Salmonella typhimurium, and of T42M-aroA mutant in strain BL21(DE3) | Escherichia coli |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D242A | site-directed mutagenesis, the mutation is responsible for the high increase in activity | Escherichia coli |
| D348A | site-directed mutagenesis, the mutation is responsible for the high increase in activity | Escherichia coli |
| additional information | construction of chimeric mutants by fusing the N-terminal part of the Escherichia coli enzyme with the C-terminal part of the enzyme from Salmonella typhimurium, the chimeric mutant shows a 6fold lower Km for phosphoenolpyruvate, a 5fold higher Ki for glyphosate, and a 9fold higher specific activity than the wild-type enzyme from Escherichia coli | Salmonella enterica subsp. enterica serovar Typhimurium |
| additional information | construction of chimeric mutants by fusing the N-terminal part of the Escherichia coli enzyme with the C-terminal part of the enzyme from Salmonella typhimurium, the chimeric mutant shows a 6fold lower Km for phosphoenolpyruvate, a 5fold higher Ki for glyphosate, and a 9fold higher specific activity than the wild-type enzyme from Escherichia coli | Escherichia coli |
| R100A | site-directed mutagenesis, the mutation is responsible for the high increase in activity | Escherichia coli |
| T42M | site-directed mutagenesis, the mutation is responsible for the glyphosate resistance and the low Km for phosphoenolpyruvate | Salmonella enterica subsp. enterica serovar Typhimurium |
| T42M | site-directed mutagenesis, the mutation is responsible for the glyphosate resistance and the low Km for phosphoenolpyruvate | Escherichia coli |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| glyphosate | competitive inhibition of the wild-type enzyme, mutant T42M is less sensitive | Escherichia coli |  |
| glyphosate | competitive inhibition of the wild-type enzyme, mutant T42M is less sensitive | Salmonella enterica subsp. enterica serovar Typhimurium | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kinetics, wild-type and mutant enzymes | Salmonella enterica subsp. enterica serovar Typhimurium | |
| additional information | - |
additional information | kinetics, wild-type and mutant enzymes | Escherichia coli | |
| 0.0018 | - |
phosphoenolpyruvate | pH 7.0, 30°C, mutant StaroA-T42M | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.0024 | - |
phosphoenolpyruvate | pH 7.0, 30°C, mutant EcaroA-T42M | Escherichia coli | |
| 0.0223 | - |
phosphoenolpyruvate | pH 7.0, 30°C, wild-type EcaroA | Escherichia coli | |
| 0.0436 | - |
phosphoenolpyruvate | pH 7.0, 30°C, wild-type StaroA | Salmonella enterica subsp. enterica serovar Typhimurium | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 45000 | - |
* 45000, wild-type and mutant enzyme, SDS-PAGE | Escherichia coli |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| phosphoenolpyruvate + 3-phosphoshikimate | Salmonella enterica subsp. enterica serovar Typhimurium | - |
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate | - |
? | |
| phosphoenolpyruvate + 3-phosphoshikimate | Escherichia coli | - |
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
- |
- |
| Salmonella enterica subsp. enterica serovar Typhimurium | - |
- |
- |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.0058 | - |
wild-type enzyme, cell exract | Salmonella enterica subsp. enterica serovar Typhimurium |
| 0.0076 | - |
wild-type enzyme, cell exract | Escherichia coli |
| 0.0357 | - |
mutant T42M, cell extract | Escherichia coli |
| 0.136 | - |
mutant T42M, cell extract | Salmonella enterica subsp. enterica serovar Typhimurium |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| phosphoenolpyruvate + 3-phosphoshikimate | - |
Salmonella enterica subsp. enterica serovar Typhimurium | phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate | - |
? | |
| phosphoenolpyruvate + 3-phosphoshikimate | - |
Escherichia coli | phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | * 45000, wild-type and mutant enzyme, SDS-PAGE | Escherichia coli |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| 5-enolpyruvylshikimate-3-phosphate synthase | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
| 5-enolpyruvylshikimate-3-phosphate synthase | - |
Escherichia coli |
| EcaroA | - |
Escherichia coli |
| EPSPS | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
| EPSPS | - |
Escherichia coli |
| StaroA | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Salmonella enterica subsp. enterica serovar Typhimurium |
| 30 | - |
assay at | Escherichia coli |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
assay at | Salmonella enterica subsp. enterica serovar Typhimurium |
| 7 | - |
assay at | Escherichia coli |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0005 | - |
glyphosate | pH 7.0, 30°C, wild-type StaroA | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.0015 | - |
glyphosate | pH 7.0, 30°C, wild-type EcaroA | Escherichia coli | |
| 0.0128 | - |
glyphosate | pH 7.0, 30°C, mutant StaroA-T42M | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.0305 | - |
glyphosate | pH 7.0, 30°C, mutant EcaroA-T42M | Escherichia coli | |
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