Cloned (Comment) | Organism |
---|---|
by PCR from genomic DNA of strain A1501 and error-prone PCR on full-length EPSPS gene, in pACYC184 for transformation and complementation analyses in aroA-inactive mutant Escherichia coli AKM4188, or in pET-28a for expression in Escherichia coli BL21 | Pseudomonas stutzeri |
Crystallization (Comment) | Organism |
---|---|
molecular modelling, two similar globular, inside-out alpha-beta-barrel domains connected by a two-stranded hinge, substrates bind at the inter-domain cleft | Pseudomonas stutzeri |
Protein Variants | Comment | Organism |
---|---|---|
N130S | glyphosate resistance enhanced to 200 mM and glyphosate affinity reduced (2.5fold increase in IC50 and 2fold increase in Ki for glyphosate compared to wild-type) but similar catalytic activity as wild-type, generated by error-prone PCR | Pseudomonas stutzeri |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
glyphosate | at 300 mM: no growth of Escherichia coli transformed with either EPSPS wild-type or mutant N130S | Pseudomonas stutzeri |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.055 | - |
phosphoenolpyruvate | +/-0.0014 mM, mutant N130S in bacterial extact | Pseudomonas stutzeri | |
0.055 | - |
phosphoenolpyruvate | +/-0.002 mM, bacterial extract | Pseudomonas stutzeri |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
46000 | - |
wild-type and mutant N130S, SDS-PAGE on bacterial extracts | Pseudomonas stutzeri |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas stutzeri | - |
- |
- |
Pseudomonas stutzeri A1501 | - |
- |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.19 | - |
bacterial extract | Pseudomonas stutzeri |
0.22 | - |
mutant N130S in bacterial extract | Pseudomonas stutzeri |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
shikimate 3-phosphate + phosphoenolpyruvate | 1 mM shikimate 3-phosphate and 0.05-1.0 mM phosphoenolpyruvate | Pseudomonas stutzeri | phosphate + 5-enolpyruvylshikimate 3-phosphate | detection of inorganic phosphate generation by Malachite Green dye assay according to Lanzetta | ? | |
shikimate 3-phosphate + phosphoenolpyruvate | 1 mM shikimate 3-phosphate and 0.05-1.0 mM phosphoenolpyruvate | Pseudomonas stutzeri A1501 | phosphate + 5-enolpyruvylshikimate 3-phosphate | detection of inorganic phosphate generation by Malachite Green dye assay according to Lanzetta | ? |
Synonyms | Comment | Organism |
---|---|---|
5-enoylpyruvylshikimate 3-phosphate synthase | - |
Pseudomonas stutzeri |
A1501 EPSPS | class II EPSPS | Pseudomonas stutzeri |
AroA(A1501) | - |
Pseudomonas stutzeri |
EPSPS | - |
Pseudomonas stutzeri |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
glyphosate | +/-0.01 mM, in presence of 1 mM shikimate 3-phosphate and 0-500 mM inhibitor, bacterial extract | Pseudomonas stutzeri | |
0.2 | - |
glyphosate | +/-0.01 mM, mutant N130S, in presence of 1 mM shikimate 3-phosphate and 0-500 mM inhibitor, bacterial extract | Pseudomonas stutzeri |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
8 | - |
+/-0.5 mM, in presence of 1 mM shikimate 3-phosphate and 0-500 mM inhibitor, bacterial extract | Pseudomonas stutzeri | glyphosate | |
20 | - |
+/-0.2 mM, mutant N130S, in presence of 1 mM shikimate 3-phosphate and 0-500 mM inhibitor, bacterial extract | Pseudomonas stutzeri | glyphosate |