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Literature summary extracted from

  • Liang, A.; Sha, J.; Lu, W.; Chen, M.; Li, L.; Jin, D.; Yan, Y.; Wang, J.; Ping, S.; Zhang, W.; Wang, Y.; Lin, M.
    A single residue mutation of 5-enoylpyruvylshikimate 3-phosphate synthase in Pseudomonas stutzeri enhances resistance to the herbicide glyphosate (2008), Biotechnol. Lett., 30, 1397-1401.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.19 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 (Commentary)

EC Number Crystallization (Comment) Organism
2.5.1.19 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

EC Number Protein Variants Comment Organism
2.5.1.19 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

EC Number Inhibitors Comment Organism Structure
2.5.1.19 glyphosate at 300 mM: no growth of Escherichia coli transformed with either EPSPS wild-type or mutant N130S Pseudomonas stutzeri

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.19 0.055
-
 phosphoenolpyruvate +/-0.0014 mM, mutant N130S in bacterial extact Pseudomonas stutzeri
2.5.1.19 0.055
-
 phosphoenolpyruvate +/-0.002 mM, bacterial extract Pseudomonas stutzeri

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.5.1.19 46000
-
 wild-type and mutant N130S, SDS-PAGE on bacterial extracts Pseudomonas stutzeri

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.19 Pseudomonas stutzeri
-
-
-
2.5.1.19 Pseudomonas stutzeri A1501
-
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.5.1.19 0.19
-
 bacterial extract Pseudomonas stutzeri
2.5.1.19 0.22
-
 mutant N130S in bacterial extract Pseudomonas stutzeri

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.19 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 ?
2.5.1.19 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

EC Number Synonyms Comment Organism
2.5.1.19 5-enoylpyruvylshikimate 3-phosphate synthase
-
 Pseudomonas stutzeri
2.5.1.19 A1501 EPSPS class II EPSPS Pseudomonas stutzeri
2.5.1.19 AroA(A1501)
-
 Pseudomonas stutzeri
2.5.1.19 EPSPS
-
 Pseudomonas stutzeri

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.5.1.19 0.1
-
 glyphosate +/-0.01 mM, in presence of 1 mM shikimate 3-phosphate and 0-500 mM inhibitor, bacterial extract Pseudomonas stutzeri
2.5.1.19 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

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
2.5.1.19 8
-
 +/-0.5 mM, in presence of 1 mM shikimate 3-phosphate and 0-500 mM inhibitor, bacterial extract Pseudomonas stutzeri glyphosate
2.5.1.19 20
-
 +/-0.2 mM, mutant N130S, in presence of 1 mM shikimate 3-phosphate and 0-500 mM inhibitor, bacterial extract Pseudomonas stutzeri glyphosate