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Literature summary for 2.5.1.19 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
Search for more literature for 2.5.1.19

Cloned(Commentary)

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 (Commentary)

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

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

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

Organism

Organism UniProt Comment Textmining
Pseudomonas stutzeri
-
-
-
Pseudomonas stutzeri A1501
-
-
-

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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 [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 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