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Literature summary for 2.5.1.19 extracted from

  • Berti, P.J.; Chindemi, P.
    Catalytic residues and an electrostatic sandwich that promote enolpyruvyl shikimate 3-phosphate synthase (AroA) catalysis (2009), Biochemistry, 48, 3699-3707.
    View publication on PubMed
Search for more literature for 2.5.1.19

Cloned(Commentary)

Cloned (Comment) Organism
expressed as a His-tagged fusion protein Escherichia coli

Protein Variants

Protein Variants Comment Organism
D313A compared to wild-type mutations of D313 causes kcat to decrease up to 30000fold while the effects on Km (3-phosphoshikimate) or Km (phosphoenolpyruvate) are modest, never more than 40fold Escherichia coli
D313C compared to wild-type mutations of D313 causes kcat to decrease. In the mutant D313C the kcat is smaller than in other mutants (1200fold). Cys is ionizable and can potentially act as an acid/base catalyst, or the thiolate form can stabilize cationic intermediates electrostatically. This accounts for the higher catalytic activity for D313C than for D313A: The effects on Km(3-phosphoshikimate) or Km (phosphoenolpyruvate) are modest Escherichia coli
D313L compared to wild-type mutations of D313 causes kcat to decrease up to 30000fold while the effects on Km (3-phosphoshikimate) or Km (phosphoenolpyruvate) are modest, never more than 40fold Escherichia coli
D313N compared to wild-type mutations of D313 causes kcat to decrease up to 30000fold while the effects on Km (3-phosphoshikimate) or Km (phosphoenolpyruvate) are modest, never more than 40fold Escherichia coli
E341A compared to wild-type mutations of D313 causes kcat to decrease up to 30000fold while the effects on Km (3-phosphoshikimate) or Km (phosphoenolpyruvate) are modest, never more than 40fold Escherichia coli
E341C compared to wild-type mutations of D313 causes kcat to decrease up to 30000fold while the effects on Km (3-phosphoshikimate) or Km (phosphoenolpyruvate) are modest, never more than 40fold Escherichia coli
E341M compared to wild-type mutations of D313 causes kcat to decrease up to 30000fold while the effects on Km (3-phosphoshikimate) or Km (phosphoenolpyruvate) are modest, never more than 40fold Escherichia coli
E341Q compared to wild-type mutations of D313 causes kcat to decrease up to 30000fold while the effects on Km (3-phosphoshikimate) or Km (phosphoenolpyruvate) are modest, never more than 40fold Escherichia coli
H385A compared to wild-type mutation of H385 causes a 78fold decrease in kcat. Mutation has a 37fold effect on Km (phosphoenolpyruvate), almost no effect on Km (3-phosphoshikimate) Escherichia coli
K22A compared to wild-type, K22A mutation causes a more than760fold increase in Km (phosphoenolpyruvate) and a more than 100fold increase in Km(3-phosphoshikimate), indicating an important role in binding both substrates Escherichia coli
K22R compared to wild-type, K22R mutation has only a 10fold effect on kcat, presumably reflecting electrostatic stabilization of the anionic leaving groups in tetrahedral intermediate breakdown or possibly general acid catalysis by the guanidinium group. K22A mutation causes a 60fold increase in Km (phosphoenolpyruvate) Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.018
-
 3-phosphoshikimate C-terminal His6-tagged AroA, 50 mM KCl Escherichia coli
0.035
-
 3-phosphoshikimate 50 mM KCl, mutant E341M Escherichia coli
0.04
-
 3-phosphoshikimate 50 mM KCl, mutant E341Q Escherichia coli
0.04
-
 phosphoenolpyruvate C-terminal His6-tagged AroA, 50 mM KCl Escherichia coli
0.05
-
 phosphoenolpyruvate 50 mM KCl, mutant E341A Escherichia coli
0.08
-
 3-phosphoshikimate 50 mM KCl, mutant E341C Escherichia coli
0.09
-
 3-phosphoshikimate 50 mM KCl, mutant H385A Escherichia coli
0.1
-
 3-phosphoshikimate 50 mM KCl, mutant E341A Escherichia coli
0.11
-
 3-phosphoshikimate C-terminal His6-tagged AroA, 100 mM KCl Escherichia coli
0.14
-
 3-phosphoshikimate wild-type AroA, 100 mM KCl Escherichia coli
0.15
-
 phosphoenolpyruvate C-terminal His6-tagged AroA, 100 mM KCl Escherichia coli
0.16
-
 phosphoenolpyruvate wild-type AroA, 100 mM KCl Escherichia coli
0.17
-
 phosphoenolpyruvate 50 mM KCl, mutant E341C Escherichia coli
0.19
-
 phosphoenolpyruvate 50 mM KCl, mutant D313N Escherichia coli
0.19
-
 3-phosphoshikimate 50 mM KCl, mutant K22R value above 0.3 Escherichia coli
0.2
-
 3-phosphoshikimate 50 mM KCl, mutant D313N Escherichia coli
0.21
-
 3-phosphoshikimate 50 mM KCl, mutant D313A Escherichia coli
0.28
-
 phosphoenolpyruvate 50 mM KCl, mutant D313A Escherichia coli
0.4
-
 3-phosphoshikimate 50 mM KCl, mutant D313C Escherichia coli
0.7
-
 3-phosphoshikimate 50 mM KCl, mutant D313L Escherichia coli
0.9
-
 phosphoenolpyruvate 50 mM KCl, mutant E341Q Escherichia coli
1.3
-
 phosphoenolpyruvate 50 mM KCl, mutant D313C Escherichia coli
1.5
-
 phosphoenolpyruvate 50 mM KCl, mutant H385A Escherichia coli
1.6
-
 phosphoenolpyruvate 50 mM KCl, mutant D313L Escherichia coli
1.9
-
 phosphoenolpyruvate 50 mM KCl, mutant E341M Escherichia coli
2
-
 3-phosphoshikimate 50 mM KCl, mutant K22A value above 0.3 Escherichia coli
2.5
-
 phosphoenolpyruvate 50 mM KCl, mutant K22R value above 0.3 Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
K+
-
 Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
phosphoenolpyruvate + 3-phosphoshikimate Escherichia coli
-
 phosphate + 5-enolpyruvylshikimate 3-phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli P0A6D3
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
phosphoenolpyruvate + 3-phosphoshikimate
-
 Escherichia coli phosphate + 5-enolpyruvylshikimate 3-phosphate
-
 ?

Synonyms

Synonyms Comment Organism
AroA
-
 Escherichia coli
EPSP synthase
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.026
-
 phosphoenolpyruvate 50 mM KCl, mutant D313C, cosubstrate: 3-phosphoshikimate Escherichia coli
0.026
-
 3-phosphoshikimate 50 mM KCl, mutant D313C, cosubstrate: phosphoenolpyruvate Escherichia coli
0.3
-
 phosphoenolpyruvate 50 mM KCl, mutant K22A value above 0.3, cosubstrate: 3-phosphoshikimate Escherichia coli
0.3
-
 3-phosphoshikimate 50 mM KCl, mutant K22A value above 0.3, cosubstrate: phosphoenolpyruvate Escherichia coli
0.41
-
 phosphoenolpyruvate 50 mM KCl, mutant H385A, cosubstrate: 3-phosphoshikimate Escherichia coli
0.41
-
 3-phosphoshikimate 50 mM KCl, mutant H385A, cosubstrate: phosphoenolpyruvate Escherichia coli
1.1
-
 phosphoenolpyruvate 50 mM KCl, mutant D313L, cosubstrate: 3-phosphoshikimate Escherichia coli
1.1
-
 3-phosphoshikimate 50 mM KCl, mutant D313L, cosubstrate: phosphoenolpyruvate Escherichia coli
1.1
-
 phosphoenolpyruvate 50 mM KCl, mutant E341Q, cosubstrate: 3-phosphoshikimate Escherichia coli
1.1
-
 3-phosphoshikimate 50 mM KCl, mutant E341Q, cosubstrate: phosphoenolpyruvate Escherichia coli
1.2
-
 phosphoenolpyruvate 50 mM KCl, mutant D313A, cosubstrate: 3-phosphoshikimate Escherichia coli
1.2
-
 3-phosphoshikimate 50 mM KCl, mutant D313A, cosubstrate: phosphoenolpyruvate Escherichia coli
1.7
-
 phosphoenolpyruvate 50 mM KCl, mutant E341M, cosubstrate: 3-phosphoshikimate Escherichia coli
1.7
-
 3-phosphoshikimate 50 mM KCl, mutant E341M, cosubstrate: phosphoenolpyruvate Escherichia coli
1.8
-
 phosphoenolpyruvate 50 mM KCl, mutant D313N, cosubstrate: 3-phosphoshikimate Escherichia coli
1.8
-
 3-phosphoshikimate 50 mM KCl, mutant D313N, cosubstrate: phosphoenolpyruvate Escherichia coli
3
-
 phosphoenolpyruvate 50 mM KCl, mutant K22R value above 0.3, cosubstrate: 3-phosphoshikimate Escherichia coli
3
-
 3-phosphoshikimate 50 mM KCl, mutant K22R value above 0.3, cosubstrate: phosphoenolpyruvate Escherichia coli
3.6
-
 phosphoenolpyruvate 50 mM KCl, mutant E341C, cosubstrate: 3-phosphoshikimate Escherichia coli
3.6
-
 3-phosphoshikimate 50 mM KCl, mutant E341C, cosubstrate: phosphoenolpyruvate Escherichia coli
4.2
-
 phosphoenolpyruvate 50 mM KCl, mutant E341A, cosubstrate: 3-phosphoshikimate Escherichia coli
4.2
-
 3-phosphoshikimate 50 mM KCl, mutant E341A, cosubstrate: phosphoenolpyruvate Escherichia coli
14
-
 phosphoenolpyruvate wild-type AroA, 100 mM KCl, cosubstrate: 3-phosphoshikimate Escherichia coli
14
-
 3-phosphoshikimate wild-type AroA, 100 mM KCl, cosubstrate: phosphoenolpyruvate Escherichia coli
32
-
 phosphoenolpyruvate C-terminal His6-tagged AroA, 50 mM KCl, cosubstrate: 3-phosphoshikimate Escherichia coli
32
-
 3-phosphoshikimate C-terminal His6-tagged AroA, 50 mM KCl, cosubstrate: phosphoenolpyruvate Escherichia coli
100
-
 phosphoenolpyruvate C-terminal His6-tagged AroA, 100 mM KCl, cosubstrate: 3-phosphoshikimate Escherichia coli
100
-
 3-phosphoshikimate C-terminal His6-tagged AroA, 100 mM KCl, cosubstrate: phosphoenolpyruvate Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0007
-
 phosphoenolpyruvate 50 mM KCl, mutant D313L Escherichia coli
0.0009
-
 phosphoenolpyruvate 50 mM KCl, mutant E341M Escherichia coli
0.0012
-
 phosphoenolpyruvate 50 mM KCl, mutant K22R Escherichia coli
0.0013
-
 phosphoenolpyruvate 50 mM KCl, mutant E341Q Escherichia coli
0.0016
-
 3-phosphoshikimate 50 mM KCl, mutant D313L Escherichia coli
0.003
-
 phosphoenolpyruvate 50 mM KCl, mutant H385A Escherichia coli
0.004
-
 phosphoenolpyruvate 50 mM KCl, mutant D313A Escherichia coli
0.005
-
 3-phosphoshikimate 50 mM KCl, mutant E341A Escherichia coli
0.009
-
 phosphoenolpyruvate 50 mM KCl, mutant E341A Escherichia coli
0.009
-
 phosphoenolpyruvate 50 mM KCl, mutant K22A Escherichia coli
0.01
-
 phosphoenolpyruvate 50 mM KCl, mutant D313N Escherichia coli
0.02
-
 phosphoenolpyruvate 50 mM KCl, mutant D313C Escherichia coli
0.021
-
 phosphoenolpyruvate 50 mM KCl, mutant E341C Escherichia coli
0.027
-
 3-phosphoshikimate 50 mM KCl, mutant E341Q Escherichia coli
0.044
-
 3-phosphoshikimate 50 mM KCl, mutant E341C Escherichia coli
0.05
-
 3-phosphoshikimate 50 mM KCl, mutant E341M Escherichia coli
0.06
-
 3-phosphoshikimate 50 mM KCl, mutant D313A Escherichia coli
0.068
-
 3-phosphoshikimate 50 mM KCl, mutant D313C Escherichia coli
0.09
-
 3-phosphoshikimate 50 mM KCl, mutant D313N Escherichia coli
4
-
 3-phosphoshikimate 50 mM KCl, mutant H385A Escherichia coli
86
-
 phosphoenolpyruvate wild-type AroA, 100 mM KCl Escherichia coli
90
-
 phosphoenolpyruvate C-terminal His6-tagged AroA, 100 mM KCl Escherichia coli
96
-
 3-phosphoshikimate wild-type AroA, 100 mM KCl Escherichia coli
150
-
 3-phosphoshikimate C-terminal His6-tagged AroA, 100 mM KCl Escherichia coli
800
-
 phosphoenolpyruvate C-terminal His6-tagged AroA, 50 mM KCl Escherichia coli
1800
-
 3-phosphoshikimate C-terminal His6-tagged AroA, 50 mM KCl Escherichia coli