Information on EC 5.4.4.2 - Isochorismate synthase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
5.4.4.2
-
RECOMMENDED NAME
GeneOntology No.
Isochorismate synthase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Chorismate = isochorismate
show the reaction diagram
-
-
-
-
Chorismate = isochorismate
show the reaction diagram
K90 is the base that activates water for nucleophilic attack at the chorismate C2 carbon
P38051
Chorismate = isochorismate
show the reaction diagram
general acid-general base catalysis with a lysine as the base and a glutamic acid as the acid, in reverse protonation states and with a conformational or binding step as the slow step
-
Chorismate = isochorismate
show the reaction diagram
general acid-general base catalysis with a lysine as the base and a glutamic acid as the acid, in reverse protonation states and with a conformational or binding step as the slow step
Pseudomonas aeruginosa PA01
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
group transfer
-
-
intramolecular
-
isomerization
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
1,4-dihydroxy-2-naphthoate biosynthesis I
-
-
1,4-dihydroxy-2-naphthoate biosynthesis II (plants)
-
-
2,3-dihydroxybenzoate biosynthesis
-
-
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
Biosynthesis of siderophore group nonribosomal peptides
-
-
enterobactin biosynthesis
-
-
Metabolic pathways
-
-
salicylate biosynthesis I
-
-
Ubiquinone and other terpenoid-quinone biosynthesis
-
-
vitamin K metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
Isochorismate hydroxymutase
Requires Mg2+. The reaction is reversible.
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Amonabactin
-
-
-
-
DhbC
Q81QQ0
-
DhbC
Bacillus anthracis Ames
Q81QQ0
-
-
EC 5.4.99.6
-
-
formerly
-
Eds16
-
-
-
-
EntC
P0AEJ2
-
ICS
P0AEJ2
-
ICS1
-
isoform
ICS1
Q9M9V6
isozyme
ICS1
Arabidopsis thaliana Col-0
Q9S7H8
-
-
ICS1 gene product
Q9S7H8
-
ICS1 gene product
-
-
ICS2
-
isoform
ICS2
Q9M9V6
isozyme
IcsI
-
-
-
-
isochorismate mutase
-
-
-
-
isochorismate synthase
Q9S7H8
-
isochorismate synthase
-
-
isochorismate synthase 1
-
-
isochorismate synthase 1
Q9S7H8
-
isochorismate synthase 1
Arabidopsis thaliana Col-0
Q9S7H8
-
-
Isochorismic synthase
-
-
-
-
menaquinone-specific isochorismate synthase
P38051
-
MenF
P38051
-
PchA
Pseudomonas aeruginosa PA01
-
-
-
Sid2
-
-
-
-
Synthase, isochorismate
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
37318-53-9
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
activity is not regulated by light-dependent changes in stromal pH value, Mg2+ or redox potential
SwissProt
Manually annotated by BRENDA team
ecotype Col-0
-
-
Manually annotated by BRENDA team
Arabidopsis thaliana Col-0
-
SwissProt
Manually annotated by BRENDA team
gene dhbC
UniProt
Manually annotated by BRENDA team
Bacillus anthracis Ames
gene dhbC
UniProt
Manually annotated by BRENDA team
duplicate isochorismate genes, menF and dhbC. Wild-type strain and mutants carrying deletions of menF or dhbC
-
-
Manually annotated by BRENDA team
isochorismate synthase involved in enterobactin biosynthesis, MenF. The isochorismate synthase involved in menaquinone biosynthesis is different from the isochorismate synthase involved in enterobactin biosynthesis
-
-
Manually annotated by BRENDA team
isochorismate synthase involved in menaquinone biosynthesis, EntC
-
-
Manually annotated by BRENDA team
menaquinone-specific enzyme
SwissProt
Manually annotated by BRENDA team
strain AN191 and strain AN92
-
-
Manually annotated by BRENDA team
strain K-12
UniProt
Manually annotated by BRENDA team
K3-15; strain 238-7
-
-
Manually annotated by BRENDA team
Flavobacterium sp. 238-7
strain 238-7
-
-
Manually annotated by BRENDA team
Flavobacterium sp. K3-15
K3-15
-
-
Manually annotated by BRENDA team
enzyme is found in anthraquinone-producing cell line, but not in a non-producing cell line
-
-
Manually annotated by BRENDA team
Pseudomonas aeruginosa PA01
gene pchA
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
-
PchA is a member of the MST, i.e. menaquinone, siderophore and tryptophan, family of enzymes
evolution
Pseudomonas aeruginosa PA01
-
PchA is a member of the MST, i.e. menaquinone, siderophore and tryptophan, family of enzymes
-
malfunction
-
the redox status of the plastoquinone pool in knockout mutant ics1 shows significant variation depending on the leaf age. Mutant plants treated with a phylloquinone precursor display symptoms of phenotypic reversion towards the wild type. The ics1 mutant also shows altered thylakoid structure with an increased number of stacked thylakoids per granum
malfunction
Arabidopsis thaliana Col-0
-
the redox status of the plastoquinone pool in knockout mutant ics1 shows significant variation depending on the leaf age. Mutant plants treated with a phylloquinone precursor display symptoms of phenotypic reversion towards the wild type. The ics1 mutant also shows altered thylakoid structure with an increased number of stacked thylakoids per granum
-
metabolism
-
key enzyme in the isochorismate pathway
physiological function
-
Arabidopsis ICS1 represents a divergent isoform for inducible salicylic acid synthesis during defense
physiological function
-
Populus ICS primarily functions in phylloquinone biosynthesis
physiological function
Q81QQ0
the enzyme is essential for the biosynthesis of the siderophore bacillibactin by the pathogenic bacterium
physiological function
-
the enzyme is required for the appropriate hypersensitive disease defence response. It also takes part in the synthesis of phylloquinone, which is incorporated into photosystem I and is an important component of photosynthetic electron transport in plants role of ICS1 in regulation of state transition. Role of ICS1 in integration of the chloroplast ultrastructure, the redox status of the plastoquinone pool, and organization of the photosystems, which all are important for optimal immune defence and light acclimatory responses
physiological function
Bacillus anthracis Ames
-
the enzyme is essential for the biosynthesis of the siderophore bacillibactin by the pathogenic bacterium
-
physiological function
Arabidopsis thaliana Col-0
-
the enzyme is required for the appropriate hypersensitive disease defence response. It also takes part in the synthesis of phylloquinone, which is incorporated into photosystem I and is an important component of photosynthetic electron transport in plants role of ICS1 in regulation of state transition. Role of ICS1 in integration of the chloroplast ultrastructure, the redox status of the plastoquinone pool, and organization of the photosystems, which all are important for optimal immune defence and light acclimatory responses
-
metabolism
-
the enzyme produces isochorismate for conversion to salicylate by isochorismate-pyruvate lyase, PchB, and incorporation into the pyochelin siderophore. Isochorismate synthase (PchA) and isochorismate-pyruvate lyase (PchB) from Pseudomonas aeurginosa are involved in the synthesis of the siderophore pyochelin
additional information
Q81QQ0
enzyme structure comparisons and analysis, active site structure, overview. Ala304 plays an important role in positioning the peptide-bond carbonyl, enabling the formation of a proper hydrogen bond to the isochorismate C2 hydroxyl
additional information
-
solvent kinetic isotope effects, overview
additional information
-
structure-function relationships of chorismate-utilizing enzymes, structure comparisons, overview. Isochorismate synthase cannot perform any pericyclic reaction. Residues K221 and E269 are general base and acid, respectively
additional information
Bacillus anthracis Ames
-
enzyme structure comparisons and analysis, active site structure, overview. Ala304 plays an important role in positioning the peptide-bond carbonyl, enabling the formation of a proper hydrogen bond to the isochorismate C2 hydroxyl
-
additional information
Pseudomonas aeruginosa PA01
-
solvent kinetic isotope effects, overview
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
P38051
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
Q51508
-
-
r
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
r
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
r
Chorismate
Isochorismate
show the reaction diagram
P38051
-
-
-
ir
Chorismate
Isochorismate
show the reaction diagram
A9LS51
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
Q9M9V6
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
P0AEJ2
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
Q81QQ0
-
-
-
r
Chorismate
Isochorismate
show the reaction diagram
Q9S7H8
-
reaction operates near equilibrium, equilibrium constant is 0.89. Enzyme does not convert chorismate directly to salicylic acid
-
r
Chorismate
Isochorismate
show the reaction diagram
-
r
-
-
Chorismate
Isochorismate
show the reaction diagram
-
r
-
-
Chorismate
Isochorismate
show the reaction diagram
-
not reversible, isochorismate synthase involved in enterobactin biosynthesis, MenF
-
-
Chorismate
Isochorismate
show the reaction diagram
-
r, isochorismate synthase involved in menaquinone biosynthesis, EntC
-
-
Chorismate
Isochorismate
show the reaction diagram
Q51508
first enzyme of pyochelin biosynthesis
-
r
Chorismate
Isochorismate
show the reaction diagram
Q9S7H8
alternative pathway to produce salicyl acid in response to pathogens
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
involved in biosynthesis of salicylic acid
-
-
?
Chorismate
Isochorismate
show the reaction diagram
Flavobacterium sp. 238-7
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
Bacillus anthracis Ames
Q81QQ0
-
-
-
r
Chorismate
Isochorismate
show the reaction diagram
Arabidopsis thaliana Col-0
Q9S7H8
-
-
-
r
Chorismate
Isochorismate
show the reaction diagram
Flavobacterium sp. K3-15
-
r
-
-
Chorismate
Isochorismate
show the reaction diagram
Flavobacterium sp. K3-15
-
-
-
-
Chorismate
Isochorismate
show the reaction diagram
Pseudomonas aeruginosa PA01
-
-
-
-
r
Chorismate
?
show the reaction diagram
-
the first enzyme involved in the biosynthesis of the powerful iron-chelating agent enterobactin
-
-
-
Chorismate
?
show the reaction diagram
-
first committed step in the biosynthesis of menaquinone
-
-
-
Chorismate
?
show the reaction diagram
-
two different isochorismate mutases, one is involved in the biosynthesis of the respiratory chain component menaquinone, MedF, and the other is involved in the synthesis of siderophore 2,3-dihydroxybenzoate, DhbC
-
-
-
Chorismate
?
show the reaction diagram
-
enzyme catalyzes the pivotal step in enterobactin and menaquinone biosynthesis
-
-
-
isochorismate
chorismate
show the reaction diagram
Q9S7H8
-
-
-
r
additional information
?
-
-
the enzyme is essential for siderohore biosynthesis, first step in salicylate production
-
-
-
additional information
?
-
A9LS51
ICS enzyme is also required for salicylic acid biosynthesis
-
-
-
additional information
?
-
Q9M9V6
ICS2 participates in the synthesis of SA, but in limited amounts that become clearly detectable only when ICS1 is lacking
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
r
Chorismate
Isochorismate
show the reaction diagram
-
-
-
-
r
Chorismate
Isochorismate
show the reaction diagram
Q81QQ0
-
-
-
r
Chorismate
Isochorismate
show the reaction diagram
Q51508
first enzyme of pyochelin biosynthesis
-
r
Chorismate
Isochorismate
show the reaction diagram
Q9S7H8
alternative pathway to produce salicyl acid in response to pathogens
-
-
?
Chorismate
Isochorismate
show the reaction diagram
-
involved in biosynthesis of salicylic acid
-
-
?
Chorismate
?
show the reaction diagram
-
the first enzyme involved in the biosynthesis of the powerful iron-chelating agent enterobactin
-
-
-
Chorismate
?
show the reaction diagram
-
first committed step in the biosynthesis of menaquinone
-
-
-
Chorismate
?
show the reaction diagram
-
two different isochorismate mutases, one is involved in the biosynthesis of the respiratory chain component menaquinone, MedF, and the other is involved in the synthesis of siderophore 2,3-dihydroxybenzoate, DhbC
-
-
-
Chorismate
?
show the reaction diagram
-
enzyme catalyzes the pivotal step in enterobactin and menaquinone biosynthesis
-
-
-
Chorismate
Isochorismate
show the reaction diagram
Bacillus anthracis Ames
Q81QQ0
-
-
-
r
Chorismate
Isochorismate
show the reaction diagram
Arabidopsis thaliana Col-0
Q9S7H8
-
-
-
r
Chorismate
Isochorismate
show the reaction diagram
Pseudomonas aeruginosa PA01
-
-
-
-
r
additional information
?
-
-
the enzyme is essential for siderohore biosynthesis, first step in salicylate production
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
-
stimulates, optimal concentration: 2-5 mM
Mg2+
-
sharp increase in activity, optimal concentration: 1 mM, inhibition above 1 mM
Mg2+
-
absolute requirement; optimal concentration for soluble enzyme: 5-20 mM. Optimal concentration for enzyme immobilized on CNBr-Sepharose or alkylamine glass: 2.5-5 mM. Activity of the soluble enzyme without Mg2+ is 5.5% of the activity without Mg2+
Mg2+
-
absolute requirement
Mg2+
Q51508
strictly required for activity
Mg2+
P38051
required
Mg2+
Q9S7H8
absolutely required
Mg2+
P38051
required for catalysis
Mg2+
P0AEJ2
the enzyme contains Mg2+
Mg2+
Q81QQ0
Mg2+-dependent catalytic mechanism
Mg2+
-
required
Mg2+
-
required, chelated by residues D310, E313, and H348
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(4R,5R)-4-hydroxy-5-(1-carboxyvinyloxy)-cyclohex-1-ene carboxylate
-
-
(4R,5R)-4-hydroxy-5-carboxymethoxy-cyclohex-1-enecarboxylate
-
-
(4R,5R)-5-(2-carboxy-allyloxy)-4-hydroxy-cyclohex-1-enecarboxylate
-
-
(4R,5R,6S)-6-ammonio-5-[(1-carboxylatoethenyl)oxy]-4-hydroxycyclohex-1-ene-1-carboxylate
-
-
(4R,5R,7R)-5-(1-carboxy-ethoxy)-4-hydroxy-cyclohex-1-enecarboxylate
-
-
(4R,5R,7S)-5-(1-carboxy-ethoxy)-4-hydroxy-cyclohex-1-enecarboxylate
-
-
(4R,5S,6S)-4-ammonio-5-[(1-carboxylatoethenyl)oxy]-6-hydroxycyclohex-1-ene-1-carboxylate
-
-
(4R,5S,6S)-5-[(1-carboxylatoethenyl)oxy]-4,6-dihydroxycyclohex-1-ene-1-carboxylate
-
-
Mg2+
-
above 1 mM
salicylic acid
-
treatment of plants suppresses the enhancement of enzyme expression by O3
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2-mercaptoethanol
-
stimulates, 2.25fold stimulation at 20-30 mM
dithiothreitol
-
stimulates, 2fold stimulation at 1 mM
O3
-
exposure to O3 enhances the accumulation of salicylic acid and increases enzyme activity, but does not affect phenylalanine ammonia lyase activity. Treatment of plants suppresses the enhancement of enzyme expression by O3
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0009
chorismate
-
pH 7.5, 25C, wild-type enzyme
0.0045
chorismate
Q51508
-
0.007
chorismate
P0AEJ2
wild type enzyme, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.012
chorismate
-
pH 7.5, 25C, mutant E313Q
0.014
chorismate
-
-
0.017
chorismate
-
pH 7.5, 25C, mutant E313A
0.021
chorismate
-
pH 7.5, 25C, mutant K221A
0.04
chorismate
P0AEJ2
mutant enzyme F327Y/F359Q, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.0415
chorismate
Q9S7H8
pH 7.0, 30C
0.066
chorismate
P0AEJ2
mutant enzyme F359Q, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.076
chorismate
P0AEJ2
mutant enzyme F327Y, in 100 mM phosphate pH 7.0, 10 mM MgCl2; mutant enzyme I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.103
chorismate
P0AEJ2
mutant enzyme A303T, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.192
chorismate
P38051
wild-type, pH 7.5, 37C
0.195
chorismate
-
-
0.261
chorismate
P0AEJ2
mutant enzyme F359Q/I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.27
chorismate
-
enzyme immobilized on CNBr-Sepharose
0.313
chorismate
-
enzyme immobilized on alkylamine glass
0.319
chorismate
P38051
mutant L255A, pH 7.5, 37C
0.35
chorismate
-
soluble enzyme
0.466
chorismate
P0AEJ2
mutant enzyme F327Y/I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.807
chorismate
-
-
0.005
isochorismate
-
-
0.076
isochorismate
-
enzyme immobilized on alkylamine glass
0.171
isochorismate
-
enzyme immobilized on CNBr-Sepharose
0.254
isochorismate
-
soluble enzyme
0.675
isochorismate
-
-
additional information
additional information
-
steady state kinetic analysis
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.031
chorismate
P38051
mutant L255A, pH 7.5, 37C
0.0455
chorismate
-
pH 7.5, 25C, mutant K221A
0.05
chorismate
P0AEJ2
mutant enzyme F327Y/I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.067
chorismate
P0AEJ2
mutant enzyme A303T, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.108
chorismate
-
pH 7.5, 25C, mutant E313A
0.13
chorismate
P0AEJ2
mutant enzyme F327Y, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.18
chorismate
P0AEJ2
mutant enzyme F359Q/I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.35
chorismate
P38051
wild-type, pH 7.5, 37C
0.43
chorismate
P0AEJ2
mutant enzyme F327Y/F359Q, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.55
chorismate
P0AEJ2
mutant enzyme I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.6
chorismate
P0AEJ2
mutant enzyme F359Q, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.645
chorismate
Q9S7H8
pH 7.0, 30C
0.67
chorismate
P0AEJ2
wild type enzyme, in 100 mM phosphate pH 7.0, 10 mM MgCl2
0.72
chorismate
Q51508
-
0.75
chorismate
-
pH 7.5, 25C, mutant E313Q
1.33
chorismate
-
in absence of 2-mercaptoethanol
2.88
chorismate
-
-
2.93
chorismate
-
in presence of 2-mercaptoethanol
6.08
chorismate
Q51508
-
6.08
chorismate
Q9S7H8
pH 7.0, 30C
50.4
chorismate
-
pH 7.5, 25C, wild-type enzyme
1.8
isochorismate
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0022
chorismate
-
pH 7.5, 25C, mutant K221A
204
0.0064
chorismate
-
pH 7.5, 25C, mutant E313A
204
0.06
chorismate
-
pH 7.5, 25C, mutant E313Q
204
0.11
chorismate
P0AEJ2
mutant enzyme F327Y/I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2
204
0.65
chorismate
P0AEJ2
mutant enzyme A303T, in 100 mM phosphate pH 7.0, 10 mM MgCl2
204
0.69
chorismate
P0AEJ2
mutant enzyme F359Q/I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2
204
1.71
chorismate
P0AEJ2
mutant enzyme F327Y, in 100 mM phosphate pH 7.0, 10 mM MgCl2
204
7.24
chorismate
P0AEJ2
mutant enzyme I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2
204
9.1
chorismate
P0AEJ2
mutant enzyme F359Q, in 100 mM phosphate pH 7.0, 10 mM MgCl2
204
10.75
chorismate
P0AEJ2
mutant enzyme F327Y/F359Q, in 100 mM phosphate pH 7.0, 10 mM MgCl2
204
57
chorismate
-
pH 7.5, 25C, wild-type enzyme
204
95.7
chorismate
P0AEJ2
wild type enzyme, in 100 mM phosphate pH 7.0, 10 mM MgCl2
204
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.03
(4R,5R)-4-hydroxy-5-(1-carboxyvinyloxy)-cyclohex-1-ene carboxylate
-
-
2
(4R,5R)-4-hydroxy-5-carboxymethoxy-cyclohex-1-enecarboxylate
-
IC50 above 2.0 mM
2
(4R,5R)-5-(2-carboxy-allyloxy)-4-hydroxy-cyclohex-1-enecarboxylate
-
IC50 above 2.0 mM
0.00005
(4R,5R,6S)-6-ammonio-5-[(1-carboxylatoethenyl)oxy]-4-hydroxycyclohex-1-ene-1-carboxylate
-
-
2
(4R,5R,7R)-5-(1-carboxy-ethoxy)-4-hydroxy-cyclohex-1-enecarboxylate
-
IC50 above 2.0 mM
2
(4R,5R,7S)-5-(1-carboxy-ethoxy)-4-hydroxy-cyclohex-1-enecarboxylate
-
IC50 above 2.0 mM
0.00045
(4R,5S,6S)-4-ammonio-5-[(1-carboxylatoethenyl)oxy]-6-hydroxycyclohex-1-ene-1-carboxylate
-
-
0.00036
(4R,5S,6S)-5-[(1-carboxylatoethenyl)oxy]-4,6-dihydroxycyclohex-1-ene-1-carboxylate
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0017
Q51508
-
0.241
Q9S7H8
pH 7.0, 30C, recombinant protein
additional information
-
-
additional information
-
HPLC assay allows a rapid and accurate determination
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.5 - 9
-
about 20% of maximal activity at pH 6.5 and pH 9.0
7.4 - 8.5
-
enzyme immobilized on alkylamine glass
7.5 - 8
-
-
7.5 - 8
Q9S7H8
-
7.6 - 8.2
-
enzyme immobilized on CNBr-Sepharose
7.8 - 7.9
-
-
8
-
soluble enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.5 - 8.5
-
pH 5.5: about 20% of maximal activity, pH 8.5: about 90% of maximal activity
5.5 - 8.8
-
about 50% of maximal activity at pH 5.5 and at pH 8.8
additional information
-
PchA pH rate profile, overview
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
42
-
soluble enzyme
60
-
enzyme immobilized on CNBr-Sepharose or alkylamine glass
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4 - 37
Q9S7H8
more than 90% of maximum activity within this range
4 - 44
Q9S7H8
more than 75% of maximum activity within this range
15 - 50
-
15C: 72% of maximal activity, 20C: about 74% of maximal activity, 30C or 40C: about 80% of maximal activity, 50C: 42% of maximal activity
70
Q9S7H8
no activity
PDB
SCOP
CATH
ORGANISM
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
42000
-
isochorismate synthase involved in menaquinone biosynthesis, EntC, gel filtration
492270
45000
-
gel filtration
492274
48000 - 50000
Q51508
gel filtration
652382
50000
Q51508
native PAGE
652382
59000
Q9S7H8
gel filtration
680876
98000
-
isochorismate synthase involved in enterobactin biosynthesis, MenF, gel filtration
492271
additional information
-
identification and sequencing of the gene menF that encodes the enzyme that is responsible for menaquinone biosynthesis. The sequence of MenF is 23.5% identical and 57.8% similar to that of EntC
492280
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 48777, isochorismate synthase involved in menaquinone biosynthesis, calculation from nucleotide sequence, x * 49000, isochorismate synthase involved in menaquinone biosynthesis, SDS-PAGE
dimer
-
2 * 48000, isochorismate synthase involved in enterobactin biosynthesis, MenF, SDS-PAGE
monomer
-
-
monomer
Q9S7H8
1 * 59000, SDS-PAGE
monomer
Q51508
1 * 50000, SDS-PAGE
monomer
-
1 * 43000, isochorismate synthase involved in menaquinone biosynthesis, EntC, SDS-PAGE
monomer
-
1 * 36240, SDS-PAGE, a protein with MW 32770 also detected by SDS-PAGE is generated from the 36240 MW protein during the purification procedure
monomer
-
in solution
monomer
Flavobacterium sp. 238-7, Flavobacterium sp. K3-15
-
1 * 36240, SDS-PAGE, a protein with MW 32770 also detected by SDS-PAGE is generated from the 36240 MW protein during the purification procedure
-
additional information
Q81QQ0
enzyme structure comparisons and analysis, overview
additional information
Bacillus anthracis Ames
-
enzyme structure comparisons and analysis, overview
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
Q9S7H8
precursor protein of 62000 Da is cleaved to 58000 Da upon import into plastid
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant detagged selenomethionine-labeled enzyme, hanging drop vapor diffusion method, mixing of 0.002 ml of protein solution containing 16 mg/ml protein in 300 mM NaCl, 10 mM HEPES, pH 7.5, 0.5 mM TCEP, with 0.002 ml of well solution containing 2 M ammonium sulfate, 2% v/v PEG 400, 100 mM HEPES. pH 7.5, 50 mM Bis-Tris, pH 5.5, X-ray diffraction structure determination and analysis at 2.4 A resolution, single-wavelength anomalous diffraction
Q81QQ0
at 2.5 A resolution
P38051
enzyme without Mg2+, hanging drop vapour diffusion method, using 0.2 M trisodium citrate, 6% glycerol, 8% PEG 3350, enzyme in complex with Mg2+, sitting drop vapour diffusion method, using 0.1 M bis-tris pH 6.5, 20% PEG MME 5000
P38051
in complex with isochorismate and Mg2+, hanging drop vapor diffusion method, using 100 mM MES buffer pH 6.5, 12% PEG (w/v) 20000
P0AEJ2
sitting-drop vapur diffusion, crystals diffrect to a maximum resolution of 1.8 A. They belong to space group O2(1)2(1)2(1) with unit-cell parameters a = 51.8 A, b = 163.4 A, c = 194.9 A
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4
-
half-life of the enzyme immobilized on alkylamine glass: 210 days. Half-life of enzyme immobilized on CNBr Sepharose: 110 days
492273
37
-
40 h, immobilized enzyme, stable
492273
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
stability of the enzyme is greatly increased by immobilization
-
no activity in presence of 20 mM ascorbate or 5 mM thiourea. 43% decrease in activity in presence of 10 mM Cys
-
glycerol, EDTA and dithiothreitol stabilize
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 40% loss of activity after 5 days. 1% bovine serum albumin, 10% glycerol, or 20% dimethyl sulfoxide stabilize for up to 10 days
-
4C or -20C, 50% loss of activity after 4 days
-
-80C, stable for at least 2 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant selenomethionine-substituted, His-tagged enzyme from Escherichia coli strain BL21-CodonPlus(DE3)-RIPL by nickel affinity chromatography, His-tag cleavage, and a second step of nickel affinity chromatography
Q81QQ0
Co2+-ion affinity chromatography
P38051
Ni2+-charged HiTrap chelating agarose column chromatography and Superdex 75 gel filtration
P0AEJ2
DEAE-Sepharose, Phenyl-Sepharose, Mono Q
Q51508
recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) pLysS by nickel affinity chromatography, gel filtration, and ultrafitration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
Q9M9V6
expression in Escherichia coli
Q9S7H8
gene dhbC, overexpression in Escherichia coli strain BL21-CodonPlus(DE3)-RIPL as His-tagged enzyme from vector pMCSG19c in fusion N-terminally with maltose binding protein and a Tobacco vein mottling virus protease cleaving site, the His-tag is cleavable by Tobacco etch virus protease, in M9 SeMET High-Yield growth medium
Q81QQ0
DNA and amino acid sequence determination and analysis, phylogenetic analysis
-
expressed in Brassica rapa subsp. oleifera
-
expressed in Escherichia coli BL21(DE3) cells
P0AEJ2
expressed in Escherichia coli strain BL21 (DE3)
P38051
overexpression in Escherichia coli BL21(DE3)
-
expressed in Arabidopsis thaliana sid2-2 mutant
-
gene pchA, overexpression of His-tagged enzyme in Escherichia coli strain BL21 (DE3) pLysS
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
prevention of NPR1 nuclear localization causes over-accumulation of ICS1 transcripts in response to pathogen infection
-
the enzyme expression is induced after wounding, salt stress, and by salicylic acid
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A303T
P0AEJ2
the kcat/Km (chorismate) for the A303T mutant is 130fold lower than that for wild type enzyme
A303T/F327Y
P0AEJ2
the mutant shows no detectable activity
A303T/F327Y/F359Q
P0AEJ2
the mutant shows no detectable activity
A303T/F327Y/F359Q/I346L
P0AEJ2
the mutant shows no detectable activity
A303T/F327Y/I346L
P0AEJ2
the mutant shows no detectable activity
A303T/F359Q
P0AEJ2
the mutant shows no detectable activity
A344T
P38051
no catalytic activity
E240Q
P38051
no catalytic activity
F327Y
P0AEJ2
the mutant displays reduced activity compared to the wild type enzyme
F327Y/F359Q
P0AEJ2
the mutant displays a reduced kcat/Km value compared to the wild type enzyme
F327Y/F359Q/I346L
P0AEJ2
the mutant shows no detectable activity
F359Q
P0AEJ2
the mutant displays a reduced kcat/Km value compared to the wild type enzyme, the mutation causes loss of catalytic activity
F359Q/I346L
P0AEJ2
the mutant displays a reduced kcat/Km value compared to the wild type enzyme
I346L
P0AEJ2
the EntC mutant has a 12fold lower kcat/Km (chorismate) than the wild type enzyme
K90A
P38051
no catalytic activity
L255A
P38051
50% increase in Km value, 90% decrease in kcat value
L304A
P0AEJ2
the mutation causes loss of catalytic activity
L304A/F359Q
P0AEJ2
the mutant shows no detectable activity
R387A
P38051
no catalytic activity
E269A
-
site-directed mutagenesis, the residue acts as general acid, inactive mutant
E313A
-
site-directed mutagenesis, the residue chelates Mg2+ and acts as general base, mutant with highly reduced activity compared to wild-type
E313Q
-
site-directed mutagenesis, the residue chelates Mg2+ and acts as general base, mutant with highly reduced activity compared to wild-type
K221A
-
site-directed mutagenesis, the residue acts as general base, mutant with highly reduced activity compared to wild-type
E269A
Pseudomonas aeruginosa PA01
-
site-directed mutagenesis, the residue acts as general acid, inactive mutant
-
E313A
Pseudomonas aeruginosa PA01
-
site-directed mutagenesis, the residue chelates Mg2+ and acts as general base, mutant with highly reduced activity compared to wild-type
-
E313Q
Pseudomonas aeruginosa PA01
-
site-directed mutagenesis, the residue chelates Mg2+ and acts as general base, mutant with highly reduced activity compared to wild-type
-
K221A
Pseudomonas aeruginosa PA01
-
site-directed mutagenesis, the residue acts as general base, mutant with highly reduced activity compared to wild-type
-
additional information
-
plants defective in enzyme activity due to mutation sid2, level of salicylic acid and the activity of enzyme do not increase in response to O3 exposure
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
agriculture
-
the enzyme can be considered in plant breeding programs for salinity tolerance as well as for pathogen resistance for the oilseed industrial medical plant Carthamus tinctorius