Information on EC 5.4.4.2 - Isochorismate synthase

New: Word Map on EC 5.4.4.2
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Search Reference ID:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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

EC NUMBER
COMMENTARY hide
5.4.4.2
-
RECOMMENDED NAME
GeneOntology No.
Isochorismate synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Chorismate = isochorismate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
group transfer
-
-
intramolecular
-
isomerization
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
1,4-dihydroxy-2-naphthoate biosynthesis
-
-
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.
CAS REGISTRY NUMBER
COMMENTARY hide
37318-53-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
Manually annotated by BRENDA team
gene dhbC
UniProt
Manually annotated by BRENDA team
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
strain 238-7
-
-
Manually annotated by BRENDA team
K3-15
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
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
-
-
-
Manually annotated by BRENDA team
fragment
UniProt
Manually annotated by BRENDA team
gene pchA
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Chorismate
?
show the reaction diagram
Chorismate
Isochorismate
show the reaction diagram
isochorismate
chorismate
show the reaction diagram
-
-
-
r
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
Chorismate
?
show the reaction diagram
Chorismate
Isochorismate
show the reaction diagram
additional information
?
-
-
the enzyme is essential for siderohore biosynthesis, first step in salicylate production
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
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 hide
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 hide
LITERATURE
IMAGE
0.0009 - 0.807
chorismate
0.005 - 0.675
isochorismate
additional information
additional information
-
steady state kinetic analysis
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.031 - 50.4
chorismate
1.8
isochorismate
Escherichia coli
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0022 - 95.7
chorismate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
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
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
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 hide
LITERATURE
0.241
pH 7.0, 30C, recombinant protein
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
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
7.6 - 8.2
-
enzyme immobilized on CNBr-Sepharose
7.8 - 7.9
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
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 hide
LITERATURE
42
-
soluble enzyme
60
-
enzyme immobilized on CNBr-Sepharose or alkylamine glass
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 37
more than 90% of maximum activity within this range
4 - 44
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
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
42000
-
isochorismate synthase involved in menaquinone biosynthesis, EntC, gel filtration
45000
-
gel filtration
48000 - 50000
gel filtration
59000
gel filtration
98000
-
isochorismate synthase involved in enterobactin biosynthesis, MenF, gel filtration
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
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
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
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
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
at 2.5 A resolution
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
in complex with isochorismate and Mg2+, hanging drop vapor diffusion method, using 100 mM MES buffer pH 6.5, 12% PEG (w/v) 20000
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 hide
LITERATURE
4
-
half-life of the enzyme immobilized on alkylamine glass: 210 days. Half-life of enzyme immobilized on CNBr Sepharose: 110 days
37
-
40 h, immobilized enzyme, stable
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
glycerol, EDTA and dithiothreitol stabilize
no activity in presence of 20 mM ascorbate or 5 mM thiourea. 43% decrease in activity in presence of 10 mM Cys
-
stability of the enzyme is greatly increased by immobilization
-
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
-
-80C, stable for at least 2 months
-
4C or -20C, 50% loss of activity after 4 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Co2+-ion affinity chromatography
DEAE-Sepharose, Phenyl-Sepharose, Mono Q
Ni2+-charged HiTrap chelating agarose column chromatography and Superdex 75 gel filtration
recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) pLysS by nickel affinity chromatography, gel filtration, and ultrafitration
-
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
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, phylogenetic analysis
-
expressed in Arabidopsis thaliana sid2-2 mutant
-
expressed in Brassica rapa subsp. oleifera
-
expressed in Escherichia coli
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli strain BL21 (DE3)
expression in Escherichia coli
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
gene pchA, overexpression of His-tagged enzyme in Escherichia coli strain BL21 (DE3) pLysS
-
overexpression in Escherichia coli BL21(DE3)
-
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 hide
LITERATURE
A303T
the kcat/Km (chorismate) for the A303T mutant is 130fold lower than that for wild type enzyme
A303T/F327Y
the mutant shows no detectable activity
A303T/F327Y/F359Q
the mutant shows no detectable activity
A303T/F327Y/F359Q/I346L
the mutant shows no detectable activity
A303T/F327Y/I346L
the mutant shows no detectable activity
A303T/F359Q
the mutant shows no detectable activity
A344T
no catalytic activity
E240Q
no catalytic activity
F327Y
the mutant displays reduced activity compared to the wild type enzyme
F327Y/F359Q
the mutant displays a reduced kcat/Km value compared to the wild type enzyme
F327Y/F359Q/I346L
the mutant shows no detectable activity
F359Q
the mutant displays a reduced kcat/Km value compared to the wild type enzyme; the mutation causes loss of catalytic activity
F359Q/I346L
the mutant displays a reduced kcat/Km value compared to the wild type enzyme
I346L
the EntC mutant has a 12fold lower kcat/Km (chorismate) than the wild type enzyme
K90A
no catalytic activity
L255A
50% increase in Km value, 90% decrease in kcat value
L304A
the mutation causes loss of catalytic activity
L304A/F359Q
the mutant shows no detectable activity
R387A
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
-
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
-
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 hide
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
Show AA Sequence (2058 entries)
Please use the Sequence Search for a certain query.