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Information on EC 5.4.4.2 - isochorismate synthase and Organism(s) Escherichia coli and UniProt Accession P0AEJ2

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EC Tree
     5 Isomerases
         5.4 Intramolecular transferases
             5.4.4 Transferring hydroxy groups
                5.4.4.2 isochorismate synthase
IUBMB Comments
Requires Mg2+. The reaction is reversible.
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This record set is specific for:
Escherichia coli
UNIPROT: P0AEJ2
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Word Map
The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Reaction Schemes
Synonyms
ics, isochorismate synthase, amonabactin, salicylate synthase, isochorismate synthase 1, eds16, atics1, isochorismate hydroxymutase, menaquinone-specific isochorismate synthase, atics2, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Amonabactin
-
-
-
-
Eds16
-
-
-
-
IcsI
-
-
-
-
isochorismate mutase
-
-
-
-
Isochorismic synthase
-
-
-
-
menaquinone-specific isochorismate synthase
-
Sid2
-
-
-
-
Synthase, isochorismate
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
chorismate = isochorismate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
isomerization
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
isochorismate hydroxymutase
Requires Mg2+. The reaction is reversible.
CAS REGISTRY NUMBER
COMMENTARY hide
37318-53-9
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Chorismate
Isochorismate
show the reaction diagram
-
-
-
?
Chorismate
?
show the reaction diagram
Chorismate
Isochorismate
show the reaction diagram
isochorismate
chorismate
show the reaction diagram
-
-
-
r
NATURAL SUBSTRATE
NATURAL 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
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
the enzyme contains Mg2+
INHIBITOR
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)-5-(carboxymethoxy)-4-hydroxycyclohex-1-ene-1-carboxylic acid
-
(4R,5R)-5-[(1-carboxyethenyl)oxy]-4-hydroxycyclohex-1-ene-1-carboxylic acid
inhibition of isochorismate synthase activity and salicylate synthase activity
(4R,5R)-5-[(1R)-1-carboxyethoxy]-4-hydroxycyclohex-1-ene-1-carboxylic acid
-
(4R,5R)-5-[(1S)-1-carboxyethoxy]-4-hydroxycyclohex-1-ene-1-carboxylic acid
-
(4R,5R)-5-[(2-carboxyprop-2-en-1-yl)oxy]-4-hydroxycyclohex-1-ene-1-carboxylic acid
-
(4R,5R,6S)-6-amino-5-[(1-carboxyethenyl)oxy]-4-hydroxycyclohex-1-ene-1-carboxylic acid
-
(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-amino-5-[(1-carboxyethenyl)oxy]-6-hydroxycyclohex-1-ene-1-carboxylic acid
-
(4R,5S,6S)-4-ammonio-5-[(1-carboxylatoethenyl)oxy]-6-hydroxycyclohex-1-ene-1-carboxylate
-
-
(4R,5S,6S)-5-[(1-carboxyethenyl)oxy]-4,6-dihydroxycyclohex-1-ene-1-carboxylic acid
-
(4R,5S,6S)-5-[(1-carboxylatoethenyl)oxy]-4,6-dihydroxycyclohex-1-ene-1-carboxylate
-
-
Mg2+
-
above 1 mM
additional information
inhibitor structure-function relationship and molecular docking
-
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
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.007 - 0.466
chorismate
0.014 - 0.319
chorismate
0.005
isochorismate
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05 - 0.67
chorismate
0.031 - 2.93
chorismate
1.8
isochorismate
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.11 - 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
-
IC50 above 2.0 mM
2
(4R,5R)-5-(2-carboxy-allyloxy)-4-hydroxy-cyclohex-1-enecarboxylate
-
IC50 above 2.0 mM
0.03 - 0.16
(4R,5R)-5-[(1-carboxyethenyl)oxy]-4-hydroxycyclohex-1-ene-1-carboxylic acid
0.000053
(4R,5R,6S)-6-amino-5-[(1-carboxyethenyl)oxy]-4-hydroxycyclohex-1-ene-1-carboxylic acid
pH and temperature not specified in the publication
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
0.00046
(4R,5S,6S)-4-amino-5-[(1-carboxyethenyl)oxy]-6-hydroxycyclohex-1-ene-1-carboxylic acid
pH and temperature not specified in the publication
0.00045
(4R,5S,6S)-4-ammonio-5-[(1-carboxylatoethenyl)oxy]-6-hydroxycyclohex-1-ene-1-carboxylate
-
-
0.00036
(4R,5S,6S)-5-[(1-carboxyethenyl)oxy]-4,6-dihydroxycyclohex-1-ene-1-carboxylic acid
pH and temperature not specified in the publication
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
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 9
-
about 20% of maximal activity at pH 6.5 and pH 9.0
7.5 - 8
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 50
-
15°C: 72% of maximal activity, 20°C: about 74% of maximal activity, 30°C or 40°C: about 80% of maximal activity, 50°C: 42% of maximal activity
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
the first committed step during the biosynthesis of siderophores, which are small molecules capable of chelating iron from the host organism, is the conversion of chorismate into isochorismate by isochorismate synthase (EC 5.4.4.2) and consequently to salicylate by isochorismate pyruvate-lyase (EC 4.2.99.21). Salicylate synthase converts chorismate into salicylate through a two-step reaction
physiological function
chorismate-utilizing enzymes (CUE) such as chorismate mutase, anthranilate synthase, chorismate pyruvate-lyase, 4-amino-4-deoxychorismate synthase, isochorismate synthase and salicylate synthase are responsible for converting chorismate into various products necessary for the survival of bacteria
additional information
enzyme three-dimensional structure analysis, the lysine residue, Lys190, might be involved in the activation of water molecules and the subsequent nucleophilic attack on the C2 carbon of chorismate without directly involving the magnesium ion, participation of the Lys residue during the activation of the substrate or nucleophilic agent
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
42000
-
isochorismate synthase involved in menaquinone biosynthesis, EntC, gel filtration
43000
-
1 * 43000, isochorismate synthase involved in menaquinone biosynthesis, EntC, SDS-PAGE
48000
-
2 * 48000, isochorismate synthase involved in enterobactin biosynthesis, MenF, SDS-PAGE
48777
-
x * 48777, isochorismate synthase involved in menaquinone biosynthesis, calculation from nucleotide sequence
49000
-
x * 49000, isochorismate synthase involved in menaquinone biosynthesis, SDS-PAGE
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
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
2 * 48000, isochorismate synthase involved in enterobactin biosynthesis, MenF, SDS-PAGE
monomer
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
in complex with isochorismate and Mg2+, hanging drop vapor diffusion method, using 100 mM MES buffer pH 6.5, 12% PEG (w/v) 20000
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
PROTEIN VARIANTS
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
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
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
L304A
the mutation causes loss of catalytic activity
L304A/F359Q
the mutant shows no detectable activity
A344T
no catalytic activity
E240Q
no catalytic activity
K190A
inactive mutant
K90A
no catalytic activity
L255A
50% increase in Km value, 90% decrease in kcat value
R387A
no catalytic activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 40% loss of activity after 5 days. 1% bovine serum albumin, 10% glycerol, or 20% dimethyl sulfoxide stabilize for up to 10 days
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni2+-charged HiTrap chelating agarose column chromatography and Superdex 75 gel filtration
Co2+-ion affinity chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
expressed in Brassica rapa subsp. oleifera
-
expressed in Escherichia coli strain BL21 (DE3)
sequence comparisons
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Liu, J.; Quinn, N.; Berchthold, G.A.; Walsh, C.T.
Overexpression, purification, and characterization of isochorismate synthase (EnrC), the first enzyme involved in the biosynthesis of Enterobactin from Chorismate
Biochemistry
29
1417-1425
1990
Escherichia coli
Manually annotated by BRENDA team
Daruwala, R.; Bhattacharyya, D.K.; Kwon, O.; Meganathan, R.
Menaquinone (vitamin K2) biosynthesis: overexpression, purification, and characterization of a new isochorismate synthase from Escherichia coli
J. Bacteriol.
179
3133-3138
1997
Escherichia coli
Manually annotated by BRENDA team
Schaaf, P.M.; Heide, L.E.; Leistner, E.W.; Tani, Y.; Karas, M.; Deutzmann, R.
Properties of isochorismate hydroxymutase from Flavobacterium K3-15
J. Nat. Prod.
56
1294-1303
1993
Klebsiella aerogenes, Escherichia coli, Flavobacterium sp., Flavobacterium sp. 238-7, Flavobacterium sp. K3-15
Manually annotated by BRENDA team
Daruwala, R.; Kwon, O.; Meganathan, R.; Hudspeth, M.E.S.
A new isochorismate synthase specifically involved in menaquinone (vitamin K2) biosynthesis encoded by the menF gene
FEMS Microbiol. Lett.
140
159-163
1996
Escherichia coli
Manually annotated by BRENDA team
Muller, R.; Dahm, C.; Schulte, G.; Leistner, E.
An isochorismate hydroxymutase isogene in Escherichia coli
FEBS Lett.
378
131-134
1996
Escherichia coli
Manually annotated by BRENDA team
Kolappan, S.; Zwahlen, J.; Zhou, R.; Truglio, J.J.; Tonge, P.J.; Kisker, C.
Lysine 190 is the catalytic base in MenF, the menaquinone-specific isochorismate synthase from Escherichia coli: implications for an enzyme family
Biochemistry
46
946-953
2007
Escherichia coli (P38051), Escherichia coli
Manually annotated by BRENDA team
Parsons, J.F.; Shi, K.M.; Ladner, J.E.
Structure of isochorismate synthase in complex with magnesium
Acta Crystallogr. Sect. D
64
607-610
2008
Escherichia coli (P38051)
Manually annotated by BRENDA team
Sridharan, S.; Howard, N.; Kerbarh, O.; B?aszczyk, M.; Abell, C.; Blundell, T.L.
Crystal structure of Escherichia coli enterobactin-specific isochorismate synthase (EntC) bound to its reaction product isochorismate: implications for the enzyme mechanism and differential activity of chorismate-utilizing enzymes
J. Mol. Biol.
397
290-300
2010
Escherichia coli (P0AEJ2), Escherichia coli
Manually annotated by BRENDA team
Payne, R.J.; Bulloch, E.M.; Toscano, M.M.; Jones, M.A.; Kerbarh, O.; Abell, C.
Synthesis and evaluation of 2,5-dihydrochorismate analogues as inhibitors of the chorismate-utilising enzymes
Org. Biomol. Chem.
7
2421-2429
2009
Escherichia coli
Manually annotated by BRENDA team
Simoh, S.; Linthorst, H.J.; Lefeber, A.W.; Erkelens, C.; Kim, H.K.; Choi, Y.H.; Verpoorte, R.
Metabolic changes of Brassica rapa transformed with a bacterial isochorismate synthase gene
J. Plant Physiol.
167
1525-1532
2010
Escherichia coli
Manually annotated by BRENDA team
Svarcova, M.; Kratky, M.; Vinsova, J.
Investigation of potential inhibitors of chorismate-utilizing enzymes
Curr. Med. Chem.
22
1383-1399
2015
no activity in Homo sapiens, Yersinia pestis, Yersinia enterocolitica, Escherichia coli (P38051), Mycobacterium tuberculosis (P9WFX1), Pseudomonas aeruginosa (Q51508)
Manually annotated by BRENDA team