Information on EC 2.5.1.19 - 3-phosphoshikimate 1-carboxyvinyltransferase

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

EC NUMBER
COMMENTARY
2.5.1.19
-
RECOMMENDED NAME
GeneOntology No.
3-phosphoshikimate 1-carboxyvinyltransferase
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
phosphoenolpyruvate + 3-phosphoshikimate = phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
-
-
-
phosphoenolpyruvate + 3-phosphoshikimate = phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
proposed mechanism
-
phosphoenolpyruvate + 3-phosphoshikimate = phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
binding of shikimate 3-phosphate leads to a saturable and stable conformational change in the isolated N-terminal domain
-
phosphoenolpyruvate + 3-phosphoshikimate = phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
catalytic mechanism, tetrahedral reaction intermediate, catalytic cycle of the enzyme, overview
-
phosphoenolpyruvate + 3-phosphoshikimate = phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
catalytic mechanism, tetrahedral reaction intermediate, active site structure, wild-type and mutant D313A
phosphoenolpyruvate + 3-phosphoshikimate = phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
enolpyruvylshikimate 3-phosphate ketal is catalytically formed as side product upon enolpyruvyl activation through protonation by an intramolecular nucleophilic addition of O4 to C2 of the enolpyruvyl moiety
-
phosphoenolpyruvate + 3-phosphoshikimate = phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
via formation of a tetrahedral intermediate (TI) after which residue His385 is in a neutral form while residues Lys22, Lys340 and Lys411 are protonated, Asp313 mediates attack of the TI C4-OH group (as oxyanion, shikimate-3-phosphate moiety) at the TI C3-methyl group (phosphoenolpyruvate moiety), Lys22 serves as general acid catalyst for TI breakdown
-
phosphoenolpyruvate + 3-phosphoshikimate = phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
ordered substrate binding with 3-phosphoshikimate being bound first
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
enolpyruvate group transfer
-
-
-
-
enolpyruvate group transfer
-
-
enolpyruvate group transfer
-
enolpyruvate group transfer
induced-fit mechanism, formation of a 2-(S) configured tetrahedral reaction intermediate by covalent linkage of the two substrates
enolpyruvate group transfer
-
induced-fit mechanism, formation of a 2-(S) configured tetrahedral reaction intermediate by covalent linkage of the two substrates
enolpyruvate group transfer
-
induced-fit mechanism, formation of a 2-(S) configured tetrahedral reaction intermediate by covalent linkage of the two substrates
-
enolpyruvate group transfer
Dunaliella salina 435
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
chorismate biosynthesis from 3-dehydroquinate
-
-
chorismate metabolism
-
-
Phenylalanine, tyrosine and tryptophan biosynthesis
-
-
Metabolic pathways
-
-
Biosynthesis of secondary metabolites
-
-
Biosynthesis of antibiotics
-
-
SYSTEMATIC NAME
IUBMB Comments
phosphoenolpyruvate:3-phosphoshikimate 5-O-(1-carboxyvinyl)-transferase
-
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
3-enolpyruvylshikimate 5-phosphate synthase
-
-
-
-
3-enolpyruvylshikimic acid-5-phosphate synthetase
-
-
-
-
3-phosphoshikimate 1-carboxyvinyl-transferase
-
-
3-phosphoshikimate 1-carboxyvinyltransferase
-
-
5'-enolpyruvylshikimate-3-phosphate synthase
-
-
-
-
5-enol-pyruvylshikimate 3-phosphate synthase
-
-
5-enolpyruvyl-3-phosphoshikimate synthase
-
-
-
-
5-enolpyruvylshikimate 3-phosphate synthase
class II, glyphosate-tolerant
5-enolpyruvylshikimate 3-phosphate synthase
class II, glyphosate-tolerant
-
5-enolpyruvylshikimate 3-phosphate synthase
-
-
5-enolpyruvylshikimate 3-phosphate synthase
-
-
5-enolpyruvylshikimate 3-phosphate synthase
-
-
5-enolpyruvylshikimate 3-phosphate synthase
-
5-enolpyruvylshikimate 3-phosphate synthase
class I, glyphosate-sensitive
5-enolpyruvylshikimate 3-phosphate synthase
-
class II, glyphosate-tolerant
5-enolpyruvylshikimate-3-phosphate synthase
-
-
-
-
5-enolpyruvylshikimate-3-phosphate synthase
-
-
5-enolpyruvylshikimate-3-phosphate synthase
-
-
5-enolpyruvylshikimate-3-phosphate synthase
-
-
5-enolpyruvylshikimate-3-phosphate synthetase
-
-
-
-
5-enolpyruvylshikimate-3-phosphoric acid synthase
-
-
-
-
5-enolpyruvylshikimate-3phosphate synthase
-
-
5-enolpyruvylshikimate-3phosphate synthase
Bacteria ATX1398
-
-
-
5-enolpyruvylshikimate-3phosphate synthase
plant
-
-
5-enolpyruvylshikimic acid-3-phosphate synthase
-
-
-
-
5-enoylpyruvylshikimate 3-phosphate synthase
-
-
5-enoylpyruvylshikimate 3-phosphate synthase
-
-
-
A1501 EPSPS
-
class II EPSPS
A1501 EPSPS
-
class II EPSPS
-
AroA
-
gene name
AroA(1398)
-
glyphosate-tolerant gene product from strain ATX1398 that lacks protein sequence hallmarks of class II EPSP synthases
AroA(1398)
Bacteria ATX1398
-
glyphosate-tolerant gene product from strain ATX1398 that lacks protein sequence hallmarks of class II EPSP synthases
-
AroA(A1501)
-
-
AroA(A1501)
-
-
-
cp4 epsps
-
gene name
DsaroA
Dunaliella salina 435
-
-
Dunaliella salina 5-enolpyruvylshikimate-3-phosphate synthase
-
Dunaliella salina 5-enolpyruvylshikimate-3-phosphate synthase
Dunaliella salina 435
-
-
Dunaliella salina EPSP synthase
-
Dunaliella salina EPSP synthase
Dunaliella salina 435
-
-
E. coli EPSPS
-
enolpyruvylshikimate 3-phosphate synthase
-
-
enolpyruvylshikimate phosphate synthase
-
-
-
-
enolpyruvylshikimate phosphate synthase
-
-
enolpyruvylshikimate phosphate synthase
Bacteria ATX1398
-
-
-
enolpyruvylshikimate-3-phosphate synthase
-
-
EPSP synthase
-
-
-
-
EPSP synthase
-
EPSP synthase
-
-
EPSP synthase
-
-
EPSP synthase
Bacteria ATX1398
-
-
-
EPSP synthase
-
EPSP synthase
-
-
EPSP synthase
-
-
EPSP synthase
-
-
EPSPS
-
-
EPSPS
-
-
G2 5-enolpyruvyl shikimate 3-phosphate synthase
glyphosate resistant
G2 5-enolpyruvyl shikimate 3-phosphate synthase
Pseudomonas fluorescens G2
glyphosate resistant
-
G2 EPSPS
Pseudomonas fluorescens G2
-
-
maize EPSP synthase
plant
-
-
Mycobacterium tuberculosis 5-enolpyruvylshikimate-3-phosphate synthase
-
Mycobacterium tuberculosis 5-enolpyruvylshikimate-3-phosphate synthase
-
-
synthase, 5-enolpyruvoylshikimate 3-phosphate
-
-
-
-
maize EPSPS
plant
-
-
additional information
-
enzyme belongs to the family of enolpyruvyl transferases
additional information
-
class II 5-enolpyruvylshikimate-3-phosphate synthases are less sensitive to glyphosate in contrary to class I 5-enolpyruvylshikimate-3-phosphate synthases which are highly sensitive to inhibition with glyphosate
CAS REGISTRY NUMBER
COMMENTARY
9068-73-9
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain CP4
UniProt
Manually annotated by BRENDA team
strain CP4
UniProt
Manually annotated by BRENDA team
cyanobacterium
-
-
Manually annotated by BRENDA team
class II enzyme
UniProt
Manually annotated by BRENDA team
strain ATX1398; selected from soil by its ability to grow in presence of the herbicide glyphosate
-
-
Manually annotated by BRENDA team
Bacteria ATX1398
strain ATX1398; selected from soil by its ability to grow in presence of the herbicide glyphosate
-
-
Manually annotated by BRENDA team
transgenic Brassica napus x Brassica juncea outcrossing are examined
-
-
Manually annotated by BRENDA team
; strain 435
SwissProt
Manually annotated by BRENDA team
Dunaliella salina 435
strain 435
SwissProt
Manually annotated by BRENDA team
goosegrass
-
-
Manually annotated by BRENDA team
gene aroA
UniProt
Manually annotated by BRENDA team
gene aroA
-
-
Manually annotated by BRENDA team
K-12; overproducing recombinant strain AB2829pKD501
-
-
Manually annotated by BRENDA team
overproducing recombinant strain AB2829pKD501
-
-
Manually annotated by BRENDA team
strain N5259
-
-
Manually annotated by BRENDA team
Escherichia coli N5259
strain N5259
-
-
Manually annotated by BRENDA team
enzyme exists in 2 forms: one form is a monofunctional 59000 Da enzyme, the other form constitutes a single domain of the multifunctional 165000 Da arom protein
-
-
Manually annotated by BRENDA team
Halothermothrix orenii H168
-
UniProt
Manually annotated by BRENDA team
formerly Aerobacter aerogenes, glyphosate sensitive and resistant strains
-
-
Manually annotated by BRENDA team
lettuce
-
-
Manually annotated by BRENDA team
strain 74-OR23-1A
-
-
Manually annotated by BRENDA team
Neurospora crassa 74-OR23-1A
strain 74-OR23-1A
-
-
Manually annotated by BRENDA team
cv. Speg et Comes, aneuploid cell line ANS-1
-
-
Manually annotated by BRENDA team
no activity in mammals
-
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
pea, cv. Onward
-
-
Manually annotated by BRENDA team
plant
maize
-
-
Manually annotated by BRENDA team
strain PAO1
-
-
Manually annotated by BRENDA team
Pseudomonas fluorescens G2
G2
UniProt
Manually annotated by BRENDA team
strain A1501
-
-
Manually annotated by BRENDA team
Rahnella aquatilis GR20
-
UniProt
Manually annotated by BRENDA team
gene aroA, class II enzyme
-
-
Manually annotated by BRENDA team
wheat, cv. Maris Dove
-
-
Manually annotated by BRENDA team
mung bean
-
-
Manually annotated by BRENDA team
isoenzymes I and II
-
-
Manually annotated by BRENDA team
maize
-
-
Manually annotated by BRENDA team
var. Black Mexican Sweet, isoenzymes I and II
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
-
transgene inheritance and quantification upon wild Brassica juncea x transgenic Brassica napus outcrossing from three backcross generations and their progenies are examined using real-time PCR. Under in vitro conditions, a stable inheritance and similar cp4 epsps copy number between different generations and their progenies are shown. A marginally higher copy number is observed when Brassica juncea is used as the paternal as compared to when it is used as maternal parent
physiological function
enzyme is able to restore the growth of the Escherichia coli mutant ER2799, with the 5-enolpyruvylshikimate-3-phosphate synthase gene deleted from its genome, on an M9 minimal medium containing 60 mM glyphosate
physiological function
Rahnella aquatilis GR20
-
enzyme is able to restore the growth of the Escherichia coli mutant ER2799, with the 5-enolpyruvylshikimate-3-phosphate synthase gene deleted from its genome, on an M9 minimal medium containing 60 mM glyphosate
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
phosphate + 5-enolpyruvylshikimate 3-phosphate
shikimate 3-phosphate + phosphoenolpyruvate
show the reaction diagram
-
reverse reaction
phosphoenolpyruvate generation coupled to pyruvate kinase and lactate dehydrogenase and monitored spectrophotometrically at 340 nm, pH7.0, in presence of ADP, NADH, MgCl2
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
transfers enolpyruvate group from phosphoenolpyruvate to 5-hydroxyl group of 3-phosphoshikimate
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
sixth reaction of chorismate pathway
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
part of shikimate pathway
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
part of shikimate pathway
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
part of shikimate pathway
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
part of shikimate pathway
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
part of shikimate pathway
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
involved in chorismate biosynthesis
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
involved in aromatic acid biosynthesis
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
involved in aromatic acid biosynthesis
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
involved in aromatic acid biosynthesis
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
critical enzyme in the shikimate pathway
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
sixth enzyme in the shikimate pathway. The enzyme is widely involved in glyphosate tolerant transgenic plants because it is the primary target of the nonselective herbicide lyphosate
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
Neurospora crassa 74-OR23-1A
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
Dunaliella salina 435
sixth enzyme in the shikimate pathway. The enzyme is widely involved in glyphosate tolerant transgenic plants because it is the primary target of the nonselective herbicide lyphosate
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
Rahnella aquatilis GR20
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
Halothermothrix orenii H168
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
Escherichia coli N5259
-
-
i.e. 5-O-(1-carboxyvinyl)-3-phosphoshikimate
r
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
6th enzyme in the shikimate pathway
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
key enzyme in the shikimate pathway
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
penultimate step in the shikimate pathway
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
penultimate step in the shikimate pathway
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
substrate binding of wild-type and mutant D313A, overview
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
the 5-enolpyruvylshikimate-3-phosphate synthase catalyses the sixth step of the shikimate pathway that is responsible for synthesizing aromatic compounds, phosphate induces conformational change on the MtEPSPS structure
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
the enzyme must promote breakdown of enolpyruvylshikimate 3-phosphate though enolpyruvyl activation, forming either the enolpyruvylshikimate 3-phosphate cation or a highly cation-like transition state
-
?
phosphoenolpyruvate + 3-phosphoshikimate
?
show the reaction diagram
-
-
-
-
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
detection of inorganic phosphate by reaction with Lanzetta reagent and change in optical density at 660 nm
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
forward reaction
phosphate generation monitored by native gel analyses and subsequent calciumphosphate precipitation upon soaking in substrate solution in presence or absence of inhibitor glyphosate
r
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
1 mM shikimate 3-phosphate and 0.05-1.0 mM phosphoenolpyruvate
detection of inorganic phosphate generation by Malachite Green dye assay according to Lanzetta
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
30 min, 25C, pH 7.5, in presence of 2 mM dithiothreitol
detection of phosphate generation with Lanzetta reagent and change in absorbance at 650 nm
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
Bacteria, plant
-
pH 7, start with phosphoenolpyruvate, with or without inhibitor glyphosate
fluorescence-based coupled enzymatic assay including xanthine oxidase, purine nucleoside phosphorylase, inosine, horseradish peroxidase and Amplex Red
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
shikimate 3-phosphate binding site primarily at the N-terminal domain of the enzyme, phosphoenolpyruvate-binding mainly a the C-terminal domain
-
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
1 mM shikimate 3-phosphate and 0.05-1.0 mM phosphoenolpyruvate
detection of inorganic phosphate generation by Malachite Green dye assay according to Lanzetta
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
Bacteria ATX1398
-
pH 7, start with phosphoenolpyruvate, with or without inhibitor glyphosate
fluorescence-based coupled enzymatic assay including xanthine oxidase, purine nucleoside phosphorylase, inosine, horseradish peroxidase and Amplex Red
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enpolpyruvylshikimate 3-phosphate
show the reaction diagram
activity independent of transit peptide (N-terminal 70 residues)
growth of transformed Escherichia coli lacking endogenous EPSP synthase on M9 minimal medium by sustaining synthesis of aromatic amino acids through introduced 5-enolpyruvylshikimate 3-phosphate synthesis, leading to 10fold increase in glyphosate tolerance in bacteria compared to controls
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enpolpyruvylshikimate 3-phosphate
show the reaction diagram
-
enolpyruvylshikimate 3-phosphate ketal formation from shikimate 3-phosphate (radiolabelled) and phosphoenolypyruvate (in excess over shikimate 3-phosphate) by hydrolysis of product 5-enpolpyruvylshikimate 3-phosphate in presence of 25% excess of enzyme, pH 7.0, 25C
analysis of products by ion-paired C18 reverse-phase HPLC and scintillation counting
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enpolpyruvylshikimate 3-phosphate
show the reaction diagram
malachite green dye assay, 28C
-
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enpolpyruvylshikimate 3-phosphate
show the reaction diagram
-
via formation of a 2-(S)-tetrahedral reaction intermediate (TI), pH 7.5, 25C, 20 min
examination by malachite green phosphate-release assay
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enpolpyruvylshikimate 3-phosphate
show the reaction diagram
-
via formation of a 2-(S)-tetrahedral reaction intermediate (TI), pH 7.5, 25C, 30 min
examination by malachite green phosphate-release assay
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enpolpyruvylshikimate 3-phosphate
show the reaction diagram
via formation of a 2-(S)-tetrahedral reaction intermediate (TI), pH 7.5, 25C, 30 min
examination by malachite green phosphate-release assay
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enpolpyruvylshikimate 3-phosphate
show the reaction diagram
Pseudomonas fluorescens G2
malachite green dye assay, 28C
-
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enpolpyruvylshikimate 3-phosphate
show the reaction diagram
Dunaliella salina 435
activity independent of transit peptide (N-terminal 70 residues)
growth of transformed Escherichia coli lacking endogenous EPSP synthase on M9 minimal medium by sustaining synthesis of aromatic amino acids through introduced 5-enolpyruvylshikimate 3-phosphate synthesis, leading to 10fold increase in glyphosate tolerance in bacteria compared to controls
?
5-enolpyruvylshikimate 3-phosphate + H2O
5-enolpyruvylshikimate 3-phosphate ketal
show the reaction diagram
-
enzymatic hydrolysis, pH 7.0, 25C, in presence of phosphate scavenging system and excess of enzyme
-
?
additional information
?
-
Dunaliella salina, Dunaliella salina 435
sixth enzyme in shikimate pathway, biosynthesis of aromatic amino acids
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
Q93VK6
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
sixth reaction of chorismate pathway
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
part of shikimate pathway
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
part of shikimate pathway
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
part of shikimate pathway
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
part of shikimate pathway
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
part of shikimate pathway
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
involved in chorismate biosynthesis
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
involved in aromatic acid biosynthesis
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
involved in aromatic acid biosynthesis
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
involved in aromatic acid biosynthesis
-
-
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
Q1M157
critical enzyme in the shikimate pathway
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
A2T9R8
sixth enzyme in the shikimate pathway. The enzyme is widely involved in glyphosate tolerant transgenic plants because it is the primary target of the nonselective herbicide lyphosate
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
P0A6D3
6th enzyme in the shikimate pathway
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
key enzyme in the shikimate pathway
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
penultimate step in the shikimate pathway
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
penultimate step in the shikimate pathway
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
-
the 5-enolpyruvylshikimate-3-phosphate synthase catalyses the sixth step of the shikimate pathway that is responsible for synthesizing aromatic compounds
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
Dunaliella salina 435
A2T9R8
sixth enzyme in the shikimate pathway. The enzyme is widely involved in glyphosate tolerant transgenic plants because it is the primary target of the nonselective herbicide lyphosate
-
?
additional information
?
-
Dunaliella salina, Dunaliella salina 435
A2T9R8
sixth enzyme in shikimate pathway, biosynthesis of aromatic amino acids
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
no known cofactor
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
-
slightly activating
CaCl2
-
stimulation at 20 mM, inhibition above 100 mM
Cs+
-
slightly activating
K+
strong activation of forward reaction, effectiveness of activation in descending order: NH4Cl, Rb+, K+
K+
-
highly activating, best at 100 mM
Na+
-
activates
NH4+
-
highly activating
Rb+
strong activation of forward reaction, effectiveness of activation in descending order: NH4Cl, Rb+, K+
MgCl2
-
stimulation at 20 mM, inhibition above 100 mM
additional information
-
not activated by Ba2+, Ca2+, Mg2+, Ni2+ or Sr2+
additional information
-
no activation by Li+ and Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
RP-TI, (R)-phosphonate analogue of the (S)-tetrahedral reaction intermediate, competitive inhibitor of EPSPS forward reaction, 460fold decrease in potency compared to glyphosate-sensitive class I EPSPS from Escherichia coli
(3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
RP-TI, (R)-phosphonate analogue of the (S)-tetrahedral reaction intermediate, very potent competitive inhibitor of EPSPS forward reaction, binding induces substantial conformational change in enzymes backbone and active site (e.g., Glu341 and Arg124)
(3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
RP-TI, (R)-phosphonate analogue of the (S)-tetrahedral reaction intermediate, competitive inhibitor of EPSPS forward reaction, 120-fold decrease in potency compared to glyphosate-sensitive class I EPSPS from Escherichia coli
(3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
SP-TI, (S)-phosphonate analogue of the (S)-tetrahedral reaction intermediate, non-potent competitive inhibitor of EPSPS forward reaction
(3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
SP-TI, (S)-phosphonate analogue of the (S)-tetrahedral reaction intermediate, moderate competitive inhibitor of EPSPS forward reaction
(3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
SP-TI, (S)-phosphonate analogue of the (S)-tetrahedral reaction intermediate, competitive inhibitor of EPSPS forward reaction
(3R,4S,5R)-5-((R)-1-carboxy-1-phosphono-ethoxy)-4-hydroxy-3-phosphonooxy-cyclohex-1-enecarboxylic acid
-
analogue of the tetrahedral reaction intermediate, competitive to both substrates, binding structure analysis, binding induces conformational changes to residues Arg124 and Glu341 within the active site, which results in structural alterations in the amino-terminal globular domain of the enzyme
(3R,4S,5R)-5-((S)-1-carboxy-1-phosphono-ethoxy)-4-hydroxy-3-phosphonooxy-cyclohex-1-enecarboxylic acid
-
analogue of the tetrahedral reaction intermediate, competitive to both substrates, binding structure analysis, binding induces no conformational changes
(3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
2F-TI, (R)-difluoromethyl analogue of the (S)-tetrahedral reaction intermediate, competitive inhibitor of EPSPS forward reaction, 8fold decrease in potency compared to glyphosate-sensitive class I EPSPS from Escherichia coli
(3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
2F-TI, (R)-difluoromethyl analogue of the (S)-tetrahedral reaction intermediate, very potent competitive inhibitor of EPSPS forward reaction
(3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
2F-TI, (R)-difluoromethyl analogue of the (S)-tetrahedral reaction intermediate, competitive inhibitor of EPSPS forward reaction, 4-fold decrease in potency compared to glyphosate-sensitive class I EPSPS from Escherichia coli
(Z)-3-fluorophosphoenolpyruvate
-
competitive vs. phosphoenolpyruvate at saturated shikimate 3-phosphate concentration and vice versa
3-Bromopyruvate
-
0.1 mM, approx. 80% inactivation after 5 min, maximum rate-constant: 0.31/min, substrates or a combination of shikimate 3-phosphat and glyphosate protect from inactivation, bromopyruvate modifies residues C408 and L411
3-Bromopyruvate
-
1 mM, 50% inactivation after 3.4 min
3-Bromopyruvate
-
glyphosate protects
5-Deoxy-shikimate 3-phosphate
-
competitive vs. shikimate 3-phosphate
5-enolpyruvylshikimate 3-phosphate
-
product inhibition
5-enolpyruvylshikimate 3-phosphate
-
product inhibition
Al3+
-
10 mM, 37% inhibition
ammonium heptamolybdate
-
1.1 mM, 50% inhibition, no inhibition below 0.1 mM
Br-
-
inhibition above 200 mM, stimulation below
CaCl2
-
above 100 mM
Carboxyallenyl phosphate
-
strong
Cl-
-
inhibition above 400 mM, stimulation below
Co3+
-
10 mM, 25% inhibition
Cu2+
-
10 mM, 90-95% inhibition
Cu2+
-
5 mM, complete inhibition
diethyldicarbonate
-
inactivation with a second-order rate constant of 220/M/min, subtstrates protect from inactivation, enzyme activity is recovered by treatment with hydroxylamine
Fe2+
-
10 mM, complete inhibition
Fe2+
-
5 mM, 42% inhibition
Fe3+
-
10 mM, 60% inhibition
Fe3+
-
5 mM, 68% inhibition
glyphosate
-
competitive inhibition of the wild-type enzyme, mutant T42M is less sensitive
glyphosate
-
slight inhibition, insensitivity is independent from cations, mechanism
glyphosate
-
glyphosate
N-phosphonomethylglycine
glyphosate
-
-
glyphosate
Kd: 350 micromolar (pH 6.9, 25C), binding induces conformational change that leads to increase in alpha helix (40% to 57%) and decrease in beta-sheet (30% to 23%) content (revealed by circular dichroism spectroscopy) and prevents access of solvent to core molecule (revealed by hydrogen/deuterium exchange, peptic digestion and LC-ESI-MS)
glyphosate
-
N-(phosphonomethyl)glycine, broad-spectrum herbicide
glyphosate
-
at 300 mM: no growth of Escherichia coli transformed with either EPSPS wild-type or mutant N130S
glyphosate
-
3-phosphoshikimate binding is prerequisite for glyphosate binding
glyphosate
-
competitor with phosphoenolpyruvate binding site, AroA(1398) transformation allows Escherichia coli DH5alpha and XL1 Blue colonies to grow on up to 200 mM glyphosate (non-transformed bacteria stop growing on 5 mM inhibitor), AroA(1398) integrated maize transgenic plant lines showed 54% survival upon glyphosate treatment compared to none of control plants
glyphosate
plant
-
plants do not survive glyphosate spray treatment (1.78 l/ha of a 480 g AE/L formulation) in the field
glyphosate
-
inhibits forward reaction only slightly at 0.2 mM but substantially at 20 mM
I-
-
inhibition above 100 mM, stimulation below
La3+
-
10 mM, complete inhibition
MgCl2
-
above 100 mM
Mn2+
-
10 mM, 16% inhibition
Mn2+
-
5 mM, 42% inhibition
N-ethylmaleimide
-
1 mM, 50% inactivation after approx. 3 min
N-ethylmaleimide
-
glyphosate protects
N-phosphonomethylglycine
-
inhibition of enolpyruvate transfer, competitive vs. phosphoenolpyruvate
N-phosphonomethylglycine
-
-
N-phosphonomethylglycine
-
0.01 mM, 50% inhibition
N-phosphonomethylglycine
-
wild-type enzyme
N-phosphonomethylglycine
-
competitive vs. phosphoenolpyruvate, noncompetitive vs. shikimate 3-phosphate, 5-enolpyruvylshikimate 3-phosphate and phosphate, not inhibited by non-herbicidal analogues of glyphosate, e.g. aminomethylphosphonic acid, glyphosine, i.e. bis-N-(phosphonomethyl)glycine, or iminodiacetic acid; pH-dependent
N-phosphonomethylglycine
-
3 mM, 50% inhibition, 25 mM, 90% inhibition; trivial name glyphosate, non-selective herbicide 'Round up', free acid or monoisopropylamine salt
N-phosphonomethylglycine
-
-
N-phosphonomethylglycine
-
-
N-phosphonomethylglycine
-
noncompetitive vs. phosphate, competitive vs. phosphoenolpyruvate, uncompetitive vs. shikimate 3-phosphate
N-phosphonomethylglycine
-
-
N-phosphonomethylglycine
-
-
N-phosphonomethylglycine
-
0.0183 mM, 50% inhibition, 0.5 mM 96% inhibition, competitive vs. phosphoenolpyruvate, uncompetitive vs. shikimate 3-phosphate at pH 7.0, mechanism; pH-dependent
N-phosphonomethylglycine
-
0.006-0.008 mM, 50% inhibition, 1 mM, complete inhibition
N-phosphonomethylglycine
-
0.25 mM, 50% inhibition of NH4Cl activated enzyme, 75 mM, 50% inhibition of nonactivated enzyme
N-phosphonomethylglycine
-
N-phosphonomethylglycine
-
mechanism of inhibition in atomic detail
N-phosphonomethylglycine
-
0.006 mM, 50% inhibition of glyphosate-sensitve strain, 0.03 mM, 50% inhibition of glyphosate-resistant strain
NH4Cl
-
inhibition above 300 mM, stimulation below
NO3-
-
inhibition above 150 mM, stimulation below
Pb2+
-
10 mM, 90-95% inhibition
Pb2+
-
5 mM, 97% inhibition
Phenylglyoxal
-
2 mM, 50% inactivation after 32 min
Phenylglyoxal
-
glyphosate protects
phosphoenolpyruvate
-
substrate inhibition
phosphoenolpyruvate
-
-
pyruvate
-
20 mM, 85% inactivation after 1 h in the presence of cyanoborhydride, no inactivation in the absence of cyanoborhydride, preincubation with 5-enolpyruvylshikimate or a combination of shikimate 3-phosphate and glyphosate prevents from inactivation
Reaction intermediate analogue
-
-
-
shikimate 3-phosphate
-
product inhibition
shikimate 3-phosphate-5-carboxymethyl-(2R)-phosphonate
-
competitive vs. 5-enolpyruvylshikimate-3-phosphate and phosphate
shikimate 3-phosphate-5-carboxymethyl-(2S)-phosphonate
-
competitive vs. 5-enolpyruvylshikimate-3-phosphate and phosphate
SO42-
-
inhibition above 50 mM, slight stimulation below
Zn2+
-
10 mM, 90-95% inhibition
Zn2+
-
5 mM, complete inhibition
molybdate
-
10 mM, complete inhibition
additional information
-
insensitive to inhibition by glyphosate.
-
additional information
-
resistant to glyphosate
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Br-
-
stimulation at low concentrations, inhibition above 200 mM
Cl-
-
activation up to 100 mM, inhibition above
Cl-
-
at about 100 mM
F-
-
stimulation
F-
-
stimulation
I-
-
stimulation at low concentrations, inhibition above 100 mM
NH4Cl
-
stimulation, inhibition above 300 mM
NH4Cl
-
maximal activation at 100 mM
NH4Cl
strong activation of forward reaction, effectiveness of activation in descending order: NH4Cl, Rb+, K+
NO3-
-
stimulation at low concentrations, inhibition above 15 mM
NO3-
-
stimulation at low concentrations, inhibition above 15 mM
SO42-
-
slight stimulation, inhibition above 50 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00036
3-phosphoshikimate
-
-
0.0025
3-phosphoshikimate
-
recombinant overproducing strain
0.0032 - 0.0036
3-phosphoshikimate
-
wild-type enzyme
0.0032 - 0.0036
3-phosphoshikimate
-
-
0.007
3-phosphoshikimate
-
-
0.0077
3-phosphoshikimate
-
-
0.018
3-phosphoshikimate
-
C-terminal His6-tagged AroA, 50 mM KCl
0.035
3-phosphoshikimate
-
50 mM KCl, mutant E341M
0.04
3-phosphoshikimate
-
50 mM KCl, mutant E341Q
0.045
3-phosphoshikimate
-
glyphosate-sensitive enzyme
0.048
3-phosphoshikimate
-
wild-type
0.071
3-phosphoshikimate
-
mutant P101S
0.077
3-phosphoshikimate
-
mutant P101S/T97I; mutant T97I
0.08
3-phosphoshikimate
-
50 mM KCl, mutant E341C
0.084
3-phosphoshikimate
mutant G96A, pH 7.0, 28C
0.086
3-phosphoshikimate
wild-type, pH 7.0, 28C
0.09
3-phosphoshikimate
-
50 mM KCl, mutant H385A
0.1
3-phosphoshikimate
-
50 mM KCl, mutant E341A
0.11
3-phosphoshikimate
-
C-terminal His6-tagged AroA, 100 mM KCl
0.14
3-phosphoshikimate
-
pH 7.0, recombinant enzyme, in presence of 100 mM KCl
0.14
3-phosphoshikimate
-
wild-type AroA, 100 mM KCl
0.19
3-phosphoshikimate
-
50 mM KCl, mutant K22R value above 0.3
0.193
3-phosphoshikimate
-
glyphosate-resistant enzyme
0.2
3-phosphoshikimate
-
50 mM KCl, mutant D313N
0.21
3-phosphoshikimate
-
50 mM KCl, mutant D313A
0.241
3-phosphoshikimate
mutant T355S, pH 7.0, 28C
0.275
3-phosphoshikimate
pH 7.0, 28C
0.286
3-phosphoshikimate
wild-type, pH 7.0, 28C
0.304
3-phosphoshikimate
pH 7.0, 28C
0.4
3-phosphoshikimate
-
50 mM KCl, mutant D313C
0.7
3-phosphoshikimate
-
50 mM KCl, mutant D313L
2
3-phosphoshikimate
-
50 mM KCl, mutant K22A value above 0.3
0.00025
5-enolpyruvylshikimate 3-phosphate
-
-
0.001
5-enolpyruvylshikimate 3-phosphate
-
-
0.0019
5-enolpyruvylshikimate 3-phosphate
-
25C, pH 7.0, mutant enzyme G99A
0.00195
5-enolpyruvylshikimate 3-phosphate
reverse reaction, in the presence of 50 mM KCl
0.002
5-enolpyruvylshikimate 3-phosphate
-
25C, pH 7.0, wild-type enzyme
0.0025
5-enolpyruvylshikimate 3-phosphate
-
25C, pH 7.0, mutant enzyme L104P
0.003
5-enolpyruvylshikimate 3-phosphate
-
recombinant overproducing strain
0.0052
5-enolpyruvylshikimate 3-phosphate
-
-
0.007
5-enolpyruvylshikimate 3-phosphate
-
H385N mutant enzyme
0.011
5-enolpyruvylshikimate 3-phosphate
-
wild-type enzyme
1.9
5-enolpyruvylshikimate 3-phosphate
-
+/-0.3 mM, mutant G99A
2.2
5-enolpyruvylshikimate 3-phosphate
-
+/-0.5 mM
0.0018
phosphate
-
-
0.0056
phosphate
-
25C, pH 7.0, wild-type enzyme
0.0073
phosphate
-
25C, pH 7.0, mutant enzyme G99A
0.0248
phosphate
-
25C, pH 7.0, mutant enzyme L104P
0.343
phosphate
reverse reaction, in the presence of 50 mM KCl
1.2
phosphate
-
H385N mutant enzyme
2.5
phosphate
-
-
3.7
phosphate
-
-
4
phosphate
-
-
4.6
phosphate
-
wild-type enzyme
5.6
phosphate
-
+/-1.6
7.3
phosphate
-
+/-0.7 mM, mutant G99A
0.0018
phosphoenolpyruvate
-
pH 7.0, 30C, mutant StaroA-T42M
0.0024
phosphoenolpyruvate
-
pH 7.0, 30C, mutant EcaroA-T42M
0.0027
phosphoenolpyruvate
-
-
0.0038
phosphoenolpyruvate
-
glyphosate-sensitive strain
0.0052
phosphoenolpyruvate
-
cosubstrate 5-enolpyruvylshikimate 3-phosphate
0.0052
phosphoenolpyruvate
-
-
0.007
phosphoenolpyruvate
-
glyphosate-resistant strain
0.008
phosphoenolpyruvate
-
-
0.009
phosphoenolpyruvate
-
glyphosate-sensitive enzyme
0.0113
phosphoenolpyruvate
-
expressed in Escherichia coli
0.015
phosphoenolpyruvate
-
wild-type enzyme
0.016
phosphoenolpyruvate
-
recombinant overproducing strain
0.017
phosphoenolpyruvate
-
in the presence of glyphosate
0.0181
phosphoenolpyruvate
plant
-
-
0.0199
phosphoenolpyruvate
wild-type enzyme
0.021 - 0.023
phosphoenolpyruvate
-
-
0.022
phosphoenolpyruvate
in the presence of 100 mM NH4Cl
0.0223
phosphoenolpyruvate
-
pH 7.0, 30C, wild-type EcaroA
0.023
phosphoenolpyruvate
in the presence of 1 mM NH4Cl and 100 mM KCl
0.025
phosphoenolpyruvate
-
-
0.025
phosphoenolpyruvate
mutant N130W
0.028
phosphoenolpyruvate
-
isoenzyme II
0.032
phosphoenolpyruvate
wild-type
0.033
phosphoenolpyruvate
-
isoenzyme I
0.034
phosphoenolpyruvate
-
-
0.035
phosphoenolpyruvate
mutant N130S
0.04
phosphoenolpyruvate
-
C-terminal His6-tagged AroA, 50 mM KCl
0.0436
phosphoenolpyruvate
-
pH 7.0, 30C, wild-type StaroA
0.045
phosphoenolpyruvate
-
wild-type
0.05
phosphoenolpyruvate
-
50 mM KCl, mutant E341A
0.055
phosphoenolpyruvate
-
+/-0.0014 mM, mutant N130S in bacterial extact; +/-0.002 mM, bacterial extract
0.06
phosphoenolpyruvate
-
+/-0.006 mM
0.07
phosphoenolpyruvate
-
+/-0.005 mM, mutant P101S
0.071
phosphoenolpyruvate
-
mutant P101S
0.078
phosphoenolpyruvate
wild-type, pH 7.0, 28C
0.08
phosphoenolpyruvate
-
+/-0.005 mM, mutant P101A
0.085
phosphoenolpyruvate
in the presence of 1 mM NH4Cl and 10 mM KCl
0.088
phosphoenolpyruvate
-
wild-type enzyme
0.0883
phosphoenolpyruvate
mutant enzyme P106L
0.09
phosphoenolpyruvate
-
wild-type enzyme
0.09
phosphoenolpyruvate
-
+/-0.010 mM, mutant P101G
0.091
phosphoenolpyruvate
in the presence of 10 mM NH4Cl
0.0952
phosphoenolpyruvate
+/-0.00573 mM, full-length G2 EPSPS, crude lysate from overexpressing Escherichia coli ER2799, malachite green dye assay
0.0968
phosphoenolpyruvate
+/-0.01101 mM, reconstituted G2-EPSPS (by intein-mediated protein complementation), crude lysate from transformed Escherichia coli ER2799, malachite green dye assay
0.1
phosphoenolpyruvate
-
wild-type enzyme
0.1
phosphoenolpyruvate
in the presence of 1 mM NH4Cl
0.1
phosphoenolpyruvate
-
in presence of 1 mM shikimate 3-phosphate
0.1
phosphoenolpyruvate
-
mutant P101S/T97I
0.106
phosphoenolpyruvate
-
H385N mutant enzyme
0.14
phosphoenolpyruvate
-
glyphosate-resistant enzyme
0.148
phosphoenolpyruvate
mutant G96A, pH 7.0, 28C
0.15
phosphoenolpyruvate
-
+/-0.020 mM, mutant P101L
0.15
phosphoenolpyruvate
-
C-terminal His6-tagged AroA, 100 mM KCl
0.16
phosphoenolpyruvate
-
wild-type AroA, 100 mM KCl
0.17
phosphoenolpyruvate
-
50 mM KCl, mutant E341C
0.18
phosphoenolpyruvate
-
pH 7.0, recombinant enzyme, in presence of 100 mM KCl
0.19
phosphoenolpyruvate
-
50 mM KCl, mutant D313N
0.195
phosphoenolpyruvate
pH 7.0, 28C
0.2
phosphoenolpyruvate
-
R24D mutant enzyme
0.23
phosphoenolpyruvate
wild-type, pH 7.0, temperature not specified in the publication
0.25
phosphoenolpyruvate
mutant E145G/N163H/N267S/P318R/M377V/M425T/P438L, pH 7.0, temperature not specified in the publication
0.28
phosphoenolpyruvate
-
50 mM KCl, mutant D313A
0.29
phosphoenolpyruvate
pH 7.0, 28C
0.292
phosphoenolpyruvate
mutant T355S, pH 7.0, 28C
0.31
phosphoenolpyruvate
mutant P318R, pH 7.0, temperature not specified in the publication
0.316
phosphoenolpyruvate
wild-type, pH 7.0, 28C
0.32
phosphoenolpyruvate
mutant M425T, pH 7.0, temperature not specified in the publication
0.38
phosphoenolpyruvate
-
mutant T97I
0.38
phosphoenolpyruvate
mutant N267S, pH 7.0, temperature not specified in the publication
0.525
phosphoenolpyruvate
mutant N267S/P318R/M425T, pH 7.0, temperature not specified in the publication
0.59
phosphoenolpyruvate
-
P105S mutant enzyme
0.8
phosphoenolpyruvate
-
mutant G96A/A183T, in presence of 1 mM shikimate 3-phosphate
0.9
phosphoenolpyruvate
-
50 mM KCl, mutant E341Q
1
phosphoenolpyruvate
-
H385L mutant enzyme
1.3
phosphoenolpyruvate
-
50 mM KCl, mutant D313C
1.5
phosphoenolpyruvate
-
50 mM KCl, mutant H385A
1.6
phosphoenolpyruvate
-
50 mM KCl, mutant D313L
1.9
phosphoenolpyruvate
-
50 mM KCl, mutant E341M
2.5
phosphoenolpyruvate
-
50 mM KCl, mutant K22R value above 0.3
2.8
phosphoenolpyruvate
-
G96A mutant enzyme
3.1
phosphoenolpyruvate
-
mutant G96A, in presence of 1 mM shikimate 3-phosphate
3.8
phosphoenolpyruvate
-
mutant A183T, in presence of 1 mM shikimate 3-phosphate
22.5
phosphoenolpyruvate
-
recombinant enzyme, in crude extracts
43.6
phosphoenolpyruvate
-
recombinant enzyme, in crude extracts
0.02
shikimate 3-phosphate
-
-
0.029
shikimate 3-phosphate
-
isoenzyme II
0.031
shikimate 3-phosphate
in the presence of 100 mM NH4Cl
0.033
shikimate 3-phosphate
-
isoenzyme I
0.049
shikimate 3-phosphate
in the presence of 1 mM NH4Cl and 10 mM KCl
0.06
shikimate 3-phosphate
-
+/-0.004 mM, mutant P101A; +/-0.006 mM; +/-0.010 mM, mutant P101G
0.07
shikimate 3-phosphate
-
H385 mutant enzyme
0.08
shikimate 3-phosphate
-
+/-0.006 mM, mutant P101S
0.1
shikimate 3-phosphate
-
+/-0.007 mM, mutant P101L
0.109
shikimate 3-phosphate
in the presence of 1 mM NH4Cl and 100 mM KCl
0.118
shikimate 3-phosphate
in the presence of 10 mM NH4Cl
0.12
shikimate 3-phosphate
-
wild-type and G96A mutant enzyme
0.135
shikimate 3-phosphate
-
wild-type enzyme
0.145
shikimate 3-phosphate
in the presence of 1 mM NH4Cl
0.16
shikimate 3-phosphate
-
H385 mutant enzyme
0.2
shikimate 3-phosphate
-
R24D mutant enzyme
0.33
shikimate 3-phosphate
-
wild-type enzyme
0.43
shikimate 3-phosphate
-
P105S mutant enzyme
2.5
5-enolpyruvylshikimate 3-phosphate
-
+/-0.4 mM, mutant L104P
additional information
additional information
-
kinetic study
-
additional information
additional information
-
kinetic parameters of native and diethyldicarbonate-inactivated enzyme
-
additional information
additional information
-
kinetic study
-
additional information
additional information
-
comparison of kinetic parameters from a variety of organism
-
additional information
additional information
-
stopped-flow kinetics
-
additional information
additional information
-
kinetics, wild-type and mutant enzymes
-
additional information
additional information
-
-
-
additional information
additional information
-
small residue substitutions for Pro101 do not alter the KM for phosphoenolpyruvate and shikimate 3-phosphate
-
24.8
phosphate
-
+/-4.8 mM, mutant L104P
additional information
phosphoenolpyruvate
-
KM/KI: 13, mutant P101S; KM/KI: 150; KM/KI: 2.3, mutant P101L; KM/KI: 4.2, mutant P101A; KM/KI: 7.5, mutant P101G
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.026
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant D313C, cosubstrate: phosphoenolpyruvate
0.3
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant K22A value above 0.3, cosubstrate: phosphoenolpyruvate
0.41
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant H385A, cosubstrate: phosphoenolpyruvate
1.1
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant D313L, cosubstrate: phosphoenolpyruvate; 50 mM KCl, mutant E341Q, cosubstrate: phosphoenolpyruvate
1.2
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant D313A, cosubstrate: phosphoenolpyruvate
1.7
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant E341M, cosubstrate: phosphoenolpyruvate
1.8
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant D313N, cosubstrate: phosphoenolpyruvate
3
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant K22R value above 0.3, cosubstrate: phosphoenolpyruvate
3.6
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant E341C, cosubstrate: phosphoenolpyruvate
4.2
3-phosphoshikimate
Staphylococcus aureus
-
pH 7.0, recombinant enzyme, in presence of 100 mM KCl
4.2
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant E341A, cosubstrate: phosphoenolpyruvate
14
3-phosphoshikimate
Escherichia coli
-
wild-type AroA, 100 mM KCl, cosubstrate: phosphoenolpyruvate
32
3-phosphoshikimate
Escherichia coli
-
C-terminal His6-tagged AroA, 50 mM KCl, cosubstrate: phosphoenolpyruvate
33.6
3-phosphoshikimate
Neurospora crassa
-
cosubstrate phosphoenolpyruvate
56.6
3-phosphoshikimate
Escherichia coli
-
-
0.000012
5-enolpyruvylshikimate 3-phosphate
Escherichia coli
-
5-enpolpyruvylshikimate 3-phosphate ketal formation by hydrolysis of radiolabelled 5-enpolpyruvylshikimate 3-phosphate, pH 7.0, 25C
0.23
5-enolpyruvylshikimate 3-phosphate
Streptococcus pneumoniae
Q9S400
reverse reaction, in the presence of 50 mM KCl
1.2
5-enolpyruvylshikimate 3-phosphate
Pseudomonas aeruginosa
-
25C, pH 7.0, mutant enzyme G99A
1.2
5-enolpyruvylshikimate 3-phosphate
Pseudomonas aeruginosa
-
+/-0.04, mutant G99A
2.7
5-enolpyruvylshikimate 3-phosphate
Pseudomonas aeruginosa
-
25C, pH 7.0, mutant strain L104P
2.7
5-enolpyruvylshikimate 3-phosphate
Pseudomonas aeruginosa
-
+/-0.1, mutant L104P
7.4
5-enolpyruvylshikimate 3-phosphate
Pseudomonas aeruginosa
-
25C, pH 7.0, wild-type enzyme
0.22
phosphate
Streptococcus pneumoniae
Q9S400
reverse reaction, in the presence of 50 mM KCl
1
phosphate
Pseudomonas aeruginosa
-
25C, pH 7.0, mutant enzyme G99A
1
phosphate
Pseudomonas aeruginosa
-
+/-0.03, mutant G99A
2.8
phosphate
Pseudomonas aeruginosa
-
25C, pH 7.0, mutant enzyme L104P
2.8
phosphate
Pseudomonas aeruginosa
-
+/-0.2, mutant L104P
5.4
phosphate
Pseudomonas aeruginosa
-
25C, pH 7.0, wild-type enzyme
0.0000045
phosphoenolpyruvate
Escherichia coli
-
5-enpolpyruvylshikimate 3-phosphate ketal formation in presence of shikimate 3-phosphate and equimolar phosphoenolpyruvate
0.000049
phosphoenolpyruvate
Escherichia coli
-
5-enpolpyruvylshikimate 3-phosphate ketal formation in presence of shikimate 3-phosphate and 6.7 equivalents of phosphoenolpyruvate
0.00008
phosphoenolpyruvate
Escherichia coli
-
5-enpolpyruvylshikimate 3-phosphate ketal formation in presence of shikimate 3-phosphate and 6.7 equivalents of phosphoenolpyruvate and 10 mM phosphate
0.026
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant D313C, cosubstrate: 3-phosphoshikimate
0.3
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant K22A value above 0.3, cosubstrate: 3-phosphoshikimate
0.41
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant H385A, cosubstrate: 3-phosphoshikimate
1.1
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant D313L, cosubstrate: 3-phosphoshikimate; 50 mM KCl, mutant E341Q, cosubstrate: 3-phosphoshikimate
1.2
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant D313A, cosubstrate: 3-phosphoshikimate
1.7
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant E341M, cosubstrate: 3-phosphoshikimate
1.8
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant D313N, cosubstrate: 3-phosphoshikimate
2
phosphoenolpyruvate
Streptococcus pneumoniae
Q9S400
in the presence of 1 mM NH4Cl and 10 mM KCl
2.1
phosphoenolpyruvate
Streptococcus pneumoniae
Q9S400
in the presence of 1 mM NH4Cl
2.8
phosphoenolpyruvate
Streptococcus pneumoniae
Q9S400
in the presence of 10 mM NH4Cl
3
phosphoenolpyruvate
Streptococcus pneumoniae
Q9S400
in the presence of 100 mM NH4Cl
3
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant K22R value above 0.3, cosubstrate: 3-phosphoshikimate
3.6
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant E341C, cosubstrate: 3-phosphoshikimate
4.2
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant E341A, cosubstrate: 3-phosphoshikimate
4.3
phosphoenolpyruvate
Staphylococcus aureus
-
pH 7.0, recombinant enzyme, in presence of 100 mM KCl
12.8
phosphoenolpyruvate
Streptococcus pneumoniae
Q9S400
in the presence of 1 mM NH4Cl and 100 mM KCl
14
phosphoenolpyruvate
Escherichia coli
-
wild-type AroA, 100 mM KCl, cosubstrate: 3-phosphoshikimate
15
phosphoenolpyruvate
Escherichia coli
-
mutant A183T, in presence of 1 mM shikimate 3-phosphate
21
phosphoenolpyruvate
Escherichia coli
-
mutant G96A/A183T, in presence of 1 mM shikimate 3-phosphate
26.2
phosphoenolpyruvate
Pseudomonas fluorescens
D7RH58
pH 7.0, 28C
26.4
phosphoenolpyruvate
Escherichia coli
-
in presence of 1 mM shikimate 3-phosphate
26.7
phosphoenolpyruvate
Ochrobactrum anthropi
A6WUV8
wild-type, pH 7.0, temperature not specified in the publication
28.3
phosphoenolpyruvate
Bacteria
-
-
29.7
phosphoenolpyruvate
Escherichia coli
-
mutant G96A, in presence of 1 mM shikimate 3-phosphate
31.7
phosphoenolpyruvate
Ochrobactrum anthropi
A6WUV8
mutant N267S, pH 7.0, temperature not specified in the publication
31.8
phosphoenolpyruvate
Bacillus cereus
F5A7Q0
pH 7.0, 28C
32
phosphoenolpyruvate
Escherichia coli
-
C-terminal His6-tagged AroA, 50 mM KCl, cosubstrate: 3-phosphoshikimate
33.6
phosphoenolpyruvate
Neurospora crassa
-
cosubstrate 3-phosphoshikimate
35
phosphoenolpyruvate
Ochrobactrum anthropi
A6WUV8
mutant M425T, pH 7.0, temperature not specified in the publication
38.4
phosphoenolpyruvate
plant
-
-
43.3
phosphoenolpyruvate
Ochrobactrum anthropi
A6WUV8
mutant P318R, pH 7.0, temperature not specified in the publication
51.7
phosphoenolpyruvate
Ochrobactrum anthropi
A6WUV8
mutant E145G/N163H/N267S/P318R/M377V/M425T/P438L, pH 7.0, temperature not specified in the publication; mutant N267S/P318R/M425T, pH 7.0, temperature not specified in the publication
100
3-phosphoshikimate
Escherichia coli
-
C-terminal His6-tagged AroA, 100 mM KCl, cosubstrate: phosphoenolpyruvate
additional information
5-enolpyruvylshikimate 3-phosphate
Pseudomonas aeruginosa
-
kat/KM: 1100000 M(-1)s(-1), mutant L104P; kcat/KM: 3400000 M(-1)s(-1); kcat/KM: 660000 M(-1)s(-1), mutant G99A
7.4
5-enolpyruvylshikimate 3-phosphate
Pseudomonas aeruginosa
-
+/-0.4
additional information
phosphate
Pseudomonas aeruginosa
-
kcat/KM: 110 M(-1)s(-1), mutant L104P; kcat/KM: 130 M(-1)s(-1), mutant G99A; kcat/KM: 960 M(-1)s(-1)
5.4
phosphate
Pseudomonas aeruginosa
-
+/-0.4
additional information
phosphoenolpyruvate
Escherichia coli
-
kcat/KM: 240000 1/M*s, mutant P101G; kcat/KM: 280000 1/M*s, mutant P101A; kcat/KM: 290000 1/M*s, mutant P101S; kcat/KM: 41000 1/M*s, mutant P101L; kcat/KM: 640000 1/M*s
100
phosphoenolpyruvate
Escherichia coli
-
C-terminal His6-tagged AroA, 100 mM KCl, cosubstrate: 3-phosphoshikimate
additional information
shikimate 3-phosphate
Escherichia coli
-
kcat/KM: 200000 1/M*s, mutant P101S; kcat/KM: 390000 1/M*s, mutant P101A; kcat/KM: 390000 1/M*s, mutant P101G; kcat/KM: 62000 1/M*s, mutant P101L; kcat/KM: 640000 1/M*s
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0016
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant D313L
1318
0.005
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant E341A
1318
0.027
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant E341Q
1318
0.044
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant E341C
1318
0.05
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant E341M
1318
0.06
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant D313A
1318
0.068
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant D313C
1318
0.09
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant D313N
1318
4
3-phosphoshikimate
Escherichia coli
-
50 mM KCl, mutant H385A
1318
96
3-phosphoshikimate
Escherichia coli
-
wild-type AroA, 100 mM KCl
1318
150
3-phosphoshikimate
Escherichia coli
-
C-terminal His6-tagged AroA, 100 mM KCl
1318
1800
3-phosphoshikimate
Escherichia coli
-
C-terminal His6-tagged AroA, 50 mM KCl
1318
74000000
3-phosphoshikimate
Escherichia coli
-
mutant P101S/T97I
1318
120000000
3-phosphoshikimate
Escherichia coli
-
mutant T97I
1318
240000000
3-phosphoshikimate
Escherichia coli
-
mutant P101S
1318
910000000
3-phosphoshikimate
Escherichia coli
-
wild-type
1318
0.0007
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant D313L
51
0.0009
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant E341M
51
0.0012
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant K22R
51
0.0013
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant E341Q
51
0.003
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant H385A
51
0.004
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant D313A
51
0.009
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant E341A; 50 mM KCl, mutant K22A
51
0.01
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant D313N
51
0.02
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant D313C
51
0.021
phosphoenolpyruvate
Escherichia coli
-
50 mM KCl, mutant E341C
51
83.3
phosphoenolpyruvate
Ochrobactrum anthropi
A6WUV8
mutant N267S, pH 7.0, temperature not specified in the publication
51
86
phosphoenolpyruvate
Escherichia coli
-
wild-type AroA, 100 mM KCl
51
90
phosphoenolpyruvate
Escherichia coli
-
C-terminal His6-tagged AroA, 100 mM KCl
51
109.3
phosphoenolpyruvate
Ochrobactrum anthropi
A6WUV8
mutant M425T, pH 7.0, temperature not specified in the publication
51
115.9
phosphoenolpyruvate
Ochrobactrum anthropi
A6WUV8
wild-type, pH 7.0, temperature not specified in the publication
51
139.8
phosphoenolpyruvate
Ochrobactrum anthropi
A6WUV8
mutant P318R, pH 7.0, temperature not specified in the publication
51
163.1
phosphoenolpyruvate
Bacillus cereus
F5A7Q0
pH 7.0, 28C
51
206.7
phosphoenolpyruvate
Ochrobactrum anthropi
A6WUV8
mutant E145G/N163H/N267S/P318R/M377V/M425T/P438L, pH 7.0, temperature not specified in the publication; mutant N267S/P318R/M425T, pH 7.0, temperature not specified in the publication
51
800
phosphoenolpyruvate
Escherichia coli
-
C-terminal His6-tagged AroA, 50 mM KCl
51
23000000
phosphoenolpyruvate
Escherichia coli
-
mutant T97I
51
57000000
phosphoenolpyruvate
Escherichia coli
-
mutant P101S/T97I
51
250000000
phosphoenolpyruvate
Escherichia coli
-
mutant P101S
51
930000000
phosphoenolpyruvate
Escherichia coli
-
wild-type
51
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0000039
(3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
+/-0.6 nM
0.00045
(3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
+/-70 nM
0.0018
(3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
+/-0.0006 mM
0.00076
(3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
+/-200 nM
0.011
(3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
+/-0.002 mM
0.076
(3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
+/-0.009 mM
0.000016
(3R,4S,5R)-5-((R)-1-carboxy-1-phosphono-ethoxy)-4-hydroxy-3-phosphonooxy-cyclohex-1-enecarboxylic acid
-
versus 3-phosphoshikimate
0.00075
(3R,4S,5R)-5-((S)-1-carboxy-1-phosphono-ethoxy)-4-hydroxy-3-phosphonooxy-cyclohex-1-enecarboxylic acid
-
versus 3-phosphoshikimate
0.0000078
(3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
+/-0.5 nM
0.00003
(3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
+/-6 nM
0.000063
(3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
+/-15 nM
0.0029
(Z)-3-fluorophosphoenolpyruvate
-
vs. shikimate 3-phosphate
0.0064
(Z)-3-fluorophosphoenolpyruvate
-
vs. phosphoenolpyruvate
0.0126
5-Deoxy-shikimate 3-phosphate
-
-
0.0002
glyphosate
plant
-
-
0.0003
glyphosate
-
wild-type
0.0004
glyphosate
-
+/-0.00006 mM
0.0005
glyphosate
-
pH 7.0, 30C, wild-type StaroA
0.0015
glyphosate
-
pH 7.0, 30C, wild-type EcaroA
0.003
glyphosate
-
mutant P101S
0.0055
glyphosate
-
+/-0.0003 mM, mutant P101S
0.012
glyphosate
-
+/-0.004 mM, mutant P101G
0.0128
glyphosate
-
pH 7.0, 30C, mutant StaroA-T42M
0.019
glyphosate
-
+/-0.003 mM, mutant P101A
0.0305
glyphosate
-
pH 7.0, 30C, mutant EcaroA-T42M
0.035
glyphosate
+/-0.00578 mM, reconstituted G2-EPSPS (by intein-mediated protein complementation), crude lysate from transformed Escherichia coli ER2799, malachite green dye assay
0.0446
glyphosate
wild-type, pH 7.0, 28C
0.049
glyphosate
wild-type, pH 7.0, 28C
0.0495
glyphosate
+/-0.00317 mM, full-length G2 EPSPS, crude lysate from overexpressing Escherichia coli ER2799, malachite green dye assay
0.0512
glyphosate
mutant T355S, pH 7.0, 28C
0.052
glyphosate
pH 7.0, 28C
0.0554
glyphosate
pH 7.0, 28C
0.066
glyphosate
-
+/-0.002 mM, mutant P101L
0.069
glyphosate
wild-type, pH 7.0, temperature not specified in the publication
0.08
glyphosate
mutant N267S, pH 7.0, temperature not specified in the publication
0.09
glyphosate
-
mutant P101S
0.091
glyphosate
mutant M425T, pH 7.0, temperature not specified in the publication
0.1
glyphosate
-
+/-0.01 mM, in presence of 1 mM shikimate 3-phosphate and 0-500 mM inhibitor, bacterial extract
0.11
glyphosate
mutant P318R, pH 7.0, temperature not specified in the publication
0.15
glyphosate
mutant N267S/P318R/M425T, pH 7.0, temperature not specified in the publication
0.16
glyphosate
mutant E145G/N163H/N267S/P318R/M377V/M425T/P438L, pH 7.0, temperature not specified in the publication
0.161
glyphosate
-
-
0.2
glyphosate
-
+/-0.01 mM, mutant N130S, in presence of 1 mM shikimate 3-phosphate and 0-500 mM inhibitor, bacterial extract
0.00008
N-phosphonomethylglycine
-
-
0.00012 - 0.00013
N-phosphonomethylglycine
-
isoenzymes I and II, vs. phosphoenolpyruvate
0.00016
N-phosphonomethylglycine
-
vs. phosphoenolpyruvate
0.0003
N-phosphonomethylglycine
in the presence of 100 mM NH4Cl
0.00076
N-phosphonomethylglycine
-
glyphosate-resistant strain
0.0008
N-phosphonomethylglycine
-
H385N mutant enzyme
0.0009
N-phosphonomethylglycine
-
vs. shikimate 3-phosphate, recombinant enzyme
0.001
N-phosphonomethylglycine
-
vs. shikimate 3-phosphate, native enzyme
0.001
N-phosphonomethylglycine
-
vs. phosphoenolpyruvate
0.001
N-phosphonomethylglycine
-
P106S mutant enzyme
0.0012
N-phosphonomethylglycine
-
wild-type enzyme
0.00125
N-phosphonomethylglycine
-
vs. phosphoenolpyruvate
0.005
N-phosphonomethylglycine
in the presence of 1 mM NH4Cl and 100 mM K+
0.008
N-phosphonomethylglycine
-
vs. 5-enolpyruvylshikimate 3-phosphate
0.0094
N-phosphonomethylglycine
-
isoenzyme I, vs. shikimate 3-phosphate
0.01
N-phosphonomethylglycine
-
-
0.012
N-phosphonomethylglycine
-
isoenzyme II, vs. shikimate 3-phosphate
0.0183
N-phosphonomethylglycine
-
vs. shikimate 3-phosphate
0.027
N-phosphonomethylglycine
-
vs. phosphate
0.029
N-phosphonomethylglycine
-
P381L mutant enzyme
0.04
N-phosphonomethylglycine
-
H385L mutant enzyme
0.048
N-phosphonomethylglycine
-
glyphosate-sensitive strain
0.05
N-phosphonomethylglycine
-
R24D mutant enzyme
0.06 - 0.08
N-phosphonomethylglycine
-
vs. shikimate 3-phosphate
0.06
N-phosphonomethylglycine
-
wild-type enzyme
0.07 - 0.1
N-phosphonomethylglycine
-
vs. phosphate
0.08 - 0.1
N-phosphonomethylglycine
-
vs. enolpyruvate
0.098
N-phosphonomethylglycine
in the presence of 1 mM NH4Cl and 10 mM K+
0.185
N-phosphonomethylglycine
in the presence of 1 mM NH4Cl
0.21
N-phosphonomethylglycine
-
P105S mutant enzyme
0.22
N-phosphonomethylglycine
-
vs. shikimate 3-phosphate
0.35
N-phosphonomethylglycine
-
-
0.54
N-phosphonomethylglycine
-
recombinant enzyme, in crude extracts
0.96
N-phosphonomethylglycine
-
recombinant enzyme, in crude extracts
0.044
phosphoenolpyruvate
-
-
0.023
shikimate 3-phosphate
-
-
0.000015
shikimate 3-phosphate-5-carboxymethyl-(2R)-phosphonate
-
vs. 5-enolpyruvylshikimate-3-phosphate
0.00009
shikimate 3-phosphate-5-carboxymethyl-(2R)-phosphonate
-
vs. phosphate
0.0011
shikimate 3-phosphate-5-carboxymethyl-(2S)-phosphonate
-
vs. 5-enolpyruvylshikimate-3-phosphate
0.0021
shikimate 3-phosphate-5-carboxymethyl-(2S)-phosphonate
-
vs. phosphate
0.0013
5-enolpyruvylshikimate 3-phosphate
-
-
additional information
glyphosate
Ki/KM (phosphoenolpyruvate): 0.36, reconstituted G2-EPSPS (by intein-mediated protein complementation), crude lysate from overexpressing Escherichia coli ER2799, malachite green dye assay; Ki/KM (phosphoenolpyruvate): 0.52, full-length G2 EPSPS, crude lysate from overexpressing Escherichia coli ER2799, malachite green dye assay
additional information
glyphosate
-
any substitutions for residue Pro101 decrease glyphosate binding affinity and inhibitor potency
additional information
glyphosate
-
KI/KM (phosphoenolpyruvate): 14
additional information
glyphosate
plant
-
KI/KM (phosphoenolpyruvate): 0.01
0.45
glyphosate
mutant G96A, pH 7.0, 28C
additional information
additional information
-
-
-
additional information
additional information
-
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00898
glyphosate
Oryza sativa
Q93VK6
wild-type enzyme
0.05
glyphosate
Escherichia coli
-
-
0.1046
glyphosate
Oryza sativa
Q93VK6
mutant enzyme P106L
0.26
glyphosate
Rahnella aquatilis
C9EGX2
wild-type, pH 7.0, 28C
0.328
glyphosate
Halothermothrix orenii
B8CWX4
wild-type, pH 7.0, 28C
0.34
glyphosate
Rahnella aquatilis
C9EGX2
mutant G96A, pH 7.0, 28C
0.398
glyphosate
Halothermothrix orenii
B8CWX4
mutant T355S, pH 7.0, 28C
0.432
glyphosate
Pseudomonas fluorescens
D7RH58
pH 7.0, 28C
0.458
glyphosate
Bacillus cereus
F5A7Q0
pH 7.0, 28C
0.61
glyphosate
Ochrobactrum anthropi
A6WUV8
wild-type, pH 7.0, temperature not specified in the publication
0.69
glyphosate
Ochrobactrum anthropi
A6WUV8
mutant N267S, pH 7.0, temperature not specified in the publication
0.73
glyphosate
Ochrobactrum anthropi
A6WUV8
mutant M425T, pH 7.0, temperature not specified in the publication
0.81
glyphosate
Ochrobactrum anthropi
A6WUV8
mutant P318R, pH 7.0, temperature not specified in the publication
0.91
glyphosate
Ochrobactrum anthropi
A6WUV8
mutant N267S/P318R/M425T, pH 7.0, temperature not specified in the publication
0.92
glyphosate
Ochrobactrum anthropi
A6WUV8
mutant E145G/N163H/N267S/P318R/M377V/M425T/P438L, pH 7.0, temperature not specified in the publication
8
glyphosate
Pseudomonas stutzeri
-
+/-0.5 mM, in presence of 1 mM shikimate 3-phosphate and 0-500 mM inhibitor, bacterial extract
8.2
glyphosate
Escherichia coli
-
mutant A183T
20
glyphosate
Pseudomonas stutzeri
-
+/-0.2 mM, mutant N130S, in presence of 1 mM shikimate 3-phosphate and 0-500 mM inhibitor, bacterial extract
additional information
glyphosate
Escherichia coli
-
more than 10 mM for mutant G96A; more than 10 mM for mutant G96A/A183T
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0058
-
recombinant enzyme, activity in crude extracts
0.0058
-
wild-type enzyme, cell exract
0.0064
-
recombinant enzyme, activity in crude extracts
0.0076
-
wild-type enzyme, cell exract
0.0357
-
mutant T42M, cell extract
0.19
-
bacterial extract
0.22
-
mutant N130S in bacterial extract
0.4
-
mutant G99A, after DEAE-Sepharose chromatography
0.57
-
mutant G99A, in crude bacterial extracts
0.62
-
mutant L104P, in crude bacterial extracts
0.63
-
mutant L104P, after DEAE-Sepharose chromatography
0.86
-
mutant G99A, after Superdex 200 gel filtration
1.26
-
reverse reaction, i.e. phosphate + 5-enolpyruvylshikimate 3-phosphate
1.34
-
mutant G99A, after ResourceQ chromatography
1.4
-
in crude bacterial extracts
1.7
-
after DEAE-sepharose chromatography
1.83
-
mutant L104P, after Superdex 200 gel filtration
1.91
-
mutant L104P, after ResourceQ chromatography
2
-
after Superdex 200 gel filtration
4.07
-
forward reaction, i.e. phosphoenolpyruvate + 3-phosphoshikimate
4.48
+/-0.79 micromol/min/mg, reconstituted G2-EPSPS (by intein-mediated protein complementation), mean specific activity, crude lysate from transformed Escherichia coli ER2799, malachite green dye assay
4.761
+/-0.92 micromol/min/mg, reconstituted G2-EPSPS (by intein-mediated protein complementation), maximum velocity, crude lysate from transformed Escherichia coli ER2799, malachite green dye assay
5.67
-
phosphate + 5-enolpyruvylshikimate 3-phosphate
5.68
-
activity of recombinant enzyme in crude extract
6.12
-
phosphate + 5-enolpyruvylshikimate 3-phosphate
7.12
+/-0.13 micromol/min/mg, full-length G2 EPSPS, mean specific activity, crude lysate from overexpressing Escherichia coli ER2799, malachite green dye assay
7.24
+/-0.16 micromol/min/mg, full-length G2 EPSPS, maximum velocity, crude lysate from overexpressing Escherichia coli ER2799, malachite green dye assay
8
-
+/-0.2 micromol/min/mg, mutant P101L
8.5
-
recombinant
8.5
-
after ResourceQ chromatography
10.03
wild-type enzyme
10.23
mutant enzyme P106L
10.86
-
glyphosate insensitve strain
11.52
-
forward reaction, i.e. phosphoenolpyruvate + 3-phosphoshikimate
12
-
phosphoenolpyruvate + 3-phosphoshikimate
18
-
phosphate + 5-enolpyruvylshikimate 3-phosphate, native enzyme
18.13
-
recombinant enzyme
18.9
-
mutant A183T, with respect to phosphoenolpyruvate
20
-
recombinant His-tagged wild-type enzyme
21.1
-
phosphate + 5-enolpyruvylshikimate 3-phosphate, recombinant enzyme
22
-
+/-0.4 micromol/min/mg, mutant P101S
24
-
recombinant wild-type enzyme
26.5
-
mutant G96A/A183T, with respect to phosphoenolpyruvate
28
-
+/-1 micromol/min/mg, mutant P101G
31
-
+/-0.5 micromol/min/mg, mutant P101A
34
-
with respect to phosphoenolpyruvate
35.9
-
Vmax
37.4
-
mutant G96A, with respect to phosphoenolpyruvate
47.5
plant
-
Vmax
50
-
+/-1 micromol/min/mg
61
-
isoenzyme I
62
-
phosphoenolpyruvate + 3-phosphoshikimate
94.7
-
isoenzyme II
additional information
-
activity of isoenzyme II increases in elicited cells
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.4
-
wild-type enzyme, 2 maxima at pH 5.4 and pH 7.0
5.6
-
phosphate + 5-enolpyruvylshikimate 3-phosphate, 2 maxima at pH 5.6 and pH 7.6
6
-
glyphosate-resistant enzyme, 2 maxima at pH 6 and pH 7.3
7
-
wild-type enzyme, 2 maxima at pH 5.4 and pH 7.0
7
-
H385N mutant enzyme
7 - 7.2
-
50% of maximal activity between pH 6.2 and pH 8.5
7.3
-
glyphosate-resistant enzyme, 2 maxima at pH 6 and pH 7.3
7.4
-
Tris-HCl buffer preferred
7.4 - 7.5
-
isoenzymes I and II
7.6
-
phosphate + 5-enolpyruvylshikimate 3-phosphate, 2 maxima, at pH 5.6 and pH 7.6
7.8
-
wild-type enzyme
additional information
-
pI: 4.1, glyphosate-resistant enzyme; pI: 4.6, wild-type enzyme
additional information
-
pI: 4.6, wild-type enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 8.5
-
approx. half-maximal activity at pH 5.0 and 8.5
5.5 - 8.7
-
approx. half-maximal activity at pH 5.5 and 8.7, phosphate + 5-enolpyruvylshikimate 3-phosphate
5.6 - 8.4
-
H385N mutantt enzyme
6 - 8.4
-
wild-type enzyme
6.2 - 8.5
-
approx. half-maximal activity at pH 6.2 and 8.5
6.5 - 8.5
-
approx. half-maximal activity at pH 6.5 and 8.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
20
-
assay at
25
-
assay at
25
-
assay at
30
-
assay at
30
-
assay at
55
-
isoenzymes I and II
60
-
wild-type enzyme
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
45 - 62
-
approx. half-maximal activity at 45C and 62C
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8.22
calculated from sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
constitutively expressed
Manually annotated by BRENDA team
constitutively expressed, low expression
Manually annotated by BRENDA team
-
shoot tissue
Manually annotated by BRENDA team
constitutively expressed
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
monofunctional enzyme form
Manually annotated by BRENDA team
-
isoenzymes I and II
Manually annotated by BRENDA team
putative N-terminal transit peptide consisting of 70 amino acids which is present in other plant EPSP synthases and lacking in the bacterial enzyme
Manually annotated by BRENDA team
Dunaliella salina 435
-
putative N-terminal transit peptide consisting of 70 amino acids which is present in other plant EPSP synthases and lacking in the bacterial enzyme
-
Manually annotated by BRENDA team
-
multifunctional enzyme form i.e. single domain of arom protein
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Agrobacterium sp. (strain CP4)
Agrobacterium sp. (strain CP4)
Agrobacterium sp. (strain CP4)
Agrobacterium sp. (strain CP4)
Agrobacterium sp. (strain CP4)
Agrobacterium sp. (strain CP4)
Agrobacterium sp. (strain CP4)
Bacillus halodurans (strain ATCC BAA-125 / DSM 18197 / FERM 7344 / JCM 9153 / C-125)
Coxiella burnetii (strain RSA 493 / Nine Mile phase I)
Coxiella burnetii (strain RSA 493 / Nine Mile phase I)
Coxiella burnetii (strain RSA 493 / Nine Mile phase I)
Coxiella burnetii (strain RSA 493 / Nine Mile phase I)
Coxiella burnetii (strain RSA 493 / Nine Mile phase I)
Coxiella burnetii (strain RSA 493 / Nine Mile phase I)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Streptococcus pneumoniae serotype 4 (strain ATCC BAA-334 / TIGR4)
Streptococcus pneumoniae serotype 4 (strain ATCC BAA-334 / TIGR4)
Streptococcus pneumoniae serotype 4 (strain ATCC BAA-334 / TIGR4)
Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
20400
C-terminal fragment (from Thr296) expressed in pKEB12, Western blot
690523
32400
-
gel filtration
638181
42000
-
gel filtration
638173, 638175
44000
-
gel filtration
638184
44000
-
SDS-PAGE
691087
46000
-
wild-type and mutant N130S, SDS-PAGE on bacterial extracts
691436
46110
-
calculated from polypeptide sequence of aroA gene product
638173
46430
by ESI-MS; deduced from primary structure
690976
47000
full-length protein, Western blot analyses
690523
48000
-
mixture of monomers and dimers in the absence of NH4Cl, only monomers in the presence of NH4Cl, gel filtration
638199
50000
-
wild-type and mutants, SDS-PAGE after induced overexpression in Escherichia coli
694929
54040
-
deduced from primary sequence
694929
54600
deduced from primary structure, 514 amino acids
681333
55000
-
gel filtration
638173, 638174
57000
-
gel filtration
638187
58100
-
isoenzyme II, gel filtration
638198
58700
-
isoenzyme I, gel filtration
638198
59000
-
monofunctional enzyme
638197
73000
overexpressed mature protein (47000 Da) with GST tag (26000 Da)
681333
91000
-
gel filtration
638204
159000
-
multifunctional enzyme called arom
638197
290000
-
aromatic multifunctional enzyme protein, sedimentation equilibrium analysis
137639
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 46425, deduced from nucleotide sequence; x * 46458, electrospray ionization mass spectrometry
?
-
* 45000, wild-type and mutant enzyme, SDS-PAGE
?
x * 55600, calculation from sequence
?
x * 54600, cylculated from sequence
?
-
x * 50000, SDS-PAGE
?
x * 46020, calculated
?
x * 46960, calculated
?
x * 47000, SDS-PAGE
?
x * 43350, calculated
?
Dunaliella salina 435
-
x * 54600, cylculated from sequence
-
?
Halothermothrix orenii H168
-
x * 46020, calculated
-
?
Rahnella aquatilis GR20
-
x * 47000, SDS-PAGE
-
dimer
-
2 * 49000, SDS-PAGE
monomer
-
1 * 46112, deduced from nucleotide sequence
monomer
-
1 * 49000
monomer
-
1 * 42500, wild-type enzyme, SDS-PAGE; 1 * 42900, glyphosate-resistant enzyme, SDS-PAGE
monomer
-
1 * 42900, glyphosate-resistant enzyme, SDS-PAGE
monomer
-
SDS-PAGE
monomer
-
1 * 50000, SDS-PAGE
monomer
-
1 * 50600-51900, 3 isozymes, SDS-PAGE
monomer
-
1 * 53600, isoenzyme II, SDS-PAGE; 1 * 54900, isoenzyme I, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
wild-type in presence of 5 mM inhibitor (3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid (2F-TI, PDB: 2PQB) or (3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid (RP-TI, PDB: 2PQC), and mutant A100G in presence of 5 mM inhibitor 2F-TI (PDB: 2PQD): 2F-TI-binding induces only slight shift of Glu354 side chain but substantial conformational change of 2F-TI molecule that is independent of residue 100 but mediated by residue Thr101 and due to rigidity of active site, RP-TI-binding induces slight movement of Glu354 side chain and occurs via salt bridge with residue Arg128 but does not alter enzymes backbone compared to the shikimate 3-phosphate bound enzyme, crystals: space group P2(1), unit cell parameters: a: 62.8-63.4, b: 44.9-45.0, c: 77.1-77.6, beta: 106.1-106.3, hanging-drop vapour-diffusion, ammonium sulfate conditions, molecular replacement using PDB: 2GG6 as model, co-crystallization with (3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid (SP-TI) not succeeded, docking of SP-TI into substrate- or inhibitor-bound structures indicates low electrostatic interactions between enzyme and inhibitor
molecular docking using known structures of EPSPS in complex with (R)-phosphonate analogue of the tetrahedral reaction intermediate
-
enzyme complexed with inhibitor (3R,4S,5R)-5-((S)-1-carboxy-1-phosphono-ethoxy)-4-hydroxy-3-phosphonooxy-cyclohex-1-enecarboxylic acid or with inhibitor (3R,4S,5R)-5-((R)-1-carboxy-1-phosphono-ethoxy)-4-hydroxy-3-phosphonooxy-cyclohex-1-enecarboxylic acid, 100 mg/ml recombinant enzyme in 2.5 M sodium formate with 10 mM inhibitor, hanging drop vapour diffusion method, 19C, X-ray diffraction at -180C using the rotation method on single flash-frozen crystals, structure determination and analysis at 1.5 and 1.9 A resolution, respectively
-
enzyme in complex with shikimate/glyphosate at 1.5 A resolution
-
in presence of 10 mM inhibitor (3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid (2F-TI, PDB: 2PQ9), 2F-TI-binding induces large conformational change of Glu341 side chain compared to TI-binding, crystals: space group P2(1)2(1)2(1), unit cell parameters: a: 58.2, b: 85.1, c: 87.6, beta: 90, hanging-drop vapour-diffusion, sodium format conditions, molecular replacement using PDB: 1G6S as model
-
mutants P101S and P101L in complex with shikimate 3-phosphate and with or without inhibitor glyphosate, long-range structural change in glyphosate binding site compared to wild-type, Gly96 and Thr97 are shifted towards the glyphosate-binding site, which gets slightly narrowed, crystals: space group: P2(1)2(1)2(1), unit cell parameters: a: 57.6-57.8, b: 85.1-85.6, c: 87.9-88.3, beta: 90, hanging-drop vapour-diffusion, protein solution (37.5 mg/ml, 5 mM inhibitor and/or 5 mM shikimate 3-phosphate), 19C, molecular replacement using PDB: 1G6S as model
-
purified recombinant wild-type and mutant enzymes in complex with the substrates, at 19C, from 4 M sodium formate in presence of 5 mM 3-phosphoshikimate and 5 mM phosphoenolpyruvate, X-ray diffraction structure determination and analysis at 1.6 A resolution
semiempirical molecular modelling using, among others, the crystal structure of EPSPS mutant D313A (PDB: 1Q36) as model for assignment of protonation states of all basic amino acids in the active site: in the enzyme-tetrahedral reaction intermediate (TI) complex is residue His385 in a neutral form (with protonated epsilon-N atom) while residues Lys22, Lys340 and Lys411 are protonated, hydrogen bonds occur between Lys22 and the carboxylate oxygen atom of the phosphoenolpyruvate moiety of the TI, Asp313 has only minor effects on TI positioning within the active site but mediates as a base the attack of the TI C4-hydroxyl group on the TI methyl group prior to EPSP formation
-
using the hanging drop, vapor-diffusion method in the presence of 5 mM 3-phosphoshikimate
-
restrained-based, molecular modelling; open conformation model of apoenzyme generated based on coordinates from PDB: 2BJB (MtEPSPS), closed conformation model of enzyme/3-phosphoshikimate/glyphosate complex generated based on PDB: 1G6S
three-dimensional structure modeling of mutations P318R and M425T. Results indicate that the enzyme's lower hemispheres upper surface is possibly another important region for glyphosate resistance
molecular modelling, two similar globular, inside-out alpha-beta-barrel domains connected by a two-stranded hinge, substrates bind at the inter-domain cleft
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
40
-
rapid inactivation above, glyphosate resistant enzyme
638180
45
-
isoenzyme I, 50% loss of activity after 20 min, isoenzyme II, 50% loss of activity after 60 min
638198
60
-
rapid inactivation above
638181
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
complete unfolding of the protein at 2.5 M guanidine hydrochloride at pH 7.8 (midpoint guanidine hydrochloride concentration of unfolding: 0.8 +/-0.1 M), enhanced flexibility, lower stability and increased proteolysis of mutants A183T and G96A/A183T compared to wild-type and mutant G96A as revealed by susceptibility to proteolytic digestion with trypsin, thermolysin, pronase (25C for various time intervals), unfolding profiles (intrinsic tryptophan fluorescence measurements at 340 nm) after treatment with guanidine hydrochloride (30 min at 25C) and dynamic quenching with acrylamide or not, and secondary structure determination by circular dichroism spectroscopy
-
dithiothreitol reverses aggregation which sometimes occurs after storage at -20C
-
dithiothreitol, 0.4 mM, stabilizes during purification
-
glycerol, 50% v/v, stabilizes
-
EDTA stabilizes
-
freezing inactivates, Polybuffer 74, operating range pH 7-4, 1 mM dithioerythritol stabilize
-
glyphosate-sensitive enzyme is more stable than wild-type enzyme
-
prolonged dialysis against 50 mM Tris-HCl buffer, pH 7.5, 0.1 mM DTT, 1 mM benzamidine hydrochloride, 5 mM EDTA, 1 mM PMSF, stable to
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 50 mM Tris-HCl, pH 7.5, 50 mM KCl, 0.4 mM dithiothreitol, 50% v/v glycerol, at least 4 months, no loss of activity
-
-20C, in 50% v/v glycerol, enzyme concentration 5 mg/ml, at least 1 year
-
-18C, in Polybuffer 74 from Pharmacia, operating range pH 7-4, 1 mM dithioerythritol, at least 9 months
-
0-4C, 2 months
-
4C, ammonium sulfate precipitate, 4 months, no loss of activity
-
-20C, more than a year
-
-20C, 50% glycerol, 10-15% loss of activity per month
-
-20C, 40% v/v glycerol, at least 1 year, no loss of activity
-
-20C, 3 months, 15% loss of activity
-
0C, 30 d, 25% loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate, DEAE-Sephacel, Mono Q, cellulose phosphate
-
ammonium sulfate, anion exchange chromatography, gel filtration, chromatofocusing
-
recombinant enzyme, ammonium sulfate
-
from bacterial lysate by single step cobalt affinity chromatography followed by dialysis
-
from supernatant from 25% (w/v) ammonium sulfate precipitation of crude bacterial extracts, Q-sepharose anion exchange chromatography (NaCl gradient elution) and FPLC (Mono-Q column, NaCl gradient elution)
-
recombinant enzyme
-
recombinant enzyme, ammonium sulfate, DEAE-Sephacel, phenyl-Sepharose, phosphocellulose chromatography
-
recombinant wild-type and G96A mutant enzyme
-
glyphosate insensitve strain, ammonium sulfate, heat treatment, DEAE-cellulose, Sephadex G-75, cellulose phosphate, chromatofocusing
-
recombinant enzyme, Q-Sepharose, one step purification
-
enzyme is part of multifunctional protein with distinct domains
-
DEAE-cellulose, CM-cellulose, partial purification
-
ammonium sulfate, DEAE-Sephacel, phenyl-Sepharose, cellulose phosphate
-
from crude cell extracts by DEAE-sepharose anion-exchange chromatography (Pharmacia XK-16/40 column, elution with linear NaCl gradient), Superdex 200 gel filtration chromatography (Pharmacia Superdex 200 HR 10/30 column coupled to FPLC, pH 6.8), Resource Q anion-exchange chromatography (elution with linear NaCl gradient, pH 6.8)
-
using Ni-NTA chromatography
ammonium sulfate, DEAE-cellulose, hydroxyapatite, phenylaggarose, Sephacryl S-200, isozymes I, II and III
-
recombinant enzyme from Escherichia coli strain BL21(DE3)
-
recombinant enzyme, ammonium sulfate, hydrophobic interaction chromatography, anion-exchange chromatography
ammonium sulfate, DEAE-Sephacel, cellulose phosphate column, isoenzymes I and II
-
using an anion exchange Q-Sepharose column, ammonium sulfate precipitation using an equilibrated phenyl Sepharose column
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
expression of wild-type and R24D, P105S, H385L mutant enzymes in Escherichia coli
-
from genomic DNA, in cosmid vector for phage packaging and transfection in Escherichia coli JM101 or in vitro transposon mutagenesis and transformation in Escherichia coli XL1 Blue MRF, in pAX306 for transformation of Escherichia coli DH5alpha and XL1 Blue MRF, in pRSF1b for inducible expression in Escherichia coli, in pSB11 shuttle vector for integration into Agrobacterium tumefaciens LBA4404 (already harbouring pSB1) and subsequent transformation of positive Agrobacterium strains in maize embryo for generation of transgenic plants
-
coding sequence of CaEPSPS gene is successfully subcloned in a plasmid-Escherichia coli system (pET-32a), and the cells containing the plasmid carrying the CaEPSPS gene exhibit enhanced tolerance to herbicide glyphosate, compared to the control
amplified by 3 and 5 RACE on total RNA according to an EST found in Dunaliella salina cDNA library, cloned into pMD18-T for sequencing, cloned in pGEX-4T-1 for expression as mature protein (lacking N-terminal 70 amino acids long transit peptide) in Escherichia coli JM109 or EPSP synthase gene deleted Escherichia coli strain ER2799; expression in Escherichia coli
Escherichia coli structural gene aroA
-
Escherichia coli structural gene aroA; subcloned from bacteriophage lambdapserC into multicopy plasmid pAT153 with subsequent transformation of Escherichia coli AB2829 CGSC2829 cells
-
expressed as a His-tagged fusion protein
-
expression of the wild-type enzyme and of the chimeric mutant aroA-M1, comprising N-terminal part of the Escherichia coli enzyme and the C-terminal part of the enzyme from Salmonella typhimurium in strain XL1-Blue
-
expression of the wild-type enzyme, of the chimeric mutant aroA-M1, comprising N-terminal part of the Escherichia coli enzyme and the C-terminal part of the enzyme from Salmonella typhimurium, and of T42M-aroA mutant in strain BL21(DE3)
-
expression of wild-type and chimeras of Salmonella and Escherichia coli mutant enzymes in Escherichia coli
-
expression of wild-type and G96A mutant enzyme in Escherichia coli
-
in pET24d for expression in Escherichia coli DE3
-
mutant enzyme G96A/A183T is transferred to Brassica napus via Agrobacterium-mediated transformation. Transgenic plants are resistant to glyphosate at a concentration of 10 mM whereas the non-transformed control plants are unable to survive 1 mM glyphosate
-
overexpressed in Escherichia coli
-
overexpression in Escherichia coli
-
overexpression of wild-type and mutant enzymes in strain BL21(DE3)
overexpression of wild-type, His-tagged wild-type and several mutant enzymes in Escherichia coli
-
wild-type and mutants in pET-24d for expression in Escherichia coli BL21(DE3)
-
expression in Escherichia coli
overexpression in Escherichia coli
-
expression in Escherichia coli
expression in Escherichia coli
-
from genomic DNA, in pUC18 for complementation studies in Escherichia coli strain AB2829 (aroA auxotroph), in pTrc99A for inducible expression in Escherichia coli AB 2829
-
expression in Escherichia coli
N-terminal fragment (until Phe295) of G2 EPSPS or the complete gene in pMEB2 (N-terminal of splicing domain of of DnaE intein, strain PCC6803) and C-terminus (from Thr296) of G2 EPSPS in pKEB12 (C-terminal of splicing domain of DnaE intein, strain PCC6803) for intein-mediated protein complementation and expression in Escherichia coli strain ER2799, complete G2 EPSPS gene, N-terminal and C-terminal fragments in pBI121 for introduction into Agrobacterium sp. strain LBA4404 and transformation in Nicotiana tabacum variant NC89 for generation of transgenic plants
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
-
overexpressed as a His-tagged fusion protein
expression of the chimeric mutant aroA-M1, comprising N-terminal part of the Escherichia coli enzyme and the C-terminal part of the enzyme from Salmonella typhimurium in Escherichia coli strain XL1-Blue
-
expression of the chimeric mutant aroA-M1, comprising N-terminal part of the Escherichia coli enzyme and the C-terminal part of the enzyme from Salmonella typhimurium, and of T42M-aroA mutant in Escherichia coli strain BL21(DE3)
-
expression of wild-type and chimeras of Salmonella typhimurium and Escherichia coli mutant enzymes in Escherichia coli
-
overexpression in Escherichia coli strain BL21(DE3)
-
expression in Escherichia coli
expressed in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A100G
glyphosate-sensitive, compared to wild-type similar inhibition constants for (3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid, (3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid, and (3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
A100G
-
glyphosate-sensitive, compared to wild-type similar inhibition constants for (3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid, (3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid, and (3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid
-
H385L
-
0.2% of wild-type activity, 2fold activation at 100 mM NH4Cl
H385N
-
6% of wild-type activity
P105S
-
69% of wild-type activity, 8fold activation at 100 mM NH4Cl
R24D
-
0.8% of wild-type activity, 2fold activation at 100 mM NH4Cl
DELTAN1-22
-
lacking N-terminal 22 residues including critical amino acid Lys20, fails to provide glyphosate tolerance compared to full-length enzyme when transformed in glyphosate-sensitive Escherichia coli DH5alpha and XL1 Blue
DELTAN1-22
Bacteria ATX1398
-
lacking N-terminal 22 residues including critical amino acid Lys20, fails to provide glyphosate tolerance compared to full-length enzyme when transformed in glyphosate-sensitive Escherichia coli DH5alpha and XL1 Blue
-
P106S
-
glyphosate-insensitive
P381L
-
similiar glyphosate sensitivity like wild-type
A183T
-
38fold decrease in phosphoenolpyruvate-binding affinity, more solvent-exposed tryptophan residues and lower stability against guanidine hydrochloride compared to wild-type and mutant G96A, midpoint guanidine hydrochloride concentration of unfolding: 0.7 M, higher structural flexibility and decrease of secondary structure (28% alpha helix, 35% beta sheet) compared to wild-type (40% alpha helix, 31% beta sheet) and lowest resistance against proteolysis, residue A183 located on the exterior in the N-terminal domain
D242A
-
site-directed mutagenesis, the mutation is responsible for the high increase in activity
D313A
-
0.42% of wild-type activity
D313A
site-directed mutagenesis, comparison of the mutant active site and substrate binding structures to those of the wild-type enzyme
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
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
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
D313N
-
5% of wild-type activity
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
D348A
-
site-directed mutagenesis, the mutation is responsible for the high increase in activity
D49A
-
41% of wild-type activity
E341A
-
0.3% of wild-type activity
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
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
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
E341Q
-
10% of wild-type activity
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
G96A
-
glyphosate-insensitive
G96A
-
glyphosate-insensitive, 31fold decrease in phosphoenolpyruvate-binding affinity, little change with respect to structural flexibility and stability compared to wild-type, lower structural flexibility than wild-type and mutants A183T and G96A/A183T, midpoint guanidine hydrochloride concentration of unfolding: 1.1 M, residue Gly96 located at interphase between two domains
G96A/A183T
-
mutant enzyme is resistant to glyphosate
G96A/A183T
-
glyphosate-insensitive, 8fold decrease in phosphoenolpyruvate-binding affinity, more solvent-exposed tryptophan residues and lower stability against guanidine hydrochloride compared to wild-type and mutant G96A, midpoint guanidine hydrochloride concentration of unfolding: 0.65 M, higher structural flexibility and decrease of secondary structure (36% alpha helix, 38% beta sheet) compared to wild-type (40% alpha helix, 31% beta sheet)
H385A
-
0.08% of wild-type activity
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)
K22A
-
0.7% of wild-type activity
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
K22R
-
3% of wild-type activity
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)
K340A
-
2.4% of wild-type activity
N94A
-
50% of wild-type activity
P101A
-
slight decrease in catalytic efficiency, decreased inhibitory potency of glyphosate
P101G
-
slight decrease in catalytic efficiency, decreased inhibitory potency of glyphosate
P101L
-
lowest catalytic efficiency, decreased inhibitory potency of glyphosate due to long-range conformational changes
P101S
-
slight decrease in catalytic efficiency, decreased inhibitory potency of glyphosate due to long-range conformational changes, flexibility of Ser101 side chain disables hydrophobic bonding with carbonyl oxygen of Thr97
P101S
-
the substrate binding affinities, as reflected by the respective Km values, are only slightly decreased for the P101S mutant. Km (3-phosphoshikimate): 0.071 mM, Km (phosphoenolpyruvate): 0.071, kcat/Km (phosphoenolpyruvate): 230000/Msec, kcat/Km (3-phosphoshikimate): 240000/Msec. Mutant P101S is moderately inhibited by glyphosate
Q171A
-
1.7% of wild-type activity
R100A
-
site-directed mutagenesis, the mutation is responsible for the high increase in activity
R100M
-
0.2% of wild-type activity
R124A
-
19.6% of wild-type activity
R27A
-
binding of shikimate 3-phosphate is abolished
R344K
-
31.7% of wild-type activity
R344M
-
16.3% of wild-type activity
R386M
-
15.8% of wild-type activity
T42M
-
site-directed mutagenesis, the mutation is responsible for the glyphosate resistance and the low Km for phosphoenolpyruvate
T971I
-
the single site T97I mutation renders the enzyme sensitive to glyphosate and causes a substantial decrease in the affinity for phosphoenolpyruvate. Km (3-phosphoshikimate): 0.077 mM, Km (phosphoenolpyruvate): 0.38, kcat/Km (phosphoenolpyruvate): 23000/Msec, kcat/Km (3-phosphoshikimate): 1200000/Msec
T97I/P101S
-
mutant is essentially insensitive to glyphosate (Ki 2.4 mM) but maintains high affinity for the substrate phosphoenolpyruvate (Km: 0.1 mM) and 3-phosphoshikimate (Km: 0.077 mM). kcat/Km (phosphoenolpyruvate): 57000/Msec, kcat/Km (3-phosphoshikimate): 740000/Msec. The crystal structure at 1.7 A resolution reveals that the dual mutation causes a shift of residue Gly96 toward the glyphosate binding site, impairing efficient binding of glyphosate, while the side chain of Ile97 points away from the substrate binding site, facilitating phosphoenolpyruvate utilization
Y200F
-
1% of wild-type activity
T355S
mutation results in a catalytically more efficient glyphosate-resistant enzyme
T355S
Halothermothrix orenii H168
-
mutation results in a catalytically more efficient glyphosate-resistant enzyme
-
G96A
-
glyphosate-insensitive
E145G/N163H/N267S/P318R/M377V/M425T/P438L
mutant is able to restore the growth of the Escherichia coli mutant ER2799, with the 5-enolpyruvylshikimate-3-phosphate synthase gene deleted from its genome, on an M9 minimal medium containing 300 mM glyphosate. The effectiveness of amino acid alterations N267S, P318R, and M245T on glyphosate tolerance is in the decreasing order P318R, M425T, N267S
M425T
mutation identified in a multiple mutant enzyme resistant to glyphosate
N267S
mutation identified in a multiple mutant enzyme resistant to glyphosate
N267S/P318R/M425T
mutant enzyme shows high tolerance against glyphosate
P318R
mutation identified in a multiple mutant enzyme resistant to glyphosate
P106L
P106L mutant has a high glyphosate resistance while retaining relatively high catalytic efficiency at low phosphoenolpyruvate concentrations. 3-fold increase in glyphosate resistance of Escherichia coli expressing the P106L mutant. Affinity of the P106L mutant for glyphosate and phosphoenolpyruvate is decreased about 70-fold and 4.6-fold, respectively, compared to wild-type
G99A
-
KM-value for 5-enolpyruvylshikimate 3-phosphate is 86% of the wild-type value, KM-value for phosphate is 1.3fold higher than wild-type value. Turnover-number for 5-enolpyruvylshikimate 3-phosphate is 6.2fold higher than wild-type value, turnover-number for phosphate is 5.4fold lower than wild-type value
G99A
-
similar substrate affinities and decreased turnover compared to wild-type, fails to complement aroA auxotrophy in transformed Escherichia coli
L104P
-
KM-value for 5-enolpyruvylshikimate 3-phosphate is 1.14fold higher than the wild-type value, KM-value for phosphate is 4.4fold higher than wild-type value. Turnover-number for 5-enolpyruvylshikimate 3-phosphate is 2.7fold lower than wild-type value, turnover-number for phosphate is 1.9fold lower than wild-type value
L104P
-
lower affinity to phosphate and decreased turnover compared to wild-type, fails to complement aroA auxotrophy in transformed Escherichia coli
M129W
no enzymatic activity
N130D
in a growth complementation assay no difference to wild-type
N130G
in a growth complementation assay no difference to wild-type
N130H
in a growth complementation assay no difference to wild-type
N130L
in a growth complementation assay no difference to wild-type
N130P
partial impaiment in growth complementation assay
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
N130S
in a growth complementation assay no difference to wild-type, Km (phosphoenolpyruvate): 0.032 mM
N130W
partial impaiment in growth complementation assay, mutant shows a significant decrease in resistance to glyphosate, Km (phosphoenolpyruvate): 0.025 mM
P128R
no enzymatic activity
R127E
no enzymatic activity
R127K
no enzymatic activity
R131E
no enzymatic activity
R131K
no enzymatic activity
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
-
G96A
mutant with improved resistance towards glyphosate
T38K/R40V/R222Q/S224V/I225V/Q226K
mutant with the c-strand of subdomain 3 and the f-strand of subdomain 5 substituted by the corresponding region of the Escherichia coli enzyme. The mutant enzyme exhibits greater sensitivity to glyphosate than the wild type with little change of affinity for substrates shikimate-3-phosphate and phosphoenolpyruvate
G96A
Rahnella aquatilis GR20
-
mutant with improved resistance towards glyphosate
-
T42M
-
site-directed mutagenesis, the mutation is responsible for the glyphosate resistance and the low Km for phosphoenolpyruvate
T103I/P107S
-
double mutant (47 kDa) which is widely used by genetic engineering to confer glyphosate tolerant properties, to various crops such as corn, cotton, canola, and soybean is assessed in terms of safety aspects: The expressed protein is innocuous. The double mutant enzyme does not possess any of the properties associated with known toxins or allergens, a rapid degradation in simulated gastric and intestinal fluids, and no adverse effects in mice after intravenousor oral administration is shown
K411A
-
10.4% of wild-type activity
additional information
-
construction of chimeric mutants by fusing the N-terminal part of the Escherichia coli enzyme with the C-terminal part of the enzyme from Salmonella typhimurium, the chimeric mutant shows a 6fold lower Km for phosphoenolpyruvate, a 5fold higher Ki for glyphosate, and a 9fold higher specific activity than the wild-type enzyme from Escherichia coli
additional information
-
construction of chimeric mutants by fusing the N-terminal part of the Escherichia coli enzyme with the C-terminal part of the enzyme from Salmonella typhimurium, the chimeric mutant shows a 6fold lower Km for phosphoenolpyruvate, a 5fold higher Ki for glyphosate, and a 9fold higher specific activity than the wild-type enzyme from Escherichia coli, construction of glyphosate-resistant transgenic Nicotiana tabacum plants expressing the chimeric mutant aroA-M1 using the Agrobacterium tumefaciens strain LBA 4404 infection system
T38K/R40V/R222Q/S224V/I225V/Q226K
Rahnella aquatilis GR20
-
mutant with the c-strand of subdomain 3 and the f-strand of subdomain 5 substituted by the corresponding region of the Escherichia coli enzyme. The mutant enzyme exhibits greater sensitivity to glyphosate than the wild type with little change of affinity for substrates shikimate-3-phosphate and phosphoenolpyruvate
-
additional information
-
construction of chimeric mutants by fusing the N-terminal part of the Escherichia coli enzyme with the C-terminal part of the enzyme from Salmonella typhimurium, the chimeric mutant shows a 6fold lower Km for phosphoenolpyruvate, a 5fold higher Ki for glyphosate, and a 9fold higher specific activity than the wild-type enzyme from Escherichia coli
additional information
-
construction of chimeric mutants by fusing the N-terminal part of the Escherichia coli enzyme with the C-terminal part of the enzyme from Salmonella typhimurium, i.e. mutant construct aroA-M1, the chimeric mutant shows a 6fold lower Km for phosphoenolpyruvate, a 5fold higher Ki for glyphosate, and a 9fold higher specific activity than the wild-type enzyme from Escherichia coli, construction of glyphosate-resistant transgenic Nicotiana tabacum plants expressing the chimeric mutant aroA-M1 using the Agrobacterium tumefaciens strain LBA 4404 infection system
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
pellet from 80% (w/v) ammonium sulfate solution, resolubilization and dialysis against 5 mM sodium phosphate, pH 6.9
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
drug development
differential inhibition of class I and class II EPSPS by tetrahedral reaction intermediate-analogues possibly due to alteration of open-close transition during catalysis and/or upon inhibitor binding but not due to energy differences during complex formation
drug development
-
differential inhibition of class I and class II EPSPS by tetrahedral reaction intermediate-analogues possibly due to alteration of open-close transition during catalysis and/or upon inhibitor binding but not due to energy differences during complex formation
-
agriculture
transgenic Arabidopsis thaliana plants expressing the enzyme from Bacillus cereus germinate well in presence of 1 mM glyphosate, while control plants do not germinate at 0.2 mM
biotechnology
-
conferring glyphosate tolerance in transgenic crop plants
drug development
-
more experimental data needed for effective structure-based drug design
biotechnology
Bacteria ATX1398
-
conferring glyphosate tolerance in transgenic crop plants
-
biotechnology
glyphosate tolerance in bacteria
biotechnology
Dunaliella salina 435
-
glyphosate tolerance in bacteria
-
drug development
-
protonated enolpyruvylshikimate 3-phosphate (cation) intermediate as potential target for inhibitor design
drug development
-
differential inhibition of class I and class II EPSPS by tetrahedral reaction intermediate-analogues possibly due to alteration of open-close transition during catalysis and/or upon inhibitor binding but not due to energy differences during complex formation
synthesis
-
enzyme is a target for development and synthesis of antimicrobial drugs
agriculture
transgenic Arabidopsis thaliana plants expressing the enzyme from Halothermothrix orenii are more resistant to glyphosate exposure than transgenic plants expressing the Escherichia coli enzyme and control plants
agriculture
Halothermothrix orenii H168
-
transgenic Arabidopsis thaliana plants expressing the enzyme from Halothermothrix orenii are more resistant to glyphosate exposure than transgenic plants expressing the Escherichia coli enzyme and control plants
-
agriculture
Arabidopsis thaliana transgenic plants expressing mutant E145G/N163H/N267S/P318R/M377V/M425T/P438L grow well in presence of 0.75 mM glyphosate, whereas Arabidopsis thaliana plants expressing wild-type enzyme grow only in presence of 0.25 mM glyphosate
agriculture
transgenic Arabidopsis thaliana plants expressing the enzyme from Pseudomonas fluorescens are more resistant to glyphosate exposure compared with wild-type
biotechnology
strain PCC6803 DnaE intein N-terminal and C-terminal splicing domains can reconstitute EPSPS activities and glyphosate resistance in plant
biotechnology
Pseudomonas fluorescens G2
-
strain PCC6803 DnaE intein N-terminal and C-terminal splicing domains can reconstitute EPSPS activities and glyphosate resistance in plant
-
agriculture
transgenic Arabidopsis thaliana plants expressing the enzyme from Rahnella aquatilis are more resistant to glyphosate exposure than plants expressing the Escherichia coli enzyme
agriculture
Rahnella aquatilis GR20
-
transgenic Arabidopsis thaliana plants expressing the enzyme from Rahnella aquatilis are more resistant to glyphosate exposure than plants expressing the Escherichia coli enzyme
-
drug development
-
differential inhibition of class I and class II EPSPS by tetrahedral reaction intermediate-analogues possibly due to alteration of open-close transition during catalysis and/or upon inhibitor binding but not due to energy differences during complex formation