Any feedback?
No. of entries
Information on EC 6.2.1.71 - 2,3-dihydroxybenzoate-[aryl-carrier protein] ligase
for references in articles please use BRENDA:EC6.2.1.71
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree 6 Ligases
6.2 Forming carbon-sulfur bonds
6.2.1 Acid-thiol ligases
6.2.1.71 2,3-dihydroxybenzoate-[aryl-carrier protein] ligase
IUBMB Comments
The adenylation domain of the enzyme catalyses the activation of 2,3-dihydroxybenzoate to (2,3-dihydroxybenzoyl)adenylate, followed by the transfer the activated compound to the free thiol of a phosphopantetheine arm of an aryl-carrier protein domain of a specific non-ribosomal peptide synthase. For example, the EntE enzyme of Escherichia coli is part of the enterobactin synthase complex, the VibE enzyme of Vibrio cholerae is part of the vibriobactin synthase complex, and the DhbE enzyme of Bacillus subtilis is part of the bacillibactin synthase complex.
The enzyme appears in viruses and cellular organisms
Reaction Schemes
show+
+
=
+
+
+
+
holo-[aryl-carrier protein]
=
+
+
Synonyms
2,3-dihydroxybenzoate-amp ligase, vibriobactin-specific 2,3-dihydroxybenzoate-amp ligase, more
topPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
2,3-dihydroxybenzoate-AMP ligase
angE
-
-
-
-
dhbE
-
-
-
-
EC 2.7.7.58
-
-
formerly
-
EntE
synthetase, (2,3-dihydroxybenzoyl)adenylate
-
-
-
-
vibE
vibriobactin-specific 2,3-dihydroxybenzoate-AMP ligase
2,3-dihydroxybenzoate-AMP ligase
-
-
-
-
EntE
-
-
-
-
vibriobactin-specific 2,3-dihydroxybenzoate-AMP ligase
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
(2,3-dihydroxybenzoyl)adenylate + holo-[aryl-carrier protein] = AMP + 2,3-dihydroxybenzoyl-[aryl-carrier protein]
(1b)
-
-
-
ATP + 2,3-dihydroxybenzoate + holo-[aryl-carrier protein] = AMP + diphosphate + 2,3-dihydroxybenzoyl-[aryl-carrier protein]
overall reaction
-
-
-
ATP + 2,3-dihydroxybenzoate = diphosphate + (2,3-dihydroxybenzoyl)adenylate
(1a)
-
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
nucleotidyl group transfer
-
-
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PATHWAY SOURCE
PATHWAYS
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SYSTEMATIC NAME
IUBMB Comments
2,3-dihydroxybenzoate:[aryl-carrier protein] ligase (AMP-forming)
The adenylation domain of the enzyme catalyses the activation of 2,3-dihydroxybenzoate to (2,3-dihydroxybenzoyl)adenylate, followed by the transfer the activated compound to the free thiol of a phosphopantetheine arm of an aryl-carrier protein domain of a specific non-ribosomal peptide synthase. For example, the EntE enzyme of Escherichia coli is part of the enterobactin synthase complex, the VibE enzyme of Vibrio cholerae is part of the vibriobactin synthase complex, and the DhbE enzyme of Bacillus subtilis is part of the bacillibactin synthase complex.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CAS REGISTRY NUMBER
COMMENTARY
122332-73-4
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(2,3-dihydroxybenzoyl)adenylate + holo-[aryl-carrier protein EntB]
AMP + 2,3-dihydroxybenzoyl-[aryl-carrier protein EntB]
Substrates: -
Products: -
Products: -
?
ATP + 2,3,4-trihydroxybenzoic acid
diphosphate + (2,3,4-trihydroxybenzoyl)adenylate
-
Substrates: -
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate
diphosphate + 2,3-dihydroxybenzoyl adenylate
-
Substrates: -
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate + holo-[aryl-carrier protein EntB]
AMP + diphosphate + 2,3-dihydroxybenzoyl-[aryl-carrier protein EntB]
Substrates: -
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate + [aryl-carrier protein domain of EntB]
AMP + diphosphate + 2,3-dihydroxybenzoyl-[aryl-carrier protein domain of EntB]
Substrates: -
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate + [aryl-carrier protein EntB]
AMP + diphosphate + 2,3-dihydroxybenzoyl-[aryl-carrier protein EntB]
ATP + 2,4-dihydroxybenzoic acid
diphosphate + (2,4-dihydroxybenzoyl)adenylate
-
Substrates: -
Products: -
Products: -
?
ATP + 2,5-dihydroxybenzoic acid
diphosphate + (2,5-dihydroxybenzoyl)adenylate
-
Substrates: i.e. gentisic acid
Products: -
Products: -
?
ATP + 2-fluorobenzoate
diphosphate + (2-fluorobenzoyl)adenylate
Substrates: low activity
Products: -
Products: -
?
ATP + 2-hydroxybenzoic acid
diphosphate + (2-hydroxybenzoyl)adenylate
-
Substrates: i.e. salicylic acid, not 3- or 4-derivative
Products: -
Products: -
?
ATP + 3-aminobenzoate
diphosphate + (3-aminobenzoyl)adenylate
Substrates: low activity
Products: -
Products: -
?
ATP + 3-bromobenzoate
diphosphate + (3-bromobenzoyl)adenylate
Substrates: low activity
Products: -
Products: -
?
ATP + 3-chlorobenzoate
diphosphate + (3-chlorobenzoyl)adenylate
Substrates: low activity
Products: -
Products: -
?
ATP + 3-ethynylbenzoate
diphosphate + (3-ethynylbenzoyl)adenylate
Substrates: low activity
Products: -
Products: -
?
ATP + 3-fluorobenzoate
diphosphate + (3-fluorobenzoyl)adenylate
Substrates: low activity
Products: -
Products: -
?
ATP + 3-hydroxybenzoate
diphosphate + (3-hydroxybenzoyl)adenylate
Substrates: low activity
Products: -
Products: -
?
ATP + 3-hydroxybenzoate + [aryl-carrier protein EntB]
AMP + diphosphate + 3-hydroxybenzoyl-[aryl-carrier protein EntB]
Substrates: -
Products: -
Products: -
?
ATP + 3-iodobenzoate
diphosphate + (3-iodobenzoyl)adenylate
Substrates: low activity
Products: -
Products: -
?
ATP + 3-methoxybenzoate
diphosphate + (3-methoxybenzoyl)adenylate
Substrates: low activity
Products: -
Products: -
?
ATP + 3-methylbenzoate
diphosphate + (3-methylbenzoyl)adenylate
Substrates: low activity
Products: -
Products: -
?
ATP + 4-aminosalicylate + [aryl-carrier protein EntB]
AMP + diphosphate + 4-aminosalicyl-[aryl-carrier protein EntB]
Substrates: -
Products: -
Products: -
?
ATP + anthranilate
diphosphate + anthranylyl-adenylate
Substrates: low activity
Products: -
Products: -
?
ATP + salicylate
diphosphate + salicyl-adenylate
Substrates: low activity
Products: -
Products: -
r
ATP + salicylate + holo-[aryl-carrier protein domain of EntB]
AMP + diphosphate + salicyl-[aryl-carrier protein domain of EntB]
Substrates: -
Products: -
Products: -
?
ATP + salicylate + holo-[aryl-carrier protein domain of SrfB1]
AMP + diphosphate + salicyl-[aryl-carrier protein domain of SrfB1]
Substrates: SrfB1, i.e. peptidyl carrier protein of surfactin synthetase
Products: -
Products: -
?
ATP + salicylate + holo-[aryl-carrier protein EntB]
AMP + diphosphate + salicyl-[aryl-carrier protein EntB]
ATP + salicylate + [aryl-carrier protein EntB]
AMP + diphosphate + salicyl-[aryl-carrier protein EntB]
Substrates: -
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
-
Substrates: -
Products: product remains enzyme-bound for further reaction in overall biosynthesis of enterobactin
Products: product remains enzyme-bound for further reaction in overall biosynthesis of enterobactin
?
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
-
Substrates: -
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
-
Substrates: activation of 2,3-dihydroxybenzoic acid in the biosynthesis of siderophore enterobactin
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
-
Substrates: two-step reaction catalyzed by EnTE of Escherichia coli: enzyme first catalyzes the formation of the DHB adenylate
Products: -
Products: -
r
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
Substrates: -
Products: -
Products: -
r
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
Substrates: -
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
Substrates: highly preferred substrate
Products: -
Products: -
r
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
-
Substrates: two-step reaction catalyzed by EnTE of Escherichia coli: enzyme first catalyzes the formation of the DHB adenylate
Products: -
Products: -
r
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
-
Substrates: involved in the biosynthesis of anguibactin
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
-
Substrates: -
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate + [aryl-carrier protein EntB]
AMP + diphosphate + 2,3-dihydroxybenzoyl-[aryl-carrier protein EntB]
Substrates: 2,3-dihydroxybenzoate binds to EntE with a 1:1 stoichiometry and a KD of 7.4 microM. The EC50 value of EntE-EntB interaction is approximately 1.5 microM
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate + [aryl-carrier protein EntB]
AMP + diphosphate + 2,3-dihydroxybenzoyl-[aryl-carrier protein EntB]
Substrates: -
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate + [aryl-carrier protein EntB]
AMP + diphosphate + 2,3-dihydroxybenzoyl-[aryl-carrier protein EntB]
Substrates: -
Products: -
Products: -
?
ATP + salicylate + holo-[aryl-carrier protein EntB]
AMP + diphosphate + salicyl-[aryl-carrier protein EntB]
Substrates: -
Products: -
Products: -
?
ATP + salicylate + holo-[aryl-carrier protein EntB]
AMP + diphosphate + salicyl-[aryl-carrier protein EntB]
Substrates: -
Products: -
Products: -
?
additional information
?
-
-
Substrates: no substrates are benzoic acid, 2,6-dihydroxybenzoic acid, 2,4,6-trihydroxybenzoic acid, anthranilic acid, thiosalicylic acid
Products: -
Products: -
?
additional information
?
-
-
Substrates: in a second half reaction, EnTE transfers the DHB moiety to the phosphopantetheine (PPant) cofactor of the EntB aryl carrier protein (ArCP) domain, corresponding to the reaction (2,3-dihydroxybenzoyl)adenylate + phosphopantetheine cofactor-ENTB = 2,3-DHB-S-EntB + AMP
Products: -
Products: -
?
additional information
?
-
Substrates: substrate specificity analysis of wild-type and mutant enzymes, overview. veryl low activity of wild-type enzyme with 3-nitrobenzoate, 3-cyanobenzoate, 2-nitrobenzoate, 2-methylbenzoate, 2-trifluoromethylbenzoate, 2-ethynylbenzoate, 2-azidobenzoate, 2-chlorobenzoate, 2-iodobenzoate, and 2-bromobenzoate
Products: -
Products: -
?
additional information
?
-
-
Substrates: in a second half reaction, EnTE transfers the DHB moiety to the phosphopantetheine (PPant) cofactor of the EntB aryl carrier protein (ArCP) domain, corresponding to the reaction (2,3-dihydroxybenzoyl)adenylate + phosphopantetheine cofactor-ENTB = 2,3-DHB-S-EntB + AMP
Products: -
Products: -
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(2,3-dihydroxybenzoyl)adenylate + holo-[aryl-carrier protein EntB]
AMP + 2,3-dihydroxybenzoyl-[aryl-carrier protein EntB]
Substrates: -
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate + holo-[aryl-carrier protein EntB]
AMP + diphosphate + 2,3-dihydroxybenzoyl-[aryl-carrier protein EntB]
Substrates: -
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
-
Substrates: activation of 2,3-dihydroxybenzoic acid in the biosynthesis of siderophore enterobactin
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
Substrates: -
Products: -
Products: -
r
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
Substrates: -
Products: -
Products: -
?
ATP + 2,3-dihydroxybenzoate
diphosphate + (2,3-dihydroxybenzoyl)adenylate
-
Substrates: involved in the biosynthesis of anguibactin
Products: -
Products: -
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2,3-dihydroxybenzohydroxamoyl adenylate
-
strong product analog inhibitor, competitive inhibitor
2-hydroxybenzohydroxamoyl adenylate
-
deletion of the catecholic 3-OH group of 2,3-dihydroxybenzohydroxamoyl adenylate reduces inhibitory activity by a factor of 3.5
5'-O-[N-(2,3-dihydroxybenzoyl)sulfamoyl]adenosine
acts as slow-onset tight-binding inhibitor
5'-O-[N-(salicyl)sulfamoyl]adenosine
i.e.Sal-AMS, acts as slow-onset tight-binding inhibitor
acetohydroxamoyl adenylate
-
replacing the entire catechol moiety of 2,3-dihydroxybenzohydroxamoyl adenylate by a hydrogen atom gives up to 10% inhibition
benzohydroxamoyl adenylate
-
deletion of both the catecholic 3-OH and 2-OH group of 2,3-dihydroxybenzohydroxamoyl adenylate reduces inhibitory activity by a factor of 2000
additional information
usage of mechanism-based inhibitors containing a vinylsulfonamide functional group appended to capture the terminal thiol group of the Ppant arm of the ArCP domain
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
addition of a 4fold excess of dihydro-2,3-dihydroxybenzoate dehydrogenase EntA to an 2,3-dihydroxybenzoate-AMP ligaseEntE assay mixture results in a 6fold stimulation of EntE activity
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
steady-state Michaelis-Menten kinetic analysis of wild-type and mutant enzymes, overview
-
0.417
2,3-Dihydroxybenzoate
pH 8.0, 37°C, mutant Y236F/S240C/V339I/N340C
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.147
2,3-Dihydroxybenzoate
pH 8.0, 37°C, mutant Y236F/S240C/V339I/N340C
0.008
salicylate
acceptor aryl-carrier protein domain of SrfB1, pH 7.5, 37°C
2.15
salicylate
acceptor aryl-carrier protein domain of EntB, pH 7.5, 37°C
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.353
2,3-Dihydroxybenzoate
pH 8.0, 37°C, mutant Y236F/S240C/V339I/N340C
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.000009
2,3-dihydroxybenzohydroxamoyl adenylate
-
product analog inhibitor, competitive inhibitor, result indicates that the catecholic 2-OH group is critical for tight binding
0.000037
2-hydroxybenzohydroxamoyl adenylate
-
product analog inhibitor, competitive inhibitor
0.0000038
5'-O-[N-(2,3-dihydroxybenzoyl)sulfamoyl]adenosine
pH 7.8, 25°C
0.013
benzohydroxamoyl adenylate
-
product analog inhibitor, competitive inhibitor
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0054
-
specific activity of mutant R437D/K473D by using complete reaction, measuring AMP production with coupled assay
0.018
-
specific activity of mutant R437D by using complete reaction, measuring AMP production with coupled assay
0.156
-
specific activity of mutant K473D by using complete reaction, measuring AMP production with coupled assay
0.165
-
specific activity of mutant R494D by using complete reaction, measuring AMP production with coupled assay
0.18
-
specific activity of wild-type EntE by using complete reaction, measuring AMP production with coupled assay
176.4
-
specific activity of mutant R437D/K473D by measuring diphosphate exchange
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Highest Expressing Human Cell Lines
Cell Line LinksPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
the NRPS codes of DHB- and Sal-activating A-domains are about 60% similar. The notable differences involve amino acid residues at positions 236, 240, 339, and 340. Tyr236, which is positioned near the aryl group of DHB in EntE, tends to be a Phe in Sal-activating A-domains. The hydroxyl group of active-site Ser240 of EntE engages in effective hydrogen-bonding interaction with the 3-hydroxyl moiety of the DHB substrate. Consequently, the Ser residue is highly conserved in DHB-activating A-domains (EntE, BasE, and VibE). Sequence alignment reveals that this Ser is replaced with a Cys residue in Sal-activating A-domains (YbtE, MbtA, and PchD). Val339 of DhbE is conserved in the DHB-activating A-domains EntE and BasE, but not VibE, whereas the corresponding residue is replaced with a relatively more sterically demanding Leu (MbtA and YbtE) or Ile (PchD) in Sal-activating A-domains. In addition to Ser240, which functions as the hydrogen-bonding donor, Val339 is likely to act as a key residue for the selective recognition of the DHB substrate in DHB-activating A-domains. Asn340 in the active site of DhbE is strictly conserved in the DHB-activating A-domains EntE, BasE, and VibE. By contrast, the residue corresponding to Asn340 is a Cys in the Sal-activating A-domains YbtE and PchD, but not MbtA
malfunction
substituting Val339 with a bulky Leu or Ile residue reduces the size of the active-site cavity, allowing for the selective activation of the Sal substrate in the active site
metabolism
physiological function
metabolism
enzyme EntE catalyzes the adenylation of DHB with ATP to form DHB-AMP. The activated DHB is transferred to the ArCP domain of EntB to generate a thioester conjugate of DHB harboring the 4'-phosphopantetheine (Ppant) functionality of the ArCP domain of EntB
metabolism
EntE only interacts efficiently with EntB in the presence of 2,3-dihydroxybenzoate. The transient complex is formed to facilitate channeling of 2,3-dihydroxybenzoate-AMP
metabolism
EntE proceeds via a Bi Uni Uni Bi ping-pong kinetic mechanism. During the first half-reaction (adenylation), dihydroxybenzoate binds first to the free enzyme, followed by ATP and the release of diphosphate to form the adenylate intermediate. During the second half-reaction (ligation), phosphopantetheinylated EntB binds to the enzyme followed by the release of products, AMP and arylated EntB
physiological function
The Escherichia coli gene cluster for the synthesis and activation of 2,3-dihydroxybenzoic acid in the formation of the catechol siderophore enterobactin shows the gene order entEBG(AC). The genes are organized as three independent transcriptional units, composed of entE, entBG, and entAC, with the entBG mRNA transcribed in a clockwise direction
physiological function
the holo-EntB ArCP domain is acylated with 2,3-dihydroxybenzoate in a reaction in which EntE catalyzes the activation of 2,3-dihydroxybenzoate by adenylation as well as the transfer of 2,3-dihydroxybenzoate to the phosphopantetheine cofactor of holo-EntB
physiological function
enterobactin, the tris-(N-(2,3-dihydroxybenzoyl)serine) trilactone siderophore of Escherichia coli, is synthesized by a three-protein (EntE, B, F) six-module nonribosomal peptide synthetase
physiological function
during biosynthesis of Vibrio cholerae siderophore vibriobactin, VibE activates 2,3-dihydroxybenzoate as the acyl adenylate and then transfers it to the free thiol of the phosphopantetheine arm of VibB's aryl carrier domain
Show AA Sequence and Transmembrane Helices (143 entries)
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PDB
SCOP
CATH
UNIPROT
ORGANISM
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
60000
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
dihydro-2,3-dihydroxybenzoate dehydrogenase EntA can form a specific complex with 2,3-dihydroxybenzoate-AMP ligaseEntE with a stoichiometry of one EntA tetramer and four EntE monomers. EntA-EntE binding behavior is sigmoidal
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
construction of a model for DHB binding to EntE (PDB ID 3RG2) through superposition of the DHB-DhbE complex structure (PDB entry 1MD9)
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K473D
-
specific activity of mutant K473D by measuring AMP production with coupled assay= 0.0156 micromol/min/mg, by measuring diphosphate exchange= 182.7 micromol/min/mg
N235A
site-directed mutagenesis, altered substrate specificity and kinetics compared to wild-type
N340C
site-directed mutagenesis, altered substrate specificity and kinetics compared to wild-type
R437D
-
specific activity of mutant R437D by measuring AMP production with coupled assay= 0.018 micromol/min/mg, by measuring diphosphate exchange= 197.4 micromol/min/mg
R437D/K473D
-
specific activity of mutant R437D/K473D by measuring AMP production with coupled assay= 0.0054 micromol/min/mg, by measuring diphosphate exchange= 176.4 micromol/min/mg
R494D
-
specific activity of mutant R494D by measuring AMP production with coupled assay= 0.0165 micromol/min/mg, by measuring diphosphate exchange= 189 micromol/min/mg
S240C
site-directed mutagenesis, altered substrate specificity and kinetics compared to wild-type
V339I
site-directed mutagenesis, altered substrate specificity and kinetics compared to wild-type
V339L
site-directed mutagenesis, altered substrate specificity and kinetics compared to wild-type
Y236F
site-directed mutagenesis, altered substrate specificity and kinetics compared to wild-type
Y236F/S240C
site-directed mutagenesis, altered substrate specificity and kinetics compared to wild-type
Y236F/S240C/N340C
site-directed mutagenesis, altered substrate specificity and kinetics compared to wild-type
Y236F/S240C/V339I/N340C
site-directed mutagenesis, altered substrate specificity and kinetics compared to wild-type
Y236F/S240C/V339L/N340C
K473D
-
specific activity of mutant K473D by measuring AMP production with coupled assay= 0.0156 micromol/min/mg, by measuring diphosphate exchange= 182.7 micromol/min/mg
-
R437D
-
specific activity of mutant R437D by measuring AMP production with coupled assay= 0.018 micromol/min/mg, by measuring diphosphate exchange= 197.4 micromol/min/mg
-
R437D/K473D
-
specific activity of mutant R437D/K473D by measuring AMP production with coupled assay= 0.0054 micromol/min/mg, by measuring diphosphate exchange= 176.4 micromol/min/mg
-
R494D
-
specific activity of mutant R494D by measuring AMP production with coupled assay= 0.0165 micromol/min/mg, by measuring diphosphate exchange= 189 micromol/min/mg
-
additional information
Y236F/S240C/V339L/N340C
site-directed mutagenesis, altered substrate specificity and kinetics compared to wild-type
Y236F/S240C/V339L/N340C
site-directed mutagenesis, altered substrate specificity and kinetics compared to wild-type, very poor activityy with 2,3-dihydroxybenzoate
additional information
-
mutagenesis studies support hypothesis that members of adenylate -forming family of enzymes adopt two distinct conformations to catalyze the two-step mechanism
additional information
detection of the role of active-site residues in aryl acid adenylation domains (A-domains) by gradually grafting the NRPS codes used for salicylic acid (Sal) into an archetypal aryl acid A domain, EntE, that is specific for the substrate 2,3-dihydroxybenzoic acid (DHB). Enzyme kinetics and modeling studies of the EntE variants demonstrate that the NRPS code residues at positions 236, 240, and 339 collectively regulate the substrate specificity toward DHB and Sal. The EntE variants exhibit the ability to activate the non-native aryl acids 3-hydroxybenzoic acid, 3-aminobenzoic acid, 3-fluorobenzoic acid, and 3-chlorobenzoic acid. The didomain structure of the ArCP domain of EntB and EntE is also determined using mechanism-based inhibitors containing a vinylsulfonamide functional group appended to capture the terminal thiol group of the the 4'-phosphopantetheine (Ppant) arm of the ArCP domain
additional information
-
mutagenesis studies support hypothesis that members of adenylate -forming family of enzymes adopt two distinct conformations to catalyze the two-step mechanism
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70°C to 2°C, 25 mM Tris-HCl buffer, pH 8.0, 5 mM DTT, 10 mM MgCl2, 50% glycerol, stable for several months
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant protein using His-tag with poor purification results, purification by conventional purification methods
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed as His-tag fusion protein in Escherichia coli origami(DE3), complementation of an enzyme deficient Escherichia coli strain
-
expression in Escherichia coli
gene entE, recobinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
structural gene entE subcloned into multi-copy plasmid pKK223-3 under control of tac-promoter, DNA sequence determination
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Nahlik, M.S.; Fleming, T.P.; McIntosh, M.A.
Cluster of genes controlling synthesis and activation of 2,3-dihydroxybenzoic acid in production of enterobactin in Escherichia coli
J. Bacteriol.
169
4163-4170
1987
Escherichia coli (P10378)
brenda
Rusnak, F.; Faraci, W.S.; Walsh, C.T.
Subcloning, expression, and purification of the enterobactin biosynthetic enzyme 2,3-dihydroxybenzoate-AMP ligase: demonstration of enzyme-bound (2,3-dihydroxybenzoyl)adenylate product
Biochemistry
28
6827-6835
1989
Escherichia coli
brenda
Staab, J.F.; Elkins, M.F.; Earhart, C.F.
Nucleotide sequence of the Escherichia coli entE gene
FEMS Microbiol. Lett.
59
15-20
1989
Escherichia coli
-
brenda
Liu, Q.; Ma, Y.; Wu, H.; Shao, M.; Liu, H.; Zhang, Y.
Cloning, identification and expression of an entE homologue angE from Vibrio anguillarum serotype 01
Arch. Microbiol.
181
287-293
2004
Vibrio anguillarum
brenda
Callahan, B.P.; Lomino, J.V.; Wolfenden, R.
Nanomolar inhibition of the enterobactin biosynthesis enzyme, EntE: synthesis, substituent effects, and additivity
Bioorg. Med. Chem. Lett.
16
3802-3805
2006
Escherichia coli
brenda
Drake, E.J.; Nicolai, D.A.; Gulick, A.M.
Structure of the EntB multidomain nonribosomal peptide synthetase and functional analysis of its interaction with the EntE adenylation domain
Chem. Biol.
13
409-419
2006
Escherichia coli, Escherichia coli JM109
brenda
Liu, X.; Wang, Z.; Zhu, D.; Wei, T.; Gu, L.; Xu, S.
Crystallization and preliminary X-ray crystallographic studies of VibE, a vibriobactin-specific 2,3-dihydroxybenzoate-AMP ligase from Vibrio cholerae
Acta Crystallogr. Sect. F
67
1563-1565
2011
Vibrio cholerae (Q9FDB2), Vibrio cholerae
brenda
Khalil, S.; Pawelek, P.D.
Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE)
Biochemistry
50
533-545
2011
Escherichia coli
brenda
Sikora, A.L.; Wilson, D.J.; Aldrich, C.C.; Blanchard, J.S.
Kinetic and inhibition studies of dihydroxybenzoate-AMP ligase from Escherichia coli
Biochemistry
49
3648-3657
2010
Escherichia coli (P10378)
brenda
Gehring, A.M.; Bradley, K.A.; Walsh, C.T.
Enterobactin biosynthesis in Escherichia coli isochorismate lyase (EntB) is a bifunctional enzyme that is phosphopantetheinylated by EntD and then acylated by EntE using ATP and 2,3-dihydroxybenzoate
Biochemistry
36
8495-8503
1997
Escherichia coli (P10378)
brenda
Keating, T.; Marshall, C.; Walsh, C.
Vibriobactin biosynthesis in Vibrio cholerae VibH is an amide synthase homologous to nonribosomal peptide synthetase condensation domains
Biochemistry
39
15513-15521
2000
Vibrio cholerae (Q9FDB2)
brenda
Ishikawa, F.; Nohara, M.; Nakamura, S.; Nakanishi, I.; Tanabe, G.
Precise probing of residue roles by NRPS code swapping mutation, enzymatic characterization, modeling, and substrate promiscuity of aryl acid adenylation domains
Biochemistry
59
351-363
2020
Escherichia coli (P10378)
brenda
Wyckoff, E.; Stoebner, J.; Reed, K.; Payne, S.
Cloning of a Vibrio cholerae vibriobactin gene cluster Identification of genes required for early steps in siderophore biosynthesis
J. Bacteriol.
179
7055-7062
1997
Vibrio cholerae serotype O1 (O07899), Vibrio cholerae serotype O1 ATCC 39315 (O07899)
brenda
Khalil, S.; Pawelek, P.D.
Ligand-induced conformational rearrangements promote interaction between the Escherichia coli enterobactin biosynthetic proteins EntE and EntB
J. Mol. Biol.
393
658-671
2009
Escherichia coli (P10378)
brenda
Ehmann, D.; Shaw-Reid, C.; Losey, H.; Walsh, C.
The EntF and EntE adenylation domains of Escherichia coli enterobactin synthetase Sequestration and selectivity in acyl-AMP transfers to thiolation domain cosubstrates
Proc. Natl. Acad. Sci. USA
97
2509-2514
2000
Escherichia coli (P10378)
brenda
EXTERNAL LINKS (specific for EC-Number 6.2.1.71)
html completed
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2026.1 (March 2026)
About BRENDA
Social media
Help
Survey
Download
Contribution
Legal
Curated at
Member of
![DSMZ Digital Diversity]()
![Global Core Biodata Resource]()
![ELIXIR Core Data Resource]()
![German Network for Bioinformatics Infrastructure]()
