Information on EC 3.1.2.23 - 4-hydroxybenzoyl-CoA thioesterase

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The expected taxonomic range for this enzyme is: Bacteria

EC NUMBER
COMMENTARY
3.1.2.23
-
RECOMMENDED NAME
GeneOntology No.
4-hydroxybenzoyl-CoA thioesterase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
4-Hydroxybenzoyl-CoA + H2O = 4-hydroxybenzoate + CoA
show the reaction diagram
mechanism
-
4-Hydroxybenzoyl-CoA + H2O = 4-hydroxybenzoate + CoA
show the reaction diagram
active site structure and ligand binding mechanism, overview
-
4-Hydroxybenzoyl-CoA + H2O = 4-hydroxybenzoate + CoA
show the reaction diagram
catalytic sites of wild-type and mutant enzymes, substrate binding modifications, reaction kinetics
-
4-Hydroxybenzoyl-CoA + H2O = 4-hydroxybenzoate + CoA
show the reaction diagram
substrate and inhibitor binding by wild-type and mutant D17N enzymes, reaction mechanism and active site structure
-
4-Hydroxybenzoyl-CoA + H2O = 4-hydroxybenzoate + CoA
show the reaction diagram
catalytic sites of wild-type and mutant enzymes, substrate binding modifications, reaction kinetics
-
-
4-Hydroxybenzoyl-CoA + H2O = 4-hydroxybenzoate + CoA
show the reaction diagram
active site structure and ligand binding mechanism, overview
-
-
4-Hydroxybenzoyl-CoA + H2O = 4-hydroxybenzoate + CoA
show the reaction diagram
mechanism, substrate and inhibitor binding by wild-type and mutant D17N enzymes, reaction mechanism and active site structure
Pseudomonas sp. CBS-3
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of thioester
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
4-chlorobenzoate degradation
-
-
4-hydroxybenzoate biosynthesis I (eukaryotes)
-
-
4-hydroxybenzoate biosynthesis V
-
-
Benzoate degradation
-
-
Biosynthesis of secondary metabolites
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
Ubiquinone and other terpenoid-quinone biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
4-hydroxybenzoyl-CoA hydrolase
This enzyme is part of the bacterial 2,4-dichlorobenzoate degradation pathway.
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Hydrolase, 4-hydroxybenzoyl coenzyme A
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
141583-19-9
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
KB-740, DSMZ 6869, formerly designated Pseudomonas sp. KB740
UniProt
Manually annotated by BRENDA team
CBS3, recombinantly expressed in Escherichia coli
SwissProt
Manually annotated by BRENDA team
strain CBS3
-
-
Manually annotated by BRENDA team
Pseudomonas sp. CBS-3
-
SwissProt
Manually annotated by BRENDA team
Pseudomonas sp. CBS-3
strain CBS-3
-
-
Manually annotated by BRENDA team
strain CBS3
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,3-dihydro-2,3-dihydroxybenzoyl-CoA + H2O
2,3-dihydro-2,3-dihydroxybenzoate + CoA
show the reaction diagram
Q84HI6
-
-
-
?
2-aminobenzoyl-CoA + H2O
2-aminobenzoate + CoA
show the reaction diagram
Q84HI6
305% relative activity with 2-aminobenzoyl-CoA compared to benzoyl-CoA as substrate
-
-
?
2-fluorobenzoyl-CoA + H2O
2-fluorobenzoate + CoA
show the reaction diagram
Q84HI6
189% relative activity with 2-fluorobenzoyl-CoA compared to benzoyl-CoA as substrate
-
-
?
2-hydroxybenzoyl-CoA + H2O
2-hydroxybenzoate + CoA
show the reaction diagram
Q84HI6
486% relative activity with 2-hydroxybenzoyl-CoA compared to benzoyl-CoA as substrate
-
-
?
2-methylbenzoyl-CoA + H2O
2-methylbenzoate + CoA
show the reaction diagram
Q84HI6
10% relative activity with 2-methylbenzoyl-CoA compared to benzoyl-CoA as substrate
-
-
?
4-Chlorobenzoyl-CoA + H2O
4-Chlorobenzoate + CoA
show the reaction diagram
-
-
-
-
4-Chlorobenzoyl-CoA + H2O
4-Chlorobenzoate + CoA
show the reaction diagram
P56653
-
-
-
?
4-Chlorobenzoyl-CoA + H2O
4-Chlorobenzoate + CoA
show the reaction diagram
Pseudomonas sp. CBS-3
-
-
-
-
4-fluorobenzoyl-CoA + H2O
4-fluorobenzoate + CoA
show the reaction diagram
P56653
-
-
-
?
4-fluorobenzoyl-CoA + H2O
4-fluorobenzoate + CoA
show the reaction diagram
Q84HI6
380% relative activity with 4-fluorobenzoyl-CoA compared to benzoyl-CoA as substrate
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
-
-
-
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
-
-
-
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
-
-
-
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
-
-
-
-
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
P56653
-
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
Q84HI6
-
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
Q04416
-
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
enzyme is part of the dehalogenation pathway of 4-chlorobenzoyl-CoA in soil bacteria
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
enzyme is part of the dehalogenation pathway of 4-chlorobenzoyl-CoA in soil bacteria
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
Q84HI6
1300% relative activity with 4-hydroxybenzoyl-CoA compared to benzoyl-CoA as substrate
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
the CoA pantothenic acid moiety is inserted into the outer part of the active site tunnel and the nucleotide moiety is anchored to the protein surface, whereas the hydroxybenzoyl group binds in an enclosed pocket. Product 4-hydroxybenzoate is released from the active site of the enzyme prior to the release of product CoA following catalytic turnover. 4-Hydroxybenzoate dissociates directly from the hydroxybenzoyl binding pocket more quickly than CoA can move out of the connecting tunnel
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
-
-
-
-
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
enzyme is part of the dehalogenation pathway of 4-chlorobenzoyl-CoA in soil bacteria
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
the CoA pantothenic acid moiety is inserted into the outer part of the active site tunnel and the nucleotide moiety is anchored to the protein surface, whereas the hydroxybenzoyl group binds in an enclosed pocket. Product 4-hydroxybenzoate is released from the active site of the enzyme prior to the release of product CoA following catalytic turnover. 4-Hydroxybenzoate dissociates directly from the hydroxybenzoyl binding pocket more quickly than CoA can move out of the connecting tunnel
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
Q04416
-
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
Arthrobacter sp. 4-CB1
-
-
-
-
-
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
Pseudomonas sp. CBS-3
-
-
-
-
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
Pseudomonas sp. CBS-3
-
-
-
-
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
Pseudomonas sp. CBS-3
-
-
-
-
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
Pseudomonas sp. CBS-3
-
enzyme is part of the dehalogenation pathway of 4-chlorobenzoyl-CoA in soil bacteria
-
?
4-Hydroxybenzoyl-CoA + H2O
?
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. CBS-3
-
the enzyme is involved in the pathway by which this organism converts 4-chlorobenzoate to 4-hydroxybenzoate
-
-
-
4-methoxybenzoyl-CoA + H2O
4-methoxybenzoate + CoA
show the reaction diagram
P56653
-
-
-
?
4-methylbenzoyl-CoA + H2O
4-methylbenzoate + CoA
show the reaction diagram
P56653
-
-
-
?
4-trifluoromethylbenzoyl-CoA + H2O
4-trifluoromethylbenzoate + CoA
show the reaction diagram
P56653
-
-
-
?
Benzoyl-CoA + H2O
Benzoate + CoA
show the reaction diagram
-
-
-
-
Benzoyl-CoA + H2O
Benzoate + CoA
show the reaction diagram
P56653
-
-
-
?
Benzoyl-CoA + H2O
Benzoate + CoA
show the reaction diagram
Q84HI6
-
-
-
?
Benzoyl-CoA + H2O
Benzoate + CoA
show the reaction diagram
Pseudomonas sp. CBS-3
-
-
-
-
additional information
?
-
Q84HI6
the intermediates of the pathway are processed as coenzyme A thioesters and the cleavage of the aromatic ring is non-xygenolytic
-
-
-
additional information
?
-
P56653
reaction proceeds via formation of an aspartyl17-(4-hydroxybenzoyl)anhydride intermediate that undergoes rate-limiting hydrolytic cleavage at the hydroxybenzoyl carbonyl carbon atom. CoA and 4-hydroxybenzoate binding is weak
-
-
-
additional information
?
-
Q04416
residue Glu73 functions in nucleophilic catalysis, Gly65 and Gln58 contribute to transition-state stabilization via hydrogen bond formation with the thioester moiety and Thr77 orients the water nucleophile for attack at the 4-hydroxybenzoyl carbon of the enzymeanhydride intermediate. Mutation E73D switches the function of the carboxylate residue from nucleophilic catalysis to base catalysis and thus, the reaction from a two-step process involving a covalent enzyme intermediate to a single-step hydrolysis reaction. The substrate nucleotide unit is bound to the enzyme surface
-
-
-
additional information
?
-
Pseudomonas sp. CBS-3
P56653
reaction proceeds via formation of an aspartyl17-(4-hydroxybenzoyl)anhydride intermediate that undergoes rate-limiting hydrolytic cleavage at the hydroxybenzoyl carbonyl carbon atom. CoA and 4-hydroxybenzoate binding is weak
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
2,3-dihydro-2,3-dihydroxybenzoyl-CoA + H2O
2,3-dihydro-2,3-dihydroxybenzoate + CoA
show the reaction diagram
Q84HI6
-
-
-
?
2-aminobenzoyl-CoA + H2O
2-aminobenzoate + CoA
show the reaction diagram
Q84HI6
305% relative activity with 2-aminobenzoyl-CoA compared to benzoyl-CoA as substrate
-
-
?
2-fluorobenzoyl-CoA + H2O
2-fluorobenzoate + CoA
show the reaction diagram
Q84HI6
189% relative activity with 2-fluorobenzoyl-CoA compared to benzoyl-CoA as substrate
-
-
?
2-hydroxybenzoyl-CoA + H2O
2-hydroxybenzoate + CoA
show the reaction diagram
Q84HI6
486% relative activity with 2-hydroxybenzoyl-CoA compared to benzoyl-CoA as substrate
-
-
?
2-methylbenzoyl-CoA + H2O
2-methylbenzoate + CoA
show the reaction diagram
Q84HI6
10% relative activity with 2-methylbenzoyl-CoA compared to benzoyl-CoA as substrate
-
-
?
4-fluorobenzoyl-CoA + H2O
4-fluorobenzoate + CoA
show the reaction diagram
Q84HI6
380% relative activity with 4-fluorobenzoyl-CoA compared to benzoyl-CoA as substrate
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
Q84HI6
-
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
enzyme is part of the dehalogenation pathway of 4-chlorobenzoyl-CoA in soil bacteria
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
enzyme is part of the dehalogenation pathway of 4-chlorobenzoyl-CoA in soil bacteria
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
Q84HI6
1300% relative activity with 4-hydroxybenzoyl-CoA compared to benzoyl-CoA as substrate
-
-
?
4-Hydroxybenzoyl-CoA + H2O
?
show the reaction diagram
-
the enzyme is involved in the pathway by which this organism converts 4-chlorobenzoate to 4-hydroxybenzoate
-
-
-
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
-
-
?
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
-
enzyme is part of the dehalogenation pathway of 4-chlorobenzoyl-CoA in soil bacteria
-
?
4-Hydroxybenzoyl-CoA + H2O
?
show the reaction diagram
Pseudomonas sp. CBS-3
-
the enzyme is involved in the pathway by which this organism converts 4-chlorobenzoate to 4-hydroxybenzoate
-
-
-
4-Hydroxybenzoyl-CoA + H2O
4-Hydroxybenzoate + CoA
show the reaction diagram
Pseudomonas sp. CBS-3
-
enzyme is part of the dehalogenation pathway of 4-chlorobenzoyl-CoA in soil bacteria
-
?
Benzoyl-CoA + H2O
Benzoate + CoA
show the reaction diagram
Q84HI6
-
-
-
?
additional information
?
-
Q84HI6
the intermediates of the pathway are processed as coenzyme A thioesters and the cleavage of the aromatic ring is non-xygenolytic
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K+
Q84HI6
recombinant enzyme tolerates up to 0.6 M KCl, 27% activity enhancement when 0.1 M KCl is added to the assay
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2,5-dihydroxybenzoyl-CoA
Q84HI6
the strongest inhibitor of the di-substituted non-substrates
4-hydroxybenzyl-CoA
-
-
4-hydroxybenzyl-CoA
-
-
4-hydroxyphenacyl-CoA
-
-
4-hydroxyphenacyl-CoA
-
binding mechanism
phenylacetyl-CoA
Q84HI6
inhibits the hydrolysis of benzoyl-CoA
acetyl-CoA
Q84HI6
inhibits the hydrolysis of benzoyl-CoA
additional information
Q84HI6
di-substituted derivatives of benzoyl-CoA, phenylacetyl-CoA and aliphatic CoA thioesters are not hydrolyzed but some act as inhibitors
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.19
4-chlorobenzoyl-CoA
-
-
550
4-chlorobenzoyl-CoA
P56653
pH 7.5, 25C
5 - 20
4-fluorobenzoyl-CoA
P56653
pH 7.5, 25C
0.00014
4-hydroxybenzoyl-CoA
-
mutant K90A, pH 7.5, 25C
0.00038
4-hydroxybenzoyl-CoA
Q04416
mutant T77D, pH 7.5, 25C
0.00077
4-hydroxybenzoyl-CoA
Q04416
mutant T77S, pH 7.5, 25C
0.0012
4-hydroxybenzoyl-CoA
Q04416
mutant T77A, pH 7.5, 25C; wild-type, pH 7.5, 25C
0.0016
4-hydroxybenzoyl-CoA
Q04416
mutant Q58A, pH 7.5, 25C; mutant S120A, pH 7.5, 25C
0.0024
4-hydroxybenzoyl-CoA
-
mutant D17E, pH 7.5, 25C
0.004
4-hydroxybenzoyl-CoA
Q04416
mutant D31A, pH 7.5, 25C; mutant Q58E, pH 7.5, 25C
0.0045
4-hydroxybenzoyl-CoA
-
mutant R88A, pH 7.5, 25C
0.0045
4-hydroxybenzoyl-CoA
-
-
0.0046
4-hydroxybenzoyl-CoA
Q04416
mutant E73D/T77A, pH 7.5, 25C
0.005
4-hydroxybenzoyl-CoA
-
mutant R126L, pH 7.5, 25C
0.0052
4-hydroxybenzoyl-CoA
-
mutant R89L, pH 7.5, 25C
0.0055
4-hydroxybenzoyl-CoA
Q04416
mutant T77V, pH 7.5, 25C
0.006
4-hydroxybenzoyl-CoA
-
wild-type enzyme, pH 7.5, 25C
0.006
4-hydroxybenzoyl-CoA
P56653
pH 7.5, 25C
0.0061
4-hydroxybenzoyl-CoA
Q04416
mutant T121A, pH 7.5, 25C
0.007
4-hydroxybenzoyl-CoA
Q84HI6
recombinant enzyme
0.0073
4-hydroxybenzoyl-CoA
-
mutant R128A, pH 7.5, 25C
0.0128
4-hydroxybenzoyl-CoA
Q04416
mutant D31N, pH 7.5, 25C
0.013
4-hydroxybenzoyl-CoA
Q04416
mutant E73D/T77S, pH 7.5, 25C
0.014
4-hydroxybenzoyl-CoA
Q04416
mutant R102A, pH 7.5, 25C
0.0142
4-hydroxybenzoyl-CoA
Q04416
mutant E73Q, pH 7.5, 25C
0.015
4-hydroxybenzoyl-CoA
Q04416
mutant R150A, pH 7.5, 25C
0.018
4-hydroxybenzoyl-CoA
Q04416
mutant E73D, pH 7.5, 25C
0.096
4-hydroxybenzoyl-CoA
Q04416
mutant E78A, pH 7.5, 25C
0.18
4-hydroxybenzoyl-CoA
Q04416
mutant Q58D, pH 7.5, 25C
0.237
4-hydroxybenzoyl-CoA
Q04416
mutant H64Q, pH 7.5, 25C
0.51
4-hydroxybenzoyl-CoA
Q04416
mutant H64A, pH 7.5, 25C
56
4-methoxybenzoyl-CoA
P56653
pH 7.5, 25C
230
4-methylbenzoyl-CoA
P56653
pH 7.5, 25C
300
4-trifluoromethylbenzoyl-CoA
P56653
pH 7.5, 25C
0.077
benzoyl-CoA
Q84HI6
recombinant enzyme
0.2
benzoyl-CoA
-
-
510
benzoyl-CoA
P56653
pH 7.5, 25C
additional information
additional information
-
kinetics and thermodynamic of enzyme-ligand binding
-
additional information
additional information
-
kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.123
4-Chlorobenzoate
-
-
1.1
4-chlorobenzoyl-CoA
P56653
pH 7.5, 25C
9.7
4-fluorobenzoyl-CoA
P56653
pH 7.5, 25C
0.0001
4-hydroxybenzoyl-CoA
Q04416
mutant E73A, pH 7.5, 25C
0.00055
4-hydroxybenzoyl-CoA
-
mutant D17N, pH 7.5, 25C
0.00084
4-hydroxybenzoyl-CoA
-
mutant D17S, pH 7.5, 25C
0.0017
4-hydroxybenzoyl-CoA
Q04416
mutant E73Q, pH 7.5, 25C
0.0103
4-hydroxybenzoyl-CoA
Q04416
mutant T77D, pH 7.5, 25C
0.0417
4-hydroxybenzoyl-CoA
Q04416
mutant D31A, pH 7.5, 25C
0.11
4-hydroxybenzoyl-CoA
-
mutant R126L, pH 7.5, 25C
0.123
4-hydroxybenzoyl-CoA
Q04416
mutant E73D, pH 7.5, 25C
0.127
4-hydroxybenzoyl-CoA
Q04416
mutant Q58A, pH 7.5, 25C
0.129
4-hydroxybenzoyl-CoA
Q04416
mutant E73D/T77S, pH 7.5, 25C
0.13
4-hydroxybenzoyl-CoA
-
mutant R88A, pH 7.5, 25C
0.13
4-hydroxybenzoyl-CoA
Q04416
mutant Q58D, pH 7.5, 25C
0.14
4-hydroxybenzoyl-CoA
-
mutant K90A, pH 7.5, 25C
0.15
4-hydroxybenzoyl-CoA
-
mutant R128A, pH 7.5, 25C
0.15
4-hydroxybenzoyl-CoA
Q04416
mutant H64A, pH 7.5, 25C
0.17
4-hydroxybenzoyl-CoA
Q04416
mutant T77V, pH 7.5, 25C
0.18
4-hydroxybenzoyl-CoA
-
wild-type enzyme and mutant R89L, pH 7.5, 25C
0.28
4-hydroxybenzoyl-CoA
Q04416
mutant H64Q, pH 7.5, 25C
0.364
4-hydroxybenzoyl-CoA
Q04416
mutant Q58E, pH 7.5, 25C
0.48
4-hydroxybenzoyl-CoA
-
mutant D17E, pH 7.5, 25C
1.08
4-hydroxybenzoyl-CoA
Q04416
mutant E78A, pH 7.5, 25C
1.15
4-hydroxybenzoyl-CoA
Q04416
mutant D31N, pH 7.5, 25C
1.54
4-hydroxybenzoyl-CoA
Q04416
mutant E73D/T77A, pH 7.5, 25C
2.09
4-hydroxybenzoyl-CoA
Q04416
mutant T77A, pH 7.5, 25C
2.4
4-hydroxybenzoyl-CoA
Q04416
wild-type, solvent D2O, pH 7.5, 25C
3.16
4-hydroxybenzoyl-CoA
Q04416
mutant T77S, pH 7.5, 25C
6.7
4-hydroxybenzoyl-CoA
Q04416
wild-type, pH 7.5, 25C
6.8
4-hydroxybenzoyl-CoA
-
-
9.5
4-hydroxybenzoyl-CoA
Q04416
mutant R150A, pH 7.5, 25C
9.7
4-hydroxybenzoyl-CoA
Q04416
mutant T121A, pH 7.5, 25C
10
4-hydroxybenzoyl-CoA
Q84HI6
recombinant enzyme
10.1
4-hydroxybenzoyl-CoA
Q04416
mutant S120A, pH 7.5, 25C
18.3
4-hydroxybenzoyl-CoA
P56653
pH 7.5, 25C
99
4-hydroxybenzoyl-CoA
Q04416
mutant R102A, pH 7.5, 25C
0.27
4-methoxybenzoyl-CoA
P56653
pH 7.5, 25C
0.047
4-methylbenzoyl-CoA
P56653
pH 7.5, 25C
0.038
4-trifluoromethylbenzoyl-CoA
P56653
pH 7.5, 25C
0.26
benzoyl-CoA
-
-
1
benzoyl-CoA
P56653
pH 7.5, 25C
3
benzoyl-CoA
Q84HI6
recombinant enzyme
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.12
4-hydroxybenzoyl-CoA
Q04416
mutant E73Q, pH 7.5, 25C
1434
0.29
4-hydroxybenzoyl-CoA
Q04416
mutant H64A, pH 7.5, 25C
1434
1.1
4-hydroxybenzoyl-CoA
Q04416
mutant E78A, pH 7.5, 25C
1434
1.2
4-hydroxybenzoyl-CoA
Q04416
mutant H64Q, pH 7.5, 25C
1434
6.8
4-hydroxybenzoyl-CoA
Q04416
mutant E73D, pH 7.5, 25C
1434
10
4-hydroxybenzoyl-CoA
Q04416
mutant D31A, pH 7.5, 25C; mutant E73D/T77S, pH 7.5, 25C
1434
27
4-hydroxybenzoyl-CoA
Q04416
mutant T77D, pH 7.5, 25C
1434
31
4-hydroxybenzoyl-CoA
Q04416
mutant T77V, pH 7.5, 25C
1434
72
4-hydroxybenzoyl-CoA
Q04416
mutant Q58D, pH 7.5, 25C
1434
79
4-hydroxybenzoyl-CoA
Q04416
mutant Q58A, pH 7.5, 25C
1434
90
4-hydroxybenzoyl-CoA
Q04416
mutant D31N, pH 7.5, 25C
1434
91
4-hydroxybenzoyl-CoA
Q04416
mutant Q58E, pH 7.5, 25C
1434
340
4-hydroxybenzoyl-CoA
Q04416
mutant E73D/T77A, pH 7.5, 25C
1434
630
4-hydroxybenzoyl-CoA
Q04416
mutant R150A, pH 7.5, 25C
1434
710
4-hydroxybenzoyl-CoA
Q04416
mutant R102A, pH 7.5, 25C
1434
1600
4-hydroxybenzoyl-CoA
Q04416
mutant T121A, pH 7.5, 25C
1434
1700
4-hydroxybenzoyl-CoA
Q04416
mutant T77A, pH 7.5, 25C
1434
4100
4-hydroxybenzoyl-CoA
Q04416
mutant T77S, pH 7.5, 25C
1434
5400
4-hydroxybenzoyl-CoA
Q04416
wild-type, pH 7.5, 25C
1434
6300
4-hydroxybenzoyl-CoA
Q04416
mutant S120A, pH 7.5, 25C
1434
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0006
4-hydroxybenzyl-CoA
-
pH 7.5, 25C
0.0000003
4-hydroxyphenacyl-CoA
-
pH 7.5, 25C
0.0011
4-hydroxyphenacyl-CoA
-
mutant R126L, pH 7.5, 25C
0.0012
4-hydroxyphenacyl-CoA
-
mutant R88A, pH 7.5, 25C
0.0014
4-hydroxyphenacyl-CoA
-
wild-type enzyme, pH 7.5, 25C
0.0018
4-hydroxyphenacyl-CoA
-
mutant D17E, pH 7.5, 25C
0.0021
4-hydroxyphenacyl-CoA
-
mutant R128A, pH 7.5, 25C
0.004
4-hydroxyphenacyl-CoA
-
mutant K90A, pH 7.5, 25C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.02
Q84HI6
extract of cells grown aerobically on benzoate, hydrolysis of benzoyl-CoA
additional information
Q84HI6
100% residual benzoyl-CoA thioesterase activity with benzoyl-CoA in the presence of the non-substrate malonyl-CoA; 10% relative activity with 2-methylbenzoyl-CoA compared to benzoyl-CoA as substrate; 1300% relative activity with 4-hydroxybenzoyl-CoA compared to benzoyl-CoA as substrate; 189% relative activity with 2-fluorobenzoyl-CoA compared to benzoyl-CoA as substrate; 305% relative activity with 2-aminobenzoyl-CoA compared to benzoyl-CoA as substrate; 380% relative activity with 4-fluorobenzoyl-CoA compared to benzoyl-CoA as substrate; 486% relative activity with 2-hydroxybenzoyl-CoA compared to benzoyl-CoA as substrate; 56% residual benzoyl-CoA thioesterase activity with benzoyl-CoA in the presence of the non-substrate phenylacetyl-CoA; 70% residual benzoyl-CoA thioesterase activity with benzoyl-CoA in the presence of the non-substrate 2,5-dihydroxybenzoyl-CoA; 76% residual benzoyl-CoA thioesterase activity with benzoyl-CoA in the presence of the non-substrate acetyl-CoA; 80% residual benzoyl-CoA thioesterase activity with benzoyl-CoA in the presence of the non-substrate 2,3-dihydroxybenzoyl-CoA; 90% residual benzoyl-CoA thioesterase activity with benzoyl-CoA in the presence of the non-substrate 3,5-difluorobenzoyl-CoA; 90% residual benzoyl-CoA thioesterase activity with benzoyl-CoA in the presence of the non-substrate 3,5-dihydroxybenzoyl-CoA; 90% residual benzoyl-CoA thioesterase activity with benzoyl-CoA in the presence of the non-substrate DL-3-hydroxy-3-methylglutaryl-CoA; 90% residual benzoyl-CoA thioesterase activity with benzoyl-CoA in the presence of the non-substrate DL-methylmalonyl-CoA; benzoyl-CoA thioesterase activity is specific for aerobic growth of Azoarcus evansii on benzoate; characterization is carried out using a 5,5-dithio-bis(2-nitrobenzoic acid)-based spectrophotometric assay performed at 30C in 100 mM Tris-HCl buffer; no benzoyl-CoA thioesterase activity can be detected in extracts of cells grown aerobically on phenylacetate and 3-hydroxybenzoate or grown anaerobically on benzoate, phenylacetate, 4-hydroxyphenylacetate and phenylalanine; the enzyme exhibits higher activity with mono-substituted derivatives of benzoyl-CoA, showing highest activity with 4-hydroxybenzoyl-CoA; the recombinant enzyme catalyzes the hydrolysis of 4-hydroxybenzoyl-CoA with a Vmax of 48 micromol/min/mg protein; the recombinant enzyme catalyzes the hydrolysis of benzoyl-CoA with a Vmax of 13 micromol/min/mg protein
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
-
assay at
8.5
Q84HI6
recombinant enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
Q84HI6
recombinant enzyme shows half maximal activity
10.5
Q84HI6
recombinant enzyme shows half maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
-
assay at
30
Q84HI6
assay at
PDB
SCOP
CATH
ORGANISM
Pseudomonas sp. (strain CBS-3)
Pseudomonas sp. (strain CBS-3)
Pseudomonas sp. (strain CBS-3)
Pseudomonas sp. (strain CBS-3)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
62000
Q84HI6
native protein, determined by gel filtration
690642
66000
-
gel filtration
780, 781
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homotetramer
Q84HI6
4 * 16000, determined by gel filtration
tetramer
-
4 * 16107, calculation from nucleotide sequence
tetramer
-
4 * 16000, SDS-PAGE
tetramer
-
dimer of dimers, structure analysis
tetramer
-
mass spectrometry
tetramer
-
dimer of dimers, structure analysis, mass spectrometry
-
tetramer
Pseudomonas sp. CBS-3
-
4 * 16107, calculation from nucleotide sequence, 4 * 16000, SDS-PAGE
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant wild-type enzyme with and without bound inhibitor, hanging drop vapour diffusion method, 4C, 18 mg/ml protein solution containing 10 mM HEPES, pH 7.5, 150 mM KCl, 1 mM 1,4-dithio-D,L-threitol, with or without 1 mM 4-hydroxyphenacyl-CoA, plus precipitant solution containing 17-20% PEG 3400, 100 mM MOPS, pH 7.0, and 200 mM LiCl, X-ray diffraction structure determination at 2.6 A resolution, and analysis
-
wild-type and mutant structures in complex with ligands 4-hydroxybenzoyl-CoA or CoA
Q04416
purified wild-type enzyme with bound inhibitors, 14 mg/ml protein solution containing 10 mM HEPES, pH 7.0, 200 mM KCl, 5 mM 4-hydroxyphenacyl-CoA or 4-hydroxybenzyl-CoA, addition of solution containing 10% polyethylene glycol 8000, 2% Me2SO4, 10 mM succinate, pH 5.0 at room temperature, larger crystals are obtained by usage of 2-45 PEG 8000, X-ray diffracion structure determination at 1.5-1.8 A and 4C, and analysis, D17N mutant with bound substrate 4-hydroxybenzoyl-CoA, macro-seeding into batch technique, 5 mg/ml protein solution containing 8-10% PEG 5000-O-methyl ether, 200 mM KCl, 100 mM MES, pH 6.0, 1 mM substrate, room temperature, X-ray diffraction structure determination at 3.3 A and 4C, and analysis
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
-20
Q84HI6
recombinant enzyme retains activity without significant loss by keeping at -20C for several months in Tris-HCl buffer at pH 8
690642
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, recombinant enzyme, Tris-HCl buffer, pH 8, several months stable
Q84HI6
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by nickel chelate affinity and gel filtration chromatography
Q84HI6
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
the gene is cloned and overexpressed in Escherichia coli to produce a C-terminal His-tag fusion protein
Q84HI6
expression in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D31A
Q04416
steady-state kinetic analysis
D31N
Q04416
steady-state kinetic analysis
E73A
Q04416
steady-state kinetic analysis, crystallization data
E73D
Q04416
mutation switches the function of the carboxylate residue from nucleophilic catalysis to base catalysis and thus, the reaction from a two-step process involving a covalent enzyme intermediate to a single-step hydrolysis reaction, steady-state kinetic analysis
E73D/T77A
Q04416
mutant regains most of the catalytic efficiency lost in the E73D single mutant
E73D/T77S
Q04416
steady-state kinetic analysis
E73Q
Q04416
steady-state kinetic analysis, crystallization data
E78A
Q04416
steady-state kinetic analysis
H64A
Q04416
steady-state kinetic analysis, crystallization data
H64Q
Q04416
steady-state kinetic analysis
Q58A
Q04416
steady-state kinetic analysis, crystallization data
Q58D
Q04416
steady-state kinetic analysis
Q58D/E73D NA
Q04416
complete loss of activity
Q58E
Q04416
steady-state kinetic analysis
Q58E/E73A NA
Q04416
complete loss of activity
Q58E/E73Q NA
Q04416
complete loss of activity
R102A
Q04416
steady-state kinetic analysis
R150A
Q04416
steady-state kinetic analysis
S120A
Q04416
steady-state kinetic analysis
T121A
Q04416
steady-state kinetic analysis
T77A
Q04416
configuration of the active site residues is unchanged. The Thr77 side chain is positioned to bind and orient a water molecule for attack at the benzoyl carbonyl carbon of the putative anhydride intermediate, steady-state kinetic analysis, crystallization data
T77D
Q04416
steady-state kinetic analysis
T77S
Q04416
Ser77 is more effective than is the Thr77 at positioning the water for attack at the 4-hydroxybenzoyl carbonyl carbon of the anhydride intermediate, steady-state kinetic analysis, crystallization data
T77V
Q04416
steady-state kinetic analysis
E73A
-
steady-state kinetic analysis, crystallization data
-
E78A
-
steady-state kinetic analysis
-
R102A
-
steady-state kinetic analysis
-
S120A
-
steady-state kinetic analysis
-
T77D
-
steady-state kinetic analysis
-
D17E
-
site-directed mutagensis, increased activity
D17N
-
site-directed mutagensis, highly reduced activity
D17N
-
site-directed mutagensis, role of Asp17 in ligand binding
D17S
-
site-directed mutagensis, highly reduced activity
K90A
-
site-directed mutagensis, enhanced sensitivity against inhibitor 4-hydroxyphenacyl-CoA
R126L
-
site-directed mutagensis, slightly reduced activity
R128A
-
site-directed mutagensis, similar to the wild-type enzyme
R88A
-
site-directed mutagensis, slightly reduced activity
R89L
-
site-directed mutagensis, similar to the wild-type enzyme
D17N
Pseudomonas sp. CBS-3
-
site-directed mutagensis, role of Asp17 in ligand binding
-
D17E
-
site-directed mutagensis, increased activity
-
D17N
-
site-directed mutagensis, highly reduced activity
-
D17S
-
site-directed mutagensis, highly reduced activity
-
K90A
-
site-directed mutagensis, enhanced sensitivity against inhibitor 4-hydroxyphenacyl-CoA
-
R88A
-
site-directed mutagensis, slightly reduced activity
-