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 ACCESSION NO.
LITERATURE
4-Hydroxybenzoyl-CoA + H2O = 4-hydroxybenzoate + CoA
show the reaction diagram
mechanism
-
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
active site structure and ligand binding mechanism, overview
-
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
mechanism; substrate and inhibitor binding by wild-type and mutant D17N enzymes, reaction mechanism and active site structure
Pseudomonas sp. CBS-3
-
-
4-Hydroxybenzoyl-CoA + H2O = 4-hydroxybenzoate + CoA
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of thioester
-
-
-
-
hydrolysis of thioester
-
-
hydrolysis of thioester
-
-
hydrolysis of thioester
-, Q84HI6
-
hydrolysis of thioester
-
-
-
PATHWAY
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 ACCESSION NO.
COMMENTARY
LITERATURE
4-HBA-CoA thioesterase
-
-
4-HBA-CoA thioesterase
-
-
-
4-HBA-CoA thioesterase
-
-
4-HBA-CoA thioesterase
P56653
-
4-HBA-CoA thioesterase
-
-
-
4-hydroxybenzoyl coenzyme A hydrolase
-
-
4-hydroxybenzoyl coenzyme A hydrolase
-
-
-
4-hydroxybenzoyl coenzyme A thioesterase
-
-
4-hydroxybenzoyl coenzyme A thioesterase
P56653
-
4-hydroxybenzoyl coenzyme A thioesterase
-
-
-
4-hydroxybenzoyl-coenzyme A thioesterase
-
-
4-hydroxybenzoyl-coenzyme A thioesterase
-
-
-
benzoyl-coenzyme A thioesterase
Q84HI6
-
Hydrolase, 4-hydroxybenzoyl coenzyme A
-
-
-
-
Hydrolase, 4-hydroxybenzoyl coenzyme A
-
-
Hydrolase, 4-hydroxybenzoyl coenzyme A
-
-
-
Hydrolase, 4-hydroxybenzoyl coenzyme A
-
-
Hydrolase, 4-hydroxybenzoyl coenzyme A
Pseudomonas sp. CBS-3, Pseudomonas sp. CBS3
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
141583-19-9
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
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
strain CBS-3
-
-
Manually annotated by BRENDA team
strain CBS3
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
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
-
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
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
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
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 ACCESSION NO.
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 ACCESSION NO.
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]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
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
0.0024
-
4-hydroxybenzoyl-CoA
-
mutant D17E, pH 7.5, 25C
0.0045
-
4-hydroxybenzoyl-CoA
-
mutant R88A, pH 7.5, 25C
0.0045
-
4-hydroxybenzoyl-CoA
-
-
0.005
-
4-hydroxybenzoyl-CoA
-
mutant R126L, pH 7.5, 25C
0.0052
-
4-hydroxybenzoyl-CoA
-
mutant R89L, 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
0.007
-
4-hydroxybenzoyl-CoA
-, Q84HI6
recombinant enzyme
0.0073
-
4-hydroxybenzoyl-CoA
-
mutant R128A, 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]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
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
0.00055
-
4-hydroxybenzoyl-CoA
-
mutant D17N, pH 7.5, 25C
0.00084
-
4-hydroxybenzoyl-CoA
-
mutant D17S, pH 7.5, 25C
0.11
-
4-hydroxybenzoyl-CoA
-
mutant R126L, pH 7.5, 25C
0.13
-
4-hydroxybenzoyl-CoA
-
mutant R88A, 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
0.18
-
4-hydroxybenzoyl-CoA
-
wild-type enzyme and mutant R89L, pH 7.5, 25C
0.48
-
4-hydroxybenzoyl-CoA
-
mutant D17E, pH 7.5, 25C
6.8
-
4-hydroxybenzoyl-CoA
-
-
10
-
4-hydroxybenzoyl-CoA
-, Q84HI6
recombinant enzyme
18.3
-
4-hydroxybenzoyl-CoA
P56653
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
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
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]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
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
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
-
-
assay at
8.5
-
-, Q84HI6
recombinant enzyme
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
-
-, Q84HI6
recombinant enzyme shows half maximal activity
10.5
-
-, Q84HI6
recombinant enzyme shows half maximal activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
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
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
62000
-
-, Q84HI6
native protein, determined by gel filtration
66000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
homotetramer
-, Q84HI6
4 * 16000, determined by gel filtration
tetramer
-
dimer of dimers, structure analysis
tetramer
-
mass spectrometry
tetramer
-
4 * 16000, SDS-PAGE; 4 * 16107, calculation from nucleotide sequence
tetramer
-
4 * 16000, SDS-PAGE
tetramer
-
dimer of dimers, structure analysis; mass spectrometry
-
tetramer
Pseudomonas sp. CBS-3
-
4 * 16000, SDS-PAGE; 4 * 16000, SDS-PAGE; 4 * 16107, calculation from nucleotide sequence
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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
-
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
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
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
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, recombinant enzyme, Tris-HCl buffer, pH 8, several months stable
-, Q84HI6
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
by nickel chelate affinity and gel filtration chromatography
-, Q84HI6
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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 ACCESSION NO.
COMMENTARY
LITERATURE
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
-