Information on EC 6.2.1.33 - 4-chlorobenzoate-CoA ligase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
6.2.1.33
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RECOMMENDED NAME
GeneOntology No.
4-chlorobenzoate-CoA ligase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
4-Chlorobenzoate + CoA + ATP = 4-chlorobenzoyl-CoA + AMP + diphosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
formation of thioester
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-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
4-chlorobenzoate degradation
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Fluorobenzoate degradation
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Metabolic pathways
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Microbial metabolism in diverse environments
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SYSTEMATIC NAME
IUBMB Comments
4-chlorobenzoate:CoA ligase
Requires Mg2+. This enzyme is part of the bacterial 2,4-dichlorobenzoate degradation pathway.
CAS REGISTRY NUMBER
COMMENTARY hide
141583-20-2
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain TM-1
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-
Manually annotated by BRENDA team
strain TM-1
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-
Manually annotated by BRENDA team
strain CBS3
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Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-Bromobenzoate + CoA + ATP
4-Bromobenzoyl-CoA + AMP + diphosphate
show the reaction diagram
4-Chlorobenzoate + CoA + ADP
4-Chlorobenzoyl-CoA + AMP + phosphate
show the reaction diagram
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reaction with ADP at 7.5% of the reaction with ATP
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-
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4-Chlorobenzoate + CoA + ATP
4-Chlorobenzoyl-CoA + AMP + diphosphate
show the reaction diagram
4-Chlorobenzoate + CoA + ATP
?
show the reaction diagram
4-Fluorobenzoate + CoA + ATP
4-Fluorobenzoyl-CoA + AMP + diphosphate
show the reaction diagram
4-Iodobenzoate + CoA + ATP
4-Iodobenzoyl-CoA + AMP + diphosphate
show the reaction diagram
4-Methoxybenzoate + CoA + ATP
4-Methoxybenzoyl-CoA + AMP + diphosphate
show the reaction diagram
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at 9% of the activity with 4-chlorobenzoate
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-
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4-Methylbenzoate + CoA + ATP
4-Methylbenzoyl-CoA + AMP + diphosphate
show the reaction diagram
4-Nitrobenzoate + CoA + ATP
4-Nitrobenzoyl-CoA + AMP + diphosphate
show the reaction diagram
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at 7% of the activity with 4-chlorobenzoate
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-
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Benzoate + CoA + ATP
Benzoyl-CoA + AMP + diphosphate
show the reaction diagram
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
4-Chlorobenzoate + CoA + ATP
4-Chlorobenzoyl-CoA + AMP + diphosphate
show the reaction diagram
4-Chlorobenzoate + CoA + ATP
?
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
-
70% of the activation with Mg2+
Ni2+
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11% of the activation with Mg2+
Zn2+
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23% of the activation with Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-chlorophenacyl-CoA
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AMP
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is a noncompetitive inhibitor versus ATP and an uncompetitive versus CoA
diphosphate
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competitive versus ATP
Fe3+
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0009 - 0.253
4-Chlorobenzoate
0.0131 - 3.5
ATP
0.03 - 6
CoA
0.1
MgATP2-
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-
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.036 - 34
4-Chlorobenzoate
0.039 - 54
ATP
0.04 - 39
CoA
additional information
additional information
Pseudomonas sp.
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
30 - 10000
4-Chlorobenzoate
1433
2 - 90
ATP
4
0.02 - 30
CoA
18
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
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inhibition kinetics, overview
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
111.6
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
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assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.8 - 8.2
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pH 5.8: about 50% of maximal activity, pH 8.2: about 80% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10 - 40
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10C: about 80% of maximal activity, 40C: about 50% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
56000
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2 * 56000, SDS-PAGE
57000
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2 * 57000, SDS-PAGE
86000 - 120000
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gel filtration
112000
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gel filtration
114000
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gel filtration
125000
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native gradient PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, 1.0 and 2.2 A. Crystal structures of the enzyme both in the unliganded state and bound to 4-chlorobenzoate. The space group is P3(1)21 or P3(2)21 with a = b = 129.3 A, c = 71.5 A
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purified recombinant CBL bound to 4-chlorobenzoyl-AMP, hanging drop vapor diffusion at 4C using 16-24% pentaerythritol propoxylate 426, and 0.1 M K+-HEPES, pH 6.5 or 6.75, and optimized via hanging drop vapor diffusion using 14-18% PEG 1000, 50-100 mM magnesium nitrate and 100 mM MOPS, pH 7.0, the cryoprotectant contains 24% pentaerythritol propoxylate 426, 24% ethylene glycol, and 0.1 M K+-HEPES, 1 mM ATP, and 1 mM 4-chlorobenzoate, X-ray diffraction structure determination and analysis at 2.0-2.25 A resolution, molecular replacement
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purified recombinant enzyme, X-ray diffraction structure determination and analysis, modelling
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mutant D402P, to 2.6 A resolution, and comparison with wild-type. The C-terminal domain rotates by about 140 degrees between the two states that catalyze the adenylation and thioester-forming half-reactions. The domain rotation is accompanied by a change in the main chain torsional angles of residue D402; mutant D402P, to 2.6 A resolution, and comparison with wild-type. The C-terminal domain rotates by about 140 degrees between the two states that catalyze the adenylation and thioester-forming half-reactions. The domain rotation is accompanied by a change in the main chain torsional angles of residue D402
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
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0.1-2.5 mg of protein per ml, half-life: 21 days
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, stable for months
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4C, 0.1-2.5 mg of protein per ml, half-life: 21 days
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme from Escherichia coli strain JM109 by nickel affinity chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA sequence determination, expression of the His-tagged enzyme in Escherichia coli strain JM109
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expression in Escherichia coli
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D385A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
E410A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
F473A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzymem but increased kcat
G408A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
H207A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
H207F
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site-directed mutagenesis,the mutant shows reduced activity compared to the wild-type enzyme
H207Q
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
H254A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
K477A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
K492A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
K492L
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
K492R
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
M203A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
N302A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
R400A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
R439A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
R475A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
R87A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
S407A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T161A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T251A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T306A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T307A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
W440A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, but increased kcat
Y304F
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D402A
100fold decrease in ratio kcat/Km for CoA
D402P
crystallization data. Mutant adopts the proposed adenylate-forming conformation with very little change to the overall structure.The ability of the mutant to catalyze the adenylate-forming half-reaction is reduced by about 3fold, catalysis of the second half-reaction is reduced by 4 orders of magnitude
E306Q
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mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
G163I
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mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
G166I
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mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
K169M
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mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
P168A
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mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
E306Q
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mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
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G163I
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mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
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G166I
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mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
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K169M
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mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
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P168A
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mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
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Show AA Sequence (141 entries)
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