BRENDA - Enzyme Database show
show all sequences of 6.2.1.33

Mechanism of 4-chlorobenzoate:coenzyme A ligase catalysis

Wu, R.; Cao, J.; Lu, X.; Reger, A.S.; Gulick, A.M.; Dunaway-Mariano, D.; Biochemistry 47, 8026-8039 (2008)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
DNA sequence determination, expression of the His-tagged enzyme in Escherichia coli strain JM109
Alcaligenes sp.
Crystallization (Commentary)
Crystallization
Organism
purified recombinant enzyme, X-ray diffraction structure determination and analysis, modelling
Alcaligenes sp.
Engineering
Amino acid exchange
Commentary
Organism
D385A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
E410A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
F473A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzymem but increased kcat
Alcaligenes sp.
G408A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
H207A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
H207F
site-directed mutagenesis,the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
H207Q
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
H254A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
K477A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
K492A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
K492L
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
K492R
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
M203A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
N302A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
R400A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
R439A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
R475A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
R87A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
S407A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
T161A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
T251A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
T306A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
T307A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
W440A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, but increased kcat
Alcaligenes sp.
Y304F
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
Inhibitors
Inhibitors
Commentary
Organism
Structure
4-chlorophenacyl-CoA
a product analogue, noncompetitive versus 4-methylbenzoate and ATP, dead-end inhibitor
Alcaligenes sp.
AMP
is a noncompetitive inhibitor versus ATP and an uncompetitive versus CoA
Alcaligenes sp.
diphosphate
competitive versus ATP
Alcaligenes sp.
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
transient-state and steady-state kinetics of wild-type and mutant enzymes, overview
Alcaligenes sp.
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
-
Alcaligenes sp.
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4-Chlorobenzoate + CoA + ATP
Alcaligenes sp.
-
4-Chlorobenzoyl-CoA + AMP + diphosphate
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Alcaligenes sp.
Q8GN86
-
-
Purification (Commentary)
Commentary
Organism
recombinant His-tagged enzyme from Escherichia coli strain JM109 by nickel affinity chromatography
Alcaligenes sp.
Reaction
Reaction
Commentary
Organism
4-Chlorobenzoate + CoA + ATP = 4-chlorobenzoyl-CoA + AMP + diphosphate
kinetic reaction mechanism, overview
Alcaligenes sp.
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4-Chlorobenzoate + CoA + ATP
-
690988
Alcaligenes sp.
4-Chlorobenzoyl-CoA + AMP + diphosphate
-
-
-
?
4-Chlorobenzoate + CoA + ATP
the rate-limiting step in CBL catalysis follows the formation of 4-chlorobenzoate-CoA
690988
Alcaligenes sp.
4-Chlorobenzoyl-CoA + AMP + diphosphate
-
-
-
?
4-Methylbenzoate + CoA + ATP
-
690988
Alcaligenes sp.
4-Methylbenzoyl-CoA + AMP + diphosphate
-
-
-
?
Subunits
Subunits
Commentary
Organism
More
domain and active site structure, overview
Alcaligenes sp.
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Alcaligenes sp.
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Alcaligenes sp.
Cofactor
Cofactor
Commentary
Organism
Structure
ATP
-
Alcaligenes sp.
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
additional information
-
additional information
inhibition kinetics, overview
Alcaligenes sp.
Cloned(Commentary) (protein specific)
Commentary
Organism
DNA sequence determination, expression of the His-tagged enzyme in Escherichia coli strain JM109
Alcaligenes sp.
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
ATP
-
Alcaligenes sp.
Crystallization (Commentary) (protein specific)
Crystallization
Organism
purified recombinant enzyme, X-ray diffraction structure determination and analysis, modelling
Alcaligenes sp.
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D385A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
E410A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
F473A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzymem but increased kcat
Alcaligenes sp.
G408A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
H207A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
H207F
site-directed mutagenesis,the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
H207Q
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
H254A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
K477A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
K492A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
K492L
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
K492R
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
M203A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
N302A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
R400A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
R439A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
R475A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
R87A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
S407A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
T161A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
T251A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
T306A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
T307A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
W440A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, but increased kcat
Alcaligenes sp.
Y304F
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Alcaligenes sp.
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
4-chlorophenacyl-CoA
a product analogue, noncompetitive versus 4-methylbenzoate and ATP, dead-end inhibitor
Alcaligenes sp.
AMP
is a noncompetitive inhibitor versus ATP and an uncompetitive versus CoA
Alcaligenes sp.
diphosphate
competitive versus ATP
Alcaligenes sp.
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
additional information
-
additional information
inhibition kinetics, overview
Alcaligenes sp.
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
transient-state and steady-state kinetics of wild-type and mutant enzymes, overview
Alcaligenes sp.
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
-
Alcaligenes sp.
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4-Chlorobenzoate + CoA + ATP
Alcaligenes sp.
-
4-Chlorobenzoyl-CoA + AMP + diphosphate
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant His-tagged enzyme from Escherichia coli strain JM109 by nickel affinity chromatography
Alcaligenes sp.
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4-Chlorobenzoate + CoA + ATP
-
690988
Alcaligenes sp.
4-Chlorobenzoyl-CoA + AMP + diphosphate
-
-
-
?
4-Chlorobenzoate + CoA + ATP
the rate-limiting step in CBL catalysis follows the formation of 4-chlorobenzoate-CoA
690988
Alcaligenes sp.
4-Chlorobenzoyl-CoA + AMP + diphosphate
-
-
-
?
4-Methylbenzoate + CoA + ATP
-
690988
Alcaligenes sp.
4-Methylbenzoyl-CoA + AMP + diphosphate
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
More
domain and active site structure, overview
Alcaligenes sp.
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Alcaligenes sp.
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Alcaligenes sp.
Other publictions for EC 6.2.1.33
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
702299
Wu
The mechanism of domain altern ...
Alcaligenes sp. AL3007
Biochemistry
48
4115-4125
2009
-
-
-
1
2
-
-
9
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
9
-
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
-
9
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
9
-
-
-
-
-
-
-
-
9
9
690987
Reger
Structural characterization of ...
Alcaligenes sp.
Biochemistry
47
8016-8025
2008
-
-
-
1
-
-
1
-
-
1
-
1
-
1
-
-
-
1
-
-
-
-
1
1
1
-
-
-
1
-
-
1
-
-
-
-
-
-
1
1
-
-
-
1
-
-
-
1
-
1
-
-
-
-
-
-
-
-
1
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
690988
Wu
Mechanism of 4-chlorobenzoate: ...
Alcaligenes sp.
Biochemistry
47
8026-8039
2008
-
-
1
1
25
-
3
1
-
1
-
1
-
1
-
-
1
1
-
-
-
-
3
1
1
-
-
-
1
-
-
1
1
-
-
-
-
1
1
1
25
-
-
3
1
1
-
1
-
1
-
-
-
1
-
-
-
-
3
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
661151
Gulick
Crystal structure of 4-chlorob ...
Alcaligenes sp.
Biochemistry
43
8670-8679
2004
-
-
-
1
-
-
-
-
-
-
-
1
-
3
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
1
-
-
-
1
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-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
661288
Zhou
The purification and character ...
Arthrobacter sp., Arthrobacter sp. TM-1
Biodegradation
15
97-109
2004
-
-
-
-
-
-
-
3
-
-
2
-
-
5
-
-
1
-
-
-
1
-
9
1
1
1
-
-
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
3
-
-
2
-
-
-
-
1
-
-
1
-
9
1
1
1
-
-
1
1
-
1
-
-
-
-
-
-
776
Chang
Acyl-adenylate motif of the ac ...
Pseudomonas sp., Pseudomonas sp. CBS3
Biochemistry
36
15650-15659
1997
-
-
-
-
5
-
-
15
-
-
-
-
-
10
-
-
1
1
-
-
-
-
2
-
-
-
-
15
-
-
-
-
-
-
-
-
-
-
-
-
5
-
-
-
-
15
-
-
-
-
-
-
-
1
-
-
-
-
2
-
-
-
-
15
-
-
-
-
-
-
-
-
-
-
777
Chang
Determination of the chemical ...
Pseudomonas sp., Pseudomonas sp. CBS3
Biochemistry
35
13478-13484
1996
-
-
-
-
-
-
-
-
-
-
-
2
-
10
-
-
-
1
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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-
2
-
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-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
781
Dunaway-Mariano
On the origins and functions o ...
Pseudomonas sp.
Biodegradation
5
259-276
1994
-
-
-
-
-
-
-
1
-
1
-
-
-
1
-
-
-
-
-
-
-
-
1
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-
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-
1
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1
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1
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-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
778
Loeffler
Purification and properties of ...
Pseudomonas sp.
Biol. Chem. Hoppe-Seyler
373
1001-1007
1992
-
-
-
-
-
-
4
3
-
6
3
-
-
2
-
-
1
-
-
-
1
2
9
1
1
-
1
-
1
-
-
-
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-
-
-
-
-
-
-
-
-
-
4
-
3
-
6
3
-
-
-
-
1
-
-
1
2
9
1
1
-
1
-
1
-
-
-
-
-
-
-
-
-
779
Loeffler
-
Characterization of 4-chlorobe ...
Pseudomonas sp.
DECHEMA Biotechnol. Conf. (Pt. A. Microbial Principles in Bioprocesses: Cell Culture Technology - Downstream Processing and Recovery)
5
25-28
1992
-
-
-
-
-
-
6
3
-
-
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1
-
1
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-
-
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9
-
1
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1
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6
-
3
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1
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-
-
9
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
780
Chang
Isolation and characterization ...
Pseudomonas sp.
Biochemistry
31
5605-5610
1992
-
-
1
-
-
-
-
3
-
3
2
-
-
3
-
-
1
-
-
-
-
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6
1
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-
-
-
-
-
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-
1
-
-
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3
-
3
2
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1
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6
1
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