Literature summary for 6.2.1.33 extracted from

Wu, R.; Cao, J.; Lu, X.; Reger, A.S.; Gulick, A.M.; Dunaway-Mariano, D.
Mechanism of 4-chlorobenzoate:coenzyme A ligase catalysis (2008), Biochemistry, 47, 8026-8039.

Cloned(Commentary)

Cloned (Comment)	Organism
DNA sequence determination, expression of the His-tagged enzyme in Escherichia coli strain JM109	Alcaligenes sp.

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme, X-ray diffraction structure determination and analysis, modelling	Alcaligenes sp.

Protein Variants

Protein Variants	Comment	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	Comment	Organism
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	Comment	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	Comment	Organism	Structure
Mg2+	-	Alcaligenes sp.

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
4-Chlorobenzoate + CoA + ATP	Alcaligenes sp.	-	4-Chlorobenzoyl-CoA + AMP + diphosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Alcaligenes sp.	Q8GN86	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain JM109 by nickel affinity chromatography	Alcaligenes sp.

Reaction

Reaction	Comment	Organism	Reaction ID
4-Chlorobenzoate + CoA + ATP = 4-chlorobenzoyl-CoA + AMP + diphosphate	kinetic reaction mechanism, overview	Alcaligenes sp.	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
4-Chlorobenzoate + CoA + ATP	-	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	Alcaligenes sp.	4-Chlorobenzoyl-CoA + AMP + diphosphate	-	?
4-Methylbenzoate + CoA + ATP	-	Alcaligenes sp.	4-Methylbenzoyl-CoA + AMP + diphosphate	-	?

Subunits

Subunits	Comment	Organism
More	domain and active site structure, overview	Alcaligenes sp.

Synonyms

Synonyms	Comment	Organism
4-Chlorobenzoate:coenzyme A ligase	-	Alcaligenes sp.
CBL	-	Alcaligenes sp.

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Alcaligenes sp.

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Alcaligenes sp.

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Alcaligenes sp.

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
additional information	-	additional information	inhibition kinetics, overview	Alcaligenes sp.