Cloned (Comment) | Organism |
---|---|
DNA sequence determination, expression of the His-tagged enzyme in Escherichia coli strain JM109 | Alcaligenes sp. |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme, X-ray diffraction structure determination and analysis, modelling | Alcaligenes sp. |
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 | Comment | 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 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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Alcaligenes sp. |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Alcaligenes sp. | Q8GN86 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain JM109 by nickel affinity chromatography | Alcaligenes sp. |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
4-Chlorobenzoate + CoA + ATP = 4-chlorobenzoyl-CoA + AMP + diphosphate | kinetic reaction mechanism, overview | Alcaligenes sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | Comment | Organism |
---|---|---|
More | domain and active site structure, overview | Alcaligenes sp. |
Synonyms | Comment | Organism |
---|---|---|
4-Chlorobenzoate:coenzyme A ligase | - |
Alcaligenes sp. |
CBL | - |
Alcaligenes sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Alcaligenes sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Alcaligenes sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Alcaligenes sp. |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | inhibition kinetics, overview | Alcaligenes sp. |