BRENDA - Enzyme Database

Acyl-adenylate motif of the acyl-adenylate/thioester-forming enzyme superfamily: a site-directed mutagenesis study with the Pseudomonas sp. strain CBS3 4-chlorobenzoate:coenzyme A ligase

Chang, K.H.; Xiang, H.; Dunaway-Mariano, D.; Biochemistry 36, 15650-15659 (1997)

Data extracted from this reference:

Engineering
EC Number
Amino acid exchange
Commentary
Organism
6.2.1.33
E306Q
mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
Pseudomonas sp.
6.2.1.33
G163I
mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
Pseudomonas sp.
6.2.1.33
G166I
mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
Pseudomonas sp.
6.2.1.33
K169M
mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
Pseudomonas sp.
6.2.1.33
P168A
mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
Pseudomonas sp.
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.2.1.33
0.014
-
4-Chlorobenzoate
wild-type
Pseudomonas sp.
6.2.1.33
0.02
-
4-Chlorobenzoate
mutant K169M
Pseudomonas sp.
6.2.1.33
0.062
-
CoA
mutant G163I
Pseudomonas sp.
6.2.1.33
0.073
-
4-Chlorobenzoate
mutant G166I
Pseudomonas sp.
6.2.1.33
0.076
-
CoA
mutant K169M
Pseudomonas sp.
6.2.1.33
0.085
-
CoA
wild-type
Pseudomonas sp.
6.2.1.33
0.105
-
4-Chlorobenzoate
mutant G163I
Pseudomonas sp.
6.2.1.33
0.108
-
CoA
mutant G166I
Pseudomonas sp.
6.2.1.33
0.168
-
CoA
mutant E306Q
Pseudomonas sp.
6.2.1.33
0.253
-
4-Chlorobenzoate
mutant E306Q
Pseudomonas sp.
6.2.1.33
0.308
-
ATP
wild-type
Pseudomonas sp.
6.2.1.33
0.321
-
ATP
mutant E306Q
Pseudomonas sp.
6.2.1.33
0.83
-
ATP
mutant K169M
Pseudomonas sp.
6.2.1.33
0.981
-
ATP
mutant G166I
Pseudomonas sp.
6.2.1.33
1.314
-
ATP
mutant G163I
Pseudomonas sp.
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
6.2.1.33
Pseudomonas sp.
-
; wild-type and mutant enzymes G163I, G166I, P168A, K169M and E306Q
-
6.2.1.33
Pseudomonas sp. CBS3
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
6.2.1.33
-
Pseudomonas sp.
Reaction
EC Number
Reaction
Commentary
Organism
6.2.1.33
4-Chlorobenzoate + CoA + ATP = 4-chlorobenzoyl-CoA + AMP + diphosphate
two-step reaction consisting of the adenylation of 4-chlorobenzoate with adenosine 5'-triphosphate followed by acyl transfer from 4-chlorobenzoyl adenosine 5'-monophosphate diester
Pseudomonas sp.
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.2.1.33
4-Chlorobenzoate + CoA + ATP
-
776
Pseudomonas sp.
4-Chlorobenzoyl-CoA + AMP + diphosphate
-
776
Pseudomonas sp.
-
6.2.1.33
4-Chlorobenzoate + CoA + ATP
-
776
Pseudomonas sp. CBS3
4-Chlorobenzoyl-CoA + AMP + diphosphate
-
776
Pseudomonas sp. CBS3
-
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
6.2.1.33
0.036
-
4-Chlorobenzoate
mutant G163I
Pseudomonas sp.
6.2.1.33
0.039
-
ATP
mutant G163I
Pseudomonas sp.
6.2.1.33
0.04
-
CoA
mutant G163I
Pseudomonas sp.
6.2.1.33
0.7
-
4-Chlorobenzoate
mutant E306Q
Pseudomonas sp.
6.2.1.33
0.8
-
ATP
mutant E306Q
Pseudomonas sp.
6.2.1.33
0.9
-
ATP
mutant E306Q
Pseudomonas sp.
6.2.1.33
2.3
-
4-Chlorobenzoate
mutant G166I
Pseudomonas sp.
6.2.1.33
2.7
-
CoA
mutant G166I
Pseudomonas sp.
6.2.1.33
3.3
-
ATP
mutant G166I
Pseudomonas sp.
6.2.1.33
9.4
-
ATP
mutant K169M
Pseudomonas sp.
6.2.1.33
9.8
-
4-Chlorobenzoate
mutant K169M
Pseudomonas sp.
6.2.1.33
16
-
ATP
mutant K169M
Pseudomonas sp.
6.2.1.33
34
-
4-Chlorobenzoate
wild-type
Pseudomonas sp.
6.2.1.33
39
-
CoA
wild-type
Pseudomonas sp.
6.2.1.33
54
-
ATP
wild-type
Pseudomonas sp.
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
6.2.1.33
E306Q
mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
Pseudomonas sp.
6.2.1.33
G163I
mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
Pseudomonas sp.
6.2.1.33
G166I
mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
Pseudomonas sp.
6.2.1.33
K169M
mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
Pseudomonas sp.
6.2.1.33
P168A
mutant enzymes G163I, G166I, P168A, K169M and E306Q with reduced catalysis of the adenylation of 4-chlorobenzoate
Pseudomonas sp.
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.2.1.33
0.014
-
4-Chlorobenzoate
wild-type
Pseudomonas sp.
6.2.1.33
0.02
-
4-Chlorobenzoate
mutant K169M
Pseudomonas sp.
6.2.1.33
0.062
-
CoA
mutant G163I
Pseudomonas sp.
6.2.1.33
0.073
-
4-Chlorobenzoate
mutant G166I
Pseudomonas sp.
6.2.1.33
0.076
-
CoA
mutant K169M
Pseudomonas sp.
6.2.1.33
0.085
-
CoA
wild-type
Pseudomonas sp.
6.2.1.33
0.105
-
4-Chlorobenzoate
mutant G163I
Pseudomonas sp.
6.2.1.33
0.108
-
CoA
mutant G166I
Pseudomonas sp.
6.2.1.33
0.168
-
CoA
mutant E306Q
Pseudomonas sp.
6.2.1.33
0.253
-
4-Chlorobenzoate
mutant E306Q
Pseudomonas sp.
6.2.1.33
0.308
-
ATP
wild-type
Pseudomonas sp.
6.2.1.33
0.321
-
ATP
mutant E306Q
Pseudomonas sp.
6.2.1.33
0.83
-
ATP
mutant K169M
Pseudomonas sp.
6.2.1.33
0.981
-
ATP
mutant G166I
Pseudomonas sp.
6.2.1.33
1.314
-
ATP
mutant G163I
Pseudomonas sp.
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
6.2.1.33
-
Pseudomonas sp.
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.2.1.33
4-Chlorobenzoate + CoA + ATP
-
776
Pseudomonas sp.
4-Chlorobenzoyl-CoA + AMP + diphosphate
-
776
Pseudomonas sp.
-
6.2.1.33
4-Chlorobenzoate + CoA + ATP
-
776
Pseudomonas sp. CBS3
4-Chlorobenzoyl-CoA + AMP + diphosphate
-
776
Pseudomonas sp. CBS3
-
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
6.2.1.33
0.036
-
4-Chlorobenzoate
mutant G163I
Pseudomonas sp.
6.2.1.33
0.039
-
ATP
mutant G163I
Pseudomonas sp.
6.2.1.33
0.04
-
CoA
mutant G163I
Pseudomonas sp.
6.2.1.33
0.7
-
4-Chlorobenzoate
mutant E306Q
Pseudomonas sp.
6.2.1.33
0.8
-
ATP
mutant E306Q
Pseudomonas sp.
6.2.1.33
0.9
-
ATP
mutant E306Q
Pseudomonas sp.
6.2.1.33
2.3
-
4-Chlorobenzoate
mutant G166I
Pseudomonas sp.
6.2.1.33
2.7
-
CoA
mutant G166I
Pseudomonas sp.
6.2.1.33
3.3
-
ATP
mutant G166I
Pseudomonas sp.
6.2.1.33
9.4
-
ATP
mutant K169M
Pseudomonas sp.
6.2.1.33
9.8
-
4-Chlorobenzoate
mutant K169M
Pseudomonas sp.
6.2.1.33
16
-
ATP
mutant K169M
Pseudomonas sp.
6.2.1.33
34
-
4-Chlorobenzoate
wild-type
Pseudomonas sp.
6.2.1.33
39
-
CoA
wild-type
Pseudomonas sp.
6.2.1.33
54
-
ATP
wild-type
Pseudomonas sp.