BRENDA - Enzyme Database

Complete reversal of coenzyme specificity of isocitrate dehydrogenase from Haloferax volcanii

Rodriguez-Arnedo, A.; Camacho, M.; Llorca, F.; Bonete, M.J.; Protein J. 24, 259-266 (2005)

Data extracted from this reference:

Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.42
R291S
involved in coenzyme specificity
Haloferax volcanii
1.1.1.42
R291S/K343D/Y344I/V350A/Y390P
switch in coenzyme specificity from NADP+ to NAD+
Haloferax volcanii
1.1.1.42
R291S/K343D/Y344I/Y390P
involved in coenzyme specificity
Haloferax volcanii
1.1.1.42
R291S/K343D/Y390P
involved in coenzyme specificity
Haloferax volcanii
1.1.1.42
R291S/Y390P
involved in coenzyme specificity
Haloferax volcanii
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.42
0.011
-
DL-isocitrate
mutant R291S/K343D/Y344I/V350A/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.018
-
DL-isocitrate
mutant R291S, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.036
-
DL-isocitrate
mutant R291S/K343D/Y390P, pH 8.0, 30C; mutant R291S/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.04
-
DL-isocitrate
mutant R291S/K343D/Y344I/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.101
-
NADP+
wild-type, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.108
-
DL-isocitrate
wild-type, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.144
-
NAD+
mutant R291S/K343D/Y344I/V350A/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.203
-
NADP+
mutant R291S/K343D/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.282
-
NADP+
mutant R291S/K343D/Y344I/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.327
-
NADP+
mutant R291S/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.35
-
NADP+
mutant R291S, pH 8.0, 30C
Haloferax volcanii
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.41
Escherichia coli
-
-
-
1.1.1.42
Haloferax volcanii
Q8X277
recombinant enzyme
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.42
DL-isocitrate + NAD+
NAD+ is only substrate for mutant R291S/K343D/Y344I/V350A/Y390P
670805
Haloferax volcanii
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.42
DL-isocitrate + NADP+
NAD+ is not substrate for mutant R291S/K343D/Y344I/V350A/Y390P
670805
Haloferax volcanii
2-oxoglutarate + CO2 + NADPH
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.42
0.0003
-
NADP+
mutant R291S/K343D/Y344I/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.0005
-
DL-isocitrate
mutant R291S/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.0007
-
NAD+
mutant R291S/K343D/Y344I/V350A/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.001
-
NADP+
mutant R291S/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.0013
-
DL-isocitrate
mutant R291S/K343D/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.002
-
DL-isocitrate
mutant R291S, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.0023
-
DL-isocitrate
wild-type, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.0024
-
NADP+
mutant R291S/K343D/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.0025
-
DL-isocitrate
mutant R291S/K343D/Y344I/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.003
-
NADP+
wild-type, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.004
-
NADP+
mutant R291S, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.005
-
DL-isocitrate
mutant R291S/K343D/Y344I/V350A/Y390P, pH 8.0, 30C
Haloferax volcanii
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.42
R291S
involved in coenzyme specificity
Haloferax volcanii
1.1.1.42
R291S/K343D/Y344I/V350A/Y390P
switch in coenzyme specificity from NADP+ to NAD+
Haloferax volcanii
1.1.1.42
R291S/K343D/Y344I/Y390P
involved in coenzyme specificity
Haloferax volcanii
1.1.1.42
R291S/K343D/Y390P
involved in coenzyme specificity
Haloferax volcanii
1.1.1.42
R291S/Y390P
involved in coenzyme specificity
Haloferax volcanii
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.42
0.011
-
DL-isocitrate
mutant R291S/K343D/Y344I/V350A/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.018
-
DL-isocitrate
mutant R291S, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.036
-
DL-isocitrate
mutant R291S/K343D/Y390P, pH 8.0, 30C; mutant R291S/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.04
-
DL-isocitrate
mutant R291S/K343D/Y344I/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.101
-
NADP+
wild-type, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.108
-
DL-isocitrate
wild-type, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.144
-
NAD+
mutant R291S/K343D/Y344I/V350A/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.203
-
NADP+
mutant R291S/K343D/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.282
-
NADP+
mutant R291S/K343D/Y344I/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.327
-
NADP+
mutant R291S/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.35
-
NADP+
mutant R291S, pH 8.0, 30C
Haloferax volcanii
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.42
DL-isocitrate + NAD+
NAD+ is only substrate for mutant R291S/K343D/Y344I/V350A/Y390P
670805
Haloferax volcanii
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.42
DL-isocitrate + NADP+
NAD+ is not substrate for mutant R291S/K343D/Y344I/V350A/Y390P
670805
Haloferax volcanii
2-oxoglutarate + CO2 + NADPH
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.42
0.0003
-
NADP+
mutant R291S/K343D/Y344I/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.0005
-
DL-isocitrate
mutant R291S/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.0007
-
NAD+
mutant R291S/K343D/Y344I/V350A/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.001
-
NADP+
mutant R291S/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.0013
-
DL-isocitrate
mutant R291S/K343D/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.002
-
DL-isocitrate
mutant R291S, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.0023
-
DL-isocitrate
wild-type, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.0024
-
NADP+
mutant R291S/K343D/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.0025
-
DL-isocitrate
mutant R291S/K343D/Y344I/Y390P, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.003
-
NADP+
wild-type, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.004
-
NADP+
mutant R291S, pH 8.0, 30C
Haloferax volcanii
1.1.1.42
0.005
-
DL-isocitrate
mutant R291S/K343D/Y344I/V350A/Y390P, pH 8.0, 30C
Haloferax volcanii