BRENDA - Enzyme Database

The role of the active site glutamate in the rearrangement of glutamate to 3-methylaspartate catalyzed by adenosylcobalamin-dependent glutamate mutase

Madhavapeddi, P.; Marsh, E.N.; Chem. Biol. 8, 1143-1149 (2001)

Data extracted from this reference:

Engineering
EC Number
Amino acid exchange
Commentary
Organism
5.4.99.1
E171A
turnover number for glutamate is reduced 27.6fold, KM-value is increased 1.1fold, Km-value for adenosylcobalamin is reduced 1.23fold
Clostridium cochlearium
5.4.99.1
E171D
turnover number for glutamate is reduced 1.8fold, KM-value is reduced 1.54fold, Km-value for adenosylcobalamin is reduced 2.7fold
Clostridium cochlearium
5.4.99.1
E171N
turnover number for glutamate is reduced 232fold, KM-value is increased by 1.8fold, Km-value for adenosylcobalamin is reduced 1.4fold
Clostridium cochlearium
5.4.99.1
E171Q
turnover number for glutamate is reduced 53fold, KM-value is reduced 2.4fold, Km-value for adenosylcobalamin is reduced 2fold, mutant enzyme is independent of pH
Clostridium cochlearium
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
5.4.99.1
0.24
-
L-glutamate
23°C, mutant enzyme E171Q
Clostridium cochlearium
5.4.99.1
0.38
-
L-glutamate
23°C, mutant enzyme E171D
Clostridium cochlearium
5.4.99.1
0.58
-
L-glutamate
23°C, wild-type enzyme
Clostridium cochlearium
5.4.99.1
0.65
-
L-glutamate
23°C, mutant enzyme E171A
Clostridium cochlearium
5.4.99.1
1.07
-
L-glutamate
23°C, mutant enzyme E171N
Clostridium cochlearium
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
5.4.99.1
Clostridium cochlearium
-
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
5.4.99.1
L-glutamate
-
650824
Clostridium cochlearium
L-threo-3-methylaspartate
-
650824
Clostridium cochlearium
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
5.4.99.1
0.025
-
L-glutamate
23°C, mutant enzyme E171N
Clostridium cochlearium
5.4.99.1
0.11
-
L-glutamate
23°C, mutant enzyme E171Q
Clostridium cochlearium
5.4.99.1
0.21
-
L-glutamate
23°C, mutant enzyme E171A
Clostridium cochlearium
5.4.99.1
3
6
L-glutamate
23°C, mutant enzyme E171D
Clostridium cochlearium
5.4.99.1
3.19
-
L-glutamate
23°C, mutant enzyme E171D
Clostridium cochlearium
5.4.99.1
5.8
-
L-glutamate
23°C, wild-type enzyme
Clostridium cochlearium
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
5.4.99.1
8
-
-
Clostridium cochlearium
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
5.4.99.1
6.5
8.7
pH 6.5: about 50% of maximal activity, pH 8.7: about 80% of maximal activity
Clostridium cochlearium
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
5.4.99.1
adenosylcobalamin
enzyme is dependent on, KM for wild-type enzyme is 0.0055 mM
Clostridium cochlearium
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
5.4.99.1
adenosylcobalamin
enzyme is dependent on, KM for wild-type enzyme is 0.0055 mM
Clostridium cochlearium
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
5.4.99.1
E171A
turnover number for glutamate is reduced 27.6fold, KM-value is increased 1.1fold, Km-value for adenosylcobalamin is reduced 1.23fold
Clostridium cochlearium
5.4.99.1
E171D
turnover number for glutamate is reduced 1.8fold, KM-value is reduced 1.54fold, Km-value for adenosylcobalamin is reduced 2.7fold
Clostridium cochlearium
5.4.99.1
E171N
turnover number for glutamate is reduced 232fold, KM-value is increased by 1.8fold, Km-value for adenosylcobalamin is reduced 1.4fold
Clostridium cochlearium
5.4.99.1
E171Q
turnover number for glutamate is reduced 53fold, KM-value is reduced 2.4fold, Km-value for adenosylcobalamin is reduced 2fold, mutant enzyme is independent of pH
Clostridium cochlearium
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
5.4.99.1
0.24
-
L-glutamate
23°C, mutant enzyme E171Q
Clostridium cochlearium
5.4.99.1
0.38
-
L-glutamate
23°C, mutant enzyme E171D
Clostridium cochlearium
5.4.99.1
0.58
-
L-glutamate
23°C, wild-type enzyme
Clostridium cochlearium
5.4.99.1
0.65
-
L-glutamate
23°C, mutant enzyme E171A
Clostridium cochlearium
5.4.99.1
1.07
-
L-glutamate
23°C, mutant enzyme E171N
Clostridium cochlearium
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
5.4.99.1
L-glutamate
-
650824
Clostridium cochlearium
L-threo-3-methylaspartate
-
650824
Clostridium cochlearium
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
5.4.99.1
0.025
-
L-glutamate
23°C, mutant enzyme E171N
Clostridium cochlearium
5.4.99.1
0.11
-
L-glutamate
23°C, mutant enzyme E171Q
Clostridium cochlearium
5.4.99.1
0.21
-
L-glutamate
23°C, mutant enzyme E171A
Clostridium cochlearium
5.4.99.1
3
6
L-glutamate
23°C, mutant enzyme E171D
Clostridium cochlearium
5.4.99.1
3.19
-
L-glutamate
23°C, mutant enzyme E171D
Clostridium cochlearium
5.4.99.1
5.8
-
L-glutamate
23°C, wild-type enzyme
Clostridium cochlearium
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
5.4.99.1
8
-
-
Clostridium cochlearium
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
5.4.99.1
6.5
8.7
pH 6.5: about 50% of maximal activity, pH 8.7: about 80% of maximal activity
Clostridium cochlearium