BRENDA - Enzyme Database

Alteration of the diastereoselectivity of 3-methylaspartate ammonia lyase by using structure-based mutagenesis

Raj, H.; Weiner, B.; Veetil, V.P.; Reis, C.R.; Quax, W.J.; Janssen, D.B.; Feringa, B.L.; Poelarends, G.J.; ChemBioChem 10, 2236-2245 (2009)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
4.3.1.2
expression in Escherichia coli
Clostridium tetanomorphum
Engineering
EC Number
Amino acid exchange
Commentary
Organism
4.3.1.2
H194A
enhanced diastereoselectivities
Clostridium tetanomorphum
4.3.1.2
K331A
enhanced diastereoselectivities
Clostridium tetanomorphum
4.3.1.2
Q329A
enhanced diastereoselectivities
Clostridium tetanomorphum
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.1.2
0.7
-
mesaconic acid
wild-type
Clostridium tetanomorphum
4.3.1.2
1
-
L-threo-3-methylaspartic acid
mutant Q329A; wild-type
Clostridium tetanomorphum
4.3.1.2
1.7
-
mesaconic acid
mutant Q329A
Clostridium tetanomorphum
4.3.1.2
1.8
-
L-threo-3-methylaspartic acid
mutant H194A
Clostridium tetanomorphum
4.3.1.2
14
-
mesaconic acid
mutant H194A
Clostridium tetanomorphum
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.3.1.2
Clostridium tetanomorphum
Q05514
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.3.1.2
L-threo-3-methylaspartic acid
assay at pH 9.0, 30°C
703197
Clostridium tetanomorphum
mesaconate + NH3
-
-
-
?
4.3.1.2
mesaconic acid + NH3
assay at pH 9.0, 30°C
703197
Clostridium tetanomorphum
L-threo-3-methylaspartate
-
-
-
r
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.3.1.2
30
-
assay at
Clostridium tetanomorphum
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.3.1.2
0.01
-
L-threo-3-methylaspartic acid
mutant Q329A
Clostridium tetanomorphum
4.3.1.2
0.25
-
mesaconic acid
mutant Q329A
Clostridium tetanomorphum
4.3.1.2
0.55
-
L-threo-3-methylaspartic acid
mutant H194A
Clostridium tetanomorphum
4.3.1.2
8.4
-
mesaconic acid
mutant H194A
Clostridium tetanomorphum
4.3.1.2
61
-
mesaconic acid
wild-type
Clostridium tetanomorphum
4.3.1.2
89
-
L-threo-3-methylaspartic acid
wild-type
Clostridium tetanomorphum
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.3.1.2
9
-
assay at
Clostridium tetanomorphum
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.1.2
expression in Escherichia coli
Clostridium tetanomorphum
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
4.3.1.2
H194A
enhanced diastereoselectivities
Clostridium tetanomorphum
4.3.1.2
K331A
enhanced diastereoselectivities
Clostridium tetanomorphum
4.3.1.2
Q329A
enhanced diastereoselectivities
Clostridium tetanomorphum
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.1.2
0.7
-
mesaconic acid
wild-type
Clostridium tetanomorphum
4.3.1.2
1
-
L-threo-3-methylaspartic acid
mutant Q329A; wild-type
Clostridium tetanomorphum
4.3.1.2
1.7
-
mesaconic acid
mutant Q329A
Clostridium tetanomorphum
4.3.1.2
1.8
-
L-threo-3-methylaspartic acid
mutant H194A
Clostridium tetanomorphum
4.3.1.2
14
-
mesaconic acid
mutant H194A
Clostridium tetanomorphum
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.3.1.2
L-threo-3-methylaspartic acid
assay at pH 9.0, 30°C
703197
Clostridium tetanomorphum
mesaconate + NH3
-
-
-
?
4.3.1.2
mesaconic acid + NH3
assay at pH 9.0, 30°C
703197
Clostridium tetanomorphum
L-threo-3-methylaspartate
-
-
-
r
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.3.1.2
30
-
assay at
Clostridium tetanomorphum
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.3.1.2
0.01
-
L-threo-3-methylaspartic acid
mutant Q329A
Clostridium tetanomorphum
4.3.1.2
0.25
-
mesaconic acid
mutant Q329A
Clostridium tetanomorphum
4.3.1.2
0.55
-
L-threo-3-methylaspartic acid
mutant H194A
Clostridium tetanomorphum
4.3.1.2
8.4
-
mesaconic acid
mutant H194A
Clostridium tetanomorphum
4.3.1.2
61
-
mesaconic acid
wild-type
Clostridium tetanomorphum
4.3.1.2
89
-
L-threo-3-methylaspartic acid
wild-type
Clostridium tetanomorphum
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.3.1.2
9
-
assay at
Clostridium tetanomorphum
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
4.3.1.2
additional information
-
L-threo-3-methylaspartic acid
mutant K331A, value below 0.000001
Clostridium tetanomorphum
4.3.1.2
additional information
-
mesaconic acid
mutant K331A, value below 0.000001
Clostridium tetanomorphum
4.3.1.2
0.01
-
L-threo-3-methylaspartic acid
mutant Q329A
Clostridium tetanomorphum
4.3.1.2
0.15
-
mesaconic acid
mutant Q329A
Clostridium tetanomorphum
4.3.1.2
0.31
-
L-threo-3-methylaspartic acid
mutant H194A
Clostridium tetanomorphum
4.3.1.2
0.6
-
mesaconic acid
mutant H194A
Clostridium tetanomorphum
4.3.1.2
87
-
mesaconic acid
wild-type
Clostridium tetanomorphum
4.3.1.2
89
-
L-threo-3-methylaspartic acid
wild-type
Clostridium tetanomorphum
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
4.3.1.2
additional information
-
L-threo-3-methylaspartic acid
mutant K331A, value below 0.000001
Clostridium tetanomorphum
4.3.1.2
additional information
-
mesaconic acid
mutant K331A, value below 0.000001
Clostridium tetanomorphum
4.3.1.2
0.01
-
L-threo-3-methylaspartic acid
mutant Q329A
Clostridium tetanomorphum
4.3.1.2
0.15
-
mesaconic acid
mutant Q329A
Clostridium tetanomorphum
4.3.1.2
0.31
-
L-threo-3-methylaspartic acid
mutant H194A
Clostridium tetanomorphum
4.3.1.2
0.6
-
mesaconic acid
mutant H194A
Clostridium tetanomorphum
4.3.1.2
87
-
mesaconic acid
wild-type
Clostridium tetanomorphum
4.3.1.2
89
-
L-threo-3-methylaspartic acid
wild-type
Clostridium tetanomorphum