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Literature summary for 4.3.1.2 extracted from

  • Raj, H.; Poelarends, G.J.
    The roles of active site residues in the catalytic mechanism of methylaspartate ammonia-lyase (2013), FEBS open bio, 3, 285-290.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 4.3.1.2

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Clostridium tetanomorphum

Protein Variants

Protein Variants Comment Organism
C361A kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 47% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type) Clostridium tetanomorphum
C361K mutant shows no reactivity. Conversion 2h: 10% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type) Clostridium tetanomorphum
F170A kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 63% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type) Clostridium tetanomorphum
L384A kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 61% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type) Clostridium tetanomorphum
Q172A kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 16% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type) Clostridium tetanomorphum
Q172N mutant shows no reactivity. Conversion 2h: 6% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type) Clostridium tetanomorphum
Q73A kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 37% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 91:9 (89:11 wild-type) Clostridium tetanomorphum
Q73N mutant shows very low-level amination activity preventing the measurement of kinetic parameters. Conversion 2h: 18% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type) Clostridium tetanomorphum
T360A kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 38% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 93:7 (89:11 wild-type) Clostridium tetanomorphum
T360S kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 76% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 86:14 (89:11 wild-type) Clostridium tetanomorphum
Y356A kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 24% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type) Clostridium tetanomorphum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.2
-
 mesaconate pH 9, 30°C, mutant Q73A Clostridium tetanomorphum
0.7
-
 mesaconate pH 9, 30°C, wild-type Clostridium tetanomorphum
0.9
-
 mesaconate pH 9, 30°C, mutant F170A Clostridium tetanomorphum
1.4
-
 mesaconate pH 9, 30°C, mutant T360S Clostridium tetanomorphum
2.9
-
 mesaconate pH 9, 30°C, mutant C361A Clostridium tetanomorphum
15
-
 mesaconate pH 9, 30°C, mutant T360A Clostridium tetanomorphum
60
-
 mesaconate pH 9, 30°C, mutant L384A Clostridium tetanomorphum
60
-
 mesaconate pH 9, 30°C, mutant Q172A Clostridium tetanomorphum
60
-
 mesaconate pH 9, 30°C, mutant Y356A Clostridium tetanomorphum

Organism

Organism UniProt Comment Textmining
Clostridium tetanomorphum Q05514
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Clostridium tetanomorphum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
mesaconate + NH3 conversion 2h: 73% (wild-type), diastereomeric product ratio (threo:erythro) after 2h: 89:11 Clostridium tetanomorphum L-threo-3-methylaspartate + L-erythro-3-methylaspartate
-
 r

Synonyms

Synonyms Comment Organism
MAL
-
 Clostridium tetanomorphum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Clostridium tetanomorphum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.6
-
 mesaconate pH 9, 30°C, mutant T360A Clostridium tetanomorphum
0.7
-
 mesaconate pH 9, 30°C, mutant Q73A Clostridium tetanomorphum
0.7
-
 mesaconate pH 9, 30°C, mutant Y356A Clostridium tetanomorphum
10
-
 mesaconate pH 9, 30°C, mutant F170A Clostridium tetanomorphum
16
-
 mesaconate pH 9, 30°C, mutant L384A Clostridium tetanomorphum
20
-
 mesaconate pH 9, 30°C, mutant Q172A Clostridium tetanomorphum
27.5
-
 mesaconate pH 9, 30°C, mutant C361A Clostridium tetanomorphum
35
-
 mesaconate pH 9, 30°C, mutant T360S Clostridium tetanomorphum
61
-
 mesaconate pH 9, 30°C, wild-type Clostridium tetanomorphum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
 assay at Clostridium tetanomorphum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.04
-
 mesaconate pH 9, 30°C, mutant T360A Clostridium tetanomorphum
0.27
-
 mesaconate pH 9, 30°C, mutant L384A Clostridium tetanomorphum
0.342
-
 mesaconate pH 9, 30°C, mutant Q172A Clostridium tetanomorphum
3.5
-
 mesaconate pH 9, 30°C, mutant Q73A Clostridium tetanomorphum
9.5
-
 mesaconate pH 9, 30°C, mutant C361A Clostridium tetanomorphum
11
-
 mesaconate pH 9, 30°C, mutant F170A Clostridium tetanomorphum
12
-
 mesaconate pH 9, 30°C, mutant Y356A Clostridium tetanomorphum
25
-
 mesaconate pH 9, 30°C, mutant T360S Clostridium tetanomorphum
87
-
 mesaconate pH 9, 30°C, wild-type Clostridium tetanomorphum