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Literature summary 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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.3.1.2 expressed in Escherichia coli Clostridium tetanomorphum

Protein Variants

EC Number Protein Variants Comment Organism
4.3.1.2 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
4.3.1.2 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
4.3.1.2 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
4.3.1.2 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
4.3.1.2 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
4.3.1.2 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
4.3.1.2 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
4.3.1.2 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
4.3.1.2 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
4.3.1.2 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
4.3.1.2 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.3.1.2 0.2
-
 mesaconate pH 9, 30°C, mutant Q73A Clostridium tetanomorphum
4.3.1.2 0.7
-
 mesaconate pH 9, 30°C, wild-type Clostridium tetanomorphum
4.3.1.2 0.9
-
 mesaconate pH 9, 30°C, mutant F170A Clostridium tetanomorphum
4.3.1.2 1.4
-
 mesaconate pH 9, 30°C, mutant T360S Clostridium tetanomorphum
4.3.1.2 2.9
-
 mesaconate pH 9, 30°C, mutant C361A Clostridium tetanomorphum
4.3.1.2 15
-
 mesaconate pH 9, 30°C, mutant T360A Clostridium tetanomorphum
4.3.1.2 60
-
 mesaconate pH 9, 30°C, mutant L384A Clostridium tetanomorphum
4.3.1.2 60
-
 mesaconate pH 9, 30°C, mutant Q172A Clostridium tetanomorphum
4.3.1.2 60
-
 mesaconate pH 9, 30°C, mutant Y356A Clostridium tetanomorphum

Organism

EC Number Organism UniProt Comment Textmining
4.3.1.2 Clostridium tetanomorphum Q05514
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.3.1.2
-
 Clostridium tetanomorphum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.3.1.2 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

EC Number Synonyms Comment Organism
4.3.1.2 MAL
-
 Clostridium tetanomorphum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment 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 Comment Organism Structure
4.3.1.2 0.6
-
 mesaconate pH 9, 30°C, mutant T360A Clostridium tetanomorphum
4.3.1.2 0.7
-
 mesaconate pH 9, 30°C, mutant Q73A Clostridium tetanomorphum
4.3.1.2 0.7
-
 mesaconate pH 9, 30°C, mutant Y356A Clostridium tetanomorphum
4.3.1.2 10
-
 mesaconate pH 9, 30°C, mutant F170A Clostridium tetanomorphum
4.3.1.2 16
-
 mesaconate pH 9, 30°C, mutant L384A Clostridium tetanomorphum
4.3.1.2 20
-
 mesaconate pH 9, 30°C, mutant Q172A Clostridium tetanomorphum
4.3.1.2 27.5
-
 mesaconate pH 9, 30°C, mutant C361A Clostridium tetanomorphum
4.3.1.2 35
-
 mesaconate pH 9, 30°C, mutant T360S Clostridium tetanomorphum
4.3.1.2 61
-
 mesaconate pH 9, 30°C, wild-type Clostridium tetanomorphum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment 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 Comment Organism Structure
4.3.1.2 0.04
-
 mesaconate pH 9, 30°C, mutant T360A Clostridium tetanomorphum
4.3.1.2 0.27
-
 mesaconate pH 9, 30°C, mutant L384A Clostridium tetanomorphum
4.3.1.2 0.342
-
 mesaconate pH 9, 30°C, mutant Q172A Clostridium tetanomorphum
4.3.1.2 3.5
-
 mesaconate pH 9, 30°C, mutant Q73A Clostridium tetanomorphum
4.3.1.2 9.5
-
 mesaconate pH 9, 30°C, mutant C361A Clostridium tetanomorphum
4.3.1.2 11
-
 mesaconate pH 9, 30°C, mutant F170A Clostridium tetanomorphum
4.3.1.2 12
-
 mesaconate pH 9, 30°C, mutant Y356A Clostridium tetanomorphum
4.3.1.2 25
-
 mesaconate pH 9, 30°C, mutant T360S Clostridium tetanomorphum
4.3.1.2 87
-
 mesaconate pH 9, 30°C, wild-type Clostridium tetanomorphum