Information on EC 4.3.1.2 - methylaspartate ammonia-lyase and Organism(s) Clostridium tetanomorphum and UniProt Accession Q05514

for references in articles please use BRENDA:EC4.3.1.2
Word Map on EC 4.3.1.2
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
This record set is specific for:
Clostridium tetanomorphum
UNIPROT: Q05514


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota


The taxonomic range for the selected organisms is: Clostridium tetanomorphum

EC NUMBER
COMMENTARY hide
4.3.1.2
-
RECOMMENDED NAME
GeneOntology No.
methylaspartate ammonia-lyase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-threo-3-methylaspartate = mesaconate + NH3
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
L-glutamate degradation VI (to pyruvate)
-
-
methylaspartate cycle
-
-
glutamate and glutamine metabolism
-
-
Glyoxylate and dicarboxylate metabolism
-
-
C5-Branched dibasic acid metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
L-threo-3-methylaspartate ammonia-lyase (mesaconate-forming)
A cobalamin protein.
CAS REGISTRY NUMBER
COMMENTARY hide
9033-26-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2E)-2-butylbut-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 0%, mutant L384A: conversion 59%
-
-
r
(2E)-2-ethylbut-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 74%, mutant L384A: conversion 76%
-
-
r
(2E)-2-hexylbut-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 0%, mutant L384A: conversion 53%
-
-
r
(2E)-2-methylbut-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 80%, mutant L384A: conversion 75%
-
-
r
(2E)-2-pentylbut-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 0%, mutant L384A: conversion 52%
-
-
r
(2E)-2-phenylbut-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 0%, mutant L384A: conversion 90%
-
-
r
(2E)-2-propylbut-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 57%, mutant L384A: conversion 67%
-
-
r
(2Z)-2-(benzyloxy)but-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 0%, mutant L384A: conversion 60%
-
-
r
(2Z)-2-(benzylsulfanyl)but-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 0%, mutant L384A: conversion 42%
-
-
r
(2Z)-2-(ethylsulfanyl)but-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 36%, mutant L384A: conversion 50%
-
-
r
(2Z)-2-(phenylsulfanyl)but-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 0%, mutant L384A: conversion 34%
-
-
r
(2Z)-2-ethoxybut-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 46%, mutant L384A: conversion 43%
-
-
r
(2Z)-2-phenoxybut-2-enedioate + NH3
?
show the reaction diagram
-
wild-type: conversion 0%, mutant L384A: conversion 44%
-
-
r
mesaconate + (aminooxy)ethane
?
show the reaction diagram
-
mutant Q73A, conversion: 100%, yield: 61%
-
-
r
mesaconate + (aminooxy)methane
?
show the reaction diagram
-
mutant Q73A, conversion: 99%, excess of threo isomer over erythro isomer: more than 95%
-
-
r
mesaconate + 1-cyclopropylmethanamine
?
show the reaction diagram
mesaconate + 2-aminoethanol
?
show the reaction diagram
-
mutant Q73A, conversion: 37%, excess of threo isomer over erythro isomer: more than 95%
-
-
r
mesaconate + 2-methoxyethanamine
?
show the reaction diagram
mesaconate + 3-aminopropan-1-ol
?
show the reaction diagram
mesaconate + aminobutane
?
show the reaction diagram
mesaconate + aniline
?
show the reaction diagram
-
mutant Q73A, conversion: 6%, excess of threo isomer over erythro isomer: more than 95%
-
-
r
mesaconate + cyclobutanamine
?
show the reaction diagram
mesaconate + cyclohexanamine
?
show the reaction diagram
-
mutant Q73A, conversion: 21%, excess of threo isomer over erythro isomer: more than 95%
-
-
r
mesaconate + cyclopentanamine
?
show the reaction diagram
-
mutant Q73A, conversion: 36%, excess of threo isomer over erythro isomer: more than 95%
-
-
r
mesaconate + cyclopropanamine
?
show the reaction diagram
mesaconate + ethanamine
?
show the reaction diagram
mesaconate + ethane-1,2-diamine
?
show the reaction diagram
mesaconate + hexanamine
?
show the reaction diagram
-
mutant Q73A, conversion: 17%, excess of threo isomer over erythro isomer: more than 95%
-
-
r
mesaconate + methanamine
?
show the reaction diagram
mesaconate + N-methylethane-1,2-diamine
?
show the reaction diagram
-
mutant Q73A, conversion: 80%, excess of threo isomer over erythro isomer: more than 95%
-
-
r
mesaconate + NH3
L-threo-3-methylaspartate
show the reaction diagram
mesaconate + NH3
L-threo-3-methylaspartate + L-erythro-3-methylaspartate
show the reaction diagram
-
conversion 2h: 73% (wild-type), diastereomeric product ratio (threo:erythro) after 2h: 89:11
-
-
r
mesaconate + pentanamine
?
show the reaction diagram
mesaconate + propan-2-amine
?
show the reaction diagram
-
mutant Q73A, conversion: 8%, excess of threo isomer over erythro isomer: more than 95%, enantiomeric excess: more than 99%
-
-
r
mesaconate + propanamine
?
show the reaction diagram
mesaconate + propane-1,3-diamine
?
show the reaction diagram
(2S)-aspartate
fumarate + NH3
show the reaction diagram
-
-
-
?
(2S,3R)-3-ethylaspartic acid
?
show the reaction diagram
-
-
-
-
-
(2S,3S)-3-ethylaspartic acid
?
show the reaction diagram
-
-
-
-
-
3-bromoaspartate
bromofumarate + NH3
show the reaction diagram
3-chloroaspartate
chlorofumarate + NH3
show the reaction diagram
3-fluoroaspartate
fluorofumarate + NH3
show the reaction diagram
-
-
not a good substrate
-
beta-ethylaspartate
?
show the reaction diagram
beta-isopropylaspartate
?
show the reaction diagram
-
-
-
-
-
beta-propylaspartate
?
show the reaction diagram
erythro-3-methylaspartate
?
show the reaction diagram
ethylfumarate + NH3
?
show the reaction diagram
-
-
-
-
-
iso-propylfumarate + NH3
?
show the reaction diagram
-
-
-
-
-
L-aspartate
fumarate + NH3
show the reaction diagram
L-erythro-3-methylaspartate
mesaconate + NH3
show the reaction diagram
L-threo-3-methylaspartate
?
show the reaction diagram
-
-
-
-
-
L-threo-3-methylaspartate
mesaconate + NH3
show the reaction diagram
L-threo-3-methylaspartic acid
mesaconate + NH3
show the reaction diagram
assay at pH 9.0, 30C
-
-
?
mesaconic acid + NH3
L-threo-3-methylaspartate
show the reaction diagram
assay at pH 9.0, 30C
-
-
r
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
L-erythro-3-methylaspartate
mesaconate + NH3
show the reaction diagram
-
i.e. (2S,3R)-3-methylaspartate, slow syn elimination
-
r
L-threo-3-methylaspartate
?
show the reaction diagram
-
-
-
-
-
L-threo-3-methylaspartate
mesaconate + NH3
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cd2+
-
activates
Fe2+
-
activates
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-bromoaspartate
-
irreversible
4-nitrophenylhydrazine
-
90% inhibition after 200 min
hydroxylamine
-
-
methylhydrazine
-
-
N-ethylmaleimide
p-mercuribenzoate
-
-
phenylhydrazine
Sr2+
-
competitive inhibition of Mg2+ activation of the enzyme
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.2 - 60
mesaconate
5.2 - 40
(2S,3R)-3-methylaspartate
0.58
L-threo-3-methylaspartate
-
recombinant 3-methylaspartase
1 - 1.8
L-threo-3-methylaspartic acid
0.7 - 14
mesaconic acid
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.6 - 61
mesaconate
240 - 467
L-threo-3-methylaspartate
0.01 - 89
L-threo-3-methylaspartic acid
0.25 - 61
mesaconic acid
additional information
additional information
-
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.04 - 87
mesaconate
0.01 - 89
L-threo-3-methylaspartic acid
0.15 - 87
mesaconic acid
additional information
L-threo-3-methylaspartic acid
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45500
-
2 * 45500
45538
-
2 * 45538, mass spectrometry
45539
-
2 * 45539, calculation from sequence of amino acid
49000
-
2 * 49000, SDS-PAGE
100000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystallized from ammonium sulfate
-
once recrystallized
-
sitting-drop vapour-diffusion, 0.0045 ml protein sample and 0.0045 ml precipitant, crystals are obtained using 20-25% polyethylene glycol 6000, 100 mM sodium acetate, pH 7.0, 25 mM Tris-HCl as precipitant solution and 16-22% ethylene glycol as additive, crystals diffract to 2.0 A
-
sitting-drop vapur diffusion against a well solution of 20-25% polyethylene glycol 6000, 100 mM sodium acetate, pH 7.0, 25 mM Tris-HCl pH 7.0 as precipitating solution and 16-22% ethylene glycol as additive, crystals diffract to 1.9 A
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
0-3C, in 0.5 M tetramethylammonium chloride, pH 6.8 for many months, with little loss of activity
-
4C, enzyme crystals retain full activity after 1 year
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native and recombinant 3-methylaspartase
-
recombinant 3-methylaspartase
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
-
expression in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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)
C361K
-
mutant shows no reactivity. Conversion 2h: 10% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
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)
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)
Q172N
-
mutant shows no reactivity. Conversion 2h: 6% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
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)
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)
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)
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)
H194A
-
enhanced diastereoselectivities
K331A
-
enhanced diastereoselectivities
Q329A
-
enhanced diastereoselectivities
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
-
enzyme is used for L-aspartic acid production