Information on EC 4.3.1.16 - threo-3-hydroxy-L-aspartate ammonia-lyase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
4.3.1.16
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RECOMMENDED NAME
GeneOntology No.
threo-3-hydroxy-L-aspartate ammonia-lyase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
threo-3-hydroxy-L-aspartate = oxaloacetate + NH3
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Deamination
SYSTEMATIC NAME
IUBMB Comments
threo-3-hydroxy-L-aspartate ammonia-lyase (oxaloacetate-forming)
A pyridoxal-phosphate protein. The enzyme, purified from the bacterium Pseudomonas sp. T62, is highly specific, and does not accept any other stereoisomer of 3-hydroxyaspartate. Different from EC 4.3.1.20, erythro-3-hydroxy-L-aspartate ammonia-lyase and EC 4.3.1.27, threo-3-hydroxy-D-aspartate ammonia-lyase. Requires a divalent cation such as Mn2+, Mg2+, or Ca2+.
CAS REGISTRY NUMBER
COMMENTARY hide
248270-70-4
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Manually annotated by BRENDA team
T62
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Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
L-threo-3-hydroxyaspartate
oxaloacetate + NH3
show the reaction diagram
additional information
?
-
none of the other 3-hydroxy amino acids tested (DL-erythro- and D-threo-3-hydroxyaspartate, D-threonine, L-threonine, DL-allo-threonine, D-serine, L-serine, and DL-phenylserine) is a substrate for this enzyme, either at 5 mM or 50 mM, as in the case of the enzyme from the original strain
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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-threo-3-hydroxyaspartate
oxaloacetate + NH3
show the reaction diagram
A4F2N8
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r
additional information
?
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A4F2N8
none of the other 3-hydroxy amino acids tested (DL-erythro- and D-threo-3-hydroxyaspartate, D-threonine, L-threonine, DL-allo-threonine, D-serine, L-serine, and DL-phenylserine) is a substrate for this enzyme, either at 5 mM or 50 mM, as in the case of the enzyme from the original strain
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyridoxal 5'-phosphate
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
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activates
Fe2+
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activates
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ATP
addition of 10 mM decreases enzyme activity to 89% of the control
CoCl2
1 mM causes inhibition enzyme activity (81% relative activity)
CuCl2
1 mM causes inhibition enzyme activity (92% relative activity)
D-serine
5 mM decrease the enzyme activity to 27%
GDP
addition of 10 mM decreases enzyme activity to 73% of the control
hydroxylamine
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L-erythro-3-hydroxyaspartate
5 mM, decrease the enzyme activity to 15%
SnCl2
1 mM causes inhibition enzyme activity (70% relative activity)
ZnCl2
1 mM causes inhibition enzyme activity (56% relative activity)
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ADP
addition of 10 mM increases enzyme activity to 106% of the control
AMP
addition of 10 mM increases enzyme activity to 144% of the control
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.54 - 0.74
L-threo-3-hydroxyaspartate
additional information
additional information
none of the other 3-hydroxy amino acids tested (DL-erythro- and D-threo-3-hydroxyaspartate, D-threonine, L-threonine, DL-allo-threonine, D-serine, L-serine, and DL-phenylserine) is a substrate for this enzyme, either at 5 mM or 50 mM, as in the case of the enzyme from the original strain
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
22.8
D-serine
non-competitive inhibition
0.2
L-erythro-3-hydroxyaspartate
strong competitive inhibition
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
39
recombinant enzyme
additional information
exhibits no detectable serine/aspartate racemase activity; K53A mutant enzyme shows no detectable activity
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36000
determined by using a MALDI-TOF mass spectrometer, measured value is lower than that determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (40 kDa), and that of the enzyme purified from the original strain (approximately 39 kDa by SDS-PAGE). The discrepancy may be due to the surface charge of the protein or some unknown factors such as the conformation of the enzyme
59000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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SDS-PAGE analysis shows 39000 subunit, either monomer or dimer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystals of D-THA DH belong to space group I4(1)22, with unit-cell parameters a = b = 157.3, c = 157.9 A. Single wavelength anomalous diffraction data are collected to a resolution of 2.0 A
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
after centrifugation of cultivated cell culture with the recombinant enzyme, resulting supernatant is applied to a Ni-sepharose column connected to a fast protein liquid chromatography system. The column is equilibrated with buffer supplemented with 20 mM imidazole. The enzyme is eluted with imidazole gradient.
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
overexpression of recombinant D-THA DH is carried out using a Rhodococcus erythropolis expression system
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the gene encoding L-THA DH is cloned and expressed in Escherichia coli
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K53A
mutant enzyme is produced in Escherichia coli JM109 cells