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Literature summary for 2.6.1.5 extracted from

  • Crismaru, C.G.; Wybenga, G.G.; Szymanski, W.; Wijma, H.J.; Wu, B.; Bartsch, S.; de Wildeman, S.; Poelarends, G.J.; Feringa, B.L.; Dijkstra, B.W.; Janssen, D.B.
    Biochemical properties and crystal structure of a beta-phenylalanine aminotransferase from Variovorax paradoxus (2013), Appl. Environ. Microbiol., 79, 185-195.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
recombinantly expressed in Escherichia coli as a His-tagged fusion protein Variovorax paradoxus

Crystallization (Commentary)

Crystallization (Comment) Organism
the crystal structures of the holoenzyme and of the enzyme in complex with the inhibitor 2-aminooxyacetate reveal structural similarity to the beta-phenylalanine aminotransferase from Mesorhizobium sp. strain LUK Variovorax paradoxus

Protein Variants

Protein Variants Comment Organism
R41A mutant is inactive when tested with (S)-beta-phenylalanine as amino donor and 2-oxoglutarate as amino acceptor Variovorax paradoxus

Inhibitors

Inhibitors Comment Organism Structure
2-aminooxyacetate
-
Variovorax paradoxus
2-oxoglutarate enzyme shows substrate inhibition Variovorax paradoxus
beta-phenylalanine enzyme shows substrate inhibition Variovorax paradoxus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.3
-
2-oxoglutarate pH 7.6, 30°C Variovorax paradoxus
1.5
-
(S)-beta-phenylalanine pH 7.6, 30°C Variovorax paradoxus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
46420
-
calculated from cDNA Variovorax paradoxus

Organism

Organism UniProt Comment Textmining
Variovorax paradoxus H8WR05
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography Variovorax paradoxus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-3-amino-5-methyl-hexanoic acid + 2-oxoglutarate relative activity: 48% Variovorax paradoxus ?
-
?
(S)-beta-phenylalanine + 2-oxoglutarate relative activity: 100% Variovorax paradoxus 3-oxo-3-phenylpropionic acid + L-glutamate
-
?
(S)-beta-phenylalanine + pyruvate the activity of VpAT with pyruvate is 85% of that with alpha-ketoglutarate as the amino acceptor Variovorax paradoxus 3-oxo-3-phenylpropionic acid + L-alanine
-
?
rac-2-fluoro-beta-phenylalanine + 2-oxoglutarate relative activity: 15% Variovorax paradoxus ?
-
?
rac-2-methyl-beta-phenylalanine + 2-oxoglutarate relative activity: 5% Variovorax paradoxus ?
-
?
rac-3-bromo-beta-phenylalanine + 2-oxoglutarate relative activity: 53% Variovorax paradoxus ?
-
?
rac-3-chloro-beta-phenylalanine + 2-oxoglutarate relative activity: 56% Variovorax paradoxus ?
-
?
rac-3-fluoro-beta-phenylalanine + 2-oxoglutarate relative activity: 48% Variovorax paradoxus ?
-
?
rac-3-methyl-beta-phenylalanine + 2-oxoglutarate relative activity: 54% Variovorax paradoxus ?
-
?
rac-4-bromo-beta-phenylalanine + 2-oxoglutarate relative activity: 62% Variovorax paradoxus ?
-
?
rac-4-ethyl-beta-phenylalanine + 2-oxoglutarate relative activity: 134% Variovorax paradoxus ?
-
?
rac-4-fluoro-beta-phenylalanine + 2-oxoglutarate relative activity: 87% Variovorax paradoxus ?
-
?
rac-4-iso-propyl-beta-phenylalanine + 2-oxoglutarate relative activity: 61% Variovorax paradoxus ?
-
?
rac-4-methyl-beta-phenylalanine + 2-oxoglutarate relative activity: 100% Variovorax paradoxus ?
-
?
rac-4-nitro-beta-phenylalanine + 2-oxoglutarate relative activity: 32% Variovorax paradoxus ?
-
?
rac-4-nitro-hydroxy-phenylalanine + 2-oxoglutarate relative activity: 61% Variovorax paradoxus ?
-
?
rac-4-propyl-beta-phenylalanine + 2-oxoglutarate relative activity: 126% Variovorax paradoxus ?
-
?
rac-4-trifluoromethyl-beta-phenylalanine + 2-oxoglutarate relative activity: 73% Variovorax paradoxus ?
-
?
rac-beta-leucine + 2-oxoglutarate relative activity: 38% Variovorax paradoxus ?
-
?
rac-beta-phenylalanine + 2-oxoglutarate relative activity: 100% Variovorax paradoxus ?
-
?

Synonyms

Synonyms Comment Organism
beta-phenylalanine aminotransferase
-
Variovorax paradoxus
VpAT
-
Variovorax paradoxus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
-
Variovorax paradoxus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
10.6
-
2-oxoglutarate pH 7.6, 30°C Variovorax paradoxus
11.8
-
(S)-beta-phenylalanine pH 7.6, 30°C Variovorax paradoxus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
assay at Variovorax paradoxus

pH Range

pH Minimum pH Maximum Comment Organism
4 11.2 enzyme shows high activity over a broad pH range Variovorax paradoxus

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate
-
Variovorax paradoxus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
40.3
-
beta-phenylalanine enzyme shows substrate inhibition, pH 7.6, 30°C Variovorax paradoxus
82.4
-
2-oxoglutarate enzyme shows substrate inhibition, pH 7.6, 30°C Variovorax paradoxus