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Literature summary extracted from

  • Fernandez, F.J.; de Vries, D.; Pena-Soler, E.; Coll, M.; Christen, P.; Gehring, H.; Vega, M.C.
    Structure and mechanism of a cysteine sulfinate desulfinase engineered on the aspartate aminotransferase scaffold (2012), Biochim. Biophys. Acta, 1824, 339-349.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.6.1.1 expressed in Escherichia coli Escherichia coli

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.6.1.1 the crystal triple mutant I33Q/Y214Q/R280Y is determined to relate the observed changes in reaction kinetics to the changes in active-site structure Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
2.6.1.1 I33Q/Y214Q/R280Y substitution of three active-site residues in Escherichia coli L-aspartateartate aminotransferase highly reduces kcat for transamination reaction. Ratio of L-cysteine sulfinate desulfinase to transaminase activity is increased by 100000fold in mutant. kcat for desulfination of L-cysteine sulfinate increases to 0.5/sec (from 0.05/sec in wild-type enzyme). kcat for beta-decarboxylation of L-aspartateartate increases from below 0.0001/sec to 0.07/sec Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.6.1.1 3.1
-
 L-aspartate pH 7.5, 25°C, mutant I33Q/Y214Q/R280Y Escherichia coli
2.6.1.1 4
-
 L-aspartate pH 7.5, 25°C, wild-type Escherichia coli
2.6.1.1 5.2
-
 L-glutamate pH 7.5, 25°C, mutant I33Q/Y214Q/R280Y Escherichia coli
2.6.1.1 37
-
 L-glutamate pH 7.5, 25°C, wild-type Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
2.6.1.1 Escherichia coli P00509
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.6.1.1 EcAspAT is purified by anion exchange chromatography in a Fractogel EMD DEAE 650-S column, pooled fractions are dialyzed against 20 mM sodium phosphate, pH 7.5, and loaded onto a ceramic hydroxyapatite column Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.6.1.1 L-aspartate + 2-oxoglutarate
-
 Escherichia coli oxaloacetate + L-glutamate
-
 r

Synonyms

EC Number Synonyms Comment Organism
2.6.1.1 EcAspAT wild-type enzyme shows weak desulfination activity and decarboxylation activity Escherichia coli
2.6.1.1 L-aspartateartate aminotransferase
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.6.1.1 25
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.6.1.1 0.03
-
 L-glutamate pH 7.5, 25°C, mutant I33Q/Y214Q/R280Y Escherichia coli
2.6.1.1 0.13
-
 L-aspartate pH 7.5, 25°C, mutant I33Q/Y214Q/R280Y Escherichia coli
2.6.1.1 530
-
 L-aspartate pH 7.5, 25°C, wild-type Escherichia coli
2.6.1.1 670
-
 L-glutamate pH 7.5, 25°C, wild-type Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.6.1.1 7.5
-
 assay at Escherichia coli

Cofactor

EC Number Cofactor Comment Organism Structure
2.6.1.1 pyridoxal 5'-phosphate
-
 Escherichia coli
2.6.1.1 pyridoxamine 5'-phosphate
-
 Escherichia coli

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.6.1.1 0.006
-
 L-glutamate pH 7.5, 25°C, mutant I33Q/Y214Q/R280Y Escherichia coli
2.6.1.1 0.042
-
 L-aspartate pH 7.5, 25°C, mutant I33Q/Y214Q/R280Y Escherichia coli
2.6.1.1 18
-
 L-glutamate pH 7.5, 25°C, wild-type Escherichia coli
2.6.1.1 133
-
 L-aspartate pH 7.5, 25°C, wild-type Escherichia coli