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Literature summary for 2.6.1.67 extracted from

  • Der Garabedian, P.A.; Vermeersch, J.J.
    Candida L-norleucine,leucine:2-oxoglutarate aminotransferase. Purification and properties (1987), Eur. J. Biochem., 167, 141-147.
    View publication on PubMed

Inhibitors

Inhibitors Comment Organism Structure
L-leucine in presence of L-norleucine: ping-pong bi-bi inhibition with alternative substrates Candida guilliermondii var. membranaefaciens
L-norvaline non-competitive inhibitor Candida guilliermondii var. membranaefaciens
L-valine competitive inhibitor Candida guilliermondii var. membranaefaciens
additional information 6-aminohexanoic acid, L-isoleucine, L-ornithine, L-lysine are not inhibitors Candida guilliermondii var. membranaefaciens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.8
-
L-norleucine pH 7.8, 35°C, in the presence of 2-oxoglutarate Candida guilliermondii var. membranaefaciens
2
-
2-oxoglutarate pH 7.8, 35°C, in the presence of L-norleucine or L-leucine Candida guilliermondii var. membranaefaciens
6.6
-
L-leucine in the presence of 2-oxoglutarate Candida guilliermondii var. membranaefaciens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
51000
-
2 * 51000, SDS-PAGE Candida guilliermondii var. membranaefaciens
100000
-
gel filtration Candida guilliermondii var. membranaefaciens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-leucine + 2-oxoglutarate Candida guilliermondii var. membranaefaciens 54.2% of activity compared to L-norleucine 4-methyl-2-oxopentanoate + L-glutamate
-
?
L-norleucine + 2-oxoglutarate Candida guilliermondii var. membranaefaciens best amino donor, classical ping pong bi-bi mechanism 2-oxohexanoate + L-glutamate
-
?

Organism

Organism UniProt Comment Textmining
Candida guilliermondii var. membranaefaciens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate partial purification and several chromatographic steps, all steps at 4°C Candida guilliermondii var. membranaefaciens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
26
-
pH 7.8, 35°C Candida guilliermondii var. membranaefaciens

Storage Stability

Storage Stability Organism
-25°C, stable in the presence of 0.01 mM pyridoxal 5'-phosphate and 0.01 mM 2-oxoglutarate Candida guilliermondii var. membranaefaciens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5-N-methylornithine + 2-oxoglutarate 10% of activity compared to L-norleucine Candida guilliermondii var. membranaefaciens 5-(methylamino)-2-oxopentanoic acid + L-glutamate
-
?
6-N-acetyllysine + 2-oxoglutarate 31.9% of activity compared to L-norleucine Candida guilliermondii var. membranaefaciens 6-(acetylamino)-2-oxohexanoic acid + L-glutamate
-
?
L-aspartate + 2-oxoglutarate 4.3% of activity compared to L-norleucine Candida guilliermondii var. membranaefaciens oxaloacetate + L-glutamate
-
?
L-isoleucine + 2-oxoglutarate 7.7% of activity compared to L-norleucine Candida guilliermondii var. membranaefaciens 3-methyl-2-oxopentanoate + L-glutamate
-
?
L-leucine + 2-oxoglutarate 54.2% of activity compared to L-norleucine Candida guilliermondii var. membranaefaciens 4-methyl-2-oxopentanoate + L-glutamate
-
?
L-norleucine + 2-oxoglutarate best amino donor, classical ping pong bi-bi mechanism Candida guilliermondii var. membranaefaciens 2-oxohexanoate + L-glutamate
-
?
L-norvaline + 2-oxoglutarate 10% of activity compared to L-norleucine Candida guilliermondii var. membranaefaciens 2-oxopentanoate + L-glutamate
-
?
additional information glycine, L-alanine, L-tyrosine, beta-alanine, epsilon-aminocaproate, delta-aminovalerate, L-arginine, L-lysine or L-ornithine are no substrates Candida guilliermondii var. membranaefaciens ?
-
?

Subunits

Subunits Comment Organism
dimer 2 * 51000, SDS-PAGE Candida guilliermondii var. membranaefaciens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
35
-
assay at Candida guilliermondii var. membranaefaciens
55
-
-
Candida guilliermondii var. membranaefaciens

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
25 55 reaction velocity increases linearly with increasing temperature at pH 7.8, above 55°C activity rapidly decreases, 2-oxoglutarate and to a lesser extent pyridoxal 5'-phosphate are effective protecting agents toward heating Candida guilliermondii var. membranaefaciens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
3 min, about 17% remaining activity, with addition of pyridoxal 5'-phosphate 70% remaining activity, with addition of 2-oxoglutarate 95% remaining activity Candida guilliermondii var. membranaefaciens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
-
assay at Candida guilliermondii var. membranaefaciens

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate absolute requirement, probably 1 mol pyridoxal 5'-phosphate binds to each subunit, apoenzyme is inactive and regains more than 92% of original enzymatic activity after addition of pyridoxal 5'-phosphate Candida guilliermondii var. membranaefaciens