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Literature summary for 1.1.1.40 extracted from

  • Kuo, C.C.; Lin, K.Y.; Hsu, Y.J.; Lin, S.Y.; Lin, Y.T.; Chang, G.G.; Chou, W.Y.
    The roles of Tyr91 and Lys162 in general acid-base catalysis in the pigeon NADP+-dependent malic enzyme (2008), Biochem. J., 411, 467-473.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid sequence determination and analysis, expression of wild-type and mutant enzymes in Escherichia coli strain JM109 Columba sp.

Protein Variants

Protein Variants Comment Organism
K162A site-directed mutagenesis, the mutation does not affect Mn2+ binding of the mutant enzyme, but kcat is 27000fold reduced compared to the wild-type enzyme, NH4Cl shows no rescue of the pyruvate reduction in the K162A mutant, while for oxaloacetate decarboxylation, ammonium chloride demonstrated a maximum restoration of 3.5fold at 1 mM, and its rescue efficiency decreases with increasing concentration Columba sp.
K162Q site-directed mutagenesis, the mutation does not affect Mn2+ binding of the mutant enzyme, but kcat is 3500fold reduced compared to the wild-type enzyme Columba sp.
K162R site-directed mutagenesis,the mutation does not affect Mn2+ binding of the mutant enzyme, but kcat is 125fold reduced compared to the wild-type enzyme Columba sp.
Y91F site-directed mutagenesis, the mutation does not affect Mn2+ binding of the mutant enzyme, the mutant shows a 25fold increase and a 3fold decrease in the Km values for (S)-malate and NADP+ respectively, and its kcat value is decreased by 200fold compared to wild-type enzyme Columba sp.

Inhibitors

Inhibitors Comment Organism Structure
(S)-malate substrate inhibition Columba sp.
NADP+ substrate inhibition Columba sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics of mutant enzymes, overview Columba sp.
0.0034
-
NADP+ pH 4.5, 25°C, wild-type enzyme Columba sp.
0.23
-
(S)-malate pH 4.5, 25°C, wild-type enzyme Columba sp.

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Columba sp. 5829
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ required for forward and reverse reaction, residues Tyr91 and Lys162 are not involved in metal ion binding Columba sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-malate + NADP+ Columba sp.
-
pyruvate + CO2 + NADPH
-
r

Organism

Organism UniProt Comment Textmining
Columba sp.
-
gene maeB encoding isozyme MaeB
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain JM109 by anion exchange chromatography and 2',5'-ADP affinity chromatography to over 95% purity Columba sp.

Reaction

Reaction Comment Organism Reaction ID
(S)-malate + NADP+ = pyruvate + CO2 + NADPH + H+ catalytic mechanism Columba sp.

Source Tissue

Source Tissue Comment Organism Textmining
liver
-
Columba sp.
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-malate + NADP+
-
Columba sp. pyruvate + CO2 + NADPH
-
r
(S)-malate + NADP+ via oxaloacetate, roles of Tyr91 and Lys162 in general acid-base catalysis in the pigeon NADP+-dependent malic enzyme, overview Columba sp. pyruvate + CO2 + NADPH
-
r

Subunits

Subunits Comment Organism
More circular dichroism structure analysis of wild-type and mutant enzymes, overview Columba sp.

Synonyms

Synonyms Comment Organism
MaeB
-
Columba sp.
NADP+-dependent malic enzyme
-
Columba sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Columba sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
140.1
-
(S)-malate pH 4.5, 25°C, wild-type enzyme Columba sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4.5
-
assay at, forward reaction Columba sp.
7.4
-
assay at, reverse reaction Columba sp.

pH Range

pH Minimum pH Maximum Comment Organism
additional information
-
pH profiles of wild-type and mutant enzymes, overview Columba sp.

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Columba sp.
NADPH
-
Columba sp.

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0089
-
NADP+ pH 4.5, 25°C, wild-type enzyme Columba sp.
0.28
-
(S)-malate pH 4.5, 25°C, wild-type enzyme Columba sp.