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Literature summary extracted from

  • Detarsio, E.; Andreo, C.S.; Drincovich, M.F.
    Basic residues play key roles in catalysis and NADP(+)-specificity in maize (Zea mays L.) photosynthetic NADP(+)-dependent malic enzyme (2004), Biochem. J., 382, 1025-1030.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.40 expression of wild-type and mutants in Escherichia coli BL21(DE3) Zea mays

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.40 K225I site-directed mutagenesis, mutation of a conserved residue involved in catalysis and substrate binding, mutant shows highly reduced activity and a 10fold higher partitioning ratio of oxaloacetate and malate compared to the wild-type enzyme, preference for reduction of oxaloacetate instead of decarboxylation Zea mays
1.1.1.40 K435L/K436L site-directed mutagenesis, mutation of residues which are important in cofactor binding, over 6fold increased Ki for 2'-AMP, and 1.7fold decreased Ki for 5'-AMP, and increased activity with NAD+ compared to the wild-type enzyme Zea mays
1.1.1.40 R237L site-directed mutagenesis, mutation of a conserved residue involved in catalysis and substrate binding, mutant shows and an over 100fold higher partitioning ratio of oxaloacetate and malate compared to the wild-type enzyme, preference for reduction of oxaloacetate instead of decarboxylation Zea mays

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.40 2'-AMP competitive Zea mays
1.1.1.40 5'-AMP competitive Zea mays

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.40 0.008
-
 NADP+ wild-type enzyme, pH 8.0, 30°C Zea mays
1.1.1.40 0.073
-
 NADP+ mutant K435L/K436L, pH 8.0, 30°C Zea mays
1.1.1.40 0.123
-
 NADP+ mutant K225I, pH 8.0, 30°C Zea mays
1.1.1.40 0.23
-
 L-malate wild-type enzyme, pH 8.0, 30°C Zea mays
1.1.1.40 0.29
-
 NADP+ mutant R237L, pH 8.0, 30°C Zea mays
1.1.1.40 0.31
-
 L-malate mutant K435L/K436L, pH 8.0, 30°C Zea mays
1.1.1.40 1.9
-
 NAD+ mutant K435L/K436L, pH 8.0, 30°C Zea mays
1.1.1.40 2.6
-
 L-malate mutant K225I, pH 8.0, 30°C Zea mays
1.1.1.40 2.9
-
 L-malate mutant R237L, pH 8.0, 30°C Zea mays
1.1.1.40 8.1
-
 NAD+ wild-type enzyme, pH 8.0, 30°C Zea mays

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.40 Mg2+
-
 Zea mays

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.40 Zea mays
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.40 recombinant wild-type and mutants from Escherichia coli to homogeneity Zea mays

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.1.40 (S)-malate + NADP+ = pyruvate + CO2 + NADPH + H+ the conserved residues Arg237 and Lys225 are involved in catalysis and substrate binding as proton acceptors Zea mays
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.40 (S)-malate + NAD+
-
 Zea mays pyruvate + CO2 + NADH
-
 ?
1.1.1.40 (S)-malate + NADP+
-
 Zea mays pyruvate + CO2 + NADPH
-
 ?
1.1.1.40 Oxaloacetate
-
 Zea mays Pyruvate + CO2
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.40 NADP+-dependent malic enzyme
-
 Zea mays
1.1.1.40 NADP+-ME
-
 Zea mays

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.40 30
-
 assay at Zea mays

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.40 0.38
-
 NADP+ mutant R237L, pH 8.0, 30°C Zea mays
1.1.1.40 1.1
-
 NADP+ mutant K225I, pH 8.0, 30°C Zea mays
1.1.1.40 13.5
-
 NAD+ wild-type enzyme, pH 8.0, 30°C Zea mays
1.1.1.40 18.4
-
 NAD+ mutant K435L/K436L, pH 8.0, 30°C Zea mays
1.1.1.40 181.1
-
 NADP+ mutant K435L/K436L, pH 8.0, 30°C Zea mays
1.1.1.40 201.3
-
 NADP+ wild-type enzyme, pH 8.0, 30°C Zea mays

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.40 8
-
 assay at Zea mays

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.40 NAD+ very low activity with Zea mays
1.1.1.40 NADP+ cofactor specificity is determined by Lys435/Lys436 interacting with the 2'-phosphate group of the ribose ring Zea mays

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.1.1.40 0.105
-
 2'-AMP wild-type enzyme, pH 8.0, 30°C Zea mays
1.1.1.40 0.256
-
 5'-AMP mutant K435L/K436L, pH 8.0, 30°C Zea mays
1.1.1.40 0.439
-
 5'-AMP wild-type enzyme, pH 8.0, 30°C Zea mays
1.1.1.40 0.688
-
 2'-AMP mutant K435L/K436L, pH 8.0, 30°C Zea mays