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Literature summary for 4.1.3.1 extracted from

  • Nakazawa, M.; Nishimura, M.; Inoue, K.; Ueda, M.; Inui, H.; Nakano, Y.; Miyatake, K.
    Characterization of a bifunctional glyoxylate cycle enzyme, malate synthase/isocitrate lyase, of Euglena gracilis (2011), J. Eukaryot. Microbiol., 58, 128-133.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
acetyl-CoA increases the substrate binding affinities isocitrate and succinate, but had little effect on the affinity for glyoxylate Euglena gracilis

Cloned(Commentary)

Cloned (Comment) Organism
His-tagged version (without signal peptide (amino acid residue 1-12)) expressed in Escherichia coli BL21(DE3) Euglena gracilis

Protein Variants

Protein Variants Comment Organism
D475N putative acetyl-CoA binding site of malate synthase, no malate synthase activity, but isocitrate lyase activity Euglena gracilis
additional information truncated mutants: 13-573 (no isocitrate lyase activity, malate synthase activity), 566-1165 (no malate synthase activity), 20-573 (no isocitrate lyase activity, malate synthase activity), 38-573 (no isocitrate lyase activity), 20-1165 (isocitrate lyase activity and malate synthase activity), 38-1165 Euglena gracilis
R187K putative acetyl-CoA binding site of malate synthase, no malate synthase activity, but isocitrate lyase activity Euglena gracilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.96
-
glyoxylate D475N mutant protein, 0.15 mM acetyl-CoA, 10 mM succinate, pH 6.5, 30°C Euglena gracilis
1.3
-
glyoxylate D475N mutant protein, 10 mM succinate, pH 6.5, 30°C Euglena gracilis
6
-
succinate D475N mutant protein, 1.5 mM glyoxylate, sharply decreased with increasing acetyl-CoA concentration, pH 6.5, 30°C Euglena gracilis
6.7
-
isocitrate when acetyl-CoA is present at concentrations higher than 5 mM, the Km value for isocitrate decreased by about one-third and the Vmax value increased about 2-fold, pH not specified in the publication, temperature not specified in the publication Euglena gracilis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
131000
-
4 * 131000, gel filtration, calculated subunit molecular mass Euglena gracilis
530000
-
gel filtration Euglena gracilis

Organism

Organism UniProt Comment Textmining
Euglena gracilis Q8LPA6
-
-

Purification (Commentary)

Purification (Comment) Organism
anion exchange chromatography (DEAE), immobilized metal ion affinity chromatography (Ni2+), gel filtration Euglena gracilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
isocitrate
-
Euglena gracilis succinate + glyoxylate
-
r

Subunits

Subunits Comment Organism
homotetramer 4 * 131000, gel filtration, calculated subunit molecular mass Euglena gracilis

Synonyms

Synonyms Comment Organism
isocitrate lyase activity of a bifunctional protein named glyoxylate cycle enzyme (malate synthase and isocitrate lyase activity) Euglena gracilis