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Literature summary 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

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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.3.9 wild type enzyme is expressed in Escherichia coli BL21(DE3) cells, mutant enzymes are expressed in Escherichia coli ArcticExpress(DE3) RP cells Euglena gracilis
4.1.3.1 His-tagged version (without signal peptide (amino acid residue 1-12)) expressed in Escherichia coli BL21(DE3) Euglena gracilis

Protein Variants

EC Number Protein Variants Comment Organism
2.3.3.9 D475N the glyoxylate cycle enzyme mutant completely loses malate synthase activity but retains isocitrate lyase activity Euglena gracilis
2.3.3.9 additional information the C-terminal domain of the glyoxylate cycle enzyme, when expressed alone (GCE(566-1165)), does not show any enzyme activity Euglena gracilis
2.3.3.9 R187K the glyoxylate cycle enzyme mutant completely loses malate synthase activity but retains isocitrate lyase activity Euglena gracilis
4.1.3.1 D475N putative acetyl-CoA binding site of malate synthase, no malate synthase activity, but isocitrate lyase activity Euglena gracilis
4.1.3.1 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
4.1.3.1 R187K putative acetyl-CoA binding site of malate synthase, no malate synthase activity, but isocitrate lyase activity Euglena gracilis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.3.9 0.065
-
 glyoxylate wild type full-length glyoxylate cycle enzyme, at 25°C, pH not specified in the publication Euglena gracilis
2.3.3.9 0.073
-
 glyoxylate N-terminal malate synthase-active domain GCE(13-573) of glyoxylate cycle enzyme, at 25°C, pH not specified in the publication Euglena gracilis
4.1.3.1 0.96
-
 glyoxylate D475N mutant protein, 0.15 mM acetyl-CoA, 10 mM succinate, pH 6.5, 30°C Euglena gracilis
4.1.3.1 1.3
-
 glyoxylate D475N mutant protein, 10 mM succinate, pH 6.5, 30°C Euglena gracilis
4.1.3.1 6
-
 succinate D475N mutant protein, 1.5 mM glyoxylate, sharply decreased with increasing acetyl-CoA concentration, pH 6.5, 30°C Euglena gracilis
4.1.3.1 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.3.3.9 62000
-
 x * 62000, the 62000 Da N-terminal domain of glyoxylate cycle enzyme provides malate synthase activity, SDS-PAGE Euglena gracilis
4.1.3.1 131000
-
 4 * 131000, gel filtration, calculated subunit molecular mass Euglena gracilis
4.1.3.1 530000
-
 gel filtration Euglena gracilis

Organism

EC Number Organism UniProt Comment Textmining
2.3.3.9 Euglena gracilis
-
-
-
4.1.3.1 Euglena gracilis Q8LPA6
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.3.9 DEAE Sepharose column chromatography, HisTrap HP column chromatography, and Superdex 200 pg gel filtration Euglena gracilis
4.1.3.1 anion exchange chromatography (DEAE), immobilized metal ion affinity chromatography (Ni2+), gel filtration Euglena gracilis

Source Tissue

EC Number Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.3.9 glyoxylate + acetyl-CoA + H2O
-
 Euglena gracilis (S)-malate + CoA
-
 ?
4.1.3.1 isocitrate
-
 Euglena gracilis succinate + glyoxylate
-
 r

Subunits

EC Number Subunits Comment Organism
2.3.3.9 ? x * 62000, the 62000 Da N-terminal domain of glyoxylate cycle enzyme provides malate synthase activity, SDS-PAGE Euglena gracilis
4.1.3.1 homotetramer 4 * 131000, gel filtration, calculated subunit molecular mass Euglena gracilis

Synonyms

EC Number Synonyms Comment Organism
2.3.3.9 GCE
-
 Euglena gracilis
2.3.3.9 GCE(13-573) N-terminal malate synthase-active domain of glyoxylate cycle enzyme Euglena gracilis
2.3.3.9 glyoxylate cycle enzyme bifunctional enzyme possessing malate synthase and isocitrate lyase activities Euglena gracilis
4.1.3.1 isocitrate lyase activity of a bifunctional protein named glyoxylate cycle enzyme (malate synthase and isocitrate lyase activity) Euglena gracilis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.3.9 14
-
 glyoxylate N-terminal malate synthase-active domain GCE(13-573) of glyoxylate cycle enzyme, at 25°C, pH not specified in the publication Euglena gracilis
2.3.3.9 17
-
 glyoxylate wild type full-length glyoxylate cycle enzyme, at 25°C, pH not specified in the publication Euglena gracilis