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Literature summary extracted from

  • Schoebel, F.; Jacobsen, I.D.; Brock, M.
    Evaluation of lysine biosynthesis as an antifungal drug target: biochemical characterization of Aspergillus fumigatus homocitrate synthase and virulence studies (2010), Eukaryot. Cell, 9, 878-893.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.3.14 expressed in Escherichia coli BL21(DE3) Rosetta 2 cells Aspergillus fumigatus

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.3.14 2-methylcitrate 89.6% residual activity at 5 mM Aspergillus fumigatus
2.3.3.14 Ca2+ 96.3% residual activity at 1 mM Aspergillus fumigatus
2.3.3.14 citrate 66.1% residual activity at 10 mM Aspergillus fumigatus
2.3.3.14 Cu2+ 0.7% residual activity at 0.025 mM Aspergillus fumigatus
2.3.3.14 dipyridyl 95% residual activity at 0.1 mM Aspergillus fumigatus
2.3.3.14 EDTA 13.4% residual activity at 5 mM Aspergillus fumigatus
2.3.3.14 K+ 91.7% residual activity at 0.1 mM Aspergillus fumigatus
2.3.3.14 L-lysine 24.4% residual activity at 50 mM Aspergillus fumigatus
2.3.3.14 Na2SO4 90.9% residual activity at 0.2 mM Aspergillus fumigatus
2.3.3.14 Zn2+ 0.8% residual activity at 0.1 mM Aspergillus fumigatus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.3.14 0.017
-
 acetyl-CoA in 200 mM HEPES, pH 7.5, at 22°C Aspergillus fumigatus
2.3.3.14 0.261
-
 2-oxoglutarate in 200 mM HEPES, pH 7.5, at 22°C Aspergillus fumigatus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.3.3.14 additional information enzyme activity is independent of any metal ions Aspergillus fumigatus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.3.3.14 50600
-
 x * 50600, deduced from amino acid sequence Aspergillus fumigatus
2.3.3.14 51000
-
 x * 51000, SDS-PAGE Aspergillus fumigatus
2.3.3.14 66000
-
 different bands are detected at 66 kDa (monomer), 148 kDa (dimer), 226 kDa (trimer), and 280 kDa (tetramer), and the dimeric and tetrameric compositions are most abundant, gel filtration Aspergillus fumigatus
2.3.3.14 148000
-
 different bands are detected at 66 kDa (monomer), 148 kDa (dimer), 226 kDa (trimer), and 280 kDa (tetramer), and the dimeric and tetrameric compositions are most abundant, gel filtration Aspergillus fumigatus
2.3.3.14 226000
-
 different bands are detected at 66 kDa (monomer), 148 kDa (dimer), 226 kDa (trimer), and 280 kDa (tetramer), and the dimeric and tetrameric compositions are most abundant, gel filtration Aspergillus fumigatus
2.3.3.14 280000
-
 different bands are detected at 66 kDa (monomer), 148 kDa (dimer), 226 kDa (trimer), and 280 kDa (tetramer), and the dimeric and tetrameric compositions are most abundant, gel filtration Aspergillus fumigatus

Organism

EC Number Organism UniProt Comment Textmining
2.3.3.14 Aspergillus fumigatus C7C437 formerly Aspergillus fumigatus
-
2.3.3.14 Aspergillus fumigatus CBS 144.89 C7C437 formerly Aspergillus fumigatus
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.3.14 nickel affinity column chromatography Aspergillus fumigatus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.3.3.14 0.8
-
 purified enzyme, in 200 mM HEPES, pH 7.5, at 22°C Aspergillus fumigatus

Storage Stability

EC Number Storage Stability Organism
2.3.3.14 -20°C or -80°C, HEPES buffer with guanidine hydrochloride, alpha-cyclodextrin, ammonium sulfate, KCl and 20% (v/v) glycerol, several months, no significant loss of activity Aspergillus fumigatus
2.3.3.14 4°C, buffer without stabilizing components, 72 h, complete loss of activity Aspergillus fumigatus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.3.14 acetyl-CoA + H2O + 2-oxoglutarate specific substrates Aspergillus fumigatus 2-hydroxybutane-1,2,4-tricarboxylate + CoA
-
 ?
2.3.3.14 acetyl-CoA + H2O + 2-oxoglutarate specific substrates Aspergillus fumigatus CBS 144.89 2-hydroxybutane-1,2,4-tricarboxylate + CoA
-
 ?

Subunits

EC Number Subunits Comment Organism
2.3.3.14 ? x * 51000, SDS-PAGE Aspergillus fumigatus
2.3.3.14 ? x * 50600, deduced from amino acid sequence Aspergillus fumigatus

Synonyms

EC Number Synonyms Comment Organism
2.3.3.14 HcsA
-
 Aspergillus fumigatus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.3.14 50 52
-
 Aspergillus fumigatus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.3.3.14
-
 60 the enzyme retains 100% activity after more than 8 h at 0-37°C, 70% residual activity after 270 min at 42°C, and 50% residual activity after 2 min at 60°C Aspergillus fumigatus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.3.14 0.68
-
 acetyl-CoA in 200 mM HEPES, pH 7.5, at 22°C Aspergillus fumigatus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.3.14 7.8 8.2
-
 Aspergillus fumigatus

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
2.3.3.14 4 11 the enzyme retains 70% residual activity after 6 h at pH 4.0, 100% activity after 6 h at pH 5.0-6.0, 80% residual activity after 6.5 h at pH 7.0-9.0, and 50% residual activity after 50 min at pH 11.0 Aspergillus fumigatus

Expression

EC Number Organism Comment Expression
2.3.3.14 Aspergillus fumigatus gene expression increases under conditions of lysine supplementation up

General Information

EC Number General Information Comment Organism
2.3.3.14 malfunction an hcsA deletion mutant is lysine auxotrophic and unable to germinate on unhydrolyzed proteins given as a sole nutrient source. The mutant grows on blood and serum, indicating the existence of high-affinity lysine uptake systems. The virulence of the mutant is strongly attenuated in murine models of bronchopulmonary aspergillosis. The mutant retains full virulence when injected intravenously Aspergillus fumigatus