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Literature summary for 2.3.3.14 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
Search for more literature for 2.3.3.14

Cloned(Commentary)

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

Inhibitors

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

KM Value [mM]

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

Metals/Ions

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

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
50600
-
 x * 50600, deduced from amino acid sequence Aspergillus fumigatus
51000
-
 x * 51000, SDS-PAGE Aspergillus fumigatus
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
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
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
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

Organism UniProt Comment Textmining
Aspergillus fumigatus C7C437 formerly Aspergillus fumigatus
-
Aspergillus fumigatus CBS 144.89 C7C437 formerly Aspergillus fumigatus
-

Purification (Commentary)

Purification (Comment) Organism
nickel affinity column chromatography Aspergillus fumigatus

Specific Activity [micromol/min/mg]

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

Storage Stability

Storage Stability Organism
-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
4°C, buffer without stabilizing components, 72 h, complete loss of activity Aspergillus fumigatus

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms Comment Organism
HcsA
-
 Aspergillus fumigatus

Temperature Optimum [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
-
 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]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8 8.2
-
 Aspergillus fumigatus

pH Stability

pH Stability pH Stability Maximum Comment Organism
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

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

General Information

General Information Comment Organism
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