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Literature summary for 2.3.3.16 extracted from

  • Arnott, M.A.; Michael, R.A.; Thompson, C.R.; Hough, D.W.; Danson, M.J.
    Thermostability and thermoactivity of citrate synthases from the thermophilic and hyperthermophilic archaea, Thermoplasma acidophilum and Pyrococcus furiosus (2000), J. Mol. Biol., 304, 657-668.
    View publication on PubMed
Search for more literature for 2.3.3.16

Cloned(Commentary)

Cloned (Comment) Organism
expression of chimeric mutant in Escherichia coli citrate synthase deficient strain MOB154 Thermoplasma acidophilum
expression of wild-type, chimeric mutant and site-directed mutants in Escherichia coli citrate synthase deficient strain MOB154 Pyrococcus furiosus

Crystallization (Commentary)

Crystallization (Comment) Organism
crystal structure analysis of chimeric mutants with exchange of large and small subunits between Thermoplasma acidophilum and Pyrococcus furiosus Pyrococcus furiosus
crystal structure analysis of chimeric mutants with exchange of large and small subunits between Thermoplasma acidophilum and Pyrococcus furiosus Thermoplasma acidophilum

Protein Variants

Protein Variants Comment Organism
D113A site-directed mutagenesis, deletion of C-terminal amino acid residues which arrange the subunit contact, slightly increased Km for acetyl-CoA and oxaloacetate, slightly increased reaction velocity, reduced thermostability Pyrococcus furiosus
D113S site-directed mutagenesis, deletion of C-terminal amino acid residues which arrange the subunit contact, slightly decreased Km for acetyl-CoA and oxaloacetate, slightly increased reaction velocity, reduced thermostability Pyrococcus furiosus
additional information construction of mutant with disrupted inter-subunit ionic network by partly eleminating the C-terminal end amino acid residues, increased Km for substrates, reduced thermostability Pyrococcus furiosus
additional information construction of chimeric enzyme mutants with mix of the large and small subunits of Thermoplasma acidophilum and Pyrococcus furiosus, functional analysis of the subunits Pyrococcus furiosus
additional information construction of chimeric enzyme mutants with mix of the large and small subunits of Thermoplasma acidophilum and Pyrococcus furiosus, functional analysis of the subunits Thermoplasma acidophilum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.002
-
 oxaloacetate wild-type Thermoplasma acidophilum
0.004
-
 acetyl-CoA wild-type Pyrococcus furiosus
0.005
-
 acetyl-CoA wild-type Thermoplasma acidophilum
0.015
-
 oxaloacetate wild-type Pyrococcus furiosus

Organism

Organism UniProt Comment Textmining
Pyrococcus furiosus
-
-
-
Thermoplasma acidophilum
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant chimeric mutants from E. coli Thermoplasma acidophilum
recombinant wild-type, chimeric mutants, C-terminal deletion mutants, and D113 mutants from E. coli Pyrococcus furiosus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetyl-CoA + oxaloacetate + H2O
-
 Pyrococcus furiosus citrate + CoA
-
 ?
acetyl-CoA + oxaloacetate + H2O
-
 Thermoplasma acidophilum citrate + CoA
-
 ?

Subunits

Subunits Comment Organism
dimer
-
 Pyrococcus furiosus
dimer
-
 Thermoplasma acidophilum
More chimeric mutants with exchanged large and small subunits between Thermoplasma acidophilum and Pyrococcus furiosus, the large subunit is responsible for subunit interaction, the small subunit is responsable for catalytic activity Pyrococcus furiosus
More chimeric mutants with exchanged large and small subunits between Thermoplasma acidophilum and Pyrococcus furiosus, the large subunit is responsible for subunit interaction, the small subunit is responsable for catalytic activity Thermoplasma acidophilum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
65
-
 enzyme mutant lacking 13 amino acid residues of the C-terminal Pyrococcus furiosus
65 70 chimeric mutant containing the large subunit of Pyrcoccus furiosus and the small subunit of Thermoplasma acidophilum Pyrococcus furiosus
65 70 chimeric mutant containing the large subunit of Pyrcoccus furiosus and the small subunit of Thermoplasma acidophilum Thermoplasma acidophilum
70 75 wild-type Thermoplasma acidophilum
75
-
 enzyme mutant lacking 2 amino acid residues of the C-terminal Pyrococcus furiosus
80
-
 mutants D113S and D113A Pyrococcus furiosus
90
-
 above, wild-type Pyrococcus furiosus
90
-
 chimeric mutant containing the large subunit of Thermoplasma acidophilum and the small subunit of Pyrcoccus furiosus Pyrococcus furiosus
90
-
 chimeric mutant containing the large subunit of Thermoplasma acidophilum and the small subunit of Pyrcoccus furiosus Thermoplasma acidophilum

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
40 90
-
 Pyrococcus furiosus
40 90
-
 Thermoplasma acidophilum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 thermostability of wild-type and chimeric mutants Pyrococcus furiosus
additional information
-
 thermostability of wild-type and chimeric mutants Thermoplasma acidophilum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 enzyme assay at Pyrococcus furiosus
8
-
 enzyme assay at Thermoplasma acidophilum