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Literature summary extracted from

  • Gerike, U.; Danson, M.J.; Hough, D.W.
    Cold-active citrate synthase: mutagenesis of active-site residues (2001), Protein Eng., 14, 655-661.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.3.16 cloning and expression of wild-type and mutants in Escherichia coli Arthrobacter sp.

Protein Variants

EC Number Protein Variants Comment Organism
2.3.3.16 A10E site-directed mutagenesis, reduced kcat, increased Km for acetyl-CoA Arthrobacter sp.
2.3.3.16 A361R site-directed mutagenesis, slightly reduced kcat , reduced Km for acetyl-CoA and increased Km for oxaloacetate, enhanced activity with propionyl-CoA Arthrobacter sp.
2.3.3.16 A361R/A10E site-directed mutagenesis, reduced kcat, reduced Km for acetyl-CoA Arthrobacter sp.
2.3.3.16 K313L site-directed mutagenesis Arthrobacter sp.
2.3.3.16 K313L/A361R site-directed mutagenesis Arthrobacter sp.
2.3.3.16 K313L/A361R/A10E site-directed mutagenesis Arthrobacter sp.
2.3.3.16 additional information
-
 Pyrococcus furiosus
2.3.3.16 additional information introduction of a loop into the active site of wild-type, mutant K313L, K313L/A361R/A10E , and mutant K313L/A361R, the latter showing increased thermostability and decreased temperature optimum for catalytic activity, Km and activities Arthrobacter sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.3.16 additional information
-
 additional information Km values for acetyl-CoA and oxaloacetate of mutants with a loop introduced into the active site Arthrobacter sp.
2.3.3.16 0.002
-
 oxaloacetate mutant A361R Arthrobacter sp.
2.3.3.16 0.004
-
 propionyl-CoA mutant A361R Arthrobacter sp.
2.3.3.16 0.006
-
 oxaloacetate mutant A361R/A10E Arthrobacter sp.
2.3.3.16 0.007
-
 oxaloacetate wild-type Arthrobacter sp.
2.3.3.16 0.008
-
 oxaloacetate mutant A10E Arthrobacter sp.
2.3.3.16 0.008
-
 propionyl-CoA mutant loop/K313L/A361R Arthrobacter sp.
2.3.3.16 0.015
-
 oxaloacetate
-
 Pyrococcus furiosus
2.3.3.16 0.015
-
 oxaloacetate mutant A361R Arthrobacter sp.
2.3.3.16 0.016
-
 propionyl-CoA wild-type Arthrobacter sp.
2.3.3.16 0.02
-
 oxaloacetate mutant mutant loop/K313L/A361R Arthrobacter sp.
2.3.3.16 0.037
-
 acetyl-CoA mutant A361R/A10E Arthrobacter sp.
2.3.3.16 0.038
-
 acetyl-CoA mutant A361R Arthrobacter sp.
2.3.3.16 0.2
-
 acetyl-CoA wild-type Arthrobacter sp.
2.3.3.16 1.24
-
 acetyl-CoA mutant A10E Arthrobacter sp.

Organism

EC Number Organism UniProt Comment Textmining
2.3.3.16 Arthrobacter sp.
-
-
-
2.3.3.16 Arthrobacter sp. DS2-3R
-
-
-
2.3.3.16 Pyrococcus furiosus
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.3.16 acetyl-CoA + oxaloacetate + H2O
-
 Pyrococcus furiosus citrate + CoA
-
 ?
2.3.3.16 acetyl-CoA + oxaloacetate + H2O
-
 Arthrobacter sp. citrate + CoA
-
 ?
2.3.3.16 acetyl-CoA + oxaloacetate + H2O
-
 Arthrobacter sp. DS2-3R citrate + CoA
-
 ?
2.3.3.16 propionyl-CoA + oxaloacetate + H2O
-
 Arthrobacter sp. 2-methylcitrate + CoA
-
 ?
2.3.3.16 propionyl-CoA + oxaloacetate + H2O
-
 Arthrobacter sp. DS2-3R 2-methylcitrate + CoA
-
 ?

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.3.3.16 additional information
-
-
 Pyrococcus furiosus
2.3.3.16 additional information
-
 thermal stability of mutants Arthrobacter sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.3.16 additional information
-
 additional information values for acetyl-CoA and oxaloacetate of mutants with a loop introduced into the active site Arthrobacter sp.
2.3.3.16 4
-
 propionyl-CoA mutant loop/K313L/A361R Arthrobacter sp.
2.3.3.16 6
-
 propionyl-CoA mutant A361R Arthrobacter sp.
2.3.3.16 7
-
 acetyl-CoA mutant A10E Arthrobacter sp.
2.3.3.16 8
-
 propionyl-CoA wild-type Arthrobacter sp.
2.3.3.16 13
-
 acetyl-CoA mutant A361R/A10E Arthrobacter sp.
2.3.3.16 15
-
 acetyl-CoA mutant A361R Arthrobacter sp.
2.3.3.16 18
-
 acetyl-CoA wild-type Arthrobacter sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.3.16 8
-
 enzyme assay at Pyrococcus furiosus
2.3.3.16 8
-
 enzyme assay at Arthrobacter sp.