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Literature summary for 2.3.3.16 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
Search for more literature for 2.3.3.16

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
A10E site-directed mutagenesis, reduced kcat, increased Km for acetyl-CoA Arthrobacter sp.
A361R site-directed mutagenesis, slightly reduced kcat , reduced Km for acetyl-CoA and increased Km for oxaloacetate, enhanced activity with propionyl-CoA Arthrobacter sp.
A361R/A10E site-directed mutagenesis, reduced kcat, reduced Km for acetyl-CoA Arthrobacter sp.
K313L site-directed mutagenesis Arthrobacter sp.
K313L/A361R site-directed mutagenesis Arthrobacter sp.
K313L/A361R/A10E site-directed mutagenesis Arthrobacter sp.
additional information
-
 Pyrococcus furiosus
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]

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

Organism

Organism UniProt Comment Textmining
Arthrobacter sp.
-
-
-
Arthrobacter sp. DS2-3R
-
-
-
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
-
 Arthrobacter sp. citrate + CoA
-
 ?
acetyl-CoA + oxaloacetate + H2O
-
 Arthrobacter sp. DS2-3R citrate + CoA
-
 ?
propionyl-CoA + oxaloacetate + H2O
-
 Arthrobacter sp. 2-methylcitrate + CoA
-
 ?
propionyl-CoA + oxaloacetate + H2O
-
 Arthrobacter sp. DS2-3R 2-methylcitrate + CoA
-
 ?

Temperature Stability [°C]

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

Turnover Number [1/s]

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

pH Optimum

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