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Literature summary for 1.1.1.169 extracted from

  • Zheng, R.; Blanchard, J.S.
    Identification of active site residues in E. coli ketopantoate reductase by mutagenesis and chemical rescue (2000), Biochemistry, 39, 16244-16251.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
bromoethylamine activation of enzyme from K176C mutant Escherichia coli
ethylamine activation of enzyme from K176A mutant Escherichia coli
formate E256A mutant activity greatly increased, twice as high at pH 7.2 as at pH 5.9 Escherichia coli
methylamine activation of enzyme from K176A mutant Escherichia coli
Propylamine activation of enzyme from K176A mutant Escherichia coli

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
D248A site-directed mutagenesis, wild-type activity Escherichia coli
E210A site-directed mutagenesis, wild-type activity Escherichia coli
E240A site-directed mutagenesis, wild-type activity Escherichia coli
E256A site-directed mutagenesis, significant reduction in catalytic efficiency of enzyme Escherichia coli
E256D wild-type activity Escherichia coli
K176A site-directed mutagenesis, significant reduction in catalytic efficiency of enzyme Escherichia coli
K176A/E256A double mutant, no activity Escherichia coli
K176C wild-type activity Escherichia coli
K72A site-directed mutagenesis, wild-type activity Escherichia coli
additional information Lys176 acts as general acid in ketopantoate reduction and is involved in catalysis and ketopantoate binding, E256A functions in D-pantoate and ketopantoate binding in ketopantoate reductase Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.002
-
NADPH +/-0.0003, mutant E256A Escherichia coli
0.0029
-
NADPH +/-0.0006, mutant E256D Escherichia coli
0.0038
-
NADPH +/-0.0003, mutant K176C, alkylated Escherichia coli
0.004
-
NADPH +/-0.0004 Escherichia coli
0.0066
-
NADPH +/-0.0016, mutant K176C Escherichia coli
0.016
-
NADPH +/-0.003, mutant K176A Escherichia coli
0.038
-
ketopantoate +/-0.009, mutant K176C, alkylated Escherichia coli
0.07
-
ketopantoate +/-0.01, mutant K176C Escherichia coli
0.12
-
ketopantoate +/-0.008 Escherichia coli
0.95
-
ketopantoate +/-0.29, mutant E256D Escherichia coli
7.5
-
ketopantoate +/-2.9, mutant K256A Escherichia coli
40
-
ketopantoate +/-6, mutant K176A Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
additional information
-
Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
34000
-
-
Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
expressed in BL21(DE3)
-

Purification (Commentary)

Purification (Comment) Organism
homogeneity in a yield of 20-60 mg from 3-6 g of cells Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
no activity for double mutant K176A/E256A Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
alpha-ketopantoate + NADPH
-
Escherichia coli D-pantoate + NADP+
-
r
additional information no activity for double mutant K176A/E256A Escherichia coli ?
-
?

Subunits

Subunits Comment Organism
monomer
-
Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
additional information
-
activity decreases with pH increase, reduction of ketopantoic acid Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Escherichia coli
NADPH
-
Escherichia coli