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 (Comment) | Organism |
---|---|
expression in Escherichia coli BL21(DE3) | Escherichia coli |
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 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 | Comment | Organism | Structure |
---|---|---|---|
additional information | - |
Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
34000 | - |
- |
Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
expressed in BL21(DE3) | - |
Purification (Comment) | Organism |
---|---|
homogeneity in a yield of 20-60 mg from 3-6 g of cells | Escherichia coli |
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 | 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 | Comment | Organism |
---|---|---|
monomer | - |
Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
activity decreases with pH increase, reduction of ketopantoic acid | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Escherichia coli | |
NADPH | - |
Escherichia coli |