Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His6-tagged enzyme in complex with NADP+ and pantoate, hanging drop vapor-diffusion technique, 20°C, 15-30 mg/ml protein with 2 mM NADP+ and 10 mM pantoate, 0.002 ml of protein solution is mixed with an equal volume of well solution containing 35% v/v dioxane, cryoprotection with 20% v/v 2-methyl-2,4-pentanediol, X-ray diffraction structure determination and analysis at 2.3 A resolution | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
E256A | site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview | Escherichia coli |
K176A | site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview | Escherichia coli |
K72A | site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview | Escherichia coli |
N98A | site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview | Escherichia coli |
R31A | site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview | Escherichia coli |
S244A | site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of recombinant wild-type and mutant enzymes, overview | Escherichia coli | |
0.007 | - |
NADPH | pH 7.5, 27°C, wild-type enzyme | Escherichia coli | |
0.03 | - |
2-dehydropantoate | pH 7.5, 27°C, wild-type enzyme | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-dehydropantoate + NADPH | Escherichia coli | an essential step for the biosynthesis of pantothenate, i.e. vitamin B5 | (R)-pantoate + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A9J4 | gene panE | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-dehydropantoate + NADPH | an essential step for the biosynthesis of pantothenate, i.e. vitamin B5 | Escherichia coli | (R)-pantoate + NADP+ | - |
r | |
2-dehydropantoate + NADPH | substrate and product binding structures, hinge bending between the N- and C-terminal domains is observed, which triggers the switch of the essential Lys176 to form a key hydrogen bond with the C2 hydroxyl of pantoate, pantoate forms additional interactions with conserved residues Ser244, Asn98, and Asn180 and with two conservatively varied residues, Asn194 and Asn241, overview | Escherichia coli | (R)-pantoate + NADP+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
ketopantoate reductase | - |
Escherichia coli |
KPR | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
27 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
25 | - |
NADPH | pH 7.5, 27°C, wild-type enzyme | Escherichia coli | |
25 | - |
2-dehydropantoate | pH 7.5, 27°C, wild-type enzyme | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Escherichia coli | |
NADPH | - |
Escherichia coli |