Cloned (Comment) | Organism |
---|---|
gene TK1968, DNA and amino acid sequence determination and analysis, sequence comparison, recombinant expression in Escherichia coli | Thermococcus kodakarensis |
Protein Variants | Comment | Organism |
---|---|---|
additional information | generation of a gene TK1968 disruption mutant | Thermococcus kodakarensis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
CoA | strong inhibition, competitive versus NADH, effect on enzyme kinetics, overview | Thermococcus kodakarensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics in both reaction directions | Thermococcus kodakarensis | |
0.00135 | - |
NADPH | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
0.003 | - |
NADH | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
0.006 | - |
2-dehydropantoate | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
0.041 | - |
NAD+ | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
0.13 | - |
(R)-pantoate | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
2.04 | - |
(R)-pantoate | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
34048 | - |
2 * 34048, sequence calculation, 2 x 30000, about, recombinant enzyme, SDS-PAGE | Thermococcus kodakarensis |
60000 | - |
recombinant enzyme, gel filtration | Thermococcus kodakarensis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-pantoate + NAD+ | Thermococcus kodakarensis | - |
2-dehydropantoate + NADH + H+ | - |
r | |
(R)-pantoate + NAD+ | Thermococcus kodakarensis ATCC BAA-918 | - |
2-dehydropantoate + NADH + H+ | - |
r | |
2-dehydropantoate + NADH + H+ | Thermococcus kodakarensis | the TK1968 protein displays reductase activity specific for 2-oxopantoate and prefers NADH as the electron donor, distinct to the bacterial/eukaryotic NADPH-dependent enzymes | (R)-pantoate + NAD+ | - |
r | |
2-dehydropantoate + NADH + H+ | Thermococcus kodakarensis ATCC BAA-918 | the TK1968 protein displays reductase activity specific for 2-oxopantoate and prefers NADH as the electron donor, distinct to the bacterial/eukaryotic NADPH-dependent enzymes | (R)-pantoate + NAD+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermococcus kodakarensis | Q5JGC2 | gene TK1968 | - |
Thermococcus kodakarensis ATCC BAA-918 | Q5JGC2 | gene TK1968 | - |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Escherichia coli by heat treatment, anion-exchange chromatography, and gel filtration | Thermococcus kodakarensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-pantoate + NAD+ | - |
Thermococcus kodakarensis | 2-dehydropantoate + NADH + H+ | - |
r | |
(R)-pantoate + NAD+ | - |
Thermococcus kodakarensis ATCC BAA-918 | 2-dehydropantoate + NADH + H+ | - |
r | |
(R)-pantoate + NADP+ | - |
Thermococcus kodakarensis | 2-dehydropantoate + NADPH + H+ | - |
r | |
(R)-pantoate + NADP+ | - |
Thermococcus kodakarensis ATCC BAA-918 | 2-dehydropantoate + NADPH + H+ | - |
r | |
(S)-pantoate + NAD+ | low activity | Thermococcus kodakarensis | 2-dehydropantoate + NADH + H+ | - |
r | |
(S)-pantoate + NADP+ | low activity | Thermococcus kodakarensis | 2-dehydropantoate + NADPH + H+ | - |
r | |
2-dehydropantoate + NADH + H+ | - |
Thermococcus kodakarensis | (R)-pantoate + NAD+ | - |
r | |
2-dehydropantoate + NADH + H+ | the TK1968 protein displays reductase activity specific for 2-oxopantoate and prefers NADH as the electron donor, distinct to the bacterial/eukaryotic NADPH-dependent enzymes | Thermococcus kodakarensis | (R)-pantoate + NAD+ | - |
r | |
2-dehydropantoate + NADH + H+ | - |
Thermococcus kodakarensis ATCC BAA-918 | (R)-pantoate + NAD+ | - |
r | |
2-dehydropantoate + NADH + H+ | the TK1968 protein displays reductase activity specific for 2-oxopantoate and prefers NADH as the electron donor, distinct to the bacterial/eukaryotic NADPH-dependent enzymes | Thermococcus kodakarensis ATCC BAA-918 | (R)-pantoate + NAD+ | - |
r | |
2-dehydropantoate + NADPH + H+ | - |
Thermococcus kodakarensis | (R)-pantoate + NADP+ | - |
r | |
additional information | D-pantoate is much more preferred over L-pantoate in the reduction reaction, suggesting that the enzyme generates D-pantoate from 2-oxopantoate in the oxidation reaction | Thermococcus kodakarensis | ? | - |
? | |
additional information | D-pantoate is much more preferred over L-pantoate in the reduction reaction, suggesting that the enzyme generates D-pantoate from 2-oxopantoate in the oxidation reaction | Thermococcus kodakarensis ATCC BAA-918 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 34048, sequence calculation, 2 x 30000, about, recombinant enzyme, SDS-PAGE | Thermococcus kodakarensis |
Synonyms | Comment | Organism |
---|---|---|
2-dehydropantoate 2-reductase | UniProt | Thermococcus kodakarensis |
ketopantoate reductase | - |
Thermococcus kodakarensis |
KPR | - |
Thermococcus kodakarensis |
Tk-KPR | - |
Thermococcus kodakarensis |
TK1968 protein | - |
Thermococcus kodakarensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
90 | - |
recombinant enzyme | Thermococcus kodakarensis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | 110 | and above, activity range, profile overview | Thermococcus kodakarensis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | 90 | purified recombinant enzyme, pH 6.4, completely stable at for at least 24 h | Thermococcus kodakarensis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.22 | - |
NADPH | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
2.41 | - |
NAD+ | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
3.17 | - |
(R)-pantoate | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
3.8 | - |
(R)-pantoate | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
19.7 | - |
NADH | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
23.1 | - |
2-dehydropantoate | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.4 | - |
recombinant enzyme | Thermococcus kodakarensis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Thermococcus kodakarensis | |
NADH | preferred cofactor compared to NADPH | Thermococcus kodakarensis | |
NADP+ | - |
Thermococcus kodakarensis | |
NADPH | - |
Thermococcus kodakarensis |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000042 | - |
CoA | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis |
General Information | Comment | Organism |
---|---|---|
malfunction | gene disruption of gene TK1968results in a strain with growth defects that are complemented by addition of pantoate | Thermococcus kodakarensis |
metabolism | ketopantoate reductase from the hyperthermophilic archaeon Thermococcus kodakarensis catalyses the second step in CoA biosynthesis, the reduction of 2-oxopantoate to pantoate. CoA biosynthesis in the archaeon is regulated by feedback inhibition of enzyme ketopantoate reductase | Thermococcus kodakarensis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2 | - |
(R)-pantoate | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
29 | - |
(R)-pantoate | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
60 | - |
NAD+ | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
2220 | - |
NADPH | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
3830 | - |
2-dehydropantoate | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis | |
6540 | - |
NADH | recombinant enzyme, pH 6.4, 70°C | Thermococcus kodakarensis |