EC Number | Cloned (Comment) | Organism |
---|---|---|
1.2.4.1 | expression of wild-type E1alpha and E1beta subunits of the pyruvate decarboxylate component and mutants in Escherichia coli strain BL21(DE3) | Geobacillus stearothermophilus |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.2.4.1 | D276A | site-directed mutagenesis, reduced activity with pyruvate compared to the wild-type enzyme | Geobacillus stearothermophilus |
1.2.4.1 | F266A | site-directed mutagenesis, reduced activity with pyruvate compared to the wild-type enzyme | Geobacillus stearothermophilus |
1.2.4.1 | R267A | site-directed mutagenesis, highly reduced activity with pyruvate compared to the wild-type enzyme | Geobacillus stearothermophilus |
1.2.4.1 | R282A | site-directed mutagenesis, reduced activity with pyruvate compared to the wild-type enzyme | Geobacillus stearothermophilus |
1.2.4.1 | S283C | site-directed mutagenesis, reduced activity with pyruvate compared to the wild-type enzyme | Geobacillus stearothermophilus |
1.2.4.1 | Y281A | site-directed mutagenesis, reduced activity with pyruvate compared to the wild-type enzyme | Geobacillus stearothermophilus |
1.2.4.1 | Y281A/R282A | site-directed mutagenesis, highly reduced activity with pyruvate compared to the wild-type enzyme | Geobacillus stearothermophilus |
1.2.4.1 | Y281A/R282A/S283A | site-directed mutagenesis, highly reduced activity with pyruvate compared to the wild-type enzyme | Geobacillus stearothermophilus |
1.2.4.1 | Y281S/R282S | site-directed mutagenesis, highly reduced activity with pyruvate compared to the wild-type enzyme | Geobacillus stearothermophilus |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.2.4.1 | additional information | - |
additional information | kinetics | Geobacillus stearothermophilus | |
1.2.4.1 | 0.00056 | - |
thiamine diphosphate | mutant R267A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.0011 | - |
pyruvate | wild-type enzyme, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.0012 | - |
pyruvate | mutant F266A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.0021 | - |
pyruvate | mutant S283C, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.0027 | - |
thiamine diphosphate | mutant Y281S/R282S, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.0029 | - |
thiamine diphosphate | mutant Y281A/R282A/S283A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.003 | - |
thiamine diphosphate | mutant Y281A/R282A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.0052 | - |
thiamine diphosphate | mutant R282A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.0055 | - |
thiamine diphosphate | mutant Y281A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.0088 | - |
pyruvate | mutant R282A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.011 | - |
pyruvate | mutant D276A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.013 | - |
thiamine diphosphate | mutant S283C, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.021 | - |
thiamine diphosphate | mutants F266A and D276A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.023 | - |
thiamine diphosphate | wild-type enzyme, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.049 | - |
pyruvate | mutant Y281A/R282A/S283A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.056 | - |
pyruvate | mutant Y281A/R282A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.063 | - |
pyruvate | mutant Y281A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.066 | - |
pyruvate | mutant Y281S/R282S, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.291 | - |
pyruvate | mutant R267A, pH 7.0, 25°C | Geobacillus stearothermophilus |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.2.4.1 | Mg2+ | - |
Geobacillus stearothermophilus |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.2.4.1 | pyruvate + CoA + NAD+ | Geobacillus stearothermophilus | first step of the overall reaction of the pyruvate dehydrogenase complex | acetyl-CoA + CO2 + NADH | - |
ir |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.2.4.1 | Geobacillus stearothermophilus | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.2.4.1 | recombinant wild-type E1alpha and E1beta subunits of the pyruvate decarboxylate component and mutants from Escherichia coli strain BL21(DE3) | Geobacillus stearothermophilus |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
1.2.4.1 | pyruvate + [dihydrolipoyllysine-residue acetyltransferase] lipoyllysine = [dihydrolipoyllysine-residue acetyltransferase] S-acetyldihydrolipoyllysine + CO2 | catalytic site is formed by the conserved residues Tyr281 and Arg282 | Geobacillus stearothermophilus |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.2.4.1 | acetaldehyde + benzaldehyde | - |
Geobacillus stearothermophilus | (R)-phenylacetylcarbinol | - |
? | |
1.2.4.1 | pyruvate + CoA + NAD+ | - |
Geobacillus stearothermophilus | acetyl-CoA + CO2 + NADH | - |
ir | |
1.2.4.1 | pyruvate + CoA + NAD+ | first step of the overall reaction of the pyruvate dehydrogenase complex | Geobacillus stearothermophilus | acetyl-CoA + CO2 + NADH | - |
ir | |
1.2.4.1 | pyruvate + CoA + oxidized 2,6-dichlorophenolindophenol | artifical electron acceptor | Geobacillus stearothermophilus | acetyl-CoA + CO2 + reduced 2,6-dichlorophenolindophenol | - |
ir |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.2.4.1 | tetramer | alpha2beta2 structure | Geobacillus stearothermophilus |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.2.4.1 | More | enzyme is a component of the pyruvate dehydrogenase multienzyme complex PDH | Geobacillus stearothermophilus |
1.2.4.1 | pyruvate decarboxylase | - |
Geobacillus stearothermophilus |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.2.4.1 | 65 | - |
mutant 276A | Geobacillus stearothermophilus |
1.2.4.1 | 70 | - |
wild-type enzyme | Geobacillus stearothermophilus |
EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.2.4.1 | additional information | - |
temperature dependence of wild-type and mutant enzymes | Geobacillus stearothermophilus |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.2.4.1 | 0.3 | - |
pyruvate | mutant F266A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.35 | - |
thiamine diphosphate | mutant F266A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.38 | - |
pyruvate | mutant S283C, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.45 | - |
thiamine diphosphate | mutant S283C, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.46 | - |
thiamine diphosphate | wild-type enzyme, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.48 | - |
pyruvate | wild-type enzyme, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.55 | - |
pyruvate | mutant R282A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.62 | - |
thiamine diphosphate | mutant R282A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.71 | - |
pyruvate | mutant D276A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.82 | - |
pyruvate | mutant Y281A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.91 | - |
thiamine diphosphate | mutant D276A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.96 | - |
pyruvate | mutant Y281A/R282A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.97 | - |
thiamine diphosphate | mutant Y281A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 0.98 | - |
thiamine diphosphate | mutant Y281A/R282A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 1.21 | - |
pyruvate | mutant Y281S/R282S, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 1.23 | - |
pyruvate | mutant Y281A/R282A/S283A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 1.25 | - |
thiamine diphosphate | mutant R267A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 1.3 | - |
thiamine diphosphate | mutant Y281S/R282S, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 1.39 | - |
thiamine diphosphate | mutant Y281A/R282A/S283A, pH 7.0, 25°C | Geobacillus stearothermophilus | |
1.2.4.1 | 1.62 | - |
pyruvate | mutant R267A, pH 7.0, 25°C | Geobacillus stearothermophilus |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.2.4.1 | 7 | - |
assay at | Geobacillus stearothermophilus |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.2.4.1 | NAD+ | - |
Geobacillus stearothermophilus | |
1.2.4.1 | thiamine diphosphate | dependent on, binding site | Geobacillus stearothermophilus |