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