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Literature summary for 4.1.1.1 extracted from
- Directed evolution of yeast pyruvate decarboxylase 1 for attenuated regulation and increased stability (2008), Biochemistry, 47, 3013-3025.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli BL21(DE3)RecA- cells | Saccharomyces cerevisiae |
| expressed in Escherichia coli strain BL21(DE3)RecA- | Saccharomyces cerevisiae |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A143T/T156A/Q367H/N396I/K478R | mutant shows improved activity for 1 mM pyruvate at pH 7.5 in the presence of phosphate, has the substrate concentration required for half-saturation reduced by almost 3fold at pH 7.5 and the phosphate inhibition reduced by 4fold at pH 6.0 compared to the wild type enzyme, the mutant can be activated by pyruvate more easily than the native enzyme | Saccharomyces cerevisiae |
| A143T/T156A/Q367H/N396I/K478R | the mutant shows improved activity for 1 mM pyruvate at pH 7.5 in the presence of phosphate. In comparison with native Pdc1, the mutant has the substrate concentration required for half-saturation reduced by almost 3fold at pH 7.5 and the phosphate inhibition reduced by 4fold at pH 6.0, the apparent cooperativity for pyruvate is also reduced since it is activated by pyruvate more easily than the native enzyme | Saccharomyces cerevisiae |
General Stability
| General Stability | Organism |
|---|---|
| native Pdc1 activity is not altered by a His-tag | Saccharomyces cerevisiae |
| with about 1 mM pyruvate, native Pdc1 only reaches a stable reaction rate after exposure to pyruvate for 1 min, the N-terminal His tag has no influence on activity of native Pdc1 | Saccharomyces cerevisiae |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| phosphate | competitive inhibition | Saccharomyces cerevisiae |  |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Saccharomyces cerevisiae | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| HiTrap chelating column chromatography | Saccharomyces cerevisiae |
| HiTrap Ni2+ chelating HP column chromatography | Saccharomyces cerevisiae |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -70°C, 10 mM PIPES at pH 6.5 with 1 mM dithiothreitol, 1 mM MgCl2, and 0.1 mM thiamine diphosphate, several weeks, no loss of activity | Saccharomyces cerevisiae |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| pyruvate | - |
Saccharomyces cerevisiae | acetaldehyde + CO2 | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| PDC1 | - |
Saccharomyces cerevisiae |
| pyruvate decarboxylase 1 | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
native Pdc1 performs optimally at 30°C in 1 mM pyruvate | Saccharomyces cerevisiae |
| 30 | - |
the optimal temperature for Pdc1 activity is dependent upon pyruvate concentration, in 1 mM pyruvate, native Pdc1 performs optimally at 30°C | Saccharomyces cerevisiae |
| 45 | - |
native activity peaks at 45°C in 25 mM pyruvate | Saccharomyces cerevisiae |
| 45 | - |
the optimal temperature for Pdc1 activity is dependent upon pyruvate concentration, in 25 mM pyruvate, native activity peaks at 45?C | Saccharomyces cerevisiae |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 45 | 65 | the activity of native Pdc1 decreases with increasing temperature from 45°C and is completely abolished at 65°C, the temperature at which half of the native Pdc1 activity is irreversibly lost in 5 min is at 52.6°C | Saccharomyces cerevisiae |
| 52.6 | - |
50% activity is lost after 5 min at 52.6°C | Saccharomyces cerevisiae |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 6.6 | - |
Saccharomyces cerevisiae |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 4.9 | 7.5 | the kcat for native Pdc1 is maximal between pH 6 and pH 6.6, dropping gradually as pH increases to 7.5 and falling rapidly as pH decreases to 4.9 | Saccharomyces cerevisiae |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| thiamine diphosphate | - |
Saccharomyces cerevisiae | |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 14.7 | - |
phosphate | wild type enzyme, at pH 6.0 and 30°C | Saccharomyces cerevisiae | |
| 14.7 | - |
phosphate | native enzyme, at pH 6.0 and 30°C | Saccharomyces cerevisiae | |
| 51 | - |
phosphate | mutant enzyme A143T/T156A/Q367H/N396I/K478R, at pH 6.0 and 30°C | Saccharomyces cerevisiae | |
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