Cloned (Comment) | Organism |
---|---|
overexpression | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D76N | site-directed mutants in which the putative metal ion ligand is modified: H95N, H97N, D76N. The mutant enzymes require exogenous metal ions for full activity. Their turnover numbers are greatly reduced whereas the Km-values are only moderately affected. Low levels of aldolase activity are observed with the H97N mutant, but not with D76N or the H95N mutants. The H97N mutant enzyme catalyzes the condensation of dihydroxyacetone and glycolaldehyde phosphate to produce a mixture of L-ribulose 5-phosphate and D-xylulose 5-phosphate | Escherichia coli |
D76N | the ratio of turnover number to Km-value is 348fold lower than that of the wild-type enzyme | Escherichia coli |
H59N | site-directed mutants in which the putative metal ion ligand is modified: H95N, H97N, D76N. The mutant enzymes require exogenous metal ions for full activity. Their turnover numbers are greatly reduced whereas the Km-values are only moderately affected. Low levels of aldolase activity are observed with the H97N mutant, but not with D76N or the H95N mutants. The H97N mutant enzyme catalyzes the condensation of dihydroxyacetone and glycolaldehyde phosphate to produce a mixture of L-ribulose 5-phosphate and D-xylulose 5-phosphate | Escherichia coli |
H95N | the ratio of turnover number to Km-value is 676fold lower than that of the wild-type enzyme | Escherichia coli |
H97N | site-directed mutants in which the putative metal ion ligand is modified: H95N, H97N, D76N. The mutant enzymes require exogenous metal ions for full activity. Their turnover numbers are greatly reduced whereas the Km-values are only moderately affected. Low levels of aldolase activity are observed with the H97N mutant, but not with D76N or the H95N mutants. The H97N mutant enzyme catalyzes the condensation of dihydroxyacetone and glycolaldehyde phosphate to produce a mixture of L-ribulose 5-phosphate and D-xylulose 5-phosphate | Escherichia coli |
H97N | mutant enzyme with low levels of aldolase activity. The ratio of turnover number to Km-value is 479fold lower than that of the wild-type enzyme | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
glycoaldehyde phosphate | competitive inhibitor of mutant enzyme H97N but not of the wild-type enzyme | Escherichia coli | |
glycolaldehyde | - |
Escherichia coli | |
glycolaldehyde phosphate | competitive inhibition of H97N mutant enzyme, no inhibition of wild type enzyme | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.06 | - |
L-ribulose 5-phosphate | wild type enzyme, Zn2+ form of enzyme plus 0.1 mM Zn2+ | Escherichia coli | |
0.06 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis + 0.1 mM Zn2+, wild-type enzyme | Escherichia coli | |
0.087 | - |
L-ribulose 5-phosphate | wild type enzyme, Zn2+ form of enzyme after dialysis | Escherichia coli | |
0.087 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis, wild-type | Escherichia coli | |
0.096 | - |
L-ribulose 5-phosphate | mutant H95N, Zn2+ form of enzyme plus 0.1 mM Zn2+ | Escherichia coli | |
0.096 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis + 0.1 mM Zn2+, mutant enzyme H95N | Escherichia coli | |
0.1 | - |
L-ribulose 5-phosphate | L-ribulose 5-phosphate, mutant H97N, Zn2+ form of enzyme after dialysis | Escherichia coli | |
0.1 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis, mutant enzyme H95N | Escherichia coli | |
0.11 | - |
L-ribulose 5-phosphate | mutant D76N, Zn2+ form of enzyme after dialysis | Escherichia coli | |
0.11 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis, mutant enzyme D76N | Escherichia coli | |
0.14 | - |
L-ribulose 5-phosphate | L-ribulose 5-phosphate, mutant H97N and D76N, Zn2+ form of enzyme plus 0.1 mM Zn2+ | Escherichia coli | |
0.14 | - |
L-ribulose 5-phosphate | mutant H95N, Zn2+ form of enzyme after dialysis | Escherichia coli | |
0.14 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis + 0.1 mM Zn2+, mutant enzyme H97N or D76N | Escherichia coli | |
0.14 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis, mutant enzyme H97N | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Cu2+ | the enzyme preparation contains a mixture of the following metals in order of decreasing abundance: Zn2+, Mn2+, Cu2+ | Escherichia coli | |
Mn2+ | the enzyme preparation contains a mixture of the following metals in order of decreasing abundance: Zn2+, Mn2+, Cu2+ | Escherichia coli | |
Zn2+ | H95N, H97N, and D76N mutant enzymes require exogenous metal ions for full activity | Escherichia coli | |
Zn2+ | the enzyme preparation contains a mixture of the following metals in order of decreasing abundance: Zn2+, Mn2+, Cu2+ | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
25522 | - |
4 * 25522, electrospray mass spectrometry | Escherichia coli |
127000 | - |
wild-type enzyme and mutant enzymes H97N, D76N and H95N, gel filtration | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Escherichia coli | - |
wild type enzyme and mutants H95N, H97N, and D76N | - |
Purification (Comment) | Organism |
---|---|
- |
Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
34 | - |
- |
Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-Ribulose 5-phosphate | - |
Escherichia coli | D-Xylulose 5-phosphate | - |
? | |
additional information | the mutant enzyme H97N catalyzes the condensation of dihydroxyacetone and glycolaldehyde to produce a mixture of ribulose-5-phosphate and D-xylulose 5-phosphate | Escherichia coli | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | 4 * 25522, electrospray mass spectrometry | Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | 55 | thermal denaturation themperature of wild-type enzyme and mutant enzymes H97N, D76N and H95N | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.047 | - |
L-ribulose 5-phosphate | mutant H95N, Zn2+ form of enzyme after dialysis | Escherichia coli | |
0.047 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis, mutant enzyme H95N | Escherichia coli | |
0.048 | - |
L-ribulose 5-phosphate | mutant H97N, Zn2+ form of enzyme after dialysis | Escherichia coli | |
0.048 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis, mutant enzyme H97N | Escherichia coli | |
0.073 | - |
L-ribulose 5-phosphate | mutant D76N, Zn2+ form of enzyme after dialysis | Escherichia coli | |
0.073 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis, mutant enzyme D76N | Escherichia coli | |
0.1 | - |
L-ribulose 5-phosphate | mutant H95N, Zn2+ form of enzyme plus 0.1 mM Zn2+ | Escherichia coli | |
0.16 | - |
L-ribulose 5-phosphate | mutant D76N, Zn2+ form of enzyme plus 0.1 mM Zn2+ | Escherichia coli | |
0.16 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis + 0.1 mM Zn2+, mutant enzyme D76N | Escherichia coli | |
7.3 | - |
L-ribulose 5-phosphate | mutant H97N, Zn2+ form of enzyme plus 0.1 mM Zn2+ | Escherichia coli | |
7.3 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis + 0.1 mM Zn2+, mutant enzyme H97N | Escherichia coli | |
20.4 | - |
L-ribulose 5-phosphate | wild type enzyme, Zn2+ form of enzyme after dialysis | Escherichia coli | |
20.4 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis, wild-type | Escherichia coli | |
20.7 | - |
L-ribulose 5-phosphate | wild type enzyme, Zn2+ form of enzyme plus 0.1 mM Zn2+ | Escherichia coli | |
22.4 | - |
L-ribulose 5-phosphate | 37°C, pH 7.6, Zn2+-form after dialysis + 0.1 mM Zn2+, wild-type enzyme | Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.37 | - |
glycolaldehyde | 37°C, pH 7.6, mutant enzyme H97N, with L-ribulose 5-phosphate as substrate | Escherichia coli |