Protein Variants | Comment | Organism |
---|---|---|
S184F | active site mutation, enhances the substrate specificity to include catalysis of retro-aldol cleavage of (4S)-2-keto-4-hydroxy-4-(2'-pyridyl)butanoate | Escherichia coli |
T161S | active site mutation, enhances the substrate specificity to include catalysis of retro-aldol cleavage of (4S)-2-keto-4-hydroxy-4-(2'-pyridyl)butanoate | Escherichia coli |
T161S/S184F | active site mutation, working additively to enhance the substrate specificity to include catalysis of retro-aldol cleavage of (4S)-2-keto-4-hydroxy-4-(2'-pyridyl)butanoate | Escherichia coli |
T161S/S184L | active site mutation, working additively to enhance the substrate specificity to include catalysis of retro-aldol cleavage of (4S)-2-keto-4-hydroxy-4-(2'-pyridyl)butanoate. Mutations improve the value of kcat/KM (4S)-2-keto-4-hydroxy-4-(2'-pyridyl)butanoate by more than 450fold, resulting in a catalytic efficiency that is comparable to that of the wild-type enzyme with the natural substrate while retaining high stereoselectivity | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | mutant T161S, pH 7.5, 25°C | Escherichia coli | |
0.1 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | wild-type, pH 7.5, 25°C | Escherichia coli | |
0.24 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | mutant T161S/S18L, pH 7.5, 25°C | Escherichia coli | |
0.69 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | mutant T161S/S184F, pH 7.5, 25°C | Escherichia coli | |
1.6 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | mutant T161S/S18L, pH 7.5, 25°C | Escherichia coli | |
1.7 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | mutant S184F, pH 7.5, 25°C | Escherichia coli | |
2 | 5 | 4-hydroxy-2-oxooctonate | mutant S184F, pH 7.5, 25°C | Escherichia coli | |
3.5 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | mutant T161S/S184F, pH 7.5, 25°C | Escherichia coli | |
4.3 | - |
4-hydroxy-2-oxooctonate | mutant T161S/S18L, pH 7.5, 25°C | Escherichia coli | |
5.7 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | mutant S184F, pH 7.5, 25°C | Escherichia coli | |
15 | - |
4-hydroxy-2-oxooctonate | mutant T161S, pH 7.5, 25°C | Escherichia coli | |
32 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | wild-type, pH 7.5, 25°C | Escherichia coli | |
43 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | mutant T161S, pH 7.5, 25°C | Escherichia coli | |
146 | - |
4-hydroxy-2-oxooctonate | wild-type, pH 7.5, 25°C | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | - |
Escherichia coli | ? | - |
r | |
2-dehydro-3-deoxy-6-phospho-D-gluconate | - |
Escherichia coli | pyruvate + D-glyceraldehyde 3-phosphate | - |
r | |
4-hydroxy-2-oxooctonate | - |
Escherichia coli | ? | - |
r | |
pyruvate + 2-pyridine carboxaldehyde | wild-type, 25% of activity with D-glyceraldehyde 3-phosphate, mutant T161S/S184L, 89% of wild-type activity with D-glyceraldehyde 3-phosphate | Escherichia coli | ? | - |
r | |
pyruvate + 3-pyridine carboxaldehyde | wild-type, 0.5% of activity with D-glyceraldehyde 3-phosphate, mutant T161S/S184L, 1% of wild-type activity with D-glyceraldehyde 3-phosphate | Escherichia coli | ? | - |
r | |
pyruvate + 4-pyridine carboxaldehyde | wild-type, 3% of activity with D-glyceraldehyde 3-phosphate, mutant T161S/S184L, 2.5% of wild-type activity with D-glyceraldehyde 3-phosphate | Escherichia coli | ? | - |
r | |
pyruvate + D-glyceraldehyde | wild-type, 4% of activity with D-glyceraldehyde 3-phosphate, mutant T161S/S184L, 1% of wild-type activity with D-glyceraldehyde 3-phosphate | Escherichia coli | 2-dehydro-3-deoxy-D-gluconate | - |
r | |
Pyruvate + D-glyceraldehyde 3-phosphate | mutant T161S/S184L, 15% of wild-type activity | Escherichia coli | 6-Phospho-2-dehydro-3-deoxy-D-gluconate | - |
r |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.5 | - |
4-hydroxy-2-oxooctonate | mutant S184F, pH 7.5, 25°C | Escherichia coli | |
1.7 | - |
4-hydroxy-2-oxooctonate | mutant T161S, pH 7.5, 25°C | Escherichia coli | |
2 | - |
4-hydroxy-2-oxooctonate | wild-type, pH 7.5, 25°C | Escherichia coli | |
3.1 | - |
4-hydroxy-2-oxooctonate | mutant T161S/S18L, pH 7.5, 25°C | Escherichia coli | |
11 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | wild-type, pH 7.5, 25°C | Escherichia coli | |
19 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | mutant S184F, pH 7.5, 25°C | Escherichia coli | |
27 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | mutant T161S, pH 7.5, 25°C | Escherichia coli | |
31 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | mutant T161S/S184F, pH 7.5, 25°C | Escherichia coli | |
40 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | mutant S184F, pH 7.5, 25°C | Escherichia coli | |
41 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | mutant T161S/S18L, pH 7.5, 25°C | Escherichia coli | |
83 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | wild-type, pH 7.5, 25°C | Escherichia coli | |
175 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | mutant T161S, pH 7.5, 25°C | Escherichia coli | |
212 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | mutant T161S/S184F, pH 7.5, 25°C | Escherichia coli | |
240 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | mutant T161S/S18L, pH 7.5, 25°C | Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.014 | - |
4-hydroxy-2-oxooctonate | wild-type, pH 7.5, 25°C | Escherichia coli | |
0.06 | - |
4-hydroxy-2-oxooctonate | mutant S184F, pH 7.5, 25°C | Escherichia coli | |
0.113 | - |
4-hydroxy-2-oxooctonate | mutant T161S, pH 7.5, 25°C | Escherichia coli | |
0.33 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | wild-type, pH 7.5, 25°C | Escherichia coli | |
0.72 | - |
4-hydroxy-2-oxooctonate | mutant T161S/S18L, pH 7.5, 25°C | Escherichia coli | |
3.3 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | mutant S184F, pH 7.5, 25°C | Escherichia coli | |
4.1 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | mutant T161S, pH 7.5, 25°C | Escherichia coli | |
24 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | mutant S184F, pH 7.5, 25°C | Escherichia coli | |
45 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | mutant T161S/S184F, pH 7.5, 25°C | Escherichia coli | |
61 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | mutant T161S/S184F, pH 7.5, 25°C | Escherichia coli | |
150 | - |
(4S)-2-oxo-4-hydroxy-4-(2'-pyridyl)butanoate | mutant T161S/S18L, pH 7.5, 25°C | Escherichia coli | |
170 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | mutant T161S/S18L, pH 7.5, 25°C | Escherichia coli | |
550 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | mutant T161S, pH 7.5, 25°C | Escherichia coli | |
830 | - |
2-dehydro-3-deoxy-6-phospho-D-gluconate | wild-type, pH 7.5, 25°C | Escherichia coli |