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Literature summary for 4.1.2.14 extracted from

  • Cheriyan, M.; Toone, E.J.; Fierke, C.A.
    Improving upon nature: active site remodeling produces highly efficient aldolase activity toward hydrophobic electrophilic substrates (2012), Biochemistry, 51, 1658-1668.
    View publication on PubMedView publication on EuropePMC

Protein Variants

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 [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

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Substrates and Products (Substrate)

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 [1/s]

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 [mM/s]

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