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Literature summary for 1.1.1.6 extracted from

  • Velasco-Lozano, S.; Roca, M.; Leal-Duaso, A.; Mayoral, J.A.; Pires, E.; Moliner, V.; Lopez-Gallego, F.
    Selective oxidation of alkyl and aryl glyceryl monoethers catalysed by an engineered and immobilised glycerol dehydrogenase (2020), Chem. Sci., 11, 12009-12020 .
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
synthesis engineering and immobilizing of glycerol dehydrogenase to accept alkyl/aryl glyceryl monoethers and catalyze their enantioselective oxidation to yield the corresponding 3-alkoxy/aryloxy-1-hydroxyacetones. The enzyme is highly enantioselective towards S-isomers (ee > 99%). Application of mutant L252A in a one-pot chemoenzymatic process to convert glycidol and ethanol into 3-ethoxy-1-hydroxyacetone and (R)-3-ethoxypropan-1,2-diol, without affecting the oxidation activity Geobacillus stearothermophilus

Protein Variants

Protein Variants Comment Organism
D123N mutation abolishes the oxidative activity. The carboxylate of D123 is the base needed for abstracting the proton to form the alkoxy intermediate that precedes the hydride transfer to the nicotinamide cofactor Geobacillus stearothermophilus
L252A mutation reduces the enzyme activity to less than 10% of wild-type activity and sculpts the active site to accommodate a productive configuration of 3-monoalkyl glycerols Geobacillus stearothermophilus
L252A the mutation sculpts the active site to accommodate a productive configuration of 3-monoalkyl glycerols. This mutation enhances the kcat 163-fold towards 3-ethoxypropan-1,2-diol, resulting in a specific activity similar to the one found for the wild-type towards glycerol. No activity with 3-phenoxypropane-1,2-diol Geobacillus stearothermophilus
V131A about 70% of wild-type activity with glycerol Geobacillus stearothermophilus
Y142A mutation reduces the enzyme activity to less than 10% of wild-type activity Geobacillus stearothermophilus

General Stability

General Stability Organism
the immobilised L252A shows good reusability and increased the operational stability Geobacillus stearothermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
7
-
3-phenoxypropane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
13.4
-
3-ethoxypropane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
30
-
3-propoxypropane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
31.2
-
3-[(propan-2-yl)oxy]propane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
45
-
3-methoxypropane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
50
-
glycerol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
78
-
3-(2,2,2-trifluoroethoxy)propane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
162
-
3-butoxypropane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
281
-
3-(2,2,2-trifluoroethoxy)propane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
371
-
3-ethoxypropane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
608
-
3-propoxypropane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
707
-
3-[(propan-2-yl)oxy]propane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
977
-
3-methoxypropane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
1880
-
glycerol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus P32816
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-(2,2,2-trifluoroethoxy)propane-1,2-diol + NAD+
-
Geobacillus stearothermophilus 1-hydroxy-3-(2,2,2-trifluoroethoxy)propan-2-one + NADH + H+
-
?
3-(2,2,2-trifluoroethoxy)propane-1,2-diol + NAD+
-
Geobacillus stearothermophilus ? + NADH + H+
-
?
3-butoxypropane-1,2-diol + NAD+
-
Geobacillus stearothermophilus 1-hydroxy-3-butoxypropan-2-one + NADH + H+
-
?
3-butoxypropane-1,2-diol + NAD+
-
Geobacillus stearothermophilus ? + NADH + H+
-
?
3-ethoxypropane-1,2-diol + NAD+
-
Geobacillus stearothermophilus 1-hydroxy-3-ethoxypropan-2-one + NADH + H+
-
?
3-ethoxypropane-1,2-diol + NAD+
-
Geobacillus stearothermophilus ? + NADH + H+
-
?
3-methoxypropane-1,2-diol + NAD+
-
Geobacillus stearothermophilus 1-hydroxy-3-methoxypropan-2-one + NADH + H+
-
?
3-methoxypropane-1,2-diol + NAD+
-
Geobacillus stearothermophilus ? + NADH + H+
-
?
3-phenoxypropane-1,2-diol + NAD+
-
Geobacillus stearothermophilus 1-hydroxy-3-phenoxypropan-2-one + NADH + H+
-
?
3-phenoxypropane-1,2-diol + NAD+
-
Geobacillus stearothermophilus ? + NADH + H+
-
?
3-propoxypropane-1,2-diol + NAD+
-
Geobacillus stearothermophilus 1-hydroxy-3-propoxypropan-2-one + NADH + H+
-
?
3-propoxypropane-1,2-diol + NAD+
-
Geobacillus stearothermophilus ? + NADH + H+
-
?
3-[(propan-2-yl)oxy]propane-1,2-diol + NAD+
-
Geobacillus stearothermophilus 1-hydroxy-3-[(propan-2-yl)oxy]propan-2-one + NADH + H+
-
?
3-[(propan-2-yl)oxy]propane-1,2-diol + NAD+
-
Geobacillus stearothermophilus ? + NADH + H+
-
?
glycerol + NAD+
-
Geobacillus stearothermophilus glycerone + NADH + H+
-
?
glycerol + NAD+
-
Geobacillus stearothermophilus D-glyceraldehyde + NADH + H+
-
?
additional information enzyme is highly enantioselective towards S-isomers of substituted glycerol derivatives Geobacillus stearothermophilus ?
-
-

Synonyms

Synonyms Comment Organism
GldA
-
Geobacillus stearothermophilus
GlyDH
-
Geobacillus stearothermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.19
-
3-ethoxypropane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
0.27
-
3-propoxypropane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
0.3
-
3-[(propan-2-yl)oxy]propane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
0.32
-
3-(2,2,2-trifluoroethoxy)propane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
0.7
-
3-phenoxypropane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
0.84
-
3-methoxypropane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
1.3
-
3-butoxypropane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
6.6
-
3-methoxypropane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
7.4
-
glycerol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
9
-
glycerol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
12
-
3-propoxypropane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
13
-
3-[(propan-2-yl)oxy]propane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
14
-
3-(2,2,2-trifluoroethoxy)propane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
31
-
3-ethoxypropane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus

General Information

General Information Comment Organism
metabolism reaction mechanism leads to a proton transfer between the substrate and the acid residue before the hydride transfer from glycerol to NAD+. The mechanism takes place in a stepwise manner, consisting in the proton abstraction of the alcohol by D123 residue followed by the hydride transfer from glycerol to NAD+. This second step is the rate-limiting one Geobacillus stearothermophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.004
-
3-(2,2,2-trifluoroethoxy)propane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
0.005
-
glycerol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
0.007
-
3-methoxypropane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
0.008
-
3-butoxypropane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
0.009
-
3-propoxypropane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
0.01
-
3-[(propan-2-yl)oxy]propane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
0.014
-
3-ethoxypropane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
0.018
-
3-[(propan-2-yl)oxy]propane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
0.019
-
3-methoxypropane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
0.019
-
3-propoxypropane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
0.051
-
3-(2,2,2-trifluoroethoxy)propane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
0.085
-
3-ethoxypropane-1,2-diol mutant L252A, pH 7.0, 30°C Geobacillus stearothermophilus
0.112
-
3-phenoxypropane-1,2-diol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus
0.148
-
glycerol wild-type, pH 7.0, 30°C Geobacillus stearothermophilus