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Literature summary extracted from

  • Williams, G.J.; Woodhall, T.; Nelson, A.; Berry, A.
    Structure-guided saturation mutagenesis of N-acetylneuraminic acid lyase for the synthesis of sialic acid mimetics (2005), Protein Eng. Des. Sel., 18, 239-246.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
4.1.3.3 D191P ratio of turnover-number to KM-value for (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid is 1.4fold higher than the wild-type ratio Escherichia coli
4.1.3.3 E192N ratio of turnover-number to KM-value for N-acetyl-D-neuraminic acid is 13.4fold lower than the wild-type ratio, ratio of turnover-number to KM-value for (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid is 48.7fold higher than the wild-type ratio Escherichia coli
4.1.3.3 E192S ratio of turnover-number to KM-value for N-acetyl-D-neuraminic acid is 3.9fold lower than the wild-type ratio, ratio of turnover-number to KM-value for (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid is 32.2fold higher than the wild-type ratio Escherichia coli
4.1.3.3 E192V ratio of turnover-number to KM-value for N-acetyl-D-neuraminic acid is 17.9fold lower than the wild-type ratio, ratio of turnover-number to KM-value for (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid is 39.3fold higher than the wild-type ratio Escherichia coli
4.1.3.3 S208G ratio of turnover-number to KM-value for N-acetyl-D-neuraminic acid is 2.5fold lower than the wild-type ratio, ratio of turnover-number to KM-value for (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid is 1.5fold higher than the wild-type ratio Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.3.3 0.35
-
 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid pH 7.5, 25°C, mutant enzyme E192V Escherichia coli
4.1.3.3 0.39
-
 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid pH 7.5, 25°C, mutant enzyme E192N Escherichia coli
4.1.3.3 0.5
-
 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid pH 7.5, 25°C, mutant enzyme E192S Escherichia coli
4.1.3.3 2.54
-
 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid pH 7.5, 25°C, mutant enzyme S208G Escherichia coli
4.1.3.3 4.4
-
 N-acetyl-D-neuraminic acid pH 7.5, 25°C, wild-type enzyme Escherichia coli
4.1.3.3 5.7
-
 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid pH 7.5, 25°C, mutant enzyme D191P Escherichia coli
4.1.3.3 10.6
-
 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid pH 7.5, 25°C, wild-type enzyme Escherichia coli
4.1.3.3 10.7
-
 N-acetyl-D-neuraminic acid pH 7.5, 25°C, mutant enzyme S208G Escherichia coli
4.1.3.3 17
-
 N-acetyl-D-neuraminic acid pH 7.5, 25°C, mutant enzyme E192S Escherichia coli
4.1.3.3 34
-
 N-acetyl-D-neuraminic acid pH 7.5, 25°C, mutant enzyme E192V Escherichia coli
4.1.3.3 38
-
 N-acetyl-D-neuraminic acid pH 7.5, 25°C, mutant enzyme E192N Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
4.1.3.3 Escherichia coli
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.3.3 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
 Escherichia coli ?
-
 ?
4.1.3.3 N-acetyl-D-neuraminic acid
-
 Escherichia coli N-acetyl-D-mannosamine + pyruvate
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.3.3 0.44
-
 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid pH 7.5, 25°C, mutant enzyme S208G Escherichia coli
4.1.3.3 0.91
-
 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid pH 7.5, 25°C, mutant enzyme D191P Escherichia coli
4.1.3.3 1.22
-
 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid pH 7.5, 25°C, wild-type enzyme Escherichia coli
4.1.3.3 1.58
-
 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid pH 7.5, 25°C, mutant enzyme E192V Escherichia coli
4.1.3.3 1.85
-
 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid pH 7.5, 25°C, mutant enzyme E192S Escherichia coli
4.1.3.3 1.97
-
 N-acetyl-D-neuraminic acid pH 7.5, 25°C, mutant enzyme E192V Escherichia coli
4.1.3.3 2.18
-
 (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid pH 7.5, 25°C, mutant enzyme E192N Escherichia coli
4.1.3.3 2.77
-
 N-acetyl-D-neuraminic acid pH 7.5, 25°C, mutant enzyme E192N Escherichia coli
4.1.3.3 4.15
-
 N-acetyl-D-neuraminic acid pH 7.5, 25°C, mutant enzyme S208G Escherichia coli
4.1.3.3 4.25
-
 N-acetyl-D-neuraminic acid pH 7.5, 25°C, mutant enzyme E192S Escherichia coli
4.1.3.3 4.3
-
 N-acetyl-D-neuraminic acid pH 7.5, 25°C, wild-type enzyme Escherichia coli