Cloned (Comment) | Organism |
---|---|
His-tagged version expressed in Escherichia coli BL21 | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
sitting drop diffusion method, PEG 3350 | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
E192A | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192C | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192D | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192F | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192G | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192H | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192I | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192K | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192L | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192M | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192N | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192P | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192Q | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192R | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192S | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192T | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192V | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192W | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
E192Y | strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(2R,3R)-2,3,4-trihydroxy-N,N-dipropylbutanamide | competitive inhibitor | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.2 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192M mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.2 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192P mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.3 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192F mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.4 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192N mutant protein, pH 7.7.5, 25°C | Escherichia coli | |
0.4 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192V mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.5 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192H mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.6 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192S mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.7 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192Q mutant protein, pH 7.5, 25°C | Escherichia coli | |
1.1 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192D mutant protein, pH 7.5, 25°C | Escherichia coli | |
3.5 | - |
N-acetylneuraminic acid | E192Q mutant protein, pH 7.5, 25°C | Escherichia coli | |
4.4 | - |
N-acetylneuraminic acid | wild type (E192), pH 7.5, 25°C | Escherichia coli | |
4.6 | - |
N-acetylneuraminic acid | E192D mutant protein, pH 7.5, 25°C | Escherichia coli | |
5 | - |
N-acetylneuraminic acid | E192M mutant protein, pH 7.5, 25°C | Escherichia coli | |
5.9 | - |
N-acetylneuraminic acid | E192F mutant protein, pH 7.5, 25°C | Escherichia coli | |
6 | - |
N-acetylneuraminic acid | E192P mutant protein, pH 7.5, 25°C | Escherichia coli | |
6 | - |
N-acetylneuraminic acid | E192S mutant protein, pH 7.5, 25°C | Escherichia coli | |
6.2 | - |
N-acetylneuraminic acid | E192H mutant protein, pH 7.5, 25°C | Escherichia coli | |
11 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | wild type (E192), pH 7.5, 25°C | Escherichia coli | |
34 | - |
N-acetylneuraminic acid | E192V mutant protein, pH 7.5, 25°C | Escherichia coli | |
38 | - |
N-acetylneuraminic acid | E192N mutant protein, pH 7.7.5, 25°C | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
immobilized metal ion affinity chromatography (Ni2+), gel filtration | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | - |
Escherichia coli | pyruvate + (2R,3S)-2,3-dihydroxy-4-oxo-N,N-dipropylbutanamide | - |
r | |
N-acetylmannosamine + pyruvate | - |
Escherichia coli | N-acetylneuraminic acid | - |
r | |
N-acetylneuraminic acid | - |
Escherichia coli | N-acetylmannosamine + pyruvate | - |
r |
Synonyms | Comment | Organism |
---|---|---|
N-Acetylneuraminic acid aldolase | - |
Escherichia coli |
N-Acetylneuraminic acid lyase | - |
Escherichia coli |
Sialic acid aldolase | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192D mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.42 | - |
N-acetylneuraminic acid | E192P mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.43 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192S mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.63 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192H mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.82 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192P mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.83 | - |
N-acetylneuraminic acid | E192S mutant protein, pH 7.5, 25°C | Escherichia coli | |
1.17 | - |
N-acetylneuraminic acid | E192M mutant protein, pH 7.5, 25°C | Escherichia coli | |
1.17 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192M mutant protein, pH 7.5, 25°C | Escherichia coli | |
1.22 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | wild type (E192), pH 7.5, 25°C | Escherichia coli | |
1.47 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192F mutant protein, pH 7.5, 25°C | Escherichia coli | |
1.58 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192V mutant protein, pH 7.5, 25°C | Escherichia coli | |
2 | - |
N-acetylneuraminic acid | E192D mutant protein, pH 7.5, 25°C | Escherichia coli | |
2 | - |
N-acetylneuraminic acid | E192H mutant protein, pH 7.5, 25°C | Escherichia coli | |
2 | - |
N-acetylneuraminic acid | E192V mutant protein, pH 7.5, 25°C | Escherichia coli | |
2.17 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192N mutant protein, pH 7.7.5, 25°C | Escherichia coli | |
2.67 | - |
N-acetylneuraminic acid | E192F mutant protein, pH 7.5, 25°C | Escherichia coli | |
2.83 | - |
N-acetylneuraminic acid | E192N mutant protein, pH 7.7.5, 25°C | Escherichia coli | |
2.83 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192Q mutant protein, pH 7.5, 25°C | Escherichia coli | |
4.33 | - |
N-acetylneuraminic acid | wild type (E192), pH 7.5, 25°C | Escherichia coli | |
5 | - |
N-acetylneuraminic acid | E192Q mutant protein, pH 7.5, 25°C | Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
1.3 | - |
(2R,3R)-2,3,4-trihydroxy-N,N-dipropylbutanamide | E192N mutant protein , pH 7.5, 25°C | Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.06 | - |
N-acetylneuraminic acid | E192V mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.07 | - |
N-acetylneuraminic acid | E192N mutant protein, pH 7.7.5, 25°C | Escherichia coli | |
0.07 | - |
N-acetylneuraminic acid | E192P mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.08 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192D mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.12 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | wild type (E192), pH 7.5, 25°C | Escherichia coli | |
0.13 | - |
N-acetylneuraminic acid | E192S mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.23 | - |
N-acetylneuraminic acid | E192M mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.32 | - |
N-acetylneuraminic acid | E192H mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.43 | - |
N-acetylneuraminic acid | E192D mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.45 | - |
N-acetylneuraminic acid | E192F mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.7 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192S mutant protein, pH 7.5, 25°C | Escherichia coli | |
0.98 | - |
N-acetylneuraminic acid | wild type (E192), pH 7.5, 25°C | Escherichia coli | |
1.33 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192H mutant protein, pH 7.5, 25°C | Escherichia coli | |
1.42 | - |
N-acetylneuraminic acid | E192Q mutant protein, pH 7.5, 25°C | Escherichia coli | |
4.17 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192Q mutant protein, pH 7.5, 25°C | Escherichia coli | |
4.33 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192P mutant protein, pH 7.5, 25°C | Escherichia coli | |
4.5 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192V mutant protein, pH 7.5, 25°C | Escherichia coli | |
5.17 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192F mutant protein, pH 7.5, 25°C | Escherichia coli | |
5.33 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192M mutant protein, pH 7.5, 25°C | Escherichia coli | |
5.67 | - |
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid | E192N mutant protein, pH 7.7.5, 25°C | Escherichia coli |