Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
I229D | the mutant shows decreased kcat compared to the wild type enzyme | Escherichia coli |
I229N | the mutant shows increased kcat compared to the wild type enzyme | Escherichia coli |
I229R | the mutant shows increased kcat compared to the wild type enzyme | Escherichia coli |
L171D | the mutant is insoluble | Escherichia coli |
L171D/L199D/I229D | the mutant is insoluble | Escherichia coli |
L171D/L199N/I229D | the mutant is insoluble | Escherichia coli |
L171N | the mutant is insoluble | Escherichia coli |
L171N/I229N | the mutant shows decreased kcat compared to the wild type enzyme | Escherichia coli |
L171R | the mutant shows decreased kcat compared to the wild type enzyme | Escherichia coli |
L171R/L199N/I229R | the mutant is insoluble | Escherichia coli |
L171R/L199R/I229R | the mutant is insoluble | Escherichia coli |
L199D | the mutant is insoluble | Escherichia coli |
L199N | the mutant shows increased kcat compared to the wild type enzyme | Escherichia coli |
L199N/I229D | the mutant shows decreased kcat compared to the wild type enzyme | Escherichia coli |
L199N/I229N | the mutant shows decreased kcat compared to the wild type enzyme | Escherichia coli |
L199N/I229R | the mutant shows increased kcat compared to the wild type enzyme | Escherichia coli |
L199R | the mutant is insoluble | Escherichia coli |
L199R/I229N | the mutant shows increased kcat compared to the wild type enzyme | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.51 | - |
N-acetylneuraminate | mutant enzyme L171N/I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
3.2 | - |
N-acetylneuraminate | mutant enzyme L199N/I229D, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
3.4 | - |
N-acetylneuraminate | mutant enzyme I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
3.42 | - |
N-acetylneuraminate | mutant enzyme L171R, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
3.93 | - |
N-acetylneuraminate | mutant enzyme I229R, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
4 | - |
N-acetylneuraminate | mutant enzyme L199N/I229R, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
4.1 | - |
N-acetylneuraminate | wild type enzyme, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
4.3 | - |
N-acetylneuraminate | mutant enzyme L199N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
4.7 | - |
N-acetylneuraminate | mutant enzyme L199N/I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
5 | - |
N-acetylneuraminate | mutant enzyme I229D, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
6.1 | - |
N-acetylneuraminate | mutant enzyme L199R/I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
35000 | - |
4 * 35000, His-tagged enzyme, SDS-PAGE | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A6L4 | - |
- |
Escherichia coli W3110 / ATCC 27325 | P0A6L4 | - |
- |
Purification (Comment) | Organism |
---|---|
HisTrap column chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
N-acetylneuraminate | - |
Escherichia coli | N-acetyl-D-mannosamine + pyruvate | - |
r | |
N-acetylneuraminate | - |
Escherichia coli W3110 / ATCC 27325 | N-acetyl-D-mannosamine + pyruvate | - |
r |
Subunits | Comment | Organism |
---|---|---|
homotetramer | 4 * 35000, His-tagged enzyme, SDS-PAGE | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
NAL | - |
Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
84 | - |
the melting temperature of the wild type enzyme is at 84°C | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.29 | - |
N-acetylneuraminate | mutant enzyme L199N/I229D, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
0.664 | - |
N-acetylneuraminate | mutant enzyme L199N/I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
3.15 | - |
N-acetylneuraminate | mutant enzyme L171N/I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
4.69 | - |
N-acetylneuraminate | mutant enzyme I229D, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
5.49 | - |
N-acetylneuraminate | mutant enzyme L171R, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
6.75 | - |
N-acetylneuraminate | wild type enzyme, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
7.4 | - |
N-acetylneuraminate | mutant enzyme I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
7.72 | - |
N-acetylneuraminate | mutant enzyme I229R, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
7.94 | - |
N-acetylneuraminate | mutant enzyme L199N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
8.9 | - |
N-acetylneuraminate | mutant enzyme L199N/I229R, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli | |
11.87 | - |
N-acetylneuraminate | mutant enzyme L199R/I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C | Escherichia coli |
General Information | Comment | Organism |
---|---|---|
physiological function | the quaternary structure of Escherichia coli N-acetylneuraminate lyase is essential for functional expression | Escherichia coli |