Application | Comment | Organism |
---|---|---|
additional information | the combined mutant luciferase, which has high luminescence intensity, will be useful for detecting bacteria with high sensitivity in production safety tests | Photinus pyralis |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli JM109 cells and Bacillus subtilis strain NBRC13719 | Photinus pyralis |
into pET28a vector | Photinus pyralis |
Protein Variants | Comment | Organism |
---|---|---|
D436G | exhibits 8.5fold higher luciferase activity than the wild type enzyme | Photinus pyralis |
D436G | shows 8.5fold increase in luminescence intensity | Photinus pyralis |
I423L | exhibits 4.7fold higher luciferase activity than the wild type enzyme | Photinus pyralis |
I423L | shows 4.7fold increase in luminescence intensity | Photinus pyralis |
I423L/D436G/L530R | exhibits 12.5fold higher luciferase activity than the wild type enzyme | Photinus pyralis |
I423L/D436G/L530R | combining the mutations results in a combined mutant luciferase with higher luminescence intensity than any of the single mutant luciferases, generates more than 12.5fold higher luminescence intensity than the wild-type enzyme. The combined mutant luciferase detects ATP at 10-18 mol, whereas wild-type luciferase is unable to detect below 10-17 mol ATP | Photinus pyralis |
L530R | exhibits 5.1fold higher luciferase activity than the wild type enzyme | Photinus pyralis |
L530R | shows 5.1fold increase in luminescence intensity | Photinus pyralis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.001 | - |
ATP | mutant enzyme I423L/D436G/L530R, in 50 mM Tris/HCl, at pH 7.4 and in the presence of 10 mM MgCl2 | Photinus pyralis | |
0.001 | - |
D-luciferin | wild type enzyme, in 50 mM Tris/HCl, at pH 7.4 and in the presence of 10 mM MgCl2 | Photinus pyralis | |
0.001 | - |
D-luciferin | mutant I423L/D436G/L530R, in the presence of 0.01 mM ATP | Photinus pyralis | |
0.002 | - |
D-luciferin | mutant enzyme D436G, in 50 mM Tris/HCl, at pH 7.4 and in the presence of 10 mM MgCl2 | Photinus pyralis | |
0.002 | - |
D-luciferin | mutant D436G, in the presence of 0.01 mM ATP | Photinus pyralis | |
0.004 | - |
D-luciferin | mutant enzyme I423L, in 50 mM Tris/HCl, at pH 7.4 and in the presence of 10 mM MgCl2 | Photinus pyralis | |
0.004 | - |
D-luciferin | mutant enzyme L530R, in 50 mM Tris/HCl, at pH 7.4 and in the presence of 10 mM MgCl2 | Photinus pyralis | |
0.004 | - |
D-luciferin | mutant I423L, in the presence of 0.01 mM ATP | Photinus pyralis | |
0.004 | - |
D-luciferin | mutant L530R, in the presence of 0.01 mM ATP | Photinus pyralis | |
0.008 | - |
ATP | mutant I423L/D436G/L530R, in the presence of 0.001 mM D-luciferin | Photinus pyralis | |
0.009 | - |
ATP | mutant enzyme D436G, in 50 mM Tris/HCl, at pH 7.4 and in the presence of 10 mM MgCl2 | Photinus pyralis | |
0.009 | - |
ATP | mutant D436G, in the presence of 0.001 mM D-luciferin | Photinus pyralis | |
0.021 | - |
D-luciferin | mutant enzyme I423L/D436G/L530R, in 50 mM Tris/HCl, at pH 7.4 and in the presence of 10 mM MgCl2 | Photinus pyralis | |
0.021 | - |
D-luciferin | wild-type, in the presence of 0.001 mM D-luciferin | Photinus pyralis | |
0.047 | - |
ATP | mutant enzyme I423L, in 50 mM Tris/HCl, at pH 7.4 and in the presence of 10 mM MgCl2 | Photinus pyralis | |
0.047 | - |
ATP | mutant I423L, in the presence of 0.001 mM D-luciferin | Photinus pyralis | |
0.054 | - |
ATP | mutant enzyme L530R, in 50 mM Tris/HCl, at pH 7.4 and in the presence of 10 mM MgCl2 | Photinus pyralis | |
0.054 | - |
ATP | mutant L530R, in the presence of 0.001 mM D-luciferin | Photinus pyralis | |
0.16 | - |
ATP | wild type enzyme, in 50 mM Tris/HCl, at pH 7.4 and in the presence of 10 mM MgCl2 | Photinus pyralis | |
0.16 | - |
ATP | wild-type, in the presence of 0.001 mM D-luciferin | Photinus pyralis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | 10 mM, required | Photinus pyralis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Photinus pyralis | - |
- |
- |
Photinus pyralis | Q27758 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-luciferin + O2 + ATP | - |
Photinus pyralis | oxidized D-luciferin + CO2 + H2O + AMP + diphosphate + hv | - |
? | |
D-luciferin + O2 + ATP | - |
Photinus pyralis | oxyluciferin + AMP + diphosphate + CO2 + light | - |
? |
Synonyms | Comment | Organism |
---|---|---|
firefly luciferase | - |
Photinus pyralis |
Luc | - |
Photinus pyralis |
luciferase | - |
Photinus pyralis |