Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli strain BL21 | Photinus pyralis |
Protein Variants | Comment | Organism |
---|---|---|
F250G | 23% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
F250S | 49% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
F250T | 37% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
G246A | 104% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
G246A/F250G | 33% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
G246A/F250S | 42% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
G246A/F250T | 40% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
V241I | 136% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
V241I/F250G | 13% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
V241I/F250S | 32% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
V241I/F250T | 64% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
V241I/G246A/F250S | 73% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
V241I/G246A/F250T | 83% specific activity at pH 7.8 compared to the wild type enzyme | Photinus pyralis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.004 | - |
D-luciferin | recombinant mutant enzyme G246A, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.006 | - |
D-luciferin | recombinant mutant enzyme V241I/G246A/F250S, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.008 | - |
D-luciferin | recombinant mutant enzyme V241I, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.009 | - |
D-luciferin | recombinant mutant enzyme V241I/G246A/F250T, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.01 | - |
D-luciferin | recombinant mutant enzyme G246A/F250S, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.011 | - |
D-luciferin | recombinant mutant enzyme G246A/F250G, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.011 | - |
D-luciferin | recombinant mutant enzyme G246A/F250T, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.012 | - |
D-luciferin | recombinant mutant enzyme V241I/F250S, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.014 | - |
D-luciferin | recombinant mutant enzyme F250T, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.015 | - |
D-luciferin | recombinant wild type enzyme, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.016 | - |
D-luciferin | recombinant mutant enzyme F250G, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.022 | - |
D-luciferin | recombinant mutant enzyme F250S, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.024 | - |
D-luciferin | recombinant mutant enzyme V241I/F250G, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.024 | - |
D-luciferin | recombinant mutant enzyme V241I/F250T, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.029 | - |
ATP | recombinant mutant enzyme G246A, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.041 | - |
ATP | recombinant mutant enzyme V241I/G246A/F250T, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.06 | - |
ATP | recombinant mutant enzyme G246A/F250S, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.06 | - |
ATP | recombinant mutant enzyme G246A/F250T, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.061 | - |
ATP | recombinant mutant enzyme G246A/F250G, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.093 | - |
ATP | recombinant mutant enzyme V241I/G246A/F250S, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.133 | - |
ATP | recombinant mutant enzyme F250S, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.151 | - |
ATP | recombinant mutant enzyme V241I/F250T, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.16 | - |
ATP | recombinant mutant enzyme V241I/F250S, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.16 | - |
ATP | recombinant wild type enzyme, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.166 | - |
ATP | recombinant mutant enzyme F250G, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.182 | - |
ATP | recombinant mutant enzyme V241I/F250G, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.215 | - |
ATP | recombinant mutant enzyme F250T, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis | |
0.283 | - |
ATP | recombinant mutant enzyme V241I, in 25 mM glycylglycine buffer, at pH 7.8, in the presence of 2 mM Mg2+, at 25°C | Photinus pyralis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required for activity | Photinus pyralis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Photinus pyralis | Q27758 | - |
- |
Purification (Comment) | Organism |
---|---|
glutathione Sepharose 4B affinity chromatography | Photinus pyralis |
Storage Stability | Organism |
---|---|
4°C, 50 mM Tris-HCl (pH 7.0) containing 150 mM NaCl, 1 mM EDTA, and 1 mM dithiothreitol, up to 6 months, 10% loss of activity | Photinus pyralis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-luciferin + O2 + ATP | - |
Photinus pyralis | oxidized D-luciferin + CO2 + H2O + AMP + diphosphate + hv | - |
? |
Synonyms | Comment | Organism |
---|---|---|
firefly luciferase | - |
Photinus pyralis |
Luc | - |
Photinus pyralis |
Ppy | recombinant Photinus pyralis luciferase containing the additional N-terminal peptide Gly-Pro-Leu-Gly-Ser | Photinus pyralis |