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Literature summary for 1.13.12.7 extracted from

  • Branchini, B.R.; Ablamsky, D.M.; Rosenman, J.M.; Uzasci, L.; Southworth, T.L.; Zimmer, M.
    Synergistic mutations produce blue-shifted bioluminescence in firefly luciferase (2007), Biochemistry, 46, 13847-13855.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli strain BL21 Photinus pyralis

Protein Variants

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 [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

Metals/Ions Comment Organism Structure
Mg2+ required for activity Photinus pyralis

Organism

Organism UniProt Comment Textmining
Photinus pyralis Q27758
-
-

Purification (Commentary)

Purification (Comment) Organism
glutathione Sepharose 4B affinity chromatography Photinus pyralis

Storage Stability

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 and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-luciferin + O2 + ATP
-
Photinus pyralis oxidized D-luciferin + CO2 + H2O + AMP + diphosphate + hv
-
?

Synonyms

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