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Literature summary for 1.13.12.7 extracted from

  • Noda, K.; Matsuno, T.; Fujii, H.; Kogure, T.; Urata, M.; Asami, Y.; Kuroda, A.
    Single bacterial cell detection using a mutant luciferase (2008), Biotechnol. Lett., 30, 1051-1054.
    View publication on PubMed
Search for more literature for 1.13.12.7

Application

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(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli JM109 cells and Bacillus subtilis strain NBRC13719 Photinus pyralis
into pET28a vector Photinus pyralis

Protein Variants

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 [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

Metals/Ions Comment Organism Structure
Mg2+ 10 mM, required Photinus pyralis

Organism

Organism UniProt Comment Textmining
Photinus pyralis
-
-
-
Photinus pyralis Q27758
-
-

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
-
 ?
D-luciferin + O2 + ATP
-
 Photinus pyralis oxyluciferin + AMP + diphosphate + CO2 + light
-
 ?

Synonyms

Synonyms Comment Organism
firefly luciferase
-
 Photinus pyralis
Luc
-
 Photinus pyralis
luciferase
-
 Photinus pyralis