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Literature summary for 1.13.12.7 extracted from

  • Pozzo, T.; Akter, F.; Nomura, Y.; Louie, A.Y.; Yokobayashi, Y.
    Firefly luciferase mutant with enhanced activity and thermostability (2018), ACS Omega, 3, 2628-2633 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.13.12.7

Application

Application Comment Organism
molecular biology Ppy luciferase can been used extensively as a reporter gene in living cells and organisms. Some biological applications are limited by the low stability of the luciferase and limited intracellular luciferin concentration. The mutant enzyme T214A/A215L/I232A/F295L/E345K/I423L/D436G/L530R exhibits both improved thermostability and brighter luminescence at low luciferin concentrations, it may be useful for reporter gene applications Photinus pyralis

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) Photinus pyralis

Protein Variants

Protein Variants Comment Organism
I423L/D436G/L530R the mutant shows a lower apparent Km value of 0.00076 mM than that of the wild-type enzyme Photinus pyralis
T214A/A215L/I232A/F295L/E345K the mutant enzyme displays high thermostability, retaining about 60% activity after 120 min at 45°C. Although the mutant shows higher maximum activity at high D-luciferin concentrations, its activity at low D-luciferin concentrations (below 0.004 mM) lags behind that of the mutantI423L/D436G/L530R Photinus pyralis
T214A/A215L/I232A/F295L/E345K/I423L/D436G/L530R the mutant enzyme exhibits both improved thermostability and brighter luminescence at low luciferin concentrations, it may be useful for reporter gene applications Photinus pyralis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00076
-
 D-firefly luciferin 25°C, pH 7.8, I423L/D436G/L530R Photinus pyralis
0.00121
-
 D-firefly luciferin 25°C, pH 7.8, wild-type enzyme Photinus pyralis
0.0042
-
 D-firefly luciferin 25°C, pH 7.8, mutant enzyme T214A/A215L/I232A/F295L/E345K/I423L/D436G/L530R Photinus pyralis
0.00881
-
 D-firefly luciferin 25°C, pH 7.8, mutant enzyme T214A/A215L/I232A/F295L/E345K Photinus pyralis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-firefly luciferin + O2 + ATP Photinus pyralis
-
 firefly oxyluciferin + CO2 + AMP + diphosphate + hv
-
 ?

Organism

Organism UniProt Comment Textmining
Photinus pyralis P08659
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Photinus pyralis

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
fluc
-
 Photinus pyralis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45
-
 the mutant enzyme T214A/A215L/I232A/F295L/E345K displays high thermostability, retaining about 60% activity after 120 min at 45°C Photinus pyralis