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Literature summary extracted from

  • Karimzadeh, S.; Moradi, M.; Hosseinkhani, S.
    Delicate balance of electrostatic interactions and disulfide bridges in thermostability of firefly luciferase (2012), Int. J. Biol. Macromol., 51, 837-844.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.13.12.7 expressed in Escherichia coli as a His-tagged fusion protein Photinus pyralis

Protein Variants

EC Number Protein Variants Comment Organism
1.13.12.7 A296C/A326C specific activity 676% (compared to wild-type 100%), Km (ATP) decreased compared to wild-type, Km (D-luciferin) decreased compared to wild-type, optimal temperature 35° (wild-type 25°C), optimal pH 8.5 (wild-type pH 8), activity remains 54% at 40°C for 5 min (compared to wild-type 0%), t1/2: 40 min at 35°C (compared to wild-type 5 min) Photinus pyralis
1.13.12.7 I232R specific activity 75% (compared to wild-type 100%), Km (ATP) increased compared to wild-type, Km (D-luciferin) increased compared to wild-type, optimal temperature 25° (wild-type 25°C), optimal pH 8.5 (wild-type pH 8), activity remains 4% at 40°C for 5 min (compared to wild-type 0%), t1/2: 20 min at 35°C (compared to wild-type 5 min) Photinus pyralis
1.13.12.7 I232R/A296C/A326C specific activity 358% (compared to wild-type 100%), Km (ATP) increased compared to wild-type, Km (D-luciferin) increased compared to wild-type, optimal temperature 35° (wild-type 25°C), optimal pH 8 (wild-type pH 8), activity remains 28% at 40°C for 5 min (compared to wild-type 0%), t1/2: 10 min at 35°C (compared to wild-type 5 min) Photinus pyralis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.13.12.7 0.1
-
ATP 35°C, pH 8.5, mutant A296C/A326C Photinus pyralis
1.13.12.7 0.1
-
D-luciferin 35°C, pH 8.5, mutant A296C/A326C Photinus pyralis
1.13.12.7 0.125
-
ATP 25°C, pH 8, wild-type Photinus pyralis
1.13.12.7 0.125
-
D-luciferin 25°C, pH 8, wild-type Photinus pyralis
1.13.12.7 0.2
-
ATP 25°C, pH 8.5, mutant I232R Photinus pyralis
1.13.12.7 0.2
-
D-luciferin 25°C, pH 8.5, mutant I232R Photinus pyralis
1.13.12.7 0.2
-
ATP 35°C, pH 8, mutant A296C/A326C/I232R Photinus pyralis
1.13.12.7 0.2
-
D-luciferin 35°C, pH 8, mutant A296C/A326C/I232R Photinus pyralis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.13.12.7 62000
-
SDS-PAGE Photinus pyralis

Organism

EC Number Organism UniProt Comment Textmining
1.13.12.7 Photinus pyralis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.13.12.7 using Ni-NTA chromatography Photinus pyralis

Substrates and Products (Substrate)

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

EC Number Synonyms Comment Organism
1.13.12.7 luciferin
-
Photinus pyralis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.13.12.7 25
-
assay at Photinus pyralis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.13.12.7 35
-
T1/2 results show that half-lives (t1/2) of wild-type, mutant A296C/A326C, mutant A296C/A326C/I232R and mutant I232R are 5, 40, 10 and 20 min at 35°C respectively Photinus pyralis
1.13.12.7 40
-
thermal stability experiments of mutant A296C/A326C, mutant A296C/A326C/I232R and mutant I232R shows that their original activity remains approximately 54%, 28%, and 4% after incubation at 40°C for 5 min, respectively (compared to wild-type 0%) Photinus pyralis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.13.12.7 7.8
-
assay at Photinus pyralis