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Literature summary extracted from

  • Amini-Bayat, Z.; Hosseinkhani, S.; Jafari, R.; Khajeh, K.
    Relationship between stability and flexibility in the most flexible region of Photinus pyralis luciferase (2012), Biochim. Biophys. Acta, 1824, 350-358.
    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 D474K D476N mutation does not have any significant effect Photinus pyralis
1.13.12.7 D476N D474K mutation destabilizes the protein. Flexibility analysis using dynamic quenching and limited proteolysis demonstrates that D474K mutation is much more flexible than wild-type Photinus pyralis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.13.12.7 0.008
-
 D-luciferin mutant D476N, pH 7, 25°C Photinus pyralis
1.13.12.7 0.015
-
 D-luciferin wild-type, pH 8, 25°C Photinus pyralis
1.13.12.7 0.016
-
 D-luciferin mutant D474K, pH 8.5, 25°C Photinus pyralis
1.13.12.7 0.1
-
 ATP mutant D476N, pH 7, 25°C Photinus pyralis
1.13.12.7 0.1
-
 ATP wild-type, pH 8, 25°C Photinus pyralis
1.13.12.7 0.15
-
 ATP mutant D474K, pH 8.5, 25°C 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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.13.12.7 additional information
-
 relative specific activity: wild-type 100%, mutant D474K 55%, mutant D476N 181% Photinus pyralis

Substrates and Products (Substrate)

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

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 15 45 when native and mutants are incubated 5 min at temperatures ranging from 15 to 45°C, D474K mutant is inactivated faster than native and D476N at all temperatures Photinus pyralis
1.13.12.7 32
-
 after 20 min at 32°C wild-type and mutant D476N retain 24% original activity whereas mutant D474K shows only 2% remaining activity Photinus pyralis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.13.12.7 7
-
 mutant D476N Photinus pyralis
1.13.12.7 8
-
 wild-type Photinus pyralis
1.13.12.7 8.5
-
 mutant D474K Photinus pyralis