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Literature summary extracted from

  • Riahi-Madvar, A.; Hosseinkhani, S.
    Design and characterization of novel trypsin-resistant firefly luciferases by site-directed mutagenesis (2009), Protein Eng. Des. Sel., 22, 655-663.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.13.12.7 expression in Escherichia coli BL21 and DH5alpha, transfection in CHO-cells Photinus pyralis

Protein Variants

EC Number Protein Variants Comment Organism
1.13.12.7 Q338P about 50% decrease in specific activity compared to wild-type luciferase Photinus pyralis
1.13.12.7 R213E same activity like wild-type luciferase Photinus pyralis
1.13.12.7 R213M 3fold activity compared to wild-type luciferase Photinus pyralis
1.13.12.7 R337Q only 60% relative specific activity compared to wild-type luciferase, higher light stability, most stable luciferase mutant against trypsin hydrolysis at 23 and 37°C Photinus pyralis

General Stability

EC Number General Stability Organism
1.13.12.7 decrease of remaining activity after trypsin hydrolysis at 23 and 37°C, mutants R213M and R337Q are more stable than the wild-type luciferase Photinus pyralis
1.13.12.7 mutant R337Q still 26% remaining activity after 15 min trypsin hydrolysis at 37°C Photinus pyralis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.13.12.7 0.01
-
 Luciferin mutant R213M Photinus pyralis
1.13.12.7 0.012
-
 Luciferin wild-type luciferase Photinus pyralis
1.13.12.7 0.013
-
 Luciferin mutant Q338P Photinus pyralis
1.13.12.7 0.014
-
 Luciferin mutant R213E Photinus pyralis
1.13.12.7 0.018
-
 Luciferin mutant R337Q Photinus pyralis
1.13.12.7 0.133
-
 ATP wild-type luciferase Photinus pyralis
1.13.12.7 0.15
-
 ATP mutant R213M Photinus pyralis
1.13.12.7 0.2
-
 ATP mutant R213E Photinus pyralis
1.13.12.7 0.35
-
 ATP mutant Q338P Photinus pyralis
1.13.12.7 0.5
-
 ATP mutant R337Q Photinus pyralis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.13.12.7 intracellular
-
 Photinus pyralis 5622
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.13.12.7 Mg2+ required 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 Ni-NTA spin column Photinus pyralis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.13.12.7 luciferin + ATP + O2 assay at 25°C, pH 7.8 Photinus pyralis ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.13.12.7 firefly luciferase
-
 Photinus pyralis

Temperature Optimum [°C]

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

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.13.12.7 7.3
-
 mutant R337Q Photinus pyralis
1.13.12.7 7.8
-
 assay at Photinus pyralis
1.13.12.7 8
-
-
 Photinus pyralis