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Literature summary for 1.13.12.7 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)

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

Protein Variants

Protein Variants Comment Organism
D474K D476N mutation does not have any significant effect Photinus pyralis
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]

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

Organism

Organism UniProt Comment Textmining
Photinus pyralis
-
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
luciferin
-
Photinus pyralis

Temperature Optimum [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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
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

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