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Literature summary for 1.13.12.7 extracted from

  • Yu, H.; Zhao, Y.; Guo, C.; Gan, Y.; Huang, H.
    The role of proline substitutions within flexible regions on thermostability of luciferase (2015), Biochim. Biophys. Acta, 1854, 65-72 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Photinus pyralis

Protein Variants

Protein Variants Comment Organism
D476P site-directed mutagenesis, the mutant shows decreased thermostability compared to the wild-type Photinus pyralis
H489P site-directed mutagenesis, the mutant shows improved thermostability while maintaining its catalytic efficiency compared to that of wild-type luciferase, the overall rigidity and local rigidity of H489Pmutant are greatly strengthened Photinus pyralis
S307P/H489P site-directed mutagenesis, the mutation is randomly chosen outside the flexible regions as a control. The mutant has decreased kinetic stability and enhanced thermodynamic stability compared to the wild-type Photinus pyralis

Organism

Organism UniProt Comment Textmining
Photinus pyralis P08659
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Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Photinus pyralis

Synonyms

Synonyms Comment Organism
firefly luciferase
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Photinus pyralis
luciferase
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Photinus pyralis
Photinus pyralis luciferase
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Photinus pyralis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
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assay at room temperature Photinus pyralis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
35
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thermostability of recombinant wild-type and mutant enzymes, overview Photinus pyralis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
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assay at Photinus pyralis

General Information

General Information Comment Organism
additional information usage of three different methods, molecular dynamics (MD) simulation, B-FITTER and framework rigidity optimized dynamics algorithm (FRODA) to determine the flexible regions of Photinus pyralis luciferase: fragment 197-207, fragment 471-481, and fragment 487-495. Introduction of proline within most flexible regions is used to rigidify these flexible regions resulting in mutants D476P and H489P. Analysis of the relationship between stability, activity and flexibility of wild-type and mutant enzymes, molecular dynamics simulations, modelling, overview Photinus pyralis