Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Photinus pyralis |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Photinus pyralis | P08659 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Photinus pyralis |
Synonyms | Comment | Organism |
---|---|---|
firefly luciferase | - |
Photinus pyralis |
luciferase | - |
Photinus pyralis |
Photinus pyralis luciferase | - |
Photinus pyralis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room temperature | Photinus pyralis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | - |
thermostability of recombinant wild-type and mutant enzymes, overview | Photinus pyralis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.8 | - |
assay at | Photinus pyralis |
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 |