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Literature summary extracted from
- Super RLuc8 A novel engineered Renilla luciferase with a red-shifted spectrum and stable light emission (2017), Enzyme Microb. Technol., 96, 60-66 .
Application
| EC Number | Application | Comment | Organism |
|---|---|---|---|
| 1.13.12.5 | analysis | Renilla luciferase is a bioluminescent enzyme which is broadly used as a reporter protein in molecularbiosensors | Renilla reniformis |
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.13.12.5 | recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Renilla reniformis |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.13.12.5 | M185V/K189V/V267I | site-directed mutagenesis,compared with the native RLuc, mutant super RLuc has a higher turnover number, increased light output upon expression in Arabidopsis thaliana and enhanced half-life of photon emission, super RLuc is a blue light emitting luciferase | Renilla reniformis |
| 1.13.12.5 | additional information | enzyme engineering of blue light emitting enzyme mutant super RLuc to construct a luciferase with desired light emission wavelength and thermostability, namely super RLuc8, which has a red-shifted spectrum and shows stable light emission. Super RLuc8 shows a 10fold increase in thermostability at 37°C after 20 min incubation, in comparison to the native enzyme. The optimum temperature of the mutant increases from 30 to 37°C. Molecular dynamics simulation analysis indicates that the increased thermostability is most probably caused by a better structural compactness and more local rigidity in the regions out of the emitter site. The mutant super RLuc8 shows increased activity compared t the wild-type. Molecular dynamics simulation, overview | Renilla reniformis |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.13.12.5 | 0.00293 | - |
coelenterazine | pH 7.2, 37°C, recombinant engineered mutant super RLuc 8 | Renilla reniformis |  |
| 1.13.12.5 | 0.00997 | - |
coelenterazine | pH 7.2, 37°C, recombinant wild-type enzyme | Renilla reniformis | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.13.12.5 | coelenterazine + O2 | Renilla reniformis | - |
coelenteramide + CO2 + hv | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.13.12.5 | Renilla reniformis | P27652 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.13.12.5 | recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Renilla reniformis |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.13.12.5 | coelenterazine + O2 | - |
Renilla reniformis | coelenteramide + CO2 + hv | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.13.12.5 | ? | x * 36000, modified recombinant His-tagged enzyme super RLuc 8, SDS-PAGE | Renilla reniformis |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.13.12.5 | Renilla luciferase | - |
Renilla reniformis |
| 1.13.12.5 | RLuc | - |
Renilla reniformis |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.13.12.5 | 30 | - |
wild-type enzyme | Renilla reniformis |
| 1.13.12.5 | 36 | 37 | engineered enzyme mutant super RLuc8 | Renilla reniformis |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.13.12.5 | 37 | - |
pH 7.8, 60 min, 4 and 10fold increases in recombinant His-tagged engineered super RLuc8 thermostability are observed at 37°C after 10 and 20 min incubation, respectively, in comparison to the recombinant His-tagged wild-type enzyme. While the wild-type enzyme is completely inactivated after 30 min, the mutant enzyme retains about 30% of its initial activity | Renilla reniformis |
| 1.13.12.5 | 45 | - |
pH 7.8, the activity of purified recombinant His-tagged engineered mutant superRLuc8 remains around 60% after incubation at 45°C, while wild-type recombinant His-tagged RLuc loses approximately 82% of its initial activity at the same temperature | Renilla reniformis |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 1.13.12.5 | additional information | molecular dynamic simulation of the native RLuc based on PDB ID 2PSF, which is the crystal structure of the mutant RLuc luciferase, overview | Renilla reniformis |
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