Literature summary extracted from
Ishibashi, M.; Kawanabe, R.; Amaba, N.; Arai, S.; Laksmi, F.; Komori, K.; Tokunaga, M.
Expression and characterization of the Renilla luciferase with the cumulative mutation (2018), Protein Expr. Purif., 145, 39-44 .
Application
EC Number |
Application |
Comment |
Organism |
---|
1.13.12.5 |
diagnostics |
the enzyme is a good research tool as a reporter protein and bioimaging probes, yielding blue light using the substrate coelenterazine. However the applications are limited since RLuc is unstable under various conditions |
Renilla reniformis |
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
1.13.12.5 |
expression in Escherichia coli BL21 Star (DE3) |
Renilla reniformis |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
1.13.12.5 |
F116/I137V |
the mutant starts to denature at 30°C, and retained its activity up to 52°C with increased solubility at 34°C and specific activity up to approximately 119% |
Renilla reniformis |
1.13.12.5 |
F116L/I137V/I75A/N178D/N264S/S287P |
the thermostability effect increases, with the mutant showing approximately 10°C higher stability. The mutant shows improved tolerance for protease digestion, e.g. trypsin and proteinase and for organic solvent. The mutant enzyme retains 100% specific activity at 45°C, while the wild type loses almost all activity, and retains activity at 55°C. The specific activity is approximately 123% higher than that of the wild type |
Renilla reniformis |
1.13.12.5 |
F116L/I137V/N264S/S287P |
thermostability of the mutant is increased. The mutant enzyme shows denaturation at 45 to 52°C and specific activity up to approximately 150% compared with the wild type enzyme |
Renilla reniformis |
1.13.12.5 |
I75A |
specific activity of I75A is 47% of that of the wild type enzyme, retains activity up to 50°C |
Renilla reniformis |
1.13.12.5 |
N178D |
mutation does not affect thermostability but increases the solubility at 34°C and specific activity up to approximately 141% |
Renilla reniformis |
1.13.12.5 |
N264S/S287P |
the mutant starts to denature at 40°C and retains its activity up to 50°C with increased solubility at 34°C and specific activity up to approximately 150% |
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 h + O2 |
Renilla reniformis |
- |
excited coelenteramide h monoanion + CO2 |
- |
? |
|
Organic Solvent Stability
EC Number |
Organic Solvent |
Comment |
Organism |
---|
1.13.12.5 |
2-propanol |
10%, residual activity of the mutant after treatment is 29.4%, residual activity of the wild type enzyme is 0.4% |
Renilla reniformis |
1.13.12.5 |
dimethylformamide |
10%, residual activity of the mutant after treatment is 24.8%, residual activity of the wild type enzyme is 0.1% |
Renilla reniformis |
1.13.12.5 |
DMSO |
10%, residual activity of the mutant after treatment is 91.3%, residual activity of the wild type enzyme is 24.3% |
Renilla reniformis |
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 |
- |
Renilla reniformis |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
1.13.12.5 |
coelenterazine h + O2 |
- |
Renilla reniformis |
excited coelenteramide h monoanion + CO2 |
- |
? |
|
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
1.13.12.5 |
RLuc |
- |
Renilla reniformis |