Literature summary for 1.13.12.5 extracted from

Rahnama, S.; Saffar, B.; Kahrani, Z.F.; Nazari, M.; Emamzadeh, R.
Super RLuc8 A novel engineered Renilla luciferase with a red-shifted spectrum and stable light emission (2017), Enzyme Microb. Technol., 96, 60-66 .

Search for more literature for 1.13.12.5

Application

Application	Comment	Organism
analysis	Renilla luciferase is a bioluminescent enzyme which is broadly used as a reporter protein in molecularbiosensors	Renilla reniformis

Cloned(Commentary)

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

Protein Variants

Protein Variants	Comment	Organism
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
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]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.00293	-	coelenterazine	pH 7.2, 37°C, recombinant engineered mutant super RLuc 8	Renilla reniformis
0.00997	-	coelenterazine	pH 7.2, 37°C, recombinant wild-type enzyme	Renilla reniformis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
coelenterazine + O2	Renilla reniformis	-	coelenteramide + CO2 + hv	-	?

Organism

Organism	UniProt	Comment	Textmining
Renilla reniformis	P27652	-	-

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
coelenterazine + O2	-	Renilla reniformis	coelenteramide + CO2 + hv	-	?

Subunits

Subunits	Comment	Organism
?	x * 36000, modified recombinant His-tagged enzyme super RLuc 8, SDS-PAGE	Renilla reniformis

Synonyms

Synonyms	Comment	Organism
Renilla luciferase	-	Renilla reniformis
RLuc	-	Renilla reniformis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	wild-type enzyme	Renilla reniformis
36	37	engineered enzyme mutant super RLuc8	Renilla reniformis

Temperature Stability [°C]

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

General Information	Comment	Organism
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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Release 2023.1 (January 2023)