Luciferase (gene Rluc) from Renilla reniformis

Enzyme Description

Species

Extremophile

EC Number

1.13.12.5 ↗

Sequence

Length: 311 amino acids

MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENAVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWFELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREIPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVKGLHFSQEDAPDEMGKYIKSFVERVLKNEQ

Matsujiro Ishibashi et al. (2018) — Expression and characterization of the Renilla luciferase with the cumulative mutation

Protein Expression and Purification · doi:10.1016/j.pep.2017.12.010 ↗ · Activity + Stability - Incubation Stability - Tm

62 measurements ▼

Database ID

UniProt: P27652 ↗

Sequence Annotation

Inferred - from protein name

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (62 measurements)

62 measurements — page 4 of 4

Mutation	Property	Assay	Solvent	Solvent Volume	Aqueous Reference	WT Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
I75A/F116L/I137V/N178D/N264S/S287P	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Ethyl Acetate	10%	55.5	—	34.2	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
I75A/F116L/I137V/N178D/N264S/S287P	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	1-Butanol	10%	55.5	—	—	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)

WT	Activity + Stability - Incubation	Activity after incubation (30 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	75.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (10 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	99.0	—	42.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (5 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	100.0	—	59.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (20 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	82.0	—	40.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (30 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	75.0	—	34.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (40 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	72.0	—	27.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (50 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	60.0	—	26.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (60 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	58.0	—	24.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (5 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	100.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (10 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	99.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (20 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	82.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (40 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	72.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (50 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	60.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (60 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	58.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (5 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	100.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (10 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	99.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (20 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	82.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (30 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	75.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (40 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	72.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (50 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	60.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (60 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	58.0	—	0.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
WT	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	1-Butanol	10%	43.9	—	—	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Ethylene Glycol	10%	43.9	—	44.7	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Diethylene Glycol	10%	43.9	—	44.0	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	43.9	—	41.6	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	1,2-Dimethoxyethane	10%	43.9	—	38.3	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Dimethylformamide (DMF)	10%	43.9	—	36.0	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Isopropanol	10%	43.9	—	34.2	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Acetonitrile	10%	43.9	—	32.1	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Ethyl Acetate	10%	43.9	—	—	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	1,4-Dioxane	10%	43.9	—	34.2	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (60 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	84.0	24.0	91.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (40 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	86.0	0.0	31.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (5 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	103.0	0.0	41.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (10 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	98.0	0.0	20.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (20 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	92.0	0.0	31.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (30 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	86.0	0.0	25.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (60 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	84.0	0.0	30.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (50 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	86.0	0.0	34.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (40 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	86.0	0.0	49.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (30 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	86.0	0.0	46.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (20 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	92.0	0.0	56.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (10 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	98.0	0.0	54.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (5 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Isopropanol	10%	103.0	0.0	52.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (60 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	84.0	0.0	24.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (50 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethylformamide (DMF)	10%	86.0	0.0	27.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (50 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	86.0	26.0	90.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (40 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	86.0	27.0	92.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (30 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	86.0	34.0	91.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (20 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	92.0	40.0	96.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (10 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	98.0	42.0	86.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Activity + Stability - Incubation	Activity after incubation (5 minutes at 35°C) in the presence of organic solvent, measured by emitted luminescence measurement	Dimethyl Sulfoxide (DMSO)	20%	103.0	59.0	97.0	%	Incubation: 15 mM Tris-HCl buffer, 1 mM EDTA, 0.2 M NaCl, Assay: 100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	Incubation: 7.8, Assay: 7.6	Incubation: 35°C, Assay: 25°C	33 µM Coelenterazine	Carbon Dioxide (CO2), coelenteramide, photon	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent
I75A/F116L/I137V/N178D/N264S/S287P	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Ethylene Glycol	10%	55.5	—	54.8	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
I75A/F116L/I137V/N178D/N264S/S287P	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Diethylene Glycol	10%	55.5	—	53.9	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
I75A/F116L/I137V/N178D/N264S/S287P	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	55.5	—	51.4	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
I75A/F116L/I137V/N178D/N264S/S287P	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	1,2-Dimethoxyethane	10%	55.5	—	47.3	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
I75A/F116L/I137V/N178D/N264S/S287P	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Dimethylformamide (DMF)	10%	55.5	—	47.0	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
I75A/F116L/I137V/N178D/N264S/S287P	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Isopropanol	10%	55.5	—	44.8	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
I75A/F116L/I137V/N178D/N264S/S287P	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Acetonitrile	10%	55.5	—	41.8	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
I75A/F116L/I137V/N178D/N264S/S287P	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	1,4-Dioxane	10%	55.5	—	45.0	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
I75A/F116L/I137V/N178D/N264S/S287P	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	Ethyl Acetate	10%	55.5	—	34.2	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)
I75A/F116L/I137V/N178D/N264S/S287P	Stability - Tm	Tm measured after 1 hour incubation (thermal shift assay with Rox dye) in the presence of organic solvent	1-Butanol	10%	55.5	—	—	°C	100 mM phosphate buffer, 1 mM EDTA, 0.5 M NaCl	7.6	—	—	—	—	—	Classical aqueous control (in °C)

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Mutations in this dataset (2)

WT I75A/F116L/I137V/N178D/N264S/S287P

Visualization : Stability — T_m

One bar per measurement. Colour = solvent, shade = solvent volume. — — — Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Visualization : Activity + Stability — Incubation

One bar per measurement. Colour = solvent, shade = solvent volume. — — — Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Structure

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Luciferase (gene Rluc) from Renilla reniformis

Enzyme Description

Sequence

Experimental Data (62 measurements)

Visualization : Stability — Tm

Mutation Effect

Visualization : Activity + Stability — Incubation

Mutation Effect

Structure

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Visualization : Stability — T_m