Mutant laccase MtLT2 from Myceliophthora thermophila

Enzyme Description

Species

Extremophile

EC Number

1.10.3.2 ↗

Sequence

Length: 559 amino acids

QQICNTPSNRACWTDGYDINTDYEVDSPDTGVVRPYTLTLTEVDNWTGPDGVVKEKVMLVNNSIIGPTIFADWGDTIQVTVINNLGTNGTSIHWHGLHQKGTNLHDGVNGITECPIPPKGGRKVYRFKAQQYGTSWYHSHFSAQYGNGVVGAIQINGPASLPYDTDLGVFPISDYYYSSADELVELTKNSGAPFSDNVLFNGTAKHPETGEGEYANVTLTPGRRHRLRLINTSVENHFQVSLVNHTMTIIAADMVPVNAMTVDSLFLGVGQRYDVVIEASRTPGNYWFNVTFGGGLLCGGSRSPYPAAIFHYAGAPGGPPTDEGKAPVDHNCLDLPNLKPVVARDVPLSGLAKRPDNTLDVTLDTMGTPLFVWKVNGSAINIDWGRPVVDYVLTQNTSFPPGHNIVEVNGADQWSYWLIENDPGAPFTLPHPMHLHGHDFYVLGRSPDEPPASKERHVFDPARDAGLLSGANPVRRDVTMLPAFGWVVLAFRADNPGAWLFHCHIAWHVSGGLGVVYLERADDLRGAVSDADADDFDRLCADWRRYWPTNPNPKSDSGL

Miren Zumárraga et al. (2007) — In Vitro Evolution of a Fungal Laccase in High Concentrations of Organic Cosolvents

Chemistry & Biology · doi:10.1016/j.chembiol.2007.08.010 ↗ · Activity - Classical Stability - Incubation

110 measurements ▼

Database ID

—

Sequence Annotation

Inferred - from protein name (11 mutations compared to PDB 6F5K, article denomination : MtLT2)

Protein Source

Recombinant, host yeast Saccharomyces cerevisiae

Experimental Data (110 measurements)

110 measurements — page 3 of 6

Mutation	Property	Assay	Solvent	Solvent Volume	Aqueous Reference	WT Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	20%	100	25	34	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	30%	100	14	22	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	40%	100	6	9	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	3	5	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	60%	100	0	1	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	10%	100	49	67	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	20%	100	25	42	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	30%	100	14	31	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	40%	100	6	8	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	3	5	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	60%	100	0	2	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	10%	100	49	78	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	20%	100	25	59	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	30%	100	14	40	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	40%	100	6	20	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	3	13	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	60%	100	0	2	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	10%	100	49	95	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	20%	100	25	82	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	30%	100	14	62	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)

WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	1703	—	58	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	1703	—	149	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	10%	100	—	49	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	20%	100	—	25	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	30%	100	—	14	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	40%	100	—	6	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	—	3	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	60%	100	—	0	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	10%	100	—	54	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	20%	100	—	23	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	—	9	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	40%	100	—	3	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	1703	—	58	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
WT	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	—	80	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
WT	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	—	86	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
L429V (6C9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	3579	58	198	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
L429V (6C9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	3579	149	449	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	2048	58	93	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	2048	149	314	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
E182K/L429V (7H9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	4767	58	222	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
E182K/L429V (7H9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	4767	149	974	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	5841	58	435	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	5841	149	600	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	4085	58	524	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	4085	149	780	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	5619	58	1106	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	5619	149	1876	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
L429V (6C9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	10%	100	49	61	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
L429V (6C9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	20%	100	25	38	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
L429V (6C9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	30%	100	14	23	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
L429V (6C9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	40%	100	6	11	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
L429V (6C9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	3	5	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
L429V (6C9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	60%	100	0	2	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	10%	100	49	68	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	20%	100	25	49	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	30%	100	14	28	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	40%	100	6	13	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	3	7	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	60%	100	0	2	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	10%	100	49	58	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	20%	100	25	34	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	30%	100	14	22	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	40%	100	6	9	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	3	5	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	60%	100	0	1	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	10%	100	49	67	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	20%	100	25	42	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	30%	100	14	31	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	40%	100	6	8	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	3	5	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	60%	100	0	2	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	10%	100	49	78	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	20%	100	25	59	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	30%	100	14	40	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	40%	100	6	20	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	3	13	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	60%	100	0	2	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	10%	100	49	95	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	20%	100	25	82	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	30%	100	14	62	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	40%	100	6	31	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	3	20	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	60%	100	0	6	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
L429V (6C9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	10%	100	54	62	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
L429V (6C9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	20%	100	23	31	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
L429V (6C9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	9	13	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
L429V (6C9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	40%	100	3	4	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	10%	100	54	58	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	20%	100	23	24	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	9	10	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S280N/L429V/N552H (2B10)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	40%	100	3	4	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V (7H9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	10%	100	54	95	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V (7H9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	20%	100	23	45	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V (7H9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	9	21	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V (7H9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	40%	100	3	9	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	10%	100	54	99	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	20%	100	23	44	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	9	15	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
S179P/L429V (5B9)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	40%	100	3	5	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	10%	100	54	84	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	20%	100	23	48	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	9	19	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/L429V/N552H (3D1)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	40%	100	3	6	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	10%	100	54	85	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	20%	100	23	58	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	9	33	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
E182K/S280N/L429V/N552H (R2)	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	40%	100	3	9	%	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in %)
L429V	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	3423	58	205	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
E182K	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	2298	58	142	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
N552H	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	2167	58	203	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
S280N	Activity - Classical	Activity measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	1999	58	188	mmol product/(mmol laccase * min)	100 mM sodium acetate buffer	4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Classical aqueous control (in mmol product/(mmol laccase * min))
L429V (6C9)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	80	84	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
L429V (6C9)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	86	86	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
S179P/L429V (5B9)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	80	125	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
S179P/L429V (5B9)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	86	155	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/L429V (7H9)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	80	110	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/L429V (7H9)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	86	138	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
S280N/L429V/N552H (2B10)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	80	93	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
S280N/L429V/N552H (2B10)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	86	96	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/L429V/N552H (3D1)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	80	95	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/L429V/N552H (3D1)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	86	92	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/S280N/L429V/N552H (R2)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	50%	100	80	99	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/S280N/L429V/N552H (R2)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	30%	100	86	100	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/S280N/L429V/N552H (R2)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Dimethylformamide (DMF)	60%	100	—	60	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/S280N/L429V/N552H (R2)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	60%	100	—	100	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/S280N/L429V/N552H (R2)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Dimethylacetamide	60%	100	—	94	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/S280N/L429V/N552H (R2)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Ethanol	60%	100	—	64	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/S280N/L429V/N552H (R2)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetone	60%	100	—	100	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/S280N/L429V/N552H (R2)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Methanol	60%	100	—	100	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)
E182K/S280N/L429V/N552H (R2)	Stability - Incubation	Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (ABTS radical cation reaction products absorbance measurement, 418 nm) in the presence of organic solvent	Acetonitrile	60%	100	—	77	%	Incubation: 100 mM phosphat buffer, Assay: 20 mM sodium acetate buffer	Incubation: 6, Assay: 4.5	Room temperature	3 mM ABTS	ABTS radical cation	—	—	Non-incubated control (in %) measured in the presence of organic solvent, assay measured in the presence of organic solvent \| Clear about assay and control conditions (both in the presence of organic solvent)

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Mutations in this dataset (11)

WT S179P/L429V (5B9) E182K E182K/L429V (7H9) E182K/L429V/N552H (3D1) E182K/S280N/L429V/N552H (R2) S280N S280N/L429V/N552H (2B10) L429V L429V (6C9) N552H

Visualization : Activity — Classical

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 : 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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Mutant laccase MtLT2 from Myceliophthora thermophila

Enzyme Description

Sequence

Experimental Data (110 measurements)

Visualization : Activity — Classical

Mutation Effect

Visualization : Stability — Incubation

Mutation Effect

Structure

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