Leucine aminopeptidase from Aquifex aeolicus

Enzyme Description

Species

Aquifex aeolicus ↗

Extremophile

Yes "hyperthermophilic" organism

EC Number

3.4.11.1 ↗

Sequence

Length: 493 amino acids

MKEMEVRAFKDDIKKYKGKSVAVFVYEGDLKPLARLSKGTSNKAKRVAELENFKGKEGEILKVPTLGTSVDFVYIVGLGKKEKVGEDTYRRASANLVKRMRRDKVESTVVVIPRRGDVSKEITKAITEGAILGNYRFDKYKSKKEDEKFEIKEVLINRGDEEGIRLGKIFAEAQNYARNLVNEPGNVINPITLAEEAKKLAEEFGLECKVYDEKQIQEMGMMALYSVGKGSATPPRFIHLIYKPSGKPKEKIALVGKGLTFDSGGLNIKPGDYMRTMKMDKSGACAVLGIMRAIAQLKPDVEVHGLIGAAENMPDGNAYRPDDVIKAKNGKYIEIDNTDAEGRVTLADVLSYASELKPDKIIDMATLTGACMVALGEYTAGLFTNAPDFAEEIKKTAKRTGERVWELPMDDERLRKKIKNTVADVLNTGGRYGGAITAAMFLEEFVGEGIKWVHLDIAGPAWSKEEYGYYTKGGTGFGVRTCLEYIMKVSSNV

Anisur Rahman Khan et al. (2000) — Characterization of a solvent resistant and thermostable aminopeptidase from the hyperthermophillic bacterium, Aquifex aeolicus

Enzyme and Microbial Technology · doi:10.1016/s0141-0229(00)00194-0 ↗ · Activity - Classical Activity - Michaelis-Menten

69 measurements ▼

Database ID

UniProt: O67868 ↗

Sequence Annotation

Inferred - from protein name

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (69 measurements)

69 measurements — page 4 of 4

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	40%	100.0	7.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine , p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	30%	100.0	11.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine , p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	10%	100.0	36.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine , p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	20%	1.2	5.0	mM	50 mM Tris-HCl	8	45°C	Leucine p-nitroanilide	L-Leucine , p-Nitroaniline	—	—	Classical aqueous control (in mM)
Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	40%	1.2	8.0	mM	50 mM Tris-HCl	8	45°C	Leucine p-nitroanilide	L-Leucine , p-Nitroaniline	—	—	Classical aqueous control (in mM)
Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	60%	1.2	9.0	mM	50 mM Tris-HCl	8	45°C	Leucine p-nitroanilide	L-Leucine , p-Nitroaniline	—	—	Classical aqueous control (in mM)
Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	20%	0.7	1.0	1/s	50 mM Tris-HCl	8	45°C	Leucine p-nitroanilide	L-Leucine , p-Nitroaniline	—	—	Classical aqueous control (in 1/s)
Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	40%	0.7	0.5	1/s	50 mM Tris-HCl	8	45°C	Leucine p-nitroanilide	L-Leucine , p-Nitroaniline	—	—	Classical aqueous control (in 1/s)
Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	60%	0.7	0.3	1/s	50 mM Tris-HCl	8	45°C	Leucine p-nitroanilide	L-Leucine , p-Nitroaniline	—	—	Classical aqueous control (in 1/s)

Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	1,4-Dioxane	20%	100.0	26.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	50%	100.0	0.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Dimethylformamide (DMF)	5%	100.0	77.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Dimethylformamide (DMF)	10%	100.0	62.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Dimethylformamide (DMF)	20%	100.0	35.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Dimethylformamide (DMF)	30%	100.0	24.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Dimethylformamide (DMF)	40%	100.0	15.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Dimethylformamide (DMF)	50%	100.0	0.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	5%	100.0	72.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	55.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	100.0	34.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	100.0	20.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100.0	16.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	50%	100.0	0.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	1,4-Dioxane	5%	100.0	71.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	1,4-Dioxane	10%	100.0	48.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	10%	100.0	100.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	1,4-Dioxane	30%	100.0	13.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	1,4-Dioxane	40%	100.0	6.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	1,4-Dioxane	50%	100.0	0.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Isopropanol	5%	100.0	62.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Isopropanol	10%	100.0	61.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Isopropanol	20%	100.0	35.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Isopropanol	30%	100.0	16.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Isopropanol	40%	100.0	3.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Isopropanol	50%	100.0	0.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	1-Propanol	5%	100.0	55.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	1-Propanol	10%	100.0	51.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	1-Propanol	20%	100.0	20.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	1-Propanol	30%	100.0	6.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	1-Propanol	40%	100.0	1.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	1-Propanol	50%	100.0	0.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	30%	100.0	9.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	5%	100.0	100.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	20%	100.0	100.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	30%	100.0	100.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	40%	100.0	62.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	50%	100.0	47.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	60%	100.0	29.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	70%	100.0	16.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	80%	100.0	8.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	5%	100.0	91.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	73.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	20%	100.0	59.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	30%	100.0	46.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	40%	100.0	28.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	50%	100.0	5.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	5%	100.0	58.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	20%	100.0	19.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	40%	100.0	0.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	20%	100.0	13.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	10%	100.0	54.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	5%	100.0	65.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	50%	100.0	0.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	40%	100.0	12.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	30%	100.0	28.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	20%	100.0	51.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	10%	100.0	86.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	5%	100.0	96.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	50%	100.0	0.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	40%	100.0	7.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	30%	100.0	11.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	10%	100.0	36.0	%	50 mM Tris-HCl	8	45°C	? mM Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in %)
Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	20%	1.2	5.0	mM	50 mM Tris-HCl	8	45°C	Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in mM)
Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	40%	1.2	8.0	mM	50 mM Tris-HCl	8	45°C	Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in mM)
Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	60%	1.2	9.0	mM	50 mM Tris-HCl	8	45°C	Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in mM)
Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	20%	0.7	1.0	1/s	50 mM Tris-HCl	8	45°C	Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in 1/s)
Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	40%	0.7	0.5	1/s	50 mM Tris-HCl	8	45°C	Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in 1/s)
Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 405 nm) in the presence of organic solvent	Methanol	60%	0.7	0.3	1/s	50 mM Tris-HCl	8	45°C	Leucine p-nitroanilide	L-Leucine, p-Nitroaniline	—	—	Classical aqueous control (in 1/s)

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Visualization : Activity — Classical

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Activity — Michaelis-Menten