β-galactosidase from Sulfolobus solfataricus

Enzyme Description

Species

Sulfolobus solfataricus ↗

Extremophile

Yes thermoacidophilic organism

EC Number

3.2.1.23 ↗

Sequence

Length: 489 amino acids

MYSFPNSFRFGWSQAGFQSEMGTPGSEDPNTDWYKWVHDPENMAAGLVSGDLPENGPGYWGNYKTFHDNAQKMGLKIARLNVEWSRIFPNPLPRPQNFDESKQDVTEVEINENELKRLDEYANKDALNHYREIFKDLKSRGLYFILNMYHWPLPLWLHDPIRVRRGDFTGPSGWLSTRTVYEFARFSAYIAWKFDDLVDEYSTMNEPNVVGGLGYVGVKSGFPPGYLSFELSRRAMYNIIQAHARAYDGIKSVSKKPVGIIYANSSFQPLTDKDMEAVEMAENDNRWWFFDAIIRGEITRGNEKIVRDDLKGRLDWIGVNYYTRTVVKRTEKGYVSLGGYGHGCERNSVSLAGLPTSDFGWEFFPEGLYDVLTKYWNRYHLYMYVTENGIADDADYQRPYYLVSHVYQVHRAINSGADVRGYLHWSLADNYEWASGFSMRFGLLKVDYNTKRLYWRPSALVYREIATNGAITDEIEHLNSVPPVKPLRH

Susan M. Hancock et al. (2005) — Developing Promiscuous Glycosidases for Glycoside Synthesis: Residues W433 and E432 in Sulfolobus solfataricus β-Glycosidase are Important Glucoside- and Galactoside-Specificity Determinants

ChemBioChem · doi:10.1002/cbic.200400341 ↗ · Activity - Michaelis-Menten

51 measurements ▼

Database ID

UniProt: P22498 ↗

Sequence Annotation

Explicit - Provided PDB Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (51 measurements)

51 measurements — page 1 of 3

Mutation	Property	Assay	Solvent	Solvent Volume	Aqueous Reference	WT Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	20%	12736.0	—	18380.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	0.56	—	6.33	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	1,4-Dioxane	30%	0.56	—	5.12	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	30%	0.56	—	2.2	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	5%	0.56	—	0.49	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	30%	0.56	—	1.84	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	5%	0.56	—	0.73	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	50%	12736.0	—	4873.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	40%	12736.0	—	7021.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	30%	12736.0	—	6061.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	40%	7.2	—	19.73	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	10%	12736.0	—	19872.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	5%	12736.0	—	23039.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethylformamide (DMF)	30%	12736.0	—	1579.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	12736.0	—	2857.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	1,4-Dioxane	30%	12736.0	—	1724.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	30%	12736.0	—	8220.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	5%	12736.0	—	28331.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	30%	12736.0	—	7973.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	5%	12736.0	—	17545.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose , p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))

WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	20%	12736.0	—	18380.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	0.56	—	6.33	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	1,4-Dioxane	30%	0.56	—	5.12	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	30%	0.56	—	2.2	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	5%	0.56	—	0.49	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	30%	0.56	—	1.84	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	5%	0.56	—	0.73	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	50%	12736.0	—	4873.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	40%	12736.0	—	7021.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	30%	12736.0	—	6061.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	40%	7.2	—	19.73	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	10%	12736.0	—	19872.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	5%	12736.0	—	23039.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethylformamide (DMF)	30%	12736.0	—	1579.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	12736.0	—	2857.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	1,4-Dioxane	30%	12736.0	—	1724.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	30%	12736.0	—	8220.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	5%	12736.0	—	28331.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	30%	12736.0	—	7973.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	5%	12736.0	—	17545.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/(s*M))
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	5%	0.56	—	0.49	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	50%	7.2	—	24.66	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	30%	7.2	—	17.58	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	20%	7.2	—	12.24	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	10%	7.2	—	11.55	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	5%	7.2	—	11.38	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethylformamide (DMF)	30%	7.2	—	8.32	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	7.2	—	18.08	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	1,4-Dioxane	30%	7.2	—	8.83	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	30%	7.2	—	18.08	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	30%	7.2	—	14.67	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetone	5%	7.2	—	12.77	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	50%	0.56	—	5.06	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	40%	0.56	—	2.81	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	30%	0.56	—	2.9	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	20%	0.56	—	2.67	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	10%	0.56	—	0.58	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethylformamide (DMF)	30%	0.56	—	5.27	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in mM)
WT	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	5%	7.2	—	13.91	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	Classical aqueous control (in 1/s)
E432C	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	30%	—	8220.0	40.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	No control (in 1/(s*M))
E432C	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	30%	—	2.2	1.65	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	No control (in mM)
E432C	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	30%	—	18.08	0.067	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	No control (in 1/s)
E432C	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	30%	—	6061.0	43.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	No control (in 1/(s*M))
E432C	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	30%	—	2.9	1.3	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	No control (in mM)
E432C	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	30%	—	17.58	0.056	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	No control (in 1/s)
W433C	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	30%	—	8220.0	15.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	No control (in 1/(s*M))
W433C	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	30%	—	2.2	12.5	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	No control (in mM)
W433C	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	2-Butanone	30%	—	18.08	0.19	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	No control (in 1/s)
W433C	Activity - Michaelis-Menten	kcat/Km (1/(s*M)) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	30%	—	6061.0	4.0	1/(s*M)	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	No control (in 1/(s*M))
W433C	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	30%	—	2.9	18.3	mM	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	No control (in mM)
W433C	Activity - Michaelis-Menten	kcat (/s) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Acetonitrile	30%	—	17.58	0.08	1/s	50 mM phosphate buffer	6.5	45°C	p-nitrophenyl galactose	D-Galactose, p-Nitrophenol	—	—	No control (in 1/s)

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Mutations in this dataset (3)

WT E432C W433C

Visualization : Activity — Michaelis-Menten

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

Mutation Effect — Michaelis-Menten

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

Structure

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