Ξ²-galactosidase from Sulfolobus solfataricus

Enzyme Description

Species
Sulfolobus solfataricus β†—
Extremophile
Yes thermoacidophilic organism
EC Number
3.2.1.23 β†—

Sequence

Length: 489 amino acids
MYSFPNSFRFGWSQAGFQSEMGTPGSEDPNTDWYKWVHDPENMAAGLVSGDLPENGPGYWGNYKTFHDNAQKMGLKIARLNVEWSRIFPNPLPRPQNFDESKQDVTEVEINENELKRLDEYANKDALNHYREIFKDLKSRGLYFILNMYHWPLPLWLHDPIRVRRGDFTGPSGWLSTRTVYEFARFSAYIAWKFDDLVDEYSTMNEPNVVGGLGYVGVKSGFPPGYLSFELSRRAMYNIIQAHARAYDGIKSVSKKPVGIIYANSSFQPLTDKDMEAVEMAENDNRWWFFDAIIRGEITRGNEKIVRDDLKGRLDWIGVNYYTRTVVKRTEKGYVSLGGYGHGCERNSVSLAGLPTSDFGWEFFPEGLYDVLTKYWNRYHLYMYVTENGIADDADYQRPYYLVSHVYQVHRAINSGADVRGYLHWSLADNYEWASGFSMRFGLLKVDYNTKRLYWRPSALVYREIATNGAITDEIEHLNSVPPVKPLRH
Susan M. Hancock et al. (2005) β€” Developing Promiscuous Glycosidases for Glycoside Synthesis: Residues W433 and E432 in Sulfolobus solfataricus Ξ²-Glycosidase are Important Glucoside- and Galactoside-Specificity Determinants
ChemBioChem  Β· doi:10.1002/cbic.200400341 β†—  Β· Activity - Michaelis-Menten
51 measurements ▼
Database ID
UniProt: P22498 β†—
Sequence Annotation
Explicit - Provided PDB Accession Number
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (51 measurements)

51 measurements β€” page 2 of 3
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
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 β€” 0.49 mM 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 24.66 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 17.58 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 12.24 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 11.55 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 11.38 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 8.32 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 18.08 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 8.83 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 18.08 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 14.67 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 12.77 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 5.06 mM 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 2.81 mM 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 2.9 mM 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 2.67 mM 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 0.58 mM 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 5.27 mM 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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 β€” 13.91 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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% β€” 8220.0 40.0 1/(s*M) 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” No control (in 1/(s*M))
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Mutations in this dataset (3)

WT E432C W433C

Visualization : Activity β€” Michaelis-Menten

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

Mutation Effect β€” 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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