Ξ²-galactosidase from Sulfolobus solfataricus

Enzyme Description

Extremophile
Yes thermoacidophilic organism
EC Number

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 3 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
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% β€” 2.2 1.65 mM 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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% β€” 18.08 0.067 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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% β€” 6061.0 43.0 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))
E432C Activity - Michaelis-Menten Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent Acetonitrile 30% β€” 2.9 1.3 mM 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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% β€” 17.58 0.056 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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% β€” 8220.0 15.0 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))
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% β€” 2.2 12.5 mM 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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% β€” 18.08 0.19 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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% β€” 6061.0 4.0 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))
W433C Activity - Michaelis-Menten Km (mM) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent Acetonitrile 30% β€” 2.9 18.3 mM 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” 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% β€” 17.58 0.08 1/s 50 mM phosphate buffer 6.5 45Β°C p-nitrophenyl galactose D-Galactose , p-Nitrophenol β€” β€” No control (in 1/s)
1 2 3
Next β€Ί

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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