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Literature summary extracted from

  • Geddie, M.L.; Matsumura, I.
    Rapid evolution of beta-glucuronidase specificity by saturation mutagenesis of an active site loop (2004), J. Biol. Chem., 279, 26462-26468.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.31 gene gusA, expression of His-tagged wild-type and mutant enzymes in Escherichia coli Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.31 D531E/S557V/N566S/G601S saturation mutagenesis, mutant 1.13, altered substrate specificity compared to the wild-type enzyme Escherichia coli
3.2.1.31 additional information rapid evolution of beta-glucuronidase specificity by saturation mutagenesis of an active site loop, DNA shuffling of point mutations, construction of diverse mutants with mutation of residues 557, 566, and 568, the mutants show increased activity with beta-D-xylopyranoside and reduced activity with beta-D-glucuronide, overview Escherichia coli
3.2.1.31 S22N/G81S/K257E/T509A/S557P/N566S/K568Q/Q598R/stop604W saturation mutagenesis, mutant 1.15, altered substrate specificity compared to the wild-type enzyme Escherichia coli
3.2.1.31 S557I/N566A/K568R/A580V saturation mutagenesis, mutant 1.16, altered substrate specificity compared to the wild-type enzyme Escherichia coli
3.2.1.31 S557Q/N566K/K568S/Q598stop saturation mutagenesis, mutant 1.2, altered substrate specificity compared to the wild-type enzyme Escherichia coli
3.2.1.31 V473A/S557P/N566S/K568Q saturation mutagenesis, mutant 4.7, altered substrate specificity compared to the wild-type enzyme Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.31 additional information
-
 additional information kinetics of mutant enzymes, overview Escherichia coli
3.2.1.31 0.2
-
 4-nitrophenyl beta-D-xylopyranoside wild-type enzyme Escherichia coli
3.2.1.31 0.36
-
 4-nitrophenyl beta-D-glucuronide wild-type enzyme Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.31 Escherichia coli
-
 gene gusA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.31 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography to homogeneity Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.31 4-nitrophenyl beta-D-glucuronide + H2O
-
 Escherichia coli 4-nitrophenol + beta-D-glucuronic acid
-
 ?
3.2.1.31 4-nitrophenyl beta-D-xylopyranoside + H2O preferred substrate Escherichia coli 4-nitrophenol + beta-D-xylopyranose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.31 GUS
-
 Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.31 0.001
-
 4-nitrophenyl beta-D-glucuronide wild-type enzyme Escherichia coli
3.2.1.31 68
-
 4-nitrophenyl beta-D-xylopyranoside wild-type enzyme Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.31 7.6
-
 assay at Escherichia coli