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Literature summary for 3.2.1.167 extracted from

  • Sasaki, K.; Taura, F.; Shoyama, Y.; Morimoto, S.
    Molecular characterization of a vovel beta-glucuronidase from Scutellaria baicalensis Georgi (2000), J. Biol. Chem., 275, 27466-27472.
    View publication on PubMed
Search for more literature for 3.2.1.167

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli. Escherichia coli having pET28a/sGUS shows much higher beta-glucuronidase activity using baicalein 7-O-beta-D-glucuronide as a substrate than Escherichia coli carrying pET28a. The minor enzyme activity observed in the latter is considered to be due to endogenous beta-glucuronidase Scutellaria baicalensis

Protein Variants

Protein Variants Comment Organism
E194A beta-glucuronidase activity of the mutant enzyme with baicalein 7-O-beta-D-glucuronide is 86% of wild-type activity Scutellaria baicalensis
E212A beta-glucuronidase activity of the mutant enzyme with baicalein 7-O-beta-D-glucuronide is 1.2% of wild-type activity Scutellaria baicalensis
E225A beta-glucuronidase activity of the mutant enzyme with baicalein 7-O-beta-D-glucuronide is 94% of wild-type activity Scutellaria baicalensis
E272A beta-glucuronidase activity of the mutant enzyme with baicalein 7-O-beta-D-glucuronide is 70% of wild-type activity Scutellaria baicalensis
E329A beta-glucuronidase activity of the mutant enzyme with baicalein 7-O-beta-D-glucuronide is 0.3% of wild-type activity Scutellaria baicalensis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0097
-
 5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate pH 6.5, 28°C, mutant enzyme E329A Scutellaria baicalensis
0.0103
-
 5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate pH 6.5, 28°C, mutant enzyme E212A Scutellaria baicalensis
0.0121
-
 5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate pH 6.5, 28°C, wild-type enzyme Scutellaria baicalensis
0.027
-
 5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate pH 6.5, 28°C, mutant enzyme Y281A Scutellaria baicalensis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
56027
-
 x * 56027, calculated from sequence Scutellaria baicalensis

Organism

Organism UniProt Comment Textmining
Scutellaria baicalensis Q9LRC8 protein sequence homology shows no significant similarity to the family 2 beta-glucuronidases
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Scutellaria baicalensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5,6,7-trihydroxyflavone-7-O-beta-D-glucuronate + H2O i.e. baicalin. The enzyme shows more than 20fold higher activity for baicalein 7-O-beta-D-glucuronide as compared with luteolin 3'-O-beta-D-glucuronide Scutellaria baicalensis 5,6,7-trihydroxyflavone + D-glucuronate i.e. baicalein ?

Subunits

Subunits Comment Organism
? x * 56027, calculated from sequence Scutellaria baicalensis

Synonyms

Synonyms Comment Organism
sGUS
-
 Scutellaria baicalensis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
28
-
 assay at Scutellaria baicalensis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.011
-
 5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate pH 6.5, 28°C, mutant enzyme E329A Scutellaria baicalensis
0.135
-
 5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate pH 6.5, 28°C, mutant enzyme E212A Scutellaria baicalensis
0.77
-
 5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate pH 6.5, 28°C, mutant enzyme Y281A Scutellaria baicalensis
639
-
 5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate pH 6.5, 28°C, wild-type enzyme Scutellaria baicalensis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
 assay at Scutellaria baicalensis

pI Value

Organism Comment pI Value Maximum pI Value
Scutellaria baicalensis calculated from sequence
-
 5.52

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.2
-
 5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate pH 6.5, 28°C, mutant enzyme E329A Scutellaria baicalensis
13.1
-
 5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate pH 6.5, 28°C, mutant enzyme E212A Scutellaria baicalensis
28.5
-
 5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate pH 6.5, 28°C, mutant enzyme Y281A Scutellaria baicalensis
52810
-
 5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate pH 6.5, 28°C, wild-type enzyme Scutellaria baicalensis