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Literature summary extracted from

  • Salleh, H.M.; Muellegger, J.; Reid, S.P.; Chan, W.Y.; Hwang, J.; Warren, R.A.; Withers, S.G.
    Cloning and characterization of Thermotoga maritima beta-glucuronidase (2005), Carbohydr. Res., 341, 49-59.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.31 gene TM1062, DNA and amino acid sequence determination and analysis, expression of His6-tagged enzyme in Escherichia coli strain GMS407 Thermotoga maritima

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.31 E383A site-directed mutagenesis, the mutant enzyme shows reduced activity and similar thermal stability at 85°C compared to the wild-type enzyme Thermotoga maritima
3.2.1.31 E383Q site-directed mutagenesis, the mutant enzyme shows reduced activity and thermal stability at 85°C compared to the wild-type enzyme Thermotoga maritima
3.2.1.31 E476A site-directed mutagenesis, the mutant enzyme shows increased thermal stability at 85°C compared to the wild-type enzyme Thermotoga maritima

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.31 2-deoxy-2-fluoro-beta-D-glucosyluronic acid fluoride inactivation the recombinant His6-tagged enzyme Thermotoga maritima
3.2.1.31 azide
-
 Thermotoga maritima
3.2.1.31 D-saccharic acid 1,4-lactone competitive, protects the enzyme against inactivation by 2-deoxy-2-fluoro-beta-D-glucosyluronic acid fluoride Thermotoga maritima

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 Thermotoga maritima
3.2.1.31 0.152
-
 4-nitrophenyl beta-D-glucuronide pH 6.5, 75°C, recombinant His6-tagged enzyme Thermotoga maritima
3.2.1.31 3.06
-
 4-nitrophenyl beta-D-galactosiduronide pH 6.5, 75°C, recombinant His6-tagged enzyme Thermotoga maritima
3.2.1.31 145
-
 4-nitrophenyl beta-D-glucoside pH 6.5, 75°C, recombinant His6-tagged enzyme Thermotoga maritima

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.31 66000
-
 x * 66000, recombinant His6-tagged enzyme, SDS-PAGE Thermotoga maritima

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.31 Thermotoga maritima Q9X0F2 gene TM1062
-
3.2.1.31 Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 Q9X0F2 gene TM1062
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.31 recombinant His6-tagged enzyme from Escherichia coli strain GMS407 by metal affinity chromatography to homogeneity Thermotoga maritima

Reaction

EC Number Reaction Comment Organism Reaction ID
3.2.1.31 a beta-D-glucuronoside + H2O = D-glucuronate + an alcohol reaction mechanism, active site with catalytic nucleophile E576, residue E383 is also involved in catalysis Thermotoga maritima
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.31 4-nitrophenyl beta-D-galactosiduronide + H2O 32% of the activity with 4-nitrophenyl beta-D-glucuronide Thermotoga maritima 4-nitrophenol + beta-D-galactosiduronic acid
-
 ?
3.2.1.31 4-nitrophenyl beta-D-galactosiduronide + H2O 32% of the activity with 4-nitrophenyl beta-D-glucuronide Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 4-nitrophenol + beta-D-galactosiduronic acid
-
 ?
3.2.1.31 4-nitrophenyl beta-D-glucoside + H2O low activity Thermotoga maritima 4-nitrophenol + beta-D-glucose
-
 ?
3.2.1.31 4-nitrophenyl beta-D-glucoside + H2O low activity Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 4-nitrophenol + beta-D-glucose
-
 ?
3.2.1.31 4-nitrophenyl beta-D-glucuronide + H2O best substrate Thermotoga maritima 4-nitrophenol + beta-D-glucuronic acid
-
 ?
3.2.1.31 4-nitrophenyl beta-D-glucuronide + H2O best substrate Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 4-nitrophenol + beta-D-glucuronic acid
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.31 ? x * 66000, recombinant His6-tagged enzyme, SDS-PAGE Thermotoga maritima

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.31 85
-
 recombinant His6-tagged enzyme, half-life is 3 h Thermotoga maritima

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.31 9
-
 4-nitrophenyl beta-D-glucoside pH 6.5, 75°C, recombinant His6-tagged enzyme Thermotoga maritima
3.2.1.31 29
-
 4-nitrophenyl beta-D-glucuronide pH 6.5, 65°C, recombinant His6-tagged enzyme Thermotoga maritima
3.2.1.31 68
-
 4-nitrophenyl beta-D-glucuronide pH 6.5, 75°C, recombinant His6-tagged enzyme Thermotoga maritima
3.2.1.31 443
-
 4-nitrophenyl beta-D-galactosiduronide pH 6.5, 75°C, recombinant His6-tagged enzyme Thermotoga maritima

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.31 6.5
-
-
 Thermotoga maritima

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.31 4.5 7.5
-
 Thermotoga maritima

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.2.1.31 additional information
-
 additional information
-
 Thermotoga maritima
3.2.1.31 0.024
-
 D-saccharic acid 1,4-lactone pH 6.5, 75°C, recombinant His6-tagged enzyme Thermotoga maritima