Literature summary for 3.2.1.31 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.

Search for more literature for 3.2.1.31

Cloned(Commentary)

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

Protein Variants

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

Inhibitors

Inhibitors	Comment	Organism	Structure
2-deoxy-2-fluoro-beta-D-glucosyluronic acid fluoride	inactivation the recombinant His6-tagged enzyme	Thermotoga maritima
azide	-	Thermotoga maritima
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]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	kinetics of mutant enzymes, overview	Thermotoga maritima
0.152	-	4-nitrophenyl beta-D-glucuronide	pH 6.5, 75°C, recombinant His6-tagged enzyme	Thermotoga maritima
3.06	-	4-nitrophenyl beta-D-galactosiduronide	pH 6.5, 75°C, recombinant His6-tagged enzyme	Thermotoga maritima
145	-	4-nitrophenyl beta-D-glucoside	pH 6.5, 75°C, recombinant His6-tagged enzyme	Thermotoga maritima

Molecular Weight [Da]

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

Organism

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

Purification (Commentary)

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

Reaction

Reaction	Comment	Organism	Reaction ID
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)

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

Subunits

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

Temperature Stability [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.5	-	-	Thermotoga maritima

pH Range

pH Minimum	pH Maximum	Comment	Organism
4.5	7.5	-	Thermotoga maritima

Ki Value [mM]

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

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Release 2023.1 (January 2023)