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G1512

Sigma-Aldrich

β-Glucuronidase from Helix pomatia

Type H-5, lyophilized powder, ≥400,000 units/g solid

Synonym(s):

β-D-Glucuronide glucuronosohydrolase

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About This Item

CAS Number:
9001-45-0
Enzyme Commission number:
3.2.1.31 (BRENDA, IUBMB)
EC Number:
232-606-8
MDL number:
MFCD00130629
UNSPSC Code:
12352204
NACRES:
NA.54

type

Type H-5

Quality Level

200

form

lyophilized powder

specific activity

≥400,000 units/g solid

secondary activity

≤40,000 units/g solid sulfatase

storage temp.

−20°C

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General description

β-Glucuronidase Type H-2 from Helix pomatia is a crude solution of enzymes derived from the Roman snail. Many β-glucuronidases derived from mollusks also contain sulfatase activity.

Application

β-Glucuronidase from Helix pomatia has been used:
  • in the enzymatic treatment of urine samples for liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analysis
  • in the quercetin extraction procedure from pig or rat tissues for high performance liquid chromatography (HPLC) analysis
  • in the enzymatic deconjugation step in urine and plasma sample preparation for gas chromatography-mass spectrometry analysis (GC-MS)

β-glucuronidase was used to hydrolyze conjugated glucuronide and sulfate metabolites.
New Technical Article Comparing Performance of Different Enzymes
Learn more
about recent application data generated by Sigma R&D to optimize hydrolysis for different drug classes using enzymes from different sources and the use of a chromatographicaly purified enzyme to reduce the effect of esterase activity resulting in conversion of 6-MAM to Morphine

Biochem/physiol Actions

β-glucuronidase (β-GIc) is an exoglycosidase that catalyzes the breakdown of complex carbohydrates. In humans it converts conjugated bilirubin into the unconjugated form, making bilirubin suitable for reabsorption.
Used for the hydrolysis of glucuronide conjugates in urinary metabolite analysis

Quality

Many β-glucuronidases derived from mollusks also contain sulfatase activity.

Linkage

A further purification of G0876 by gel permeation chromatography.

Unit Definition

One Sigma or modified Fishman unit will liberate 1.0 μg of phenolphthalein from phenolphthalein glucuronide per hr at 37 °C at pH 5.0 (30 min assay).
Sulfatase Unit Definition: One unit of sulfatase will hydrolyze 1.0 μmole p-nitrocatechol sulfate per hr at pH 5.0 at 37 °C.

Pictograms

Health hazard
GHS08

Signal Word

Danger

Hazard Statements

H317,H334

Hazard Classifications

Resp. Sens. 1 - Skin Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Denis M Callewaert et al.
Methods in molecular biology (Clifton, N.J.), 610, 435-449 (2009-12-17)
Isoprostanes are a group of prostaglandin-like compounds that are derived in vivo primarily by free radical-mediated peroxidation of arachidonic acid within phospholipids. The resulting isoprostane moieties are rapidly hydrolyzed, metabolized, and excreted. It is now well recognized that isoprostane levels
Measurement of intact sulfate and glucuronide phytoestrogen conjugates in human urine using isotope dilution liquid chromatography-tandem mass spectrometry with [13C3] isoflavone internal standards
Clarke DB, et al.
Analytical Biochemistry, 309(1), 158-172 (2002)
Tissue distribution of quercetin in rats and pigs
de Boer VCJ, et al.
The Journal of Nutrition, 135(7), 1718-1725 (2005)
GC-MS methods for metabolic profiling of microbial fermentation products of dietary polyphenols in human and in vitro intervention studies
Grun CH, et al.
Journal of Chromatography. B, Biomedical Sciences and Applications, 871(2), 212-219 (2008)
Maaike M Appeldoorn et al.
The Journal of nutrition, 139(8), 1469-1473 (2009-06-06)
Intervention studies with procyanidin (PC)-rich extracts and products such as cocoa and wine suggest protective effects of PC against cardiovascular diseases. However, there is no consensus on the absorption and metabolism of PC dimers. Interestingly, nothing is known about the
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Articles

Evaluation of Different Enzymes on Hydrolysis Efficiencies of Glucuronide Drug Metabolites in Urine

β-glucuronidase (GUS) enzymes are utilized to hydrolyze glucuronide (gluc) drug metabolites to the parent drug, facilitating analysis by LC-MS/MS.

Protocols

Enzymatic Assay of ß-Glucuronidase (EC 3.2.1.31) from Helix Pomatia and Bovine Liver

Enzymatic Assay of ß-Glucuronidase (EC 3.2.1.31) from Helix Pomatia and Bovine Liver

Using UHPLC/MS (TOF) for Detection of Drugs and Metabolites in Urine Following Optimized Enzymatic Hydrolysis Conditions

To optimize hydrolysis using β-glucuronidase, factors such as incubation time, temperature, hydrolysis pH, enzyme source, and enzyme concentration must be evaluated for each glucuronide metabolite to be analyzed.

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