Sigma G8132-5X1MU Beta-Glucuronidase from Limpets (Patella Vulgata), Type L-II Lyphilized Powder (5 x 1MU)

Sigma-Aldrich G8132-5X1MU
Item Number:SIG-G8132-5X1MU
Manufacturer Part No:SIAL-G8132-5X1MU
Manufacturer:Sigma-Aldrich
Units:Packaging: 5 x 1MU

Details

  • Type L-II, Lyophilized Powder, 1,000,000-3,000,000 units/g solid
  • Synonym: β-D-Glucuronide glucuronosohydrolase
  • CAS Number: 9001-45-0
  • Enzyme Commission (EC) Number: 3.2.1.31
  • Shipped in Wet Ice. Storage Temperature: -20°C
  • Packaging: 5 x 1MU

Product Description

Glucuronidation, conjugation with glucuronic acid, by the human UDP-glucuronosyltransferase (UGT) family of enzymes plays an important role in the metabolic fate of many drugs and other xenobiotics.  This biosynthetic reaction also has a role in the conjugation and excretion of endogenous substrates, such as steroids, bilirubin, and bile acids.1 UGT activity results in the conjugation of glucuronic acid to substrates containing sulfhydryl, hydroxyl, aromatic amino, or carboxylic acid moieties. The glucuronides formed are more polar (water soluble) than the parent organic substrate and are generally excreted through the kidney.

 

b-glucuronidase catalyzes the reaction:

 

b-D-glucuronoside + H2O « an alcohol + D-glucuronate

 

b-Glucuronidase Type L-II has been shown to be a superior enzyme for the hydrolysis of drug-glucuronides from urine.  This enzyme preparation was found to be cost-effective and thermostable; it could be used at a temperature high enough to allow for a shorter incubation time as compared to the enzyme from snail or bovine. Although the exact amount needed will depend on the specific conditions used and must be determined empirically, complete hydrolysis of morphine glucuronide was reported following a 3-hour incubation at 65 °C with 5,000 units of the enzyme per ml of urine.  Another report found the optimal conditions for hydrolysis of conjugated steroid metabolites to be a one hour  incubation at 55 °C at pH 5.2 using 600 units of enzyme per ml of urine.

 

b-Glucuronidase Type L-II from keyhole limpet is a crude solution of enzymes. Many b-glucuronidase preparations may also contain sulfatase activity. For this reason, sulfatase activity is also determined.

 

 

Optimal pH:

Glucuronidase activity:   4.5 to 5.0

Sulfatase activity:             ~6.2

Inhibitors:              D-glucuronic acid (Catalog No. SIG-G5269)

D-galacturonic acid

D-glucaro-1,4-lactone

Substrates:

5-Bromo-6-chloro-3-indolyl b-D-glucuronide (SIG-B4532)

6-Bromo-2-naphthyl b-D-glucuronide (SIG-B6519)

5-Bromo-4-chloro-3-indolyl b-D-glucuronidesodium salt tablet (SIG-B8174)

8-Hydroxyquinoline glucuronide (SIG-H1254)

4-Methylumbelliferyl b-D-glucuronide (SIG-M5664)

4-Nitrophenyl b-D-glucuronide (SIG-73677)

 

Glucuronidase Activity:                1,000,000–3,000,000 units per gram solid.

 

Unit Definition:               One Sigma or modified Fishman unit will liberate 1.0 mg of phenolphthalein from phenolphthalein glucuronide per hour at 37 °C at pH 3.8 (30 minute assay).

 

Sulfatase Activity:                 Reported on the certificate of analysis.

 

Unit Definition:                      One unit of sulfatase will hydrolyze 1.0 mmole of p-nitrocatechol sulfate per hour at pH 5.0 at 37 °C.

 

Precautions and Disclaimer:  This product is for R&D use only, not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices.

 

Storage/Stability:                   Store the product at –20 °C. When stored at –20 °C, the enzyme retains activity for at least 3 years.

Documents & Videos

Product Description

Glucuronidation, conjugation with glucuronic acid, by the human UDP-glucuronosyltransferase (UGT) family of enzymes plays an important role in the metabolic fate of many drugs and other xenobiotics.  This biosynthetic reaction also has a role in the conjugation and excretion of endogenous substrates, such as steroids, bilirubin, and bile acids.1 UGT activity results in the conjugation of glucuronic acid to substrates containing sulfhydryl, hydroxyl, aromatic amino, or carboxylic acid moieties. The glucuronides formed are more polar (water soluble) than the parent organic substrate and are generally excreted through the kidney.

 

b-glucuronidase catalyzes the reaction:

 

b-D-glucuronoside + H2O « an alcohol + D-glucuronate

 

b-Glucuronidase Type L-II has been shown to be a superior enzyme for the hydrolysis of drug-glucuronides from urine.  This enzyme preparation was found to be cost-effective and thermostable; it could be used at a temperature high enough to allow for a shorter incubation time as compared to the enzyme from snail or bovine. Although the exact amount needed will depend on the specific conditions used and must be determined empirically, complete hydrolysis of morphine glucuronide was reported following a 3-hour incubation at 65 °C with 5,000 units of the enzyme per ml of urine.  Another report found the optimal conditions for hydrolysis of conjugated steroid metabolites to be a one hour  incubation at 55 °C at pH 5.2 using 600 units of enzyme per ml of urine.

 

b-Glucuronidase Type L-II from keyhole limpet is a crude solution of enzymes. Many b-glucuronidase preparations may also contain sulfatase activity. For this reason, sulfatase activity is also determined.

 

 

Optimal pH:

Glucuronidase activity:   4.5 to 5.0

Sulfatase activity:             ~6.2

Inhibitors:              D-glucuronic acid (Catalog No. SIG-G5269)

D-galacturonic acid

D-glucaro-1,4-lactone

Substrates:

5-Bromo-6-chloro-3-indolyl b-D-glucuronide (SIG-B4532)

6-Bromo-2-naphthyl b-D-glucuronide (SIG-B6519)

5-Bromo-4-chloro-3-indolyl b-D-glucuronidesodium salt tablet (SIG-B8174)

8-Hydroxyquinoline glucuronide (SIG-H1254)

4-Methylumbelliferyl b-D-glucuronide (SIG-M5664)

4-Nitrophenyl b-D-glucuronide (SIG-73677)

 

Glucuronidase Activity:                1,000,000–3,000,000 units per gram solid.

 

Unit Definition:               One Sigma or modified Fishman unit will liberate 1.0 mg of phenolphthalein from phenolphthalein glucuronide per hour at 37 °C at pH 3.8 (30 minute assay).

 

Sulfatase Activity:                 Reported on the certificate of analysis.

 

Unit Definition:                      One unit of sulfatase will hydrolyze 1.0 mmole of p-nitrocatechol sulfate per hour at pH 5.0 at 37 °C.

 

Precautions and Disclaimer:  This product is for R&D use only, not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices.

 

Storage/Stability:                   Store the product at –20 °C. When stored at –20 °C, the enzyme retains activity for at least 3 years.