Enzyme Explorer

β-Glucuronidase

Our β-Glucuronidases are routinely used for the enzymatic hydrolysis of glucuronides from urine1,2, plasma3,4, and other fluids5 prior to analysis by enzyme immunoassay, mass spectrometry, gas chromatography, high performance liquid chromatography, or other means. Typically, between 1 and 20 units of glucuronidase is used per µl of plasma, urine, or bile for the enzymatic hydrolysis of glucuronides present in these samples.1–5 The exact amount needed will depend on the specific conditions used and must be determined empirically.

The enzyme from patella vulgata is reported to be much more effective in hydrolyzing opiod-glucuronides.6,7 Whereas the Helix pomatia and E. coli enzymes are reported to be slightly more effective in hydrolyzing steroid-glucuronides.8

New Technical Article Comparing Performance of Different Enzymes

Learn More about recent application data generated by 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.

On this page:

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.9 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.

β-Glucuronidase (EC 3.2.1.31) catalyzes the reaction:

β-D-glucuronoside + H2O <--> D-glucuronate + an alcohol

&beta;-Glucuronidase

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 (pH 6.8 for the E. coli source) (30 min assay).


Molluskan Source β-Glucuronidases

β-Glucuronidase preparations isolated from mollusks also contain sulfatase activity. For this reason, the sulfatase activity of these preparations is also reported.

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

    Optimal pH    
    glucuronidase activity: 4.5 to 5.0
    sulfatase activity: ~6.2

 

Molluskan Source Products
Product Type Form Activity Product No.
Type H-5 from Patella vulgata solution minimum 85,000 units/mL,
up to 7,500 units/mL sulfatase
G2174
Type H-5 from Patella vulgata lyophilized powder 1,000,000-3,000,000 units/g solid G8132
Type HA-4 from Helix aspersa powder 250,000-500,000 units/g solid,
up to 7,500 units/g solid sulfatase
G4259
Type HP-2 from Helix pomatia solution minimum 100,000 units/mL,
up to 7,500 units/mL sulfatase
G7017
Type H-2 from Helix pomatia solution minimum 85,000 units/mL,
up to 7,500 units/mL sulfatase
G0876
Type HP-2S from Helix pomatia sterile-filtered solution 1,000-5,000 units/mL,
1,000-5,000 units/mL sulfatase
G7770
Type H-1 from Helix pomatia lyophilized powder minimum 300,000 units/g solid,
minimum 100,000 units/g solid sulfatase
G0751
Type H-3 from Helix pomatia solution ~ 100,000 units/mL,
up to 1,000 units/mL sulfatase
G8885
Type H-5 from Helix pomatia lyophilized powder minimum 400,000 units/g solid
up to 40,000 units/g solid sulfatase
(A further purification of G0876)
G1512
Type H-3AF from Helix pomatia solution minimum 60,000 units/mL
(A further purification of Type H-3 to remove agglutinin)
G0762
Purified from Abalone lyophilized powder minimum 20,000,000 units/g protein and up to 4 units/g protein sulfatase SRE0022
Purified Aqueous Solution from Abalone solution minimum 100,000 units/mL, up to 7,500 units/mL sulfatase
SRE0023
Aqueous Solution from Abalone solution minimum 100,000 units/mL, up to 8,000 units/mL sulfatase
SRE0037
Crude Powder from abalone lyophilized powder minimum 1,500,000 units/g solid, up to 100,000 units/g solid sulfatase SRE0038
β-Glucuronidase from limpets
(Patella vulgata)
aqueous solution 100,000-200,000 U/mL SRE0093
Fast β-Glucuronidase, Recombinant from limpets (Patella Vulgata) aqueous solution ≥300,000 U/mL SRE0095

 


Bovine Liver Source β-Glucuronidases

Bovine β-Glucuronidase is a 290 kD protein with an isoelectric point of 5.1.10

Bovine preparations typically contain small amounts of sulfatase activity, usually less than 0.5%.

 

    Optimal pH    
    glucuronidase activity: 4.4
    sulfatase activity: 4.4


Bovine Liver Source Products
Product Type Form Activity Product No.
Type B-1 lyophilized powder minimum 1,000 units/mg solid,
~85% protein
G0251
Type B-3 lyophilized powder ~3,000 units/mg solid,
~50% protein
G0376
Type B-10 lyophilized powder ~10,000 units/mg solid G0501

 


E. coli Source β-Glucuronidases

β-Glucuronidase is a ~290 kDa tetrameric protein with an isoelectric point of 4.8.11 Unlike the enzyme preparations from mollusks that naturally contain β-glucuronidase and sulfatase activities in almost equal amounts, the preparation of β-glucuronidase from E. coli is essentially free of sulfatase activity. The enzyme from E. coli has a high rate of hydrolytic activity and it retains this activity during hydrolysis better than similar enzymes that are more sensitive to changes in the concentration of β-glucuronide conjugates. The enzyme preparation from E. coli has been shown to be useful for determining the presence of androsterone, 17-hydroxycorticosteroids, and estriol in urine.12 The E. coli enzyme has also been shown to be more active against estrogen conjugates than other sources of the enzyme.13

 

    Optimal pH: 6-7


E. coli Source Products
Product Type Form Activity Product No.
Recombinant, from overexpressing E. coli lyophilized powder minimum 10,000,000 units/g protein G8295
Recombinant, from overexpressing E. coli lyophilized powder minimum 20,000 units/mg protein G8420
Type IX-A lyophilized powder 1,000,000-5,000,000 units/g protein,
~50% protein
G7396
Type VII-A lyophilized powder 5,000,000-20,000,000 units/g protein,
~25% protein
G7646
Type X-A lyophilized powder 20,000,000-60,000,000 units/g protein,
~30% protein
G7896
Aqueous Glycerol Solution aqueous glycerol solution 20,000,000-60,000,000 units/g protein G8162
Preweighed Vials lyophilized powder,
(min. 1,000 units per vial)
5,000,000-20,000,000 units/g protein,
when reconstituted with 10 mL water, solution will contain ~4 mM phosphate buffer, pH 6.8.
G8396
Preweighed Vials lyophilized powder,
(min. 1,000 units per vial)
5,000,000-20,000,000 units/g protein G8271

 


Related Products


Inhibitors
Product Name Product No.
D-Glucuronic acid G5269
D-Glucaro-δ-lactam potassium salt G7166
ψ-Tectorigenin T9165


Substrates
Product Name Product No.
5-Bromo-6-chloro-3-indolyl β-D-glucuronide B4532
5-Bromo-4-chloro-3-indolyl β-D-glucuronide B8174
8-Hydroxyquinoline glucuronide H1254
4-Methylumbelliferyl β-D-glucuronide M5664
4-Nitrophenyl β-D-glucopyranoside N1627
Phenolphthalein β-D-glucuronide P0501
Phenolphthalein β-D-glucuronide sodium salt P0376

 


References

  1. Xu, X., et al., J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci., 780, 315-330 (2002).
  2. Staimer, N., et al., Anal. Chim. Acta, 444, 27-36 (2001).
  3. Zhai, P., et al., Am. J. Physiol. Heart Circ. Physiol., 281, H1223-H1232 (2001).
  4. Anderson, A. et al., Cancer Chemother. Pharmacol., 44, 422-426 (1999).
  5. Nobilis, M., J. Chromatogr. A., 1031, 229-236 (2004).
  6. Combie, J. et al., Clin. Chem., 28, 83-88 (1982)
  7. Combie, J. et al., Res. Commun. Chem. Pathol. Pharmacol., 35, 27-41 (1982)
  8. Wakabayashi, M., et al., J. Biol. Chem., 236, 996-1001 (1960)
  9. Tephly, T.R., et al., Adv. Pharmacol., 42, 343-346 (1998).
  10. Himeno, et al., J. Biochem.(Tokyo), 76, 1243 (1974)
  11. Kim, D-H, et al., Biol. Pharm. Bull., 18, 1184-1188 (1995).
  12. Graef, V., et al., Clin. Chem., 23, 532-535 (1977).
  13. Jayle, M.F., Scholler, R., Jarrige, P. & Mtay, S. Bull. Soc. Chim. Biol. 41, 1593. (1959)