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β-Glucuronidase

Sigma 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
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).

View the complete list of Cerilliant® glucuronide drug standards

New Technical Article Comparing Performance of Different Enzymes
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)

Bovine Liver Source

Bovine Liver Source ß-Glucuronidases
Bovine ß-Glucuronidase is a 290 kD protein with an isoelectric point of 5.1.1
Bovine preparations typically contain small amounts of sulfatase activity, usually less than 0.5%.
Optimal pH
glucuronidase activity: 4.4
sulfatase activity: 4.4

References
1. Himeno, et al., J. Biochem.(Tokyo), 76, 1243 (1974)

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G0251 β-Glucuronidase from bovine liver Type B-1, ≥1,000,000 units/g solid
G0376 β-Glucuronidase from bovine liver Type B-3, ≥2,000,000 units/g solid
G0501 β-Glucuronidase from bovine liver Type B-10, ~10,000 units/mg solid

E. coli Source

E. coli Source ß-Glucuronidases
ß-Glucuronidase is a ~290 kDa tetrameric protein with an isoelectric point of 4.8.1 Unlike the enzyme preparations from mollusks that naturally contain ß-glucuronidase and sulfatase activities in almost equal amounts, the preparation of ß-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 ß-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.2 The E. coli enzyme has also been shown to be more active against estrogen conjugates than other sources of the enzyme.3
Optimal pH: 6-7

References
1. Kim, D-H, et al., Biol. Pharm. Bull., 18, 1184-1188 (1995)
2. Graef, V., et al., Clin. Chem., 23, 532-535 (1977)
3. Jayle, M.F., Scholler, R., Jarrige, P. & Mtay, S. Bull. Soc. Chim. Biol. 41, 1593. (1959)

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BGALS-RO β-Glucuronidase from E. coli K 12
G7396 β-Glucuronidase from Escherichia coli Type IX-A, lyophilized powder, 1,000,000-5,000,000 units/g protein (30 min assay)
G7646 β-Glucuronidase from Escherichia coli Type VII-A, lyophilized powder, 5,000,000-20,000,000 units/g protein, pH 6.8 (30 min assay)
G8162 β-Glucuronidase from Escherichia coli aqueous glycerol solution, ≥5,000,000 units/g protein, pH 6.8 (biuret)
G8295 β-Glucuronidase from Escherichia coli ≥10,000,000 units/g protein (30 min assay), recombinant, expressed in E. coli overproducing strain, lyophilized powder
G8420 β-Glucuronidase from Escherichia coli ≥20,000 units/mg protein, recombinant, expressed in E. coli overproducing strain, lyophilized powder

Molluskan Source

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

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SRE0022 β-Glucuronidase from abalone Chromatographically purified, lyophilized powder, β-glucuronidase ≥20,000,000 units/g protein
SRE0023 β-Glucuronidase from abalone Purified, aqueous solution, β-glucuronidase 150,000-250,000 units/mL, β-glucuronidase ≥20,000,000 units/g protein
SRE0037 β-Glucuronidase from abalone aqueous solution, β-glucuronidase ≥100,000 units/mL
SRE0038 β-Glucuronidase from abalone lyophilized powder, β-glucuronidase ≥1,500,000 units/g solid
G4259 β-Glucuronidase from Helix aspersa (garden snail) Type HA-4
G7017 β-Glucuronidase from Helix pomatia Type HP-2, aqueous solution, ≥100,000 units/mL
G0876 β-Glucuronidase from Helix pomatia Type H-2, aqueous solution, ≥85,000 units/mL
G7770 β-Glucuronidase from Helix pomatia Type HP-2S, aqueous solution, ≥90,000 units/mL
G0751 β-Glucuronidase from Helix pomatia Type H-1, partially purified powder, ≥300,000 units/g solid
G8885 β-Glucuronidase from Helix pomatia Type H-3, aqueous solution, ≥90,000 units/mL
G1512 β-Glucuronidase from Helix pomatia Type H-5, lyophilized powder, ≥400,000 units/g solid
G0762 β-Glucuronidase from Helix pomatia Type H-3AF, aqueous solution, ≥60,000 units/mL
G8132 β-Glucuronidase from limpets (Patella vulgata) Type L-II, lyophilized powder, 1,000,000-3,000,000 units/g solid
G2174 β-Glucuronidase from limpets (Patella vulgata) aqueous solution, ≥85,000 units/mL

Recobinant β-Glucuronidase

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SRE0093 β-Glucuronidase from limpets (Patella vulgata) aqueous solution, 100,000-200,000 U/mL

Related Products

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B8174 5-Bromo-4-chloro-3-indolyl β-D-glucuronide sodium salt tablet
B4532 5-Bromo-6-chloro-3-indolyl β-D-glucuronide cyclohexylammonium salt ≥98%
G7166 D-Glucaro-δ-lactam potassium salt solid, from microbial
G5269 D-Glucuronic acid ≥98% (GC)
N1627 4-Nitrophenyl β-D-glucuronide ≥98% (TLC)
P0501 Phenolphthalein β-D-glucuronide β-glucuronidase substrate
P0376 Phenolphthalein β-D-glucuronide sodium salt β-glucuronidase substrate