Beta-glucuronidase (Escherichia coli)

The deficiency of Beta-glucuronidase causes mucopolysaccharidosis type VII (MPSVII), leading to lysosomal storage in the brain^[1].
Product Information
Molecular weight: ~290 kDa (tetramer), 68 kDa (monomer)
Optimum pH: 6.0 - 7.0

Activity Assay
1. Reconstitution:
Dissolve the enzyme at a concentration of 2 mg/mL in cold 0.15 M NaCl. Dilute the enzyme to a concentration range of 0.25 to 2 mg/mL for the assay.
2. Procedure
1) Transfer the following reagents into a series of numbered test tubes: (including one test tube as a blank).
0.1 M Sodium acetate, pH4.5 0.8 mL
0.01 M Phenolphthalein glucuronidate, pH6.8 0.1 mL
2) Incubate in a 37°C water bath for 5 minutes to achieve temperature equilibrium. At intervals, add 0.1 mL of enzyme dilution to each test tube. Add 0.1 mL of water to the blank and incubate for 30 minutes.
3) Add 5.0 mL 0.2 M glycine, 0.2 M NaCl, pH 10 to terminate the reaction.
4) Remove from the water bath, cool to room temperature and read the A540 value and the blank value.
3. Calculation
A standard curve was drawn, where 1U was the amount of enzyme (mg) required to release 1 μmoL phenolphthalein from phenolphthalein glucuronide per hour.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.