A fluorescent assay amenable to measuring production of beta-D-glucuronides produced from recombinant UDP-glycosyl transferase enzymes.
Beta-glucuronidase cleavage of 4-methylumbelliferyl beta-D-glucuronide generates the highly fluorescent compound, 4-methylumbelliferone. We show that other beta-D-glucuronide compounds act as competitors in this assay. The 4-methylumbelliferyl beta-D-glucuronide cleavage assay can easily be adapted to high throughput formats to detect the presence of beta-D glucuronides generated using recombinant glycosyl transferase preparations.
Trubetskoy
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OV
PanVera Corporation, Madison, Wisconsin 53711, USA. olgat@panvera.com
Shaw
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PM
D001667:Binding, Competitive; D005453:Fluorescence; D005965:Glucuronates / Q000032:analysis* / Q000378:metabolism*; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D014453:Glucuronosyltransferase / Q000235:genetics / Q000378:metabolism*; D006923:Hymecromone / Q000031:analogs & derivatives / Q000378:metabolism; D007700:Kinetics; D011994:Recombinant Proteins / Q000378:metabolism*
D005965:Glucuronates; D011994:Recombinant Proteins; D006923:Hymecromone; C034888:4-methylumbelliferyl glucuronide; C110655:UDP-glucuronosyltransferase, UGT1A6; D014453:Glucuronosyltransferase; D005966:Glucuronidase