A sensitive fluorimetric rate assay for biotinidase using a new derivative of biotin, biotinyl-6-aminoquinoline.
The previously reported method for the estimation of biotinidase (EC 3.5.1.12) is an endpoint colorimetric assay based on the hydrolysis of biotinyl-4-aminobenzoate, followed by diazotization, and is not suitable for our studies of biotinidase. A fluorimetric rate assay of biotinidase which uses a newly synthesized derivative biotinyl-6-aminoquinoline is described here.
D000581:Amidohydrolases / Q000097:blood / Q000378:metabolism*; D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D043523:Biotinidase; D003124:Colorimetry; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity; D013552:Swine
D001710:Biotin; C042785:biotinyl-6-aminoquinoline; D000581:Amidohydrolases; D043523:Biotinidase