Preparation and application of a fluorogenic substrate for endo-beta-xylosidase |
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| Authors: | K Takagaki A Kon H Kawasaki T Nakamura M Endo |
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| Affiliation: | Department of Biochemistry, Hirosaki University School of Medicine, Japan. |
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| Abstract: | The present paper describes a fluorometric assay for galactosaminoglycan-degrading endo-beta-xylosidase, utilizing glycosaminoglycan chains bearing a 4-methylumbelliferyl group at the reducing terminus as a substrate. This fluorogenic substrate is synthesized by human skin fibroblasts cultured in the presence of a fluorogenic xyloside, 4-methylumbelliferyl-beta-D-xyloside. The assay is based on measurement of the fluorescence of 4-methylumbelliferone, enzymatically liberated from the synthetic substrate by endo-beta-xylosidase. We examined the applicability of the assay for analysis of endo-beta-xylosidase activity. |
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