Fluorogenic substrates, methods of making the substrates, and methods of detecting glycosidase activities

Abstract

Fluorogenic substrates, methods of making the fluorogenic substrates, and methods of detecting glycosidase activities (EC3.2.1) are provided. The fluorogenic substrates are composed of a sugar covalently bonded to the 7-hydroxy position of a substituted coumarin sulfonated fluorophore is provided. The fluorogenic substrates include sodium β-D-cellobioside 6,8-difluoro-7-hydroxycoumarin-4-methanesulfonate; sodium β-D-glucoside 6,8-difluoro-7-hydroxycoumarin-4-methanesulfonate; sodium β-D-xyloside 6,8-difluoro-7-hydroxycoumarin-4-methanesulfonate; sodium β-D-xylobioside 6,8-difluoro-7-hydroxycoumarin-4-methanesulfonate; and sodium β-D-xylopolyoside 6,8-difluoro-7-hydroxycoumarin-4-methanesulfonate. The fluorogenic substrates may be made by covalently bonding the sugar moiety to the 7-hydroxy position of the substituted coumarin sulfonated fluorophore by reaction in DMF. Glycosidase activities can be detected in enzymatic extracts, microorganisms, and cells by contacting the extracts, microorganisms, or cells with one of the fluorogenic substrates.

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