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Far‐Red Fluorogenic Probes for Esterase and Lipase Detection
Author(s) -
Tallman Katie R.,
Beatty Kimberly E.
Publication year - 2015
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201402548
Subject(s) - esterase , lipase , fluorophore , enzyme , biochemistry , chemistry , hydrolysis , molecular probe , fluorescence , enzymatic hydrolysis , dna , physics , quantum mechanics
Abstract Fluorogenic enzyme probes go from a dark to a bright state following hydrolysis and can provide a sensitive, real‐time readout of enzyme activity. They are useful for examining enzymatic activity in bacteria, including the human pathogen Mycobacterium tuberculosis . Herein, we describe two fluorogenic esterase probes derived from the far‐red fluorophore 7‐hydroxy‐9 H ‐(1,3‐dichloro‐9,9‐dimethylacridin‐2‐one) (DDAO). These probes offer enhanced optical properties compared to existing esterase probes because the hydrolysis product, DDAO, excites above 600 nm while retaining a good quantum yield ( ϕ =0.40). We validated both probes with a panel of commercially available enzymes alongside known resorufin‐ and fluorescein‐derived esterase substrates. Furthermore, we used these probes to reveal esterase activity in protein gel‐resolved mycobacterial lysates. These probes represent new tools for esterase detection and characterization and should find use in a variety of applications.