Far-red fluorogenic probes for esterase and lipase detection

Katie R. Tallman, Kimberly E. Beatty

Research output: Contribution to journalArticle

17 Scopus citations

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-9H-(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 (π = 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.

Original languageEnglish (US)
Pages (from-to)70-75
Number of pages6
JournalChemBioChem
Volume16
Issue number1
DOIs
StatePublished - 2015

Keywords

  • Esterases
  • Fluorescence
  • Fluorogenic probes
  • Lipases
  • Tuberculosis

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry

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