Far-red fluorogenic probes for esterase and lipase detection

Katie R. Tallman, Kimberly Beatty

Research output: Contribution to journalArticle

16 Citations (Scopus)

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

Fingerprint

Esterases
Lipase
Hydrolysis
Enzymes
Fluorescein
Mycobacterium tuberculosis
Fluorophores
Enzyme activity
Quantum yield
Pathogens
Gels
Bacteria
Optical properties
Proteins
Substrates
dodecyldimethylamine oxide

Keywords

  • Esterases
  • Fluorescence
  • Fluorogenic probes
  • Lipases
  • Tuberculosis

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry
  • Molecular Medicine
  • Molecular Biology

Cite this

Far-red fluorogenic probes for esterase and lipase detection. / Tallman, Katie R.; Beatty, Kimberly.

In: ChemBioChem, Vol. 16, No. 1, 2015, p. 70-75.

Research output: Contribution to journalArticle

Tallman, Katie R. ; Beatty, Kimberly. / Far-red fluorogenic probes for esterase and lipase detection. In: ChemBioChem. 2015 ; Vol. 16, No. 1. pp. 70-75.
@article{e94fcf77218a4b04a6109efdfbae9a99,
title = "Far-red fluorogenic probes for esterase and lipase detection",
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.",
keywords = "Esterases, Fluorescence, Fluorogenic probes, Lipases, Tuberculosis",
author = "Tallman, {Katie R.} and Kimberly Beatty",
year = "2015",
doi = "10.1002/cbic.201402548",
language = "English (US)",
volume = "16",
pages = "70--75",
journal = "ChemBioChem",
issn = "1439-4227",
publisher = "Wiley-VCH Verlag",
number = "1",

}

TY - JOUR

T1 - Far-red fluorogenic probes for esterase and lipase detection

AU - Tallman, Katie R.

AU - Beatty, Kimberly

PY - 2015

Y1 - 2015

N2 - 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.

AB - 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.

KW - Esterases

KW - Fluorescence

KW - Fluorogenic probes

KW - Lipases

KW - Tuberculosis

UR - http://www.scopus.com/inward/record.url?scp=84919777537&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84919777537&partnerID=8YFLogxK

U2 - 10.1002/cbic.201402548

DO - 10.1002/cbic.201402548

M3 - Article

C2 - 25469918

AN - SCOPUS:84919777537

VL - 16

SP - 70

EP - 75

JO - ChemBioChem

JF - ChemBioChem

SN - 1439-4227

IS - 1

ER -