Development of a sensitive long-wavelength fluorogenic probe for nitroreductase: A new fluorimetric indictor for analyte determination by dehydrogenase-coupled biosensors

Hun Chung Huang, Kun Li Wang, Sheng Tung Huang, Hsin Yi Lin, Chun Mao Lin

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Nitroreductase (NTR) is a flavin-containing enzyme that uses NADH as the electron source to reduce nitroaromatic compounds to the corresponding amines. Previous studies have shown that nitroreductase-targeted latent fluorophores exhibit low solubility in the aqueous media and fluoresce at lower wavelengths upon uncloaking, thus limiting their effective applications. Here, we have prepared a new switch-on long-wavelength latent fluorogenic substrate, NTRLF (4), for NTR. In the presence of NADH, NTR catalyzes the reduction of the nitroaromatic moiety in NTRLF (4), followed by the cascade reaction, 1,6-rearrangement-elimination reaction, cyclic urea formation, and concomitant ejects a long-wavelength fluorescence coumarin (8). However, this reaction was inhibited in the presence of nitroaromatic analogues. The fluorescence signal generated by the cascade reaction was specific and insensitive to various reductants. Accordingly, we propose that NTRLF and NTR in the presences of NADH constitute a useful switch-off high-throughput fluorescence sensor for screening nitroaromatic compounds. Furthermore, NTRLF in the NTR-coupled 3-hydroxybutyrate dehydrogenase and aldehyde dehydrogenase assay reactions was a sensitive fluorimetric indicator for the quantitatively measurement of 3-hydroxybutyrate and propionaldehyde, respectively within micromolar range. Our novel NTRLF and NTR-coupled dehydrogenase assay platform may thus be effectively applied for the quantitative estimation of a broad range of analytes.

Original languageEnglish
Pages (from-to)3511-3516
Number of pages6
JournalBiosensors and Bioelectronics
Volume26
Issue number8
DOIs
Publication statusPublished - Apr 15 2011

Fingerprint

Nitroreductases
Biosensing Techniques
Biosensors
Oxidoreductases
Fluorescence
Wavelength
Assays
Switches
Electron sources
NAD
Fluorophores
Aldehydes
Urea
Amines
Screening
Enzymes
Solubility
Throughput
Hydroxybutyrate Dehydrogenase
Sensors

Keywords

  • 3-Hydroxybutyrate
  • Aldehyde
  • Latent fluorogenic substrate
  • Nitroaromatic compounds
  • Nitroreductase

ASJC Scopus subject areas

  • Biophysics
  • Biomedical Engineering
  • Biotechnology
  • Electrochemistry

Cite this

Development of a sensitive long-wavelength fluorogenic probe for nitroreductase : A new fluorimetric indictor for analyte determination by dehydrogenase-coupled biosensors. / Huang, Hun Chung; Wang, Kun Li; Huang, Sheng Tung; Lin, Hsin Yi; Lin, Chun Mao.

In: Biosensors and Bioelectronics, Vol. 26, No. 8, 15.04.2011, p. 3511-3516.

Research output: Contribution to journalArticle

@article{d9aa1e27e6084acb83b6de60f0bccd77,
title = "Development of a sensitive long-wavelength fluorogenic probe for nitroreductase: A new fluorimetric indictor for analyte determination by dehydrogenase-coupled biosensors",
abstract = "Nitroreductase (NTR) is a flavin-containing enzyme that uses NADH as the electron source to reduce nitroaromatic compounds to the corresponding amines. Previous studies have shown that nitroreductase-targeted latent fluorophores exhibit low solubility in the aqueous media and fluoresce at lower wavelengths upon uncloaking, thus limiting their effective applications. Here, we have prepared a new switch-on long-wavelength latent fluorogenic substrate, NTRLF (4), for NTR. In the presence of NADH, NTR catalyzes the reduction of the nitroaromatic moiety in NTRLF (4), followed by the cascade reaction, 1,6-rearrangement-elimination reaction, cyclic urea formation, and concomitant ejects a long-wavelength fluorescence coumarin (8). However, this reaction was inhibited in the presence of nitroaromatic analogues. The fluorescence signal generated by the cascade reaction was specific and insensitive to various reductants. Accordingly, we propose that NTRLF and NTR in the presences of NADH constitute a useful switch-off high-throughput fluorescence sensor for screening nitroaromatic compounds. Furthermore, NTRLF in the NTR-coupled 3-hydroxybutyrate dehydrogenase and aldehyde dehydrogenase assay reactions was a sensitive fluorimetric indicator for the quantitatively measurement of 3-hydroxybutyrate and propionaldehyde, respectively within micromolar range. Our novel NTRLF and NTR-coupled dehydrogenase assay platform may thus be effectively applied for the quantitative estimation of a broad range of analytes.",
keywords = "3-Hydroxybutyrate, Aldehyde, Latent fluorogenic substrate, Nitroaromatic compounds, Nitroreductase",
author = "Huang, {Hun Chung} and Wang, {Kun Li} and Huang, {Sheng Tung} and Lin, {Hsin Yi} and Lin, {Chun Mao}",
year = "2011",
month = "4",
day = "15",
doi = "10.1016/j.bios.2011.01.036",
language = "English",
volume = "26",
pages = "3511--3516",
journal = "Biosensors",
issn = "0956-5663",
publisher = "Elsevier Limited",
number = "8",

}

TY - JOUR

T1 - Development of a sensitive long-wavelength fluorogenic probe for nitroreductase

T2 - A new fluorimetric indictor for analyte determination by dehydrogenase-coupled biosensors

AU - Huang, Hun Chung

AU - Wang, Kun Li

AU - Huang, Sheng Tung

AU - Lin, Hsin Yi

AU - Lin, Chun Mao

PY - 2011/4/15

Y1 - 2011/4/15

N2 - Nitroreductase (NTR) is a flavin-containing enzyme that uses NADH as the electron source to reduce nitroaromatic compounds to the corresponding amines. Previous studies have shown that nitroreductase-targeted latent fluorophores exhibit low solubility in the aqueous media and fluoresce at lower wavelengths upon uncloaking, thus limiting their effective applications. Here, we have prepared a new switch-on long-wavelength latent fluorogenic substrate, NTRLF (4), for NTR. In the presence of NADH, NTR catalyzes the reduction of the nitroaromatic moiety in NTRLF (4), followed by the cascade reaction, 1,6-rearrangement-elimination reaction, cyclic urea formation, and concomitant ejects a long-wavelength fluorescence coumarin (8). However, this reaction was inhibited in the presence of nitroaromatic analogues. The fluorescence signal generated by the cascade reaction was specific and insensitive to various reductants. Accordingly, we propose that NTRLF and NTR in the presences of NADH constitute a useful switch-off high-throughput fluorescence sensor for screening nitroaromatic compounds. Furthermore, NTRLF in the NTR-coupled 3-hydroxybutyrate dehydrogenase and aldehyde dehydrogenase assay reactions was a sensitive fluorimetric indicator for the quantitatively measurement of 3-hydroxybutyrate and propionaldehyde, respectively within micromolar range. Our novel NTRLF and NTR-coupled dehydrogenase assay platform may thus be effectively applied for the quantitative estimation of a broad range of analytes.

AB - Nitroreductase (NTR) is a flavin-containing enzyme that uses NADH as the electron source to reduce nitroaromatic compounds to the corresponding amines. Previous studies have shown that nitroreductase-targeted latent fluorophores exhibit low solubility in the aqueous media and fluoresce at lower wavelengths upon uncloaking, thus limiting their effective applications. Here, we have prepared a new switch-on long-wavelength latent fluorogenic substrate, NTRLF (4), for NTR. In the presence of NADH, NTR catalyzes the reduction of the nitroaromatic moiety in NTRLF (4), followed by the cascade reaction, 1,6-rearrangement-elimination reaction, cyclic urea formation, and concomitant ejects a long-wavelength fluorescence coumarin (8). However, this reaction was inhibited in the presence of nitroaromatic analogues. The fluorescence signal generated by the cascade reaction was specific and insensitive to various reductants. Accordingly, we propose that NTRLF and NTR in the presences of NADH constitute a useful switch-off high-throughput fluorescence sensor for screening nitroaromatic compounds. Furthermore, NTRLF in the NTR-coupled 3-hydroxybutyrate dehydrogenase and aldehyde dehydrogenase assay reactions was a sensitive fluorimetric indicator for the quantitatively measurement of 3-hydroxybutyrate and propionaldehyde, respectively within micromolar range. Our novel NTRLF and NTR-coupled dehydrogenase assay platform may thus be effectively applied for the quantitative estimation of a broad range of analytes.

KW - 3-Hydroxybutyrate

KW - Aldehyde

KW - Latent fluorogenic substrate

KW - Nitroaromatic compounds

KW - Nitroreductase

UR - http://www.scopus.com/inward/record.url?scp=79952818304&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79952818304&partnerID=8YFLogxK

U2 - 10.1016/j.bios.2011.01.036

DO - 10.1016/j.bios.2011.01.036

M3 - Article

C2 - 21398106

AN - SCOPUS:79952818304

VL - 26

SP - 3511

EP - 3516

JO - Biosensors

JF - Biosensors

SN - 0956-5663

IS - 8

ER -