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Related Experiment Video

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Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay
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Small-Molecule Analysis Based on DNA Strand Displacement Using a Bacteriorhodopsin Photoelectric Transducer: Taking

Hsiu-Mei Chen1, Wen-Chang Wang2, Hong-Ren Chen1

  • 1Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 10607, Taiwan.

Sensors (Basel, Switzerland)
|September 9, 2023
PubMed
Summary
This summary is machine-generated.

This study developed a novel biosensor using purple membrane (PM) for sensitive detection of adenosine triphosphate (ATP) and bacteria. The PM-based sensor offers rapid, selective, and quantitative analysis of small molecules and microbial populations.

Keywords:
ATPbacteriorhodopsinphotoelectric biosensorpurple membranesmall moleculestrand displacement

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Area of Science:

  • Biophotonics
  • Biosensor Technology
  • Molecular Diagnostics

Background:

  • Purple membrane (PM) with photoactive bacteriorhodopsin is a sensitive photoelectric transducer.
  • Existing PM applications focus on proteins and microbes; extension to small molecules is explored.

Purpose of the Study:

  • To develop a PM-based biosensor for quantitative detection of small molecules, using adenosine triphosphate (ATP) as a model.
  • To adapt the PM biosensor for bacterial population analysis.

Main Methods:

  • A reverse detection strategy utilizing gold nanoparticle (AuNP) labeling and DNA strand displacement was employed.
  • A PM monolayer electrode was functionalized with an ATP-specific aptamer and hybridized with an AuNP-labeled DNA strand.
  • Photocurrent modulation by ATP binding was measured for quantitative analysis.

Main Results:

  • The biosensor achieved direct, single-step ATP detection within 15 minutes.
  • Detection limits for ATP were as low as 5 nM, with a dynamic range of 5 nM–0.1 mM.
  • The sensor demonstrated high selectivity, stability, and successfully quantified Bacillus subtilis and Escherichia coli at low CFU/mL levels.

Conclusions:

  • The developed PM-based biosensor effectively quantifies small molecules like ATP.
  • This technology enables simultaneous quantification of various small molecules and microbial detection.
  • PM-based biosensors represent a versatile platform for diverse quantitative assays.