ArticleAnalytical chemistry2022
7-Aminocoumarin-4-acetic Acid as a Fluorescent Probe for Detecting Bacterial Dipeptidyl Peptidase Activities in Water-in-Oil Droplets and in Bulk.
Article in Analytical chemistry, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 20 citations in OpenAlex.
- Microfluidics to Meet Antibiotic Resistance: A Growing Research Frontier.Antibiotics (Basel, Switzerland) · 2025Review
- Ultrahigh-throughput screening of environmental bacteria for proteolytic activity using droplet-based microfluidics.Applied and environmental microbiology · 2025Article
- Droplet microfluidic screening to engineer angiotensin-converting enzyme 2 (ACE2) catalytic activity.Journal of biological engineering · 2025Article
- Coumarin-Synthetic Methodologies, Pharmacology, and Application as Natural Fluorophore.Topics in current chemistry (Cham) · 2024Review
- Clear zone formation in microdroplets for high-throughput screening for lactic acid bacteria.Frontiers in microbiology · 2024Article
- Ultrahigh-Throughput Enzyme Engineering and Discovery inChemical reviews · 2023Review
- Water-in-oil droplet-mediated method for detecting and isolating infectious bacteriophage particles via fluorescent staining.Frontiers in microbiology · 2023Article
- Back to the Drawing Board: A Unifying First-Principle Model for Correlating Sample UV-Vis Absorption and Fluorescence Emission.Analytical chemistry · 2022Article
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Authors and funding
8 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Droplet-based microfluidic systems are a powerful tool for biological assays with high throughput. Water-in-oil droplets (WODLs) are typically used in droplet-based microfluidic systems to culture microorganisms and perform enzyme assays. However, because of the oil surrounding the nanoliter and picoliter volumes of WODLs, availability of suitable substrates is limited. For instance, although 7-amino-4-methylcoumarin (AMC) is commonly used as a fluorescent probe of the substrate to detect peptidase activity, AMC leaks from WODLs to the oil phase due to its high hydrophobicity. Thus, AMC substrates cannot be used in droplet-based microfluidic systems with WODLs. In this study, we developed a peptidase substrate consisting of a dipeptide and 7-aminocoumarin-4-acetic acid (ACA), an AMC-derived fluorogenic compound. ACA was retained in the WODL for more than 7 days, and the dipeptidyl ACA substrate detected dipeptidyl peptidase (DPP) activity in the WODL. Compared to AMC substrates, the substrate specificity constants of DPPs for ACA substrates increased up to 4.7-fold. Fluorescence-activated droplet sorting made high-throughput screening of microorganisms based on DPP activity using the dipeptidyl ACA substrate possible. Since ACA could be applied to various substrates as a fluorescent probe, detectable microbial enzyme activities for droplet-based microfluidic systems can be largely expanded.
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