ArticleScientific reports2016
Culture-independent method for identification of microbial enzyme-encoding genes by activity-based single-cell sequencing using a water-in-oil microdroplet platform.
Article in Scientific reports, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 41 citations in OpenAlex.
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- 7-Aminocoumarin-4-acetic Acid as a Fluorescent Probe for Detecting Bacterial Dipeptidyl Peptidase Activities in Water-in-Oil Droplets and in Bulk.Analytical chemistry · 2022Article
- Comprehensive health risk assessment of microbial indoor air quality in microenvironments.PloS one · 2022Article
- Exploring the Diversity and Biotechnological Potential of Cultured and Uncultured Coral-Associated Bacteria.Microorganisms · 2021Article
- Bioprospecting of Novel Extremozymes From Prokaryotes-The Advent of Culture-Independent Methods.Frontiers in microbiology · 2021Review
- Session 2SHP report-decoding intracellular architecture using visualizing device development and mathematical modeling.Biophysical reviews · 2020Article
- Simultaneous Enhancement of Thermostability and Catalytic Activity of a Metagenome-Derived β-Glucosidase Using Directed Evolution for the Biosynthesis of Butyl Glucoside.International journal of molecular sciences · 2019Article
- Size-Dependent and Property-Independent Passive Microdroplet Sorting by Droplet Transfer on Dot Rails.Micromachines · 2018Article
- Construction of integrated gene logic-chip.Nature nanotechnology · 2018Article
- Development of a facile droplet-based single-cell isolation platform for cultivation and genomic analysis in microorganisms.Scientific reports · 2017Article
- Screening and identification of novel biologically active natural compounds.F1000Research · 2017Review
- Development of Droplet Microfluidics Enabling High-Throughput Single-Cell Analysis.Molecules (Basel, Switzerland) · 2016Review
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Authors and funding
11 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Environmental microbes are a great source of industrially valuable enzymes with potent and unique catalytic activities. Unfortunately, the majority of microbes remain unculturable and thus are not accessible by culture-based methods. Recently, culture-independent metagenomic approaches have been successfully applied, opening access to untapped genetic resources. Here we present a methodological approach for the identification of genes that encode metabolically active enzymes in environmental microbes in a culture-independent manner. Our method is based on activity-based single-cell sequencing, which focuses on microbial cells showing specific enzymatic activities. First, at the single-cell level, environmental microbes were encapsulated in water-in-oil microdroplets with a fluorogenic substrate for the target enzyme to screen for microdroplets that contain microbially active cells. Second, the microbial cells were recovered and subjected to whole genome amplification. Finally, the amplified genomes were sequenced to identify the genes encoding target enzymes. Employing this method, we successfully identified 14 novel β-glucosidase genes from uncultured bacterial cells in marine samples. Our method contributes to the screening and identification of genes encoding industrially valuable enzymes.
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