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Ultrahigh-throughput screening of environmental bacteria for proteolytic activity using droplet-based microfluidics.

Akihiro Nakamura1, Yoshiyuki Suzuki1, Nobuyuki Homma2

  • 1Department of Science of Technology Innovation, Nagaoka University of Technology, Nagaoka-shi, Niigata, Japan.

Applied and Environmental Microbiology
|June 13, 2025
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Summary
This summary is machine-generated.

This study introduces droplet-based microfluidics for efficient screening of environmental bacteria, discovering novel enzymes for industrial use. The new method rapidly identified a highly active peptidase, advancing biomanufacturing capabilities.

Keywords:
bacterial enzymeemulsionscreeningwater-in-oil droplet

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Area of Science:

  • Microbiology
  • Biotechnology
  • Biochemical Engineering

Background:

  • Industrial applications require novel enzymes like peptidases/proteases from diverse microbial sources.
  • Conventional screening methods are inefficient for large-scale microbial discovery.
  • Biomanufacturing of bio-based products demands efficient screening of environmental bacteria for essential functions.

Purpose of the Study:

  • To develop and demonstrate a droplet-based microfluidic system for ultra-high throughput functional screening of environmental bacteria.
  • To identify novel microbial strains with high peptidase activity for industrial applications.
  • To showcase the efficacy of microfluidics for advancing bio-based production.

Main Methods:

  • Utilized droplet-based microfluidics and fluorescence-activated droplet sorting for screening.
  • Screened approximately 630,000 microorganisms for endopeptidase activity.
  • Isolated bacteria and characterized a novel Asp-specific endopeptidase.

Main Results:

  • Successfully screened 630,000 microorganisms in 6 hours, identifying four species with high peptidase activity.
  • Isolated an Asp-specific endopeptidase from *Lysobacter soli*.
  • The novel enzyme exhibited 2.4-fold higher activity than a comparable commercial enzyme.

Conclusions:

  • Droplet-based microfluidics offers a revolutionary and efficient approach for function-based microbial screening.
  • This technology enables rapid isolation of high-value enzymes from environmental bacteria.
  • The developed platform demonstrates scalability for industrial biomanufacturing and bio-based production.