Agarose-based microwell array chip for high-throughput screening of functional microorganisms.

Area of Science:

• Microbiology
• Biotechnology
• Biocatalysis

Background:

• Natural bacterial communities are valuable sources of biocatalysts.
• High-throughput screening for functional bacteria is a significant challenge in microbiology.

Purpose of the Study:

• To develop and validate a novel agarose-based microwell array chip for high-throughput functional screening and isolation of microorganisms.
• To assess the chip's efficiency, specificity, and sensitivity in identifying bacteria with desired enzyme activities.

Main Methods:

• Loading single bacterial cells into an agarose microwell array for incubation.
• Real-time monitoring of single-cell growth and assessment of enzyme activity at the microwell level.
• Application to screen lipase-producing bacteria using rhodamine B as a substrate indicator.

Main Results:

• Screened over 310,000 isolates from pond water within 13 hours, demonstrating high throughput and reduced reagent consumption.
• Identified 12 microcolonies with the target lipase-producing phenotype.
• Discovered two new bacterial species through 16S rRNA sequencing from the screened isolates.

Conclusions:

• The developed agarose microwell array chip enables efficient, high-throughput, and specific functional screening of microorganisms.
• This method facilitates the discovery of novel biocatalysts and microbial species.
• The technology holds potential for applications in microbiology, synthetic biology, and biocatalyst discovery.