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Updated: Jul 14, 2025
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An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
Published on: May 23, 2018
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Analysing Megasynthetase Mutants at High Throughput Using Droplet Microfluidics.
Farzaneh Pourmasoumi1, Sundar Hengoju2, Katharina Beck3
1Junior Research Group Biosynthetic Design of Natural Products, Leibniz Institute for Natural Product Research and Infection Biology (HKI), Beutenbergstr. 11a, 07745, Jena, Germany.
Chembiochem : a European Journal of Chemical Biology
|October 7, 2023
Summary
Scientists developed a microfluidic platform to detect gramicidin S production by nonribosomal peptide synthetases (NRPSs). This tool accelerates antibiotic discovery by enabling faster screening of NRPS enzyme activity.
Area of Science:
- Biochemistry
- Synthetic Biology
- Microfluidics
Background:
- Nonribosomal peptide synthetases (NRPSs) are crucial for producing valuable natural products like antibiotics.
- Developing new antibiotics necessitates efficient methods for redesigning NRPS pathways.
- Current screening platforms lack the robustness and speed required for NRPS engineering.
Purpose of the Study:
- To establish a microfluidic platform for detecting nonribosomal peptide production.
- To enable rapid sorting and screening of NRPS mutants.
- To facilitate the engineering of NRPS pathways for novel antibiotic discovery.
Main Methods:
- A microfluidic platform was developed utilizing calcein-filled sensor liposomes.
- Liposome fluorescence increases upon permeabilization, indicating gramicidin S production.
- The platform was used to sort a library of NRPS mutants.
Main Results:
- The platform reliably detected gramicidin S production.
- Enrichment of gramicidin S producers by 14.5-fold was achieved.
- A 250-fold decrease in internal stop codons and correlation with enzyme activity were observed.
Conclusions:
- The microfluidic platform provides a robust and rapid method for screening NRPS activity.
- This technology can accelerate the discovery of new antibiotics and facilitate NRPS engineering.
- Future applications include advanced structure-activity relationship studies and pathway engineering.
