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Updated: Mar 14, 2026
13:16
Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles
Published on: December 31, 2019
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Functional mining of transporters using synthetic selections
Hans J Genee1,2, Anne P Bali1,2, Søren D Petersen1
1The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, Denmark.
Nature Chemical Biology
|October 4, 2016
Summary
Researchers developed a novel biosensor method to discover new bacterial membrane transporters. This approach successfully identified novel thiamine (PnuT) and xanthine transporters from metagenomic data.
Area of Science:
- Synthetic Biology
- Microbial Ecology
- Molecular Biology
Background:
- Functional annotation of bacterial membrane transporters is limited, with only 25% characterized due to experimental and predictive challenges.
- Understanding transporter function is crucial for various biological processes and biotechnological applications.
Purpose of the Study:
- To develop a sequence-independent, high-throughput method for discovering novel bacterial membrane transporters.
- To functionally mine metagenomes for previously uncharacterized transporter systems.
Main Methods:
- Utilized ligand-responsive biosensor systems enabling selective cell growth based on specific importer activity.
- Developed a synthetic selection system for thiamine pyrophosphate, applied to soil and gut metagenomes.
- Demonstrated modularity by adapting the biosensor system for xanthine transport discovery.
Main Results:
- Identified a novel class of thiamine transporters, designated PnuT, distributed across diverse bacterial phyla.
- Discovered xanthine permeases from gut and soil metagenomic samples using the adaptable biosensor approach.
- Validated the effectiveness of the synthetic biology method for functional metagenomic mining.
Conclusions:
- Synthetic biology offers a powerful strategy for functional metagenomic exploration of transporter systems.
- The developed method effectively elucidates sequence-function relationships for bacterial small-molecule transporters.
- This approach significantly expands the capacity for discovering novel transporter functions in uncultured bacteria.
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