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On-chip analysis, indexing and screening for chemical producing bacteria in a microfluidic static droplet array
Sungho Jang1, Byungjin Lee, Heon-Ho Jeong
1Department of Chemical Engineering, Pohang University of Science and Technology, 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk 790-784, Korea. gyjung@postech.ac.kr.
Lab on a Chip
|April 23, 2016
Summary
This study introduces a novel microfluidic screening platform using artificial riboswitches to measure intracellular l-tryptophan in single microbial cells. The technology successfully identified high-producing strains, boosting productivity by 145%.
Area of Science:
- Biotechnology
- Metabolic Engineering
- Synthetic Biology
Background:
- Efficient screening is vital for microbial strain improvement, particularly for chemical production.
- Current high-throughput screening methods struggle with assessing diverse intracellular metabolites at the single-cell level.
Purpose of the Study:
- To develop a novel screening platform for analyzing intracellular metabolite concentrations in single microbial cells.
- To enable efficient screening of high-producing microbial strains for chemical synthesis.
Main Methods:
- Coupling a microfluidic static droplet array (SDA) with an artificial riboswitch.
- Entrapping single Escherichia coli cells in SDA to measure intracellular l-tryptophan concentrations via fluorescence.
- Screening a mutagenized library to identify improved strains.
Main Results:
- Demonstrated successful evaluation of intracellular l-tryptophan concentration using riboswitch-induced fluorescence.
- Successfully screened high-producing microbial strains with up to 145% increased productivity compared to the parental strain.
- Validated the platform's capability for single-cell metabolite analysis.
Conclusions:
- The developed platform offers a powerful tool for high-throughput screening of intracellular metabolites at the single-cell level.
- This technology is widely applicable to microbial strain improvement for various metabolites by developing new artificial riboswitches.
- Facilitates economic chemical production through enhanced microbial strain development.
