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Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System
Published on: August 8, 2016
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A Cell-Cell Communication-Based Screening System for Novel Microbes with Target Enzyme Activities
Haseong Kim1, Eugene Rha1, Wonjae Seong1,2
1Synthetic Biology & Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology , 125 Gwahak-ro, Yuseong-gu, Daejeon, South Korea.
ACS Synthetic Biology
|July 26, 2016
Summary
This study introduces an improved high-throughput genetic enzyme screening system using phenol-mediated cell-cell communication. The novel sensor cells efficiently detect microbes with target enzyme activities on solid media, enhancing biological discovery.
Area of Science:
- Synthetic biology
- Biotechnology
- Microbiology
Background:
- Synthetic biological devices offer increasing practical benefits.
- Previous high-throughput genetic enzyme screening systems can be improved for efficiency and practicality.
- Flow cytometry analysis can be a limitation in high-throughput screening.
Purpose of the Study:
- To enhance a high-throughput genetic enzyme screening system.
- To investigate device-compatible biological components and phenol-mediated cell-cell communication.
- To develop a screening device that does not require flow cytometry analysis.
Main Methods:
- Designed sensor cells to detect novel microbes with target enzyme activities on solid media.
- Utilized pre-effector phenolic substrate treatment and phenol-mediated cell-cell communication.
- Applied sensor cells to screen soil bacteria for phosphatase activity using phenyl phosphate.
Main Results:
- Achieved detection of target enzyme activities via clear, circular colonies with fluorescence around microbes.
- Successfully visualized phosphatase activity in unknown soil microbes identified by 16S rRNA analysis.
- Identified 23 positive clones out of 33 selected colonies, confirming screening technique efficacy.
Conclusions:
- The improved screening device enhances efficiency and practicality without flow cytometry.
- The technique has broad applications for screening novel microbes with target enzymes from phenol-derived substrates.
- This method aids in identifying novel enzymes lacking existing screening assays.
