Abstract
This protocol describes a single-cell high-throughput genetic enzyme screening system (GESS) in which GFP fluorescence is used to detect the production of phenolic compounds from a given substrate by metagenomic enzyme activity. One of the important features of this single-cell genetic circuit is that it can be used to screen more than 200 different types of enzymes that produce phenolic compounds from phenyl group-containing substrates. The highly sensitive and quantitative nature of the GESS, combined with flow cytometry techniques, will facilitate rapid finding and directed evolution of valuable new enzymes such as glycosidases, cellulases, and lipases from metagenomic and other genetic libraries.
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Acknowledgment
This research was supported by grants from the Intelligent Synthetic Biology Center of Global Frontier Project (2011–0031944) and the KRIBB Research Initiative Program.
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Kim, H., Kwon, K.K., Rha, E., Lee, SG. (2015). Genetic Enzyme Screening System: A Method for High-Throughput Functional Screening of Novel Enzymes from Metagenomic Libraries. In: McGenity, T., Timmis, K., Nogales, B. (eds) Hydrocarbon and Lipid Microbiology Protocols. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8623_2015_65
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DOI: https://doi.org/10.1007/8623_2015_65
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