earticle

영어

Metagenomics promises to provide new enzymes with diverse functions, however, current methods for the screening and cloning of novel metagenome-derived genes are limited to the examination of halo on solid media or well-plate assay. In this study, we developed a new platform for the screening of diverse enzyme genes from metagenome using a synthetic genetic circuit, based on the recognition of phenol derivatives, which are produced from aromatic substrates by many enzyme reactions. The phenol derivatives activates a transcriptional factor, DmpR, to express a fluorescent reporter gene, resulting in fluorescence emission. In cells harboring this genetic circuits, enzyme activities, such as those of tyrosine phenol-lyase, lipase, cellulase, and methyl parathion hydrolase, were detected by the fluorescence emitted. Finally, by applying the screening method to metagenome libraries, a novel phosphatase was successfully isolated using flow cytometry. These results show that phenol-mediated genetic enzyme screening system (GESS) is a widely applicable tool for high throughput and quantitative screening of diverse enzymes from the genetic pools.