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Enabling tools for high-throughput detection of metabolites: Metabolic engineering and directed evolution applications
- Lin, Jyun-Liang, Wagner, James M., Alper, Hal S.
- Biotechnology advances 2017 v.35 no.8 pp. 950-970
- absorbance, biosensors, chemical reactions, directed evolution, dyes, enzymatic reactions, fluorescence, metabolic engineering, metabolites, transcription (genetics), transcription factors
- Within the Design-Build-Test Cycle for strain engineering, rapid product detection and selection strategies remain challenging and limit overall throughput. Here we summarize a wide variety of modalities that transduce chemical concentrations into easily measured absorbance, luminescence, and fluorescence signals. Specifically, we cover protein-based biosensors (including transcription factors), nucleic acid-based biosensors, coupled enzyme reactions, bioorthogonal chemistry, and fluorescent and chromogenic dyes and substrates as modalities for detection. We focus on the use of these methods for strain engineering and enzyme discovery and conclude with remarks on the current and future state of biosensor development for application in the metabolic engineering field.