This technology is an in vitro platform that detects and quantifies the production of small-molecule metabolites in a high-throughput manner to screen mutagenized microorganism strains.
Metabolic engineering is a promising avenue for low-cost, sustainable production of metabolites. However, achieving viable yields remains challenging due to bottlenecks in assessing the ability of a library of mutant cells to produce the metabolite of interest. Current methods, such as gas/liquid chromatography or fluorescence-activated cell sorting (FACS) are some combination of low-throughput, expensive, or non-generalizable beyond a specific set of metabolites. No platform provides the sensitivity, specificity, adaptability, or throughput necessary to make metabolic engineering viable in a wider range of production contexts.
The fluorescence polarization (FP) assay utilizes a competitive reporter molecule with a fluorescent tag that will bind a target receptor of the metabolite of interest. When the metabolite is present in higher concentrations, the reporter will bind with the receptor less frequently, leading to a weaker fluorescent signal that can be quantified. By applying the FP assay to mutagenized strains in 96-well microtiter plates, this technology describes a high-throughput screen for the production of FK506, an immunosuppressant drug used to prevent organ rejection. The system can also be used to screen for similar compounds (rapamycin, meridamycin, ascomycin) and serves as a proof of concept for the application of this technique to other metabolites of interest.
IR CU16161
Licensing Contact: Joan Martinez