Transporter function characterization via continuous-exchange cell-free synthesis and solid supported membrane-based electrophysiology.

Functional characterization of transporters is impeded by the high cost and technical challenges of current transporter assays. Thus, in this work, we developed a new characterization workflow that combines cell-free protein synthesis (CFPS) and solid supported membrane-based electrophysiology (SSME). For this, membrane protein synthesis was accomplished in a continuous exchange cell-free system (CECF) in the presence of nanodiscs.

Screening of metabolite transporters by C isotope substrate labeling.

The transportome of comprises approximately 340 membrane-bound proteins, of which very few are well-characterized. Elucidating transporter proteins' function is essential not only for understanding central cellular processes in metabolite exchange with the external milieu but also for optimizing the production of value-added compounds in microbial cell factories. Here, we describe the application of C-labeled stable isotopes and detection by targeted LC-MS/MS as a screening tool for identifying metabolite transporters.