<i>In vitro</i> transcription-based biosensing of glycolate for prototyping of a complex enzyme cascade.

ABSTRACT: In vitro metabolic systems allow the reconstitution of natural and new-to-nature pathways outside of their cellular context and are of increasing interest in bottom-up synthetic biology, cell-free manufacturing, and metabolic engineering. Yet, the analysis of the activity of such in vitro networks is very often restricted by time- and cost-intensive methods. To overcome these limitations, we sought to develop an in vitro transcription (IVT)-based biosensing workflow that is compatible with the complex conditions of in vitro metabolism, such as the crotonyl-CoA/ethylmalonyl-CoA/hydroxybutyryl-CoA (CETCH) cycle, a 27-component in vitro metabolic system that converts CO₂ into glycolate. As proof of concept, we constructed a novel glycolate sensor

SUBMITTER: Barthel S 

PROVIDER: S-EPMC11470758 | biostudies-literature | 2024

REPOSITORIES: biostudies-literature