Search

[Display omitted] • Cell-free systems enable biochemical reactions in the absence of cell viability constraints. • Cell-free metabolic engineering presents opportunities for biomanufacturing diverse chemical products. • Native metabolism in cell extracts can be coupled with exogenous or artificial pathways to use renewable carbon sources. • The open, cell-free reaction environment is amenable to electrochemical and photosynthetic energy regeneration modules. • Increasing cell-free reactions beyond the prototyping scale will enable sustainable biomanufacturing. Industrial biotechnology is an attractive approach to address the need for low-cost fuels and products from sustainable resources. Unfortunately, cells impose inherent limitations on the effective synthesis and release of target products. One key constraint is that cellular survival objectives often work against the production objectives of biochemical engineers. Additionally, industrial strains release CO 2 and struggle to utilize sustainable, potentially profitable feedstocks. Cell-free biotechnology, which uses biological machinery harvested from cells, can address these challenges with advantages including: (i) shorter development times, (ii) higher volumetric production rates, and (iii) tolerance to otherwise toxic molecules. In this review, we highlight recent advances in cell-free technologies toward the production of non-protein products beyond lab-scale demonstrations and describe guiding principles for designing cell-free systems. Specifically, we discuss carbon and energy sources, reaction homeostasis, and scale-up. Expanding the scope of cell-free biomanufacturing practice could enable innovative approaches for the industrial production of green chemicals. [ABSTRACT FROM AUTHOR]